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Whole-genome resequencing analysis of 20 Micro-pigs
Da‑Hye Son,Nam‑Hyun Hwang,Won‑Hyong Chung,Ha‑Seung Seong,Hyungbum Lim,Eun‑Seok Cho,Jung‑Woo Choi,Kyung‑Soo Kang,Yong‑Min Kim 한국유전학회 2020 Genes & Genomics Vol.42 No.3
Background Miniature pigs have been increasingly used as mammalian model animals for biomedical research because of their similarity to human beings in terms of their metabolic features and proportional organ sizes. However, despite their importance, there is a severe lack of genome-wide studies on miniature pigs. Objective In this study, we performed whole-genome sequencing analysis of 20 Micro-pigs obtained from Medi Kinetics to elucidate their genomic characteristics. Results Approximately 595 gigabase pairs (Gb) of sequence reads were generated to be mapped to the swine reference genome assembly (Sus scrofa 10.2); on average, the sequence reads covered 99.15% of the reference genome at an average of 9.6-fold coverage. We detected a total of 19,518,548 SNPs, of which 8.7% were found to be novel. With further annotation of all of the SNPs, we retrieved 144,507 nonsynonymous SNPs (nsSNPs); of these, 5968 were found in all 20 individuals used in this study. SIFT prediction for these SNPs identified that 812 nsSNPs in 402 genes were deleterious. Among these 402 genes, we identified some genes that could potentially affect traits of interest in Micro-pigs, such as RHEB and FRAS1. Furthermore, we performed runs of homozygosity analysis to locate potential selection signatures in the genome, detecting several loci that might be involved in phenotypic characteristics in Micro-pigs, such as MSTN, GDF5, and GDF11. Conclusion In this study, we identified numerous nsSNPs that could be used as candidate genetic markers with involvement in traits of interest. Furthermore, we detected putative selection footprints that might be associated with recent selection applied to miniature pigs.
Purification and Properties of Phenylalanine Ammonia-lyase from Chinese Cabbage
Lim, Chang-Jin,Lim, Hye-Won,Sa, Jae-Hoon,Kim, Tae-Soo,Park, Eun-Hee,Park, Soo-Sun The Korea Science and Technology Center 1998 BMB Reports Vol.31 No.1
Phenylalanine ammonia-lyase (PAL; EC 4.3.1.5), the first enzyme in the phenylpropanoid biosynthesis, catalyzes the elimination reaction of ammonium ion from L-phenylalanine. PAL was purified from the cytosolic fraction of Chinese cabbage (Brassica campestris ssp. napus var. pekinensis) through ammonium sulfate fractionation, DEAE-cellulose chromatography, Sephadex G-200 chromatography, and Q-Sepharose chromatography. It consists of four identical subunits, the molecular mass of which was estimated to be about 38,000 daltons on SDS-PAGE. The optimal pH and temperature of the purified enzyme are 8∼9 and 45℃, respectively. Its activity is greatly inhibited by Zn²?? ion, and strongly activated by caffeic acid. The purified PAL has some different characteristics compared to those obtained with other PALs.
Won-Sun Choi,Eun-Soo Lim,반가영,Ji-Hye Kim,Yoo Seob Shin,Hae Sim Park,Young Min Ye 대한내과학회 2018 The Korean Journal of Internal Medicine Vol.33 No.1
Background/Aims: Chronic urticaria (CU) is a common skin disorder characterized by wheals and pruritus lasting more than 6 weeks. Due to its long duration and changeable symptoms, the quality of life (QOL) of patients with CU can be impaired substantially. We evaluated the CU-QOL, a previously validated CU-specific QOL measure, and investigated factors influencing QOL in chronic spontaneous urticaria (CSU) patients. Methods: A hospital-based cross-sectional study was performed on 390 adult patients diagnosed with CSU from March 2009 to December 2012 at the Allergy and Clinical Immunology Clinic at Ajou University Hospital. The CU-QOL questionnaire, urticaria activity score (UAS), combined angioedema, and serum total immunoglobulin E (IgE) levels were investigated. Results: The average CU-QOL score obtained from the questionnaire was 70.6 (of 100 points). The CU-QOL scores correlated significantly with the UAS, particularly with the 15-point UAS (UAS-15; coefficient –0.532, p < 0.01) rather than the 6-point UAS (–0.502, p < 0.01). The patients presenting with angioedema and urticaria had poorer scores in the urticaria symptom domain than those with urticaria alone (37.4 vs. 46.9, p = 0.004). Log-transformed serum total IgE levels correlated significantly with CU-QOL (–0.131, p < 0.05). Multivariate regression models indicated that severe CU (UAS-15 score ≥ 13), log (total IgE), and the presence of angioedema were significant predictors of impaired CU-QOL (< 85 points). Conclusions: CU has a substantial negative impact on QOL. The assessment of UAS-15, total IgE, and the presence of angioedema can be useful to predict QOL of the patients with CSU.
Direct Reduction of DTNB by E . coli Thioredoxin Reducatse
Hye Won Lim,Chagn Jin Lim 생화학분자생물학회 1995 BMB Reports Vol.28 No.1
Thioredoxin reductase is a flavoprotein oxidoreductase catalyzing the reduction of a cystine disulfide in thioredoxin. Thioredoxin, in turn, can reduce disulfide bonds in other proteins and serves as a reducing agent in enzymatic reactions such as those of ribonucleotide reductase and methionine sulfoxide reductase. In this work thioredoxin reductase was found to directly reduce DTNB in the absence of thioredoxin. This new reactivity of E. coli thioredoxin reductase was produced by relatively high concentrations of univalent cations such as Na^+, K^+, Li^+, and NH₄^+, and it appeared with the oxidation of NADPH. These results indicate that E. coli thioredoxin reductase may be slightly modified by univalent canons, and the modified enzyme directly reacts with DTNB. This DTNB-reducing activity offers a new assay method for E. coli thioredoxin reductase.
( Won Hee Jang ),( Kyoung Mi Jung ),( Hye Ri Yang ),( Mi Ri Lee ),( Haeng Sun Jung ),( Su Hyon Lee ),( Mi Young Park ),( Kyung Min Lim ) 한국응용약물학회 2015 Biomolecules & Therapeutics(구 응용약물학회지) Vol.23 No.4
The eye irritation potential of drug candidates or pharmaceutical ingredients should be evaluated if there is a possibility of ocular exposure. Traditionally, the ocular irritation has been evaluated by the rabbit Draize test. However, rabbit eyes are more sensitive to irritants than human eyes, therefore substantial level of false positives are unavoidable. To resolve this species difference, several three-dimensional human corneal epithelial (HCE) models have been developed as alternative eye irritation test methods. Recently, we introduced a new HCE model, MCTT HCETM which is reconstructed with non-transformed human corneal cells from limbal tissues. Here, we examined if MCTT HCETM can be employed to evaluate eye irritation potential of solid substances. Through optimization of washing method and exposure time, treatment time was established as 10 min and washing procedure was set up as 4 times of washing with 10 mL of PBS and shaking in 30 mL of PBS in a beaker. With the established eye irritation test protocol, 11 solid substances (5 non-irritants, 6 irritants) were evaluated which demonstrated an excellent predictive capacity (100% accuracy, 100% specificity and 100% sensitivity). We also compared the performance of our test method with rabbit Draize test results and in vitro cytotoxicity test with 2D human corneal epithelial cell lines.