• Title/Summary/Keyword: Blood enzyme

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Inhibitory Substance Produced by Aspergillus sp. on the Snake Venom Proteinase - Isolation of Microorganism and Biological Activities of the Inhibitor - (Aspergillus 속 균주가 생성되는 사독 Proteinase에 대한 저해물질 - 균의 분리 및 저해물질의 생물학적 작용상 -)

  • Hyun, Nam-Joo;Seu, Jung-Hwn
    • Microbiology and Biotechnology Letters
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    • v.15 no.2
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    • pp.129-134
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    • 1987
  • Aspergillus sp. (MK-24) producing a biological active substance that inhibited the venom proteinase activity was isolated from soil. The substance also inhibited the activity of trypsin and coagulation of blood, but did not inhibit papain, $\alpha$-chymotrypsin and pepsin. The substance was partially purified from culture filtrate by precipitaion with acetone, and by chromatography of DEAE-Sepadex A-50 column and Amberlite IRC-50 ion exchange. The inhibitory substance was stable in the wide pH range from 2.0 to 12.0 at 37$^{\circ}C$, but not stable at $65^{\circ}C$ in the alkaline pH. Only 12% of the activity was decreased by the heat treatment at 10$0^{\circ}C$ for two hours. The inhibition on venom proteinase (Agkistrodon bromohoffi brevicaudus) was a mixed type. The inhibitory activity depended on the preincubation time and completely depressed by cupric, zinc and cobalt ions. The inhibition on the venom proteinase was appeared strongly on casein but not on ovalbumin or hemoglobin as a substrate.

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Radioprotective Effect of Quercetin Post-Treatment against γ-Irradiation-Induced Hepatocellular and Hematopoiectic System Damage in Mice (감마선 조사로 유도된 간세포와 조혈계 손상 마우스에서 퀘르세틴 투여 후의 방사선방호 효과)

  • Kang, Jung Ae;Yoon, Seon Hye;Rho, Jong Kook;Choi, Dae Seong;Jang, Beom-Su;Park, Sang Hyun
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.44 no.7
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    • pp.970-974
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    • 2015
  • This study was designed to evaluate the therapeutic effect of quercetin against radiation-induced hepatocellular and hematopoiectic damage in BALB/c mice. Mice were exposed to 6 Gy of ${\gamma}$-radiation and orally administered quercetin (25, 50 mg/kg b.w.) for 7 consecutive days. ${\gamma}$-Irradiation caused marked elevation of serum aspartate aminotransferase and alanine aminotransferase, levels as well as reduction of spleen index, thymus index, and the number of white blood cells. In addition, ${\gamma}$-irradiation induced significant elevation of lipid peroxidation as well as reduction of antioxidant enzyme activities, including superoxide dismutase, catalase, and glutathione peroxidase. However, post-treatment with quercetin resulted in a significant recovery of all of these parameters. These results suggest that quercetin acts as a potent radioprotector against irradiation-induced cellular damage in mice.

Pectinase-treated Panax ginseng protects against chronic intermittent heat stress-induced testicular damage by modulating hormonal and spermatogenesis-related molecular expression in rats

  • Kopalli, Spandana Rajendra;Cha, Kyu-Min;Lee, Sang-Ho;Ryu, Ji-Hoon;Hwang, Seock-Yeon;Jeong, Min-Sik;Sung, Jong-Hwan;Kim, Si-Kwan
    • Journal of Ginseng Research
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    • v.41 no.4
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    • pp.578-588
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    • 2017
  • Background: Elevated testicular temperature disrupts spermatogenesis and causes infertility. In the present study, the protective effect of enzymatically biotransformed Panax ginseng Meyer by pectinase (GINST) against chronic intermittent heat stress-induced testicular damage in rats was investigated. Methods: Male Sprague-Dawley rats (4 wk old, 60-70 g) were divided into four groups: normal control (NC), heat-stress control (HC), heat-stress plus GINST-100 mg/kg (HG100), and heat-stress plus GINST-200 mg/kg (HG200) treatment groups. Each dose of GINST (100 mg/kg and 200 mg/kg) was mixed separately with a regular pellet diet and was administered orally for 24 wk. For inducing heat stress, rats in the NC group were maintained at $25^{\circ}C$, whereas rats in the HC, HG100, and HG200 groups were exposed to $32{\pm}1^{\circ}C$ for 2 h daily for 6 mo. At week 25, the testes and serum from each animal were analyzed for various parameters. Results: Significant (p < 0.01) changes in the sperm kinematic values and blood chemistry panels were observed in the HC group. Furthermore, spermatogenesis-related molecules, sex hormone receptors, and selected antioxidant enzyme expression levels were also altered in the HC group compared to those in the NC group. GINST (HS100 and HS200) administration significantly (p < 0.05) restored these changes when compared with the HC group. For most of the parameters tested, the HG200 group exhibited potent effects compared with those exhibited by the HG100 group. Conclusion: GINST may be categorized as an important medicinal herb and a potential therapeutic for the treatment of male subfertility or infertility caused by hyperthermia.

