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.

Fimasartan attenuates renal ischemia-reperfusion injury by modulating inflammation-related apoptosis

Cho, Jang-Hee,Choi, Soon-Youn,Ryu, Hye-Myung,Oh, Eun-Joo,Yook, Ju-Min,Ahn, Ji-Sun,Jung, Hee-Yeon,Choi, Ji-Young,Park, Sun-Hee,Kim, Chan-Duck,Kim, Yong-Lim The Korean Society of Pharmacology 2018 The Korean Journal of Physiology & Pharmacology Vol.22 No.6

Fimasartan, a new angiotensin II receptor antagonist, reduces myocyte damage and stabilizes atherosclerotic plaque through its anti-inflammatory effect in animal studies. We investigated the protective effects of pretreatment with fimasartan on ischemia-reperfusion injury (IRI) in a mouse model of ischemic renal damage. C57BL/6 mice were pretreated with or without 5 (IR-F5) or 10 (IR-F10) mg/kg/day fimasartan for 3 days. Renal ischemia was induced by clamping bilateral renal vascular pedicles for 30 min. Histology, pro-inflammatory cytokines, and apoptosis assays were evaluated 24 h after IRI. Compared to the untreated group, blood urea nitrogen and serum creatinine levels were significantly lower in the IR-F10 group. IR-F10 kidneys showed less tubular necrosis and interstitial fibrosis than untreated kidneys. The expression of F4/80, a macrophage infiltration marker, and tumor necrosis factor $(TNF)-{\alpha}$, decreased in the IR-F10 group. High-dose fimasartan treatment attenuated the upregulation of $TNF-{\alpha}$, interleukin $(IL)-1{\beta}$, and IL-6 in ischemic kidneys. Fewer TUNEL positive cells were observed in IR-F10 compared to control mice. Fimasartan caused a significant decrease in caspase-3 activity and the level of Bax, and increased the Bcl-2 level. Fimasartan preserved renal function and tubular architecture from IRI in a mouse ischemic renal injury model. Fimasartan also attenuated upregulation of inflammatory cytokines and decreased apoptosis of renal tubular cells. Our results suggest that fimasartan inhibited the process of tubular injury by preventing apoptosis induced by the inflammatory pathway.

림프절외 NK/T세포 림프종에서 Epstein-Barr Virus에 대한 연구

조민선,우소연 이화여자대학교 의과대학 2004 EMJ (Ewha medical journal) Vol.27 No.2

목적 : NK/T 세포 림프종의 발생은 EBV와 연관되어 있으며, EBV 아형과 LMP-1유전자의 30염기쌍 결손 돌연변이가 EBV의 종양유발성과 관련 있다는 연구 결과가 있는 바, 본원에서 NK/T 세포 림프종으로 진단된 예를 대상으로 이에 대해 알아보고자 하였다. 대상과 방법 : 본원에서 진단된 NK/T세포 림프종 16예를 대상으로 EBER-1 in situ hybridization으로 EBV 양성율을 검사하고, PCR방법을 통해 EBV아형과 결손 돌연변이 유무를 관찰하였다. 결과 : 16예 모두 EBER-1 in situ hybridization에서 양성반응을 보였다. EBV는 모두 A 아형이었고 LMP-1의 30염기쌍 결손은 전체 16예 중 10예(62.5%)에서 있었고 나머지 6예는 wild type이었다. 결론 : 본원에서 진단된 NK/T세포 림프종 모든 예어서 EBV를 발견할 수 있었으며, EBV는 모두 A아형이었다. LMP-1 결손 돌연변이는 62.5%에서 관찰되었으며 예후와의 연관성은 보다 많은 예에 대한 연구를 통해 밝혀져야 할 것이다. Objectives: Epstein-Barr virus(EBV) is associated with development of various types of lymphoma, especially NK/T cell lymphoma. Recently, its subtypes and LMP-1, major oncoprotein of EBV, have been studied. We investigated the frequency of EBV, its subtypes, and LMP-1 status on the cases diagnosed at Ewha university hospital between 1993 and 2002. Material and Methods: Sixteen cases of NK/T cell lymphomas were studied. In situ hybridization for EBER-1 mRNA and PCR for EBV subtypes and 30 base pair deletion of LMP-1 were done. Results: All cases showed EBV positivity by EBER in situ hybridization. All cases contained Type A viruses and 10 cases(62.5%) revealed LMP-1 30bp deletion. Conclusion: EBV act as a causative role in the development of NK/T cell lymphoma. The exact role of LMP-1 30bp deletion variant in the lymphomatogenesis should be studied with larger number of cases.

