• 제목/요약/키워드: HSP-60

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Screening and Identification of Antigenic Proteins from the Hard Tick Dermacentor silvarum (Acari: Ixodidae)

  • Zhang, Tiantian;Cui, Xuejiao;Zhang, Jincheng;Wang, Hui;Wu, Meng;Zeng, Hua;Cao, Yuanyuan;Liu, Jingze;Hu, Yonghong
    • Parasites, Hosts and Diseases
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    • v.53 no.6
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    • pp.789-793
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    • 2015
  • In order to explore tick proteins as potential targets for further developing vaccine against ticks, the total proteins of unfed female Dermacentor silvarum were screened with anti-D. silvarum serum produced from rabbits. The results of western blot showed that 3 antigenic proteins of about 100, 68, and 52 kDa were detected by polyclonal antibodies, which means that they probably have immunogenicity. Then, unfed female tick proteins were separated by 12% SDS-PAGE, and target proteins (100, 68, and 52 kDa) were cut and analyzed by LC-MS/MS, respectively. The comparative results of peptide sequences showed that they might be vitellogenin (Vg), heat shock protein 60 (Hsp60), and fructose-1, 6-bisphosphate aldolase (FBA), respectively. These data will lay the foundation for the further validation of antigenic proteins to prevent infestation and diseases transmitted by D. silvarum.

Changing Proteins in Granulosa Cells during Follicular Development in Pig (돼지 난포 발달 시 과립막 세포에서 발현되는 단백질의 변화)

  • Chae, In-Soon;Jang, Dong-Min;Cheong, Hee-Tae;Yang, Boo-Keun;Park, Choon-Keun
    • Reproductive and Developmental Biology
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    • v.33 no.3
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    • pp.183-187
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    • 2009
  • This study analyzed change of proteins in granulosa cells during the porcine follicuar development by proteomics techniques. Granulosa cells of the follicles, of which the diameter is $2{\sim}4\;mm$ and $6{\sim}10\;mm$, were collected from ovary of slaughtered pig that each follicle of diameter $1{\sim}4\;mm$ and $6{\sim}10\;mm$. We extracted glanulosa cell proteins by M-PER Mammalian Protein Extraction Reagent. Proteins were refined by clean-up kit and quantified by Bradford method until total protein was $200{\mu}l$. Immobilized pH gradient(IPG) strip used 18 cm, $3{\sim}10\;NL$. SDS-PAGE used 10% acrylamide gel. After silver staining, Melanie 7 and naked eye test were used for spot analyzation. Increasing proteins in glanulosa cell of $6{\sim}10\;mm$ follicle were 7 spots. This spots were analyzed by MALDI-TOF MS and searched on NCBInr. In results, 7 spots were similar to zinc/ling finger protein 3 precursor (RING finger protein 203), angiomotin, heat shock 60 kDa protein 1 (chaperonin) isoform 1 (HSP60), similar to transducin-like enhancer protein 1 (TLE 1), SH3 and PX domains 2A (SH3PXD2A). Those proteins were related with transfer between cells. Increase of proteins has an effect on follicular development.

Correlation between Clinicomorphologic Findings and Clinical Outcome in Childhood $Henoch-Sch\"{o}nlein$ Purpura Nephritis (소아 $Henoch-Sch\"{o}nlein$ 자반병 신염의 임상양상 및 병리소견과 임상경과)

  • Huh Yun-Jung;Shin Jae-Il;Park Jee-Min;Jeong Hyeon-Joo;Lee Jae-Seung
    • Childhood Kidney Diseases
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    • v.7 no.1
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    • pp.30-37
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    • 2003
  • Purpose : $Henoch-Sch\"{o}nlein$ purpura(HSP) is usually a self-limited disease with a good eventual outcome. The prognosis of HSP is mainly determined by the renal involvement. In this study, We evaluated children with biopsy-proven $Henoch-Sch\"{o}nlein$ purpura nephritis about the clinical outcome correlated with renal manifestation and morphologic findings. Methods : The clinical features, initial laboratory and pathologic findings, and clinical out-come were evaluated in 60 children with biopsy-proven $Henoch-Sch\"{o}nlein$ purpura nephritis at Yonsei University Severance Hospital during the period from Jan. 1990 to Dec. 2002. Results : The ratio of male to female patients was 1.2:1. The interval between the onset of $Henoch-Sch\"{o}nlein$ purpura and renal manifestation was less than 3 months in 81% of the patients. Initial renal manifestation was microscopic hematuria in 100% of patients, isolated hematuria in 15%, acute nephritic syndrome in 7%, nephrotic syndrome In 22% of patients. Renal manifestation correlated with clinical outcome. Grade II and III were the most common in histologic grades of ISKDC. Renal pathologic finding correlated with clinical outcome. Conclusion : Renal manifestation and pathologic findings correlated with the clinical out-come. It is necessary to evaluate the correlation between pathologic findings and treatment.

