• Journal of Microbiology and Biotechnology
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• 제15권5호
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• pp.1022-1027
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• 2005

Based on plasmid display technology by the complexes of fusion protein and the encoding plasmid DNA, an in vitro selection method for high affinity DNA-binding protein was developed and experimentally demonstrated. The GAL4 DNA-binding domain (GAL4 DBD) was selected as a model DNA-binding protein, and enhanced green fluorescent protein (EGFP) was used as an expression reporter for the selection of target proteins. Error prone PCR was conducted to construct a mutant library of the model. Based on the affinity decrease with increased salt concentration, mutants of GAL4 DBD having high affinity were selected from the mutant protein library of protein-encoding plasmid complex by this method. Two mutants of (Lys33Glu, Arg123Lys, Ile127Lys) and (Ser47Pro, Ser85Pro) having high affinity were obtained from the first generation mutants. This method can be used for rapid in vitro selection of high affinity DNA-binding proteins, and has high potential for the screening of high affinity DNA-binding proteins in a sequence-specific manner.

• 생물정신의학
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• 제13권3호
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• pp.152-161
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• 2006

Objective : The relationship between the total daily dose of clozapine given and the plasma concentrations of clozapine and its metabolites(N-desmethylclozapine and clozapine N-oxide) and the effect of Glu158Lys (wild-type : Glu, 'H' ; variant : Lys, 'h') and Glu308Gly(wild-type : Glu, 'D' ; variant : Lys, 'd') variation in FMO3 gene on plasma concentrations of clozapine and its metabolites was studied in schizophrenic patients. Methods : Trough plasma concentrations of clozapine and its metabolites were measured in 34 schizophrenic patients receiving clozapine. The genetic variation of 'h' and 'd' in FMO3 were analyzed in 21 among 34 patients. Results : A linear relationship between the total daily dose of clozapine given(mg/kg body weight per day) and the plasma concentrations(nM) of clozapine was revealed by regression analysis(p<0.001) in the 23 patients receiving a constant daily dose of clozapine for 8 days. The plasma molar concentration ratios of clozapine N-oxide/clozapine in 8 subjects with 'hh' or 'Hh' alleles were not different from those in 6 subjects with 'HH' alleles and the plasma molar concentration ratios in 6 subjects with 'dd' or 'Dd' alleles were not different from those in 8 subjects with 'DD' alleles. Conclusion : The effect of Glu158Lys and Glu308Gly variation in FMO3 gene on clozapine metabolism could not be shown.

• Asian Pacific Journal of Cancer Prevention
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• 제13권7호
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• pp.3465-3469
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• 2012

Aim: Platinum agents have shown to be effective in the treatment of colorectal cancer. We assessed whether single nucleotide polymorphisms (SNPs) in GSTP1, ERCC1 Asn118Asn and ERCC2 Lys751Gln might predict the overall survival in patients receiving oxaliplatin-based chemotherapy in a Chinese population. Methods: SNPs of GSTP1, ERCC1 Asn118Asn and ERCC2 Lys751Gln in 335 colorectal cancer patients were assessed using TaqMan nuclease assays. Results: At the time of final analysis on Nov. 2011, the median follow-up period was 37.7 months (range from 1 to 60 months). A total of 229 patients died during follow-up. Our study showed GSTP1 Val/Val (HR=0.44, 95% CI=0.18-0.98), ERCC1 C/C (HR=0.20, 95% CI=0.10-0.79) and ERCC2 G/G (HR=0.48, 95% CI=0.19-0.97) to be significantly associated with better survival of colorectal cancer. GSTP1 Val/Val, ERCC1 C/C and ERCC2 G/G were also related to longer survival among patients with colon cancer, with HRs (95% CIs) of 0.41 (0.16-0.91), 0.16 (0.09-0.74) and 0.34 (0.16-0.91), respectively. Conclusion: GSTP1, GSTP1, ERCC1 Asn118Asn and ERCC2 Lys751Gln genotyping might facilitate tailored oxaliplatin-based chemotherapy for colorectal cancer patients.