Epstein-Barr 바이러스 형질전환법을 이용한 파상풍 톡소이드에 대한 사람 단세포군 항체의 생산

유승민,조정제,호순태,하윤문 大韓免疫學會 1993 大韓免疫學會誌 Vol.15 No.-

For production of tetanus toxoid(TT) specific human monoclonal antibodies, anti-TT antibody secreting peripheral B lymphocytes were separated by rosetting with TT-coated SRBC. And, B lymphoblastoid cell line(BLCL) was established by Epstein-Barr Virus(EBV) transformation methods. Stable BLCLs were established after three times of cloning by limiting dilution. To establish the maximal antibody producing condition, several environmental factors were tested. The optimal condition for maximum antibody prodiction was 4 day culture of 1 x 105cells/ml concentration in RPMI 1640 complete culture media supplemented with 20% fetal calf serum, and the maximum concentration of secreted antibody was about 900ng/ml. The antibody production of BLCL was decreased during long-term culture after establishment of cell line, but antibody production was maintained by repeated cloning by limiting dilution.

Ca^(2+)-감수성 증가기전과 평활근 수축

이연리,김보경,최원호,김태경,이창권,배영민,조성일 건국대학교 의과학연구소 2004 건국의과학학술지 Vol.14 No.-

Smooth muscle contraction is regulated by intracellular Ca^(2+) ([Ca^(2+)]_(i)) and the phosphorylation of myosin light chain (MLC). However, various kinds of vasoconstrictors induce a further contraction at a given [Ca^(2+)]_(i), and elicit a sustained contraction under Ca^(2+) -depleted conditions, referred to as "Ca^(2+) -sensitization", in intact and membrane-pen-neabilized smooth muscle. Previously, several molecules, including protein kinase C (PKC), RhoA, and mitogen-activated protein kinases (MAPK), have been suggested as candidate regulators of Ca^(2+) -sensitization. In the present review, we describe the role of PKC, RhoA, and MAPK in the regulation of Ca^(2+) -sensitization, and suggest a new model in research for the regulation of smooth muscle contraction.

VRSA에 대한 항생물질을 생산하는 방선균의 분리

이지연,양시용,고은지,이승구,김시관,조왕식,송민동 建國大學校 自然科學硏究所 2002 建國自然科學硏究誌 Vol.13 No.2

본 연구가 VRSA에 대한 신규 항생물질 생산 균주를 얻기 위하여 실시하였다. 국내외에 채취한 토양시료에서 분리한 방선균 중, 임상분리 VRSA와 돌연변이 유발 VRSA에 대해 항균성을 나타내는 JY-24를 선별하였다. 선발 균주는 원통형의 포자 모양을 갖고 있으며, Bennett's 배지에 배양시 aerial mycelium color는 흰색을 나타냈으며, reverse side color는 대체로 노란색을 나타내었다. 항균물질 생산을 위한 탄소원으로는 fructose, mannose, 질소원으로는 yeast extract가 가장 우수하였으며 glucose의 첨가로 항균 물질 생산성이 증가하는 것으로 나타났다. 최적 초기 pH는 pH 7로 나타났으며, 28 ℃, 37℃, 45 ℃의 온도에서 균이 성장하는 것으로 나타났다. 또한 배양기간에 따라 항균물질 생산성을 조사한 결과, 임상분리 균주에 대해 5일 배양 시 가장 높았으나, 3일 배양과 7일 배양 시 동일한 생산성을 나타내 생산속도가 빠르며, 안정한 물질인 것이 파악되었다. 향후 균주동정 및 항균물질 정제에 관한 연구를 수행하고자 한다. An antibiotic substance producing strain JY-24, effective to the VRSA (Vancomycin-resistant Staphylococcus aureus), was isolated from soil. The spore chain of JY-24 was cylindrical, and its sulface was smooth. The aerial mass color of the strain was white, and its reverse side color was yellow. The strain did not produce soluble pigment. For the production antibiotics, fructose, mannose and yeast extract were favorable as carbon and nitrogen sources. Optimal initial pH for the production of the antibiotic was pH 7. Accumulation of the antibiotic in the culture broth reached at maximal level after 5 days cultivation in Bennett's medium.

Fimasartan attenuates renal ischemia-reperfusion injury by modulating inflammation-related apoptosis

Jang-Hee Cho,Soon-Youn Choi,Hye-Myung Ryu,Eun-Joo Oh,Ju-Min Yook,Ji-Sun Ahn,Hee-Yeon Jung,Ji-Young Choi,Sun -Hee Park,Chan-Duck Kim,Yong-Lim Kim 대한생리학회-대한약리학회 2018 The Korean Journal of Physiology & Pharmacology Vol.22 No.6