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A Study of Equipment Accuracy and Test Precision in Dual Energy X-ray Absorptiometry (골밀도검사의 올바른 질 관리에 따른 임상적용과 해석 -이중 에너지 방사선 흡수법을 중심으로-)

  • Dong, Kyung-Rae;Kim, Ho-Sung;Jung, Woon-Kwan
    • Journal of radiological science and technology
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    • v.31 no.1
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    • pp.17-23
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    • 2008
  • Purpose : Because there is a difference depending on the environment as for an inspection equipment the important part of bone density scan and the precision/accuracy of a tester, the management of quality must be made systematically. The equipment failure caused by overload effect due to the aged equipment and the increase of a patient was made frequently. Thus, the replacement of equipment and additional purchases of new bonedensity equipment caused a compatibility problem in tracking patients. This study wants to know whether the clinical changes of patient's bonedensity can be accurately and precisely reflected when used it compatiblly like the existing equipment after equipment replacement and expansion. Materials and methods : Two equipments of GE Lunar Prodigy Advance(P1 and P2) and the Phantom HOLOGIC Spine Road(HSP) were used to measure equipment precision. Each device scans 20 times so that precision data was acquired from the phantom(Group 1). The precision of a tester was measured by shooting twice the same patient, every 15 members from each of the target equipment in 120 women(average age 48.78, 20-60 years old)(Group 2). In addition, the measurement of the precision of a tester and the cross-calibration data were made by scanning 20 times in each of the equipment using HSP, based on the data obtained from the management of quality using phantom(ASP) every morning (Group 3). The same patient was shot only once in one equipment alternately to make the measurement of the precision of a tester and the cross-calibration data in 120 women(average age 48.78, 20-60 years old)(Group 4). Results : It is steady equipment according to daily Q.C Data with $0.996\;g/cm^2$, change value(%CV) 0.08. The mean${\pm}$SD and a %CV price are ALP in Group 1(P1 : $1.064{\pm}0.002\;g/cm^2$, $%CV=0.190\;g/cm^2$, P2 : $1.061{\pm}0.003\;g/cm^2$, %CV=0.192). The mean${\pm}$SD and a %CV price are P1 : $1.187{\pm}0.002\;g/cm^2$, $%CV=0.164\;g/cm^2$, P2 : $1.198{\pm}0.002\;g/cm^2$, %CV=0.163 in Group 2. The average error${\pm}$2SD and %CV are P1 - (spine: $0.001{\pm}0.03\;g/cm^2$, %CV=0.94, Femur: $0.001{\pm}0.019\;g/cm^2$, %CV=0.96), P2 - (spine: $0.002{\pm}0.018\;g/cm^2$, %CV=0.55, Femur: $0.001{\pm}0.013\;g/cm^2$, %CV=0.48) in Group 3. The average error${\pm}2SD$, %CV, and r value was spine : $0.006{\pm}0.024\;g/cm^2$, %CV=0.86, r=0.995, Femur: $0{\pm}0.014\;g/cm^2$, %CV=0.54, r=0.998 in Group 4. Conclusion: Both LUNAR ASP CV% and HOLOGIC Spine Phantom are included in the normal range of error of ${\pm}2%$ defined in ISCD. BMD measurement keeps a relatively constant value, so showing excellent repeatability. The Phantom has homogeneous characteristics, but it has limitations to reflect the clinical part including variations in patient's body weight or body fat. As a result, it is believed that quality control using Phantom will be useful to check mis-calibration of the equipment used. A value measured a patient two times with one equipment, and that of double-crossed two equipment are all included within 2SD Value in the Bland - Altman Graph compared results of Group 3 with Group 4. The r value of 0.99 or higher in Linear regression analysis(Regression Analysis) indicated high precision and correlation. Therefore, it revealed that two compatible equipment did not affect in tracking the patients. Regular testing equipment and capabilities of a tester, then appropriate calibration will have to be achieved in order to calculate confidential BMD.

