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Purification and Characterization of Helicobacter pylori ${\gamma}$-Glutamyltranspeptidase
Song, Jae-Young,Choi, Yeo-Jeong,Kim, Jeong-Min,Kim, Yoo-Ree,Jo, Jin-Seong,Park, Jin-Sik,Park, Hee-Jin,Song, Yun-Gyu,Lee, Kon-Ho,Kang, Hyung-Lyun,Baik, Seung-Chul,Youn, Hee-Shang,Cho, Myung-Je,Rhee, Kw The Korean Society for Microbiology 2011 Journal of Bacteriology and Virology Vol.41 No.4
Gamma-glutamyltranspeptidase (GGT) was purified to electrophoretic homogeneity from the cell extract of H. pylori. The purified enzyme consisted of heavy and light subunits with molecular weights of 38 kDa and 21 kDa, respectively. N-terminal amino acid sequence of heavy and light subunits revealed that H. pylori GGT was processed into 3 parts for a signal peptide of 27 amino acid residues, a heavy subunit of 352 residues, and a light subunit of 188 residues during translation. The reaction rate for hydrolysis of ${\gamma}$-GpNA was 84.4 ${\mu}mol/min$ per milligram of protein, and that for the ${\gamma}$-glutamyl transfer from ${\gamma}$-GpNA to gly-gly was 23.8 ${\mu}mol/min$ per milligram of protein. The apparent Km values of H. pylori GGT for ${\gamma}$-glutamyl compounds were on the order of $10^{-3}$ to $10^{-4}$ M and those for acceptor peptides and amino acids were on the order of $10^{-1}$ to $10^{-2}$ M. The GGT protein kept approximately 80% of the initial enzymatic activity on incubation at $60^{\circ}C$ for 15 min. The optimum temperature and pH for reactions of both hydrolysis and transpeptidation were $40^{\circ}C$ and 9.0, respectively. The transpeptidation and hydrolysis reactions catalyzed by H. pylori GGT were strongly inhibited by L-Gln and moderately inhibited by L-Ala, L-Ser, ${\beta}$-chloro-L-Ala, and L-Glu. These results demonstrated that the biochemical properties of H. pylori GGT are different from those of other bacterial GGTs. Further, H. pylori GGT might degrade glutathione in the gastric mucous layer of humans if the enzyme could be secreted in the bacterial niches.
Yang, Kyeong Eun,Jang, Hyun‐,Jin,Hwang, In‐,Hu,Chung, Young‐,Ho,Choi, Jong‐,Soon,Lee, Tae‐,Hoon,Chung, Yun‐,Jo,Lee, Min‐,Seung,Lee, Mi Young,Yeo, Eui‐,J BLACKWELL PUBLISHING 2016 AGING CELL Vol.15 No.2
<P><B>Summary</B></P><P>Phenyl‐2‐pyridyl ketoxime (PPKO) was found to be one of the small molecules enriched in the extracellular matrix of near‐senescent human diploid fibroblasts (HDFs). Treatment of young HDFs with PPKO reduced the viability of young HDFs in a dose‐ and time‐dependent manner and resulted in senescence‐associated β‐galactosidase (SA‐β‐gal) staining and G2/M cell cycle arrest. In addition, the levels of some senescence‐associated proteins, such as phosphorylated ERK1/2, caveolin‐1, p53, p16<SUP>ink4a</SUP>, and p21<SUP>waf1</SUP>, were elevated in PPKO‐treated cells. To monitor the effect of PPKO on cell stress responses, reactive oxygen species (ROS) production was examined by flow cytometry. After PPKO treatment, ROS levels transiently increased at 30 min but then returned to baseline at 60 min. The levels of some antioxidant enzymes, such as catalase, peroxiredoxin II and glutathione peroxidase I, were transiently induced by PPKO treatment. SOD II levels increased gradually, whereas the SOD I and III levels were biphasic during the experimental periods after PPKO treatment. Cellular senescence induced by PPKO was suppressed by chemical antioxidants, such as N‐acetylcysteine, 2,2,6,6‐tetramethylpiperidinyloxy, and L‐buthionine‐(<I>S</I>,<I>R</I>)‐sulfoximine. Furthermore, PPKO increased nitric oxide (NO) production via inducible NO synthase (iNOS) in HDFs. In the presence of NOS inhibitors, such as L‐NG‐nitroarginine methyl ester and L‐NG‐monomethylarginine, PPKO‐induced transient NO production and SA‐β‐gal staining were abrogated. Taken together, these results suggest that PPKO induces cellular senescence in association with transient ROS and NO production and the subsequent induction of senescence‐associated proteins<B>.</B></P>
Yun, Yeo-Min,Song, Junghan,Ji, Misuk,Kim, Jeong-Ho,Kim, Yongkang,Park, Taesung,Song, Sang Hoon,Park, Seungman,Kim, Min Jin,Nho, Sun Jin,Oh, Kyung Won The Korean Society for Laboratory Medicine 2017 Annals of Laboratory Medicine Vol.37 No.1