Fimasartan, a new angiotensin II receptor antagonist, reduces myocyte damage and stabilizes atherosclerotic plaque through its anti-inflammatory effect in animal studies. We investigated the protective effects of pretreatment with fimasartan on ischemia-reperfusion injury (IRI) in a mouse model of ischemic renal damage. C57BL/6 mice were pretreated with or without 5 (IR-F5) or 10 (IR-F10) mg/kg/day fimasartan for 3 days. Renal ischemia was induced by clamping bilateral renal vascular pedicles for 30 min. Histology, pro-inflammatory cytokines, and apoptosis assays were evaluated 24 h after IRI. Compared to the untreated group, blood urea nitrogen and serum creatinine levels were significantly lower in the IR-F10 group. IR-F10 kidneys showed less tubular necrosis and interstitial fibrosis than untreated kidneys. The expression of F4/80, a macrophage infiltration marker, and tumor necrosis factor (TNF)-α, decreased in the IR-F10 group. High-dose fimasartan treatment attenuated the upregulation of TNF-α, interleukin (IL)-1β, and IL-6 in ischemic kidneys. Fewer TUNEL positive cells were observed in IR-F10 compared to control mice. Fimasartan caused a significant decrease in caspase-3 activity and the level of Bax, and increased the Bcl-2 level. Fimasartan preserved renal function and tubular architecture from IRI in a mouse ischemic renal injury model. Fimasartan also attenuated upregulation of inflammatory cytokines and decreased apoptosis of renal tubular cells. Our results suggest that fimasartan inhibited the process of tubular injury by preventing apoptosis induced by the inflammatory pathway.

연령에 따른 한국인의 음향지표 변화와 특성

김형태,조승호,윤성문,선동일,김민식 대한이비인후과학회 2000 대한이비인후과학회지 두경부외과학 Vol.43 No.1

Gitelman씨 증후군 1예

차영학,송명준,신요식,김성민,김영욱,김윤권,김소연,김영중,조민구,이권전 全南大醫大 2001 전남의대학술지 Vol.37 No.3

제 2형 콜라겐에 의해 경구관용 유도된 DBA/1 mice에서의 세포면역반응

양형인 ( Hyung In Yang ),김완욱 ( Wan Uk Kim ),민도준 ( Do Jun Min ),박성환 ( Sung Hwan Park ),홍연식 ( Yeon Sik Hong ),이상헌 ( Sang Heon Lee ),조철수 ( Chul Soo Cho ),김호연 ( Ho Youn Kim ) 대한류마티스학회 2000 대한류마티스학회지 Vol.7 No.4

Objective: To investigate the dosage of bovine type II collagen (BnCII) for the induction of oral tolerance in CIA animals, and to verify the changes of immune response and TGF-β production of mesenteric lymph node cells in tolerized CIA animals. Methods: Oral tolerance was induced by feeding of variable doses (5㎍, 10㎍, 20㎍ and 40㎍) of BnCII to DBA/1 mice 4 times per week during 2 weeks, and control mice were given ovalbumin (1000㎍), before immunization. We examed clinical assessment; incidence of arthritis, severity of arthritis, arthritic limb by visual analysis. IgG antibodies to BnCII were measured by ELISA, T cell responses to BnCII and PHA were quantified by antigen (CII)-induced 3H-thymidine incorporation into lymphocytes of mesenteric lymph node, draining lymph node, and spleen. TGF-β in supernatants obtained from lymph node culture medium was measured by ELISA. Results: Arthritis limbs were observed in 100% of control at 5 weeks after subcutaneous BnCII injection. The incidences of CIA in all tolerized group were significantly lower than that in control 5 weeks after immunization (control 100% vs. 5㎍ feeding group: 50%, 10㎍ feeding group: 50%, 20㎍ feeding group: 50%, 40㎍ feeding group: 55.5%, P<0.01). In comparison to control, mean articular indices were lower in all tolerized groups (control 5.13: 5㎍ feeding group 3.50, 10㎍ feeding group 2.75, 20㎍ feeding group 2.87, 40㎍ feeding group 2.63, P<0.05). Arthritic limbs were also significantly lower in tolerized groups (control 58.3: 5㎍ feeding group 20.8, 10㎍ feeding group 16.7, 20㎍ feeding group 20.8, 40㎍ feeding group 20.8, P<0.05). The titers of IgG antibody to CII were lower in tolerized group than that in control [tolerized group; median 10 (min. 0, max. 48), control; median 33 (min. 8.6, max. 101), P<0.05]. The proliferative responses to BnCII were significantly suppressed in tolerized (control 8010±2319cpm, tolerized group 4500±2060cpm, P<0.01). High TGF-β production was noted in tolerized group (control; 28pg/ml, BnCII feeding group; 73pg/ml). Conclusion: Oral tolerance in DBA/1 mice was successfully induced from low doses of BnCII (5㎍) and suppressed T and B cell function in conjunction with increased TGF-β production may play an important role for the induction of CII induced oral tolerance in DBA/1 mice.