• 약학회지
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• 제58권1호
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• pp.21-27
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• 2014

We have developed 8 peptide derivatives as potential MC1R antagonists and their inhibitory effects on ${\alpha}$-MSH induced cell growth in cultured normal human melanocytes (NHM) were investigated. From these experiments, the two most potent peptide derivatives, 5-phenylvaleric acid-(D)His-Arg-Trp-$(Lys)_6NH_2$ (P 6) and 5-phenylvaleric acid-(D)His-Arg-Trp-$(Lys)_9NH_2$ (P 7) were selected for further studies. In ${\alpha}$-MSH depleted NHM cells, we have found that the treatment with 1 ${\mu}M$ of these two peptide derivatives, P 6 and P 7, inhibited the cell proliferation induced by the addition of 1 nM ${\alpha}$- MSH by 70% and 72%, respectively. In NHM cells without previous ${\alpha}$-MSH depletion, 1 ${\mu}M$ treatment in the presence of 10 nM ${\alpha}$-MSH resulted in 70% (P 6) and 80% (P 7) decrease in cell growth and 64% (P 6) and 71% (P 7) reduction in melanin synthesis, respectively. The peptide derivatives P 6 and P 7 were proved to have no apparent cytotoxicity and inhibited the elevation of intracellular cAMP concentration triggered by ${\alpha}$-MSH. In conclusion, our data suggest that the peptide derivatives reported in this study, 5-phenylvaleric acid-(D)His-Arg-Trp-$(Lys)_6NH_2$ (P 6) and 5-phenylvaleric acid-(D)His- Arg-Trp-$(Lys)_9NH_2$ (P 7) strongly antagonize ${\alpha}$-MSH, inhibit cell proliferation and melanin synthesis, and lower the intracellular cAMP concentration, hence have a promising potential as a novel skin lightening agent.

• Journal of Preventive Medicine and Public Health
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• 제34권4호
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• pp.372-378
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• 2001

Objectives : This study introduced the healthy life-year(HeaLY), a composite indicator of disease burden, and used it to estimate the burden of major cancers in Korea. Methods : We collected data from the national death certificate database, the national health insurance claims database and the abridged life table. This data was used to create a spreadsheet and estimate the burden of major cancers by sex in terms of HeaLYs. Results : The burden of 10 major cancers for males was 2,248.97 person-year in terms of HeaLYs. Stomach cancer, liver cancer, and lung cancer were responsible for 75.2% of the burden of 10 major cancers. The disease burden of 10 major cancers for females was estimated to be 1,567.58 person-years. About two thirds of HeaLYs lost were from stomach cancer, liver cancer, lung cancer, colorectal cancer, and breast cancer. The rankings among 10 major cancers were somewhat different in terms of both HeaLYs and deaths as the HeaLY method considers both mortality and morbidity. Conclusions : Despite the limitations of the data sources, we conclude that HeaLY can aid in setting policy priorities concerning major cancers by estimating the disease burden of these cancers. Time-series analysis of the disease burden using HeaLY and DALY will elucidate the strengths and weaknesses of both methods.

• Asian Pacific Journal of Cancer Prevention
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• 제15권8호
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• pp.3667-3673
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• 2014

Background: colorectal cancer (CRC) is the third most common type of cancers and the fourth leading cause of death worldwide. In Saudi Arabia, CRC accounts for 8.5% of all tumors; it ranks first among all cancers in males and third among females. The aim of this study was to link between different PARP-1 mutations and risk of CRC in Saudi population and to determine common variants of PARP-1 in Saudi CRC patients and normal individuals. Materials and Methods: DNA samples were isolated from fifty CRC patients and from a comparable number of control subjects then sequenced to detect different variations present in exons 3, 17, and 21 of the PARP-1 gene. Results and Conclusions: When comparing the genotype and allele frequencies of all detected SNPs in CRC patients with those in controls, we found none were significantly different for all variants even the most common SNP in PARP-1 gene (Val762Ala). However, two novel alterations in exon 21 were found to be associated with increased risk of CRC. The variants identified as (1) Lys933Asn [p-value 0.0318] and (2) Lys945Asn [p-value 0.0257]. Our results suggest that PARP-1 Lys933Asn and Lys945Asn alterations could be associated with increased risk of CRC in the Saudi population.

• 대한화장품학회지
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• 제30권3호
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• pp.399-404
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• 2004

새로운 Gly-His-Lys (약자로 GHK)가 결합된 키토산을 $N^{\alpha}-Fmoc$ 아미노산과 BOP 커플링 시약을 사용한 고상법에 의해 제조하였다. 이를 위해 키토산 마이크로 비드를 W/O 에멀젼 상분리법으로 평균 입자크기 70 마이크로미터로 얻었다. GHK 펩타이드는 순차적으로 고상법에 의해 키토산 마이크로비드 위에 커플링 하였다. 아미노산 분석을 실시한 결과 Gly, His, kys의 비율이 1.02:1.13:0.96의 비율로 나타나 이론치와 거의 일치함을 확인할 수 있었다. GHK 펩타이드가 결합된 마이크로비드의 세포증식 효과는 MTT 분석으로 측정하였다. 측정결과 GHK 펩타이드가 결합된 키토산 마이크로비드는 대조군인 펩타이드가 결합되지 않은 키토산 마이크로비드 자체에 비해 높은 세포증식 효과를 보였다.