<P><B>Background</B></P><P>For correct interpretation of the high-density lipoprotein cholesterol (HDL-C) data from the Korea National Health and Nutrition Examination Survey (KNHANES), the values should be comparable to reference values. We aimed to suggest a way to calibrate KNHANES HDL-C data from 2008 to 2015 to the Centers for Disease Control and Prevention (CDC) reference method values.</P><P><B>Methods</B></P><P>We derived three calibration equations based on comparisons between the HDL-C values of the KNHANES laboratory and the CDC reference method values in 2009, 2012, and 2015 using commutable frozen serum samples. The selection of calibration equation for correcting KNHANES HDL-C in each year was determined by the accuracy-based external quality assurance results of the KNHANES laboratory.</P><P><B>Results</B></P><P>Significant positive biases of HDL-C values were observed in all years (2.85-9.40%). We created the following calibration equations: standard HDL-C=0.872×[original KNHANES HDL-C]+2.460 for 2008, 2009, and 2010; standard HDL-C=0.952×[original KNHANES HDL-C]+1.096 for 2012, 2013, and 2014; and standard HDL-C=1.01×[original KNHANES HDL-C]-3.172 for 2011 and 2015. We calibrated the biases of KNHANES HDL-C data using the calibration equations.</P><P><B>Conclusions</B></P><P>Since the KNHANES HDL-C values (2008-2015) showed substantial positive biases compared with the CDC reference method values, we suggested using calibration equations to correct KNHANES data from these years. Since the necessity for correcting the biases depends on the characteristics of research topics, each researcher should determine whether to calibrate KNHANES HDL-C data or not for each study.</P>
Sudden Cardiac Arrest Postoperative Day due to Pulmonary Embolism
Yeo-Ul Yun,Sang-Min Shim,Yun-Sook Kim 순천향대학교 순천향의학연구소 2015 Journal of Soonchunhyang Medical Science Vol.21 No.2
Cardiac arrest one day after cesarean section is extremely rare. Obstetrical clinicians have low experience to these serious situations necessitating immediate first aid and knowledge of its differential diagnosis. A 33-year-old woman underwent elective repeat cesarean section at 38 weeks of gestation under spinal anesthesia. The patient underwent uneventful course on that day. Loss of consciousness occurred one day after cesarean section during her first ambulation. Immediate cardiac compression was performed and eventually resulted in good recovery of her heartbeat. Her condition was suitable disseminated intravascular coagulation (DIC). She developed acute ischemic pancreatitis after cardiac arrest. We describe the consideration of amniotic fluid embolism with DIC as most appropriate in this case. To our knowledge, our case is one of the most dangerous conditions after the cesarean section. Here, we report our case with a review of literatures.
Validation of QTLs associated with spikelets per panicle and grain weight in rice
Yeo, Sang-Min,Yun, Yeo-Tae,Kim, Dong-Min,Chung, Chong-Tae,Ahn, Sang-Nag Cambridge University Press 2014 Plant genetic resources Vol.12 No.1
<P>In this study, a near-isogenic line (BC4F10) CR572 developed by introgressing a chromosomal segment from <I>Oryza rufipogon</I> (accession no. 105491) into the <I>Oryza sativa</I> subsp. <I>japonica</I> cv. Hwaseong was found to exhibit a significant increase in the number of spikelets per panicle (SPP) and grain weight compared with the recurrent parent Hwaseong. Quantitative trait locus (QTL) analysis in F2 generation derived from the cross between CR572 and Hwaseong revealed that two QTLs, <I>qSPP1</I> and <I>qTGW1</I>, were linked to a simple sequence repeat marker, RM283, on chromosome 1. The additive effect of the <I>O. rufipogon</I> allele at <I>qSPP1</I> was 13 SPP, and 21.6% of the phenotypic variance was explained by the segregation of RM283. The <I>qTGW1</I> QTL explained 19.1% of the phenotypic variance for grain weight. Substitution mapping was carried out with five F3 lines derived from F2 plants having informative recombination breakpoints within the target region. Substitution mapping indicated the linkage of <I>qSPP1</I> and <I>qTGW1</I>. The grain yield of CR572 was 18.2 and 15.8% higher than that of Hwaseong at two locations, respectively, mainly due to the increase in 1000-grain weight and SPP. These results are very useful for QTL cluster transfer by molecular marker-assisted selection in rice breeding programmes and for QTL gene cloning by map-based cloning.</P>
Sang-Min Yeo,Yeo-Tae Yun,Chong-Tae Chung,Jung-Phil Seo,Hae-Hwang Kim,Sang-Nag Ahn 한국육종학회 2012 한국육종학회 심포지엄 Vol.2012 No.07
The objective of this study were to identify QTLs for agronomic traits using a set of introgression lines carrying wild rice (Oryza rufipogon) segment in cultivated rice (ssp. japonica cv. Hwaseongbyeo). Ninety-six ILs were evaluated for seven agronomic traits, amylose and protein contents. The proportion of the recurrent genome in ILs ranged from 87.8 to 100%, with an average of 96.7%. The mean number of homozygous and heterozygous donor segments were 2 (ranging 0-7) and 1.7 (ranging 0-6), respectively, and the majority of these segments had size less than 10 cM. A total of 22 quantitative trait loci were identified for 9 traits and each QTL explained 7.2% to 56.6% of the phenotypic variance. Some QTLs were clustered in a few chromosomal regions. A first cluster was located near RM527 on chromosome 6 with QTLs for culm length, panicle length, days to heading, 1000-grain weight and protein content. Three ILs with high spikelets per panicle compared to the recurrent parent were selected to detect and fine map the wild segments responsible for this variation. The results will be discussed.