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LC-MS/MS Analysis of Surface Layer Proteins as a Useful Method for the Identification of Lactobacilli from the Lactobacillus acidophilus Group

  • Podlesny, Marcin;Jarocki, Piotr;Komon, Elwira;Glibowska, Agnieszka;Targonski, Zdzislaw
    • Journal of Microbiology and Biotechnology
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    • v.21 no.4
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    • pp.421-429
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    • 2011
  • For precise identification of a Lactobacillus K1 isolate, LC-MS/MS analysis of the putative surface layer protein was performed. The results obtained from LTQ-FT-ICR mass spectrometry confirmed that the analyzed protein spot is the surface layer protein originating from Lb. helveticus species. Moreover, the identified protein has the highest similarity with the surface layer protein from Lb. helveticus R0052. To evaluate the proteomic study, multilocus sequence analysis of selected housekeeping gene sequences was performed. Combination of 16S rRNA sequencing with partial sequences for the genes encoding the RNA polymerase alpha subunit (rpoA), phenylalanyl-tRNA synthase alpha subunit (pheS), translational elongation factor Tu (tuf), and Hsp60 chaperonins (groEL) also allowed to classify the analyzed isolate as Lb. helveticus. Further classification at the strain level was achieved by sequencing of the slp gene. This gene showed 99.8% identity with the corresponding slp gene of Lb. helveticus R0052, which is in good agreement with data obtained by nano-HPLC coupled to an LTQ-FT-ICR mass spectrometer. Finally, LC-MS/MS analysis of surface layer proteins extracted from three other Lactobacillus strains proved that the proposed method is the appropriate molecular tool for the identification of S-layer-possessing lactobacilli at the species and even strain levels.

Influence of Oxygen Consumption on Pregnancy Rates of Hanwoo Calves following Embryo Transfer

  • Kim, Hyun;Bok, Nan-Hee;Kim, Sung-Woo;Do, Yoon-Jung;Seong, Hwan-Hoo;Kim, Jae-Hwan;Kim, Dong Hun;Kim, Min-Kyu;Ko, Yeoung-Gyu
    • Journal of Embryo Transfer
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    • v.29 no.3
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    • pp.257-264
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    • 2014
  • Recently, several approaches have been used to measure the oxygen consumption rates of individual embryos, but relationship between oxygen consumption and pregnancy rates of Hanwoo following embryo transfer has not yet been reported. In this study, we investigated the correlation between oxygen consumption rate and pregnancy rates of Hanwoo embryo using a SECM. In addition to, the expression of apoptosis-related genes was determined using real-time PCR by extracting RNA according to the oxygen consumption of in vivo embryo. First, we found that the oxygen consumption significantly increased in blastocyst-stage embryos (blastocyst) compared to early blastocyst stage embryos, indicating that oxygen consumption reflects the embryo quality (Grade I). The oxygen consumption or GI blastocysts were significantly higher than those of GII blastocysts ($10.2{\times}10^{14}/mol\;s^{-1}$ versus $6.4{\times}10^{14}/mol\;s^{-1}$, p<0.05). Pregnant rate in recipient cow was 0, 60 and 80% in the transplantation of embryo with the oxygen consumption of below 10.0, 10.0~12.0 and over $12.0{\times}10^{14}/mol\;s^{-1}$, respectively. Apoptosis regulatory genes, Hsp-70.1 were significantly increased in over-10.0 group than below 10.0 group but in Caspase-3, Bax and P53 gene, there was no significant difference. In conclusion, These results suggest that measurement of oxygen consumption maybe help increase the pregnant rate of Hanwoo embryos.