• Plant Resources
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• 제8권1호
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• pp.60-66
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• 2005

Lysine is the first essential amino acid for optimal nutrient quality in rice grain. For the narrow genetic diversities of lysine contents in rice, somaclonal variation was the source of mutation in our breeding program. Biochemical selection was conducted using 1 mM S-(2-aminoethyl) cysteine followed by two passages of 5 mM lysine plus threonine in the callus subculture medium. The lysine contents in endosperm of all progenies recovered from the biochemical selection were higher than those of their donor cultivar 'Hwayeongbyeo'. These elevated lysine levels of mutants were successfully transmitted to $M_4$ generation. The lysine contents in endosperm varied 3.85 to $4.80\%$ compare to their donor cultivar 'Hwayeongbyeo' was $3.85\%$. Three of high-lysine germplasms, Lys-l, Lys-2 and Lys-7 were selected by biochemical selection and rapid screening methods. DNA analysis showed that a new insertion of Tos 17 which mapped to rice chromosome 11 on the high-lysine mutant, Lys-2.

• 생명과학회지
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• 제24권2호
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• pp.128-136
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• 2014

알긴산은 ${\alpha}$-L-guluronic acid와 ${\beta}$-D-mannuronic acid가 (1-4) 결합한 선형 산성다당류이다. 알긴산은 다양한 알긴산 분해효소들에 의하여 분해되는데 ${\beta}$-제거반응으로 비환원 말단에 이중결합이 있는 불포화 우론산 올리고머가 생산된다. 본 연구실에서는 이전에 Stenotrophomonas maltophilia KJ-2로부터 새로운 구조를 가진 polyMG lyase를 재조합하였다. KJ-2 polyMG lyase의 단백질구조를 예측하기 위하여 상동성 모델링을 한 결과 Azotobacter vinelandii가 생산하는 세 종류의 polyMG lyase들이 모두 PL7 family에 속하는 반면 KJ-2 polyMG lyase는 PL6 family에 속하였다. 또한 $^1H$-NMR spectra를 분석한 결과 polyMG lyase는 M-${\beta}$(1-4)-G 당쇄결합을 분해하고 G-${\alpha}$(1-4)-M 결합은 거의 분해하지 못하는 것으로 나타났다. 예측된 polyMG lyase 모델을 기초로 하여 14군데 아미노산 잔기를 선택하였으며 17개의 돌연변이체를 만들어 알긴산 분해효소의 활성을 측정하였다. Lys220Ala, Arg241Ala, Arg241Lys및 Arg265Ala 돌연변이체들은 완전히 알긴산 분해효소의 활성을 잃었으며 Arg155Ala, Gly303Glu 및 Tyr304Phe 돌연변이체들의 분해활성은 19.1-39.3%까지 감소하였다. 이러한 결과들로부터 Arg155, Lys220, Arg241, Arg265, Gly303 및 Tyr304 들은 알긴산 분해효소의 촉매활성과 기질결합에 중요한 잔기들임을 알 수 있다.

• Macromolecular Research
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• 제13권1호
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• pp.39-44
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• 2005

Ion-exchange polymeric stationary phases presenting amino acid and amino groups were prepared by the surface grafting of glycidyl methacrylate onto a silica gel surface and subsequent amination. Three kinds of amino acids-L-arginine (Arg), D-lysine (Lys), and D-histine (His)-were used in this study. An ion-exchange polymeric stationary phase presenting ethylene diamine (EDA) was also prepared by surface graft polymerization. Separation of the model proteins bovine serum albumin (BSA), chick egg albumin (CEA), and hemoglobin (Hb) was performed using the amino acid- and amine-derived columns. In separating the CEA/BSA mixture, the resolution time of BSA was longer than that of CEA when using the EDA column, whereas the resolution time of BSA was shorter than that of CEA when using the Arg, Lys, and His columns. In the separation of the Hb/BSA mixture, the resolution time of BSA was longer than that of Hb in the EDA column, whereas the resolution time of BSA was shorter than that of Hb in the amino acid columns (D-Lys, L-Arg, and D-His).