Gene Expression Analyses of Mutant Flammulina velutipes (Enokitake Mushroom) with Clogging Phenomenon

  • Ju-Ri Woo;Doo-Ho Choi;Muhammed Taofiq Hamza;Kyung-Oh Doh;Chang-Yoon Lee;Yeon-Sik Choo;Sangman Lee;Jong-Guk Kim;Heeyoun Bunch;Young-Bae Seu
    • Mycobiology
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    • v.50 no.5
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    • pp.366-373
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    • 2022
  • Regulation of proper gene expression is important for cellular and organismal survival, maintenance, and growth. Abnormal gene expression, even for a single critical gene, can thwart cellular integrity and normal physiology to cause diseases, aging, and death. Therefore, gene expression profiling serves as a powerful tool to understand the pathology of diseases and to cure them. In this study, the difference in gene expression in Flammulina velutipes was compared between the wild type (WT) mushroom and the mutant one with clogging phenomenon. Differentially expressed transcripts were screened to identify the candidate genes responsible for the mutant phenotype using the DNA microarray analysis. A total of 88 genes including 60 upregulated and 28 downregulated genes were validated using the real-time quantitative PCR analysis. In addition, proteomic differences between the WT and mutant mushroom were analyzed using two-dimensional gel electrophoresis and matrix-assisted laser desorption/ionization-time of flight (MALDI-TOF). Interestingly, the genes identified by these genomic and proteomic analyses were involved in stress response, translation, and energy/sugar metabolism, including HSP70, elongation factor 2, and pyruvate kinase. Together, our data suggest that the aberrant expression of these genes attributes to the mutant clogging phenotype. We propose that these genes can be targeted to foster normal growth in F. velutipes.

Change of Protein Patterns in Granulosa Cells and Corpus Luteum during the Ovarian Cycle in Cows (소의 난소주기 동안 과립막세포와 황체조직에서 단백질 패턴의 변화)

  • Song, Eun-Ji;Lee, Yong-Seung;Lee, Sang-Hee;Yoo, Han-Jun;Park, Joung-Jun;Cheong, Hee-Tae;Yang, Boo-Keun;Park, Choon-Keun
    • Reproductive and Developmental Biology
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    • v.37 no.3
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    • pp.149-154
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    • 2013
  • The aim of this study was to evaluate the changes of protein patterns in granulosa cells and corpus luteum in ovaries during the estrus cycle in cows. The estrus cycle was devided into five steps of follicular, ovulatory, early-luteal, mid-luteal and late-luteal phases. In results, 61 spots of total 85 spots were repeated on follicular phase and 51 spots of total 114 spots were repeated on ovulatory phase. The 40 spots of total 129 spots were repeated on early-luteal phase and 49 spots of total 104 spots were repeated on mid-luteal phase. Also 41 spots of total 60 spots were repeated on late-luteal phase. On the other hands, the 16 spots were indicated difference in follicular phase and ovulation phase had a difference 10 spots. It was showed difference No. 103 spot in ovulation phase, No. 135 spot in early-luteal phase and No. 175 and 176 spots in mid-luteal phase. Also, the 11 spots were expressed specifically in mid-luteal phase and No. 178 and 179 spots were difference of expression in late-luteal phase. We confirmed that there were 7 spots for ovulation, 4 spots for luteinization and 2 spots for luteolysis. Spot No. 89~93 in ovulation phase were transferrin, and spot No.94~98 were HSP60. Spot No. 103 was Dusty PK, spot No. 135 was OGDC-E2, and spot No. 175 and 176 were Rab GDI beta from luteinization. Spot No. 178 and 179 in luteolysis were vimentin. This results suggest that will be help to basic data about infertility.

Effects of Short-term Acute Heat Stress on Physiological Responses and Heat Shock Proteins of Hanwoo Steer (Korean Cattle)

  • Baek, Youl-Chang;Kim, Minseok;Jeong, Jin-Young;Oh, Young-Kyoon;Lee, Sung-Dae;Lee, Yoo-Kyung;Ji, Sang-Yun;Choi, Hyuck
    • Journal of Animal Reproduction and Biotechnology
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    • v.34 no.3
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    • pp.173-182
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    • 2019
  • This study was performed to evaluate the effect of heat stress on the status of physiological responses, blood parameter, serum T3 and cortisol, and heat shock proteins (HSP 27, 70, and 90) of Hanwoo cattle. Six Hanwoo steers (242.8 ± 7.2 kg of BW) were housed in the climate-controlled respiration chambers. The experiment consisted of 7 days (control; 0 day) at thermoneutral (air temperature (Ta) of 15℃ and relative humidity (RH) of 60%; temperature-humidity index (THI) = 64), and by 3 and 6 days (treatment groups) at heat stress (Ta of 35℃ and RH of 60%; THI = 87). Body temperature of each parts (frank, rump, perineum and foot) and rectal temperature elevated in heat stress groups (3 days and 6 days) than the control group (0 day). Respiration rates increased in 3 days and 6 days (88.5 ± 0.96 bpm and 86.3 ± 0.63 bpm, respectively) from 0 days (39.5 ± 0.65 bpm). Feed intake significantly decreased in heat stress groups (3 days and 6 days, 3.7 ± 0.14 kg and 4.0 ± 0.15 kg, respectively) than the control group (0 day, 5.0 ± 0.00 kg). In addition, final BW significantly decreased in heat stress groups (3 days and 6 days, 211.8 ± 4.75 kg and 215.5 ± 3.50 kg, respectively) than the control group (0 day, 240.0 ± 25.00 kg). However, heat stress has no significant effect on blood parameter, serum T3 and cortisol. Nevertheless, heat stress increased HSPs mRNA expression in liver tissue, and serum concentration of HSPs. Despite Hanwoo cattle may have high adaptive ability to heat stress, our results suggested that heat stress directly effect on body temperature and respiration rate as well as serum and tissue HSPs. Therefore, we are recommended that HSPs could be the most appropriate indicators of Hanwoo cattle response to heat stress.

The Effects of Chungganhaeju-tang(Qingganjiejiu-tang) on Alcoholic Liver Damages by Applying Proteomics (청간해주탕(淸肝解酒湯)이 알코올 유발 간섬유화와 단백질 발현에 미치는 영향)

  • Jun, Jae-Hyun;Kim, Young-Chul;Lee, Jang-Hoon;Woo, Hong-Jung
    • The Journal of Internal Korean Medicine
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    • v.29 no.2
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    • pp.469-489
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    • 2008
  • Objectives : The purpose of this study was to investigate the effects of Chungganhaeju-tang(Qingganjiejiu-tang) on alcoholic liver damaged by applying proteomics. Materials and Methods : Sprague-Dawley rats were used in this experiment the rats were divided into the normal group, the control group(alcohol) and the sample group(CGHJT +alcohol). The ethanol was orally administered twice a day for 6 weeks in the control and sample groups. Water instead of ethanol was orally administered twice a day for 6 weeks in the normal group. CGHJT extract was orally administered once a day for 6 weeks in the sample group. The livers of each group were processed and assessed by histology, Western Blot, $Oxyblot^{TM}$, CBB and 2-dimensional electrophoresis. Results : In the histological findings of the liver, CGHJT inhibited hepatic fibrogenesis induced by alcohol. TIMP-1 decreased in the sample group assessed by western blot and statistical significance was noted by dot blotting(p<0.05). In the $Oxyblot^{TM}$, protein oxidation induced by alcohol treatment decreased with CGHJT. In the 2-dimensional electrophoresis finding, increased proteins alcohol such as HSP 60, 60kDa heat shock protein, 3-mercaptopyruvate sulfurtransferase were normalized by CGHJT. CGHJT was considered to normalize the anti-oxidation activity elevated by alcohol. In the 2-dimensional electrophoresis finding, increased oxidized proteins such as actin, prolyl 4-hydroxylase beta polypeptide, 94kDa glucose regulated protein(GRP94), heat shock protein 90-alpha(HSC86), calreticulin precursor(CRP55), ATP synthase beta chain mitochondrial precursor, caspase-8 precursor, and dihydrolipoamide succinyltransferase(E2) decreased with CGHJT. CGHJT was considered to reduce the oxidative stress of alcohol. Conclusion : Chungganhaeju-tang(Qingganjiejiu-tang) exerts an inhibitory effect against the fibrosis and protein oxidation induced by alcohol treatment of rat liver. CGHJT was considered to normalize the elevated anti-oxidation activity by alcohol and to reduce the level of oxidative stress due to alcohol.

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