The Effect of Poria Cocos Pharmacopuncture at SP6(Sameumgyo) and ST36(Joksamni) on Chronic Stress Model(CSM) Rats (삼음교(三陰交)(SP6)·족삼리(足三里)(ST36) 백복령약침(白茯笭藥鍼)이 만성 스트레스 모델 흰쥐에 미치는 영향)

  • Kim, Seon Wook;Kim, Jae Hong;Cho, Myoung Rae;Shin, Jeong Cheol
    • Journal of Acupuncture Research
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    • v.32 no.2
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    • pp.187-195
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    • 2015
  • Objectives : The purpose of this study is evaluating the effect of Poria cocos pharmacopuncture (PCP) at $SP_6$(Sameumgyo) and $ST_{36}$(Joksamni) on chronic stress model(CSM) rats. Methods : The experimental rats were assigned to five groups: normal, CSM(control), PCP1X, PCP2.5X and PCP5X. The experimental rats in CSM(control), PCP1X, PCP2.5X and PCP5X were put through a timetable to cause CSM. PCP($20{\mu}l$) was administered at $SP_6$, $ST_{36}$ to groups PCP1X, PCP2.5X and PCP5X once every three days for a total of six times. The weight of the rats was measured by an electronic scale after PCP injection, once every three days for a total of six times. Blood samples were taken from the rats for analysis of aspartate aminotransferase(AST) and alanine aminotransferase(ALT). Serotonin and norepinephrine were measured by enzyme-linked immunosorbent assay(ELISA) kit. Results : The experimental groups(PCP1X, 2.5X, 5X) showed significant body weight increase compared to the control group. Regarding AST and ALT changes in serum, the experimental groups(PCP2.5X, 5X) showed a significant decrease compared to the control group. For Serotonin changes in serum, the experimental group(PCP5X) showed a significant increase compared to the control group. For Norepinephrine changes in serum, the experimental groups(PCP2.5X) showed a significant decrease compared to the control group. Conclusions : These data suggest that PCP at $SP_6$, $ST_{36}$ has therapeutic effects on CSM rats.

Effect of Glycyrrhizae Radix on the Immune Responses(II) - Immuno-regulatory Action of Glycyrrhizin and Glycyrrhetinic Acid - (감초가 면역반응에 미치는 영향(II) - Glycyrrhizin 및 Glycyrrhetinic acid의 면역조절작용 -)

  • 한종현;오찬호;은재순
    • YAKHAK HOEJI
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    • v.35 no.3
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    • pp.174-181
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    • 1991
  • These experiments were conducted to investigate the effects of glycyrrhizin(GL) and glycyrrhetinic acid(GA) on histamine synthesis, lymphocyte blastogenesis in C57BL/6J mice splenocytes, IL-1 production, $Ca^{2+}$ uptake by macrophage-like P388D$_{1}$ cells and plaque forming cell assay against SRBC. Histamine contents, lymphocyte blastogenesis, IL-1 activity, $Ca^{2+}$ uptake and plaque forming cell were determined by enzyme isotope method, [sup 3/H]-thymidine incorporation, C3H/HeJ mouse thymocytes proliferation, the addition of 5 $\mu$Ci/ml $^{45}$Ca$^{2+}$ to P388D$_{1}$, cell suspension and assay to sheep red blood cell, respectively. Cytotoxicity, which was expressed as 50% mortality, was occurred by the addition of GL(10$^{-3}$M) and GA(10$^{-4}$M). Histamine production in mouse spleen cell culture was significantly increased by the addition of 0.25 $\mu\textrm{g}$/ml of Con A, after 48 hour incubation. Con A dependent T-lymphocyte proliferation was also enhanced by the addition of 0.25 .mu.g/ml of Con A. The effects of GL on histamine contents and T-lymphocyte proliferation were significantly decreased at high dose (10$^{-5}$M), while IL-1 activity was remarkably suppressed by 10$^{-8}$~10$^{-4}$M of GL. $Ca^{2+}$ uptake was not changed, but antibody production was increased by GL(10 mg/kg). GA inhibited histamine contents at 10$^{-9}$~10$^{-7}$ and depressed Con A (0.25 $\mu\textrm{g}$/ml) dependent T-lymphocyte proliferation at 10$^{-7}$~10$^{-5}$M of GA, but increased suboptimal dose (Con A 0.1 $\mu\textrm{g}$/ml) at 10$^{-9}$~10$^{-7}$M of GA. IL-1 activity was suppressed by 10$^{-8}$~10$^{-4}$M of GA and $Ca^{2+}$ uptake was enhanced by 10$^{-9}$~10$^{-6}$ of GA, but antibody production was not changed by GA. From the above results, it is suggested that GL and GA have immuno-regulatory action. GL decreased cell-mediated immune response, and increased humoral immune response at high dose. On the other hand, low dose of GA enhanced cell-mediated immune response, while high doses of GA decreased humoral immune reaction.

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Suppression Effects of Sopunghwalhyeol-tang(Shufenghuoxie-tang) on the Monosodium Iodoacetate-induced Osteoarthritis in Rats (소풍활혈탕(疎風活血湯)이 Monosodium Iodoacetate로 유발한 흰쥐의 골관절염 억제에 미치는 영향)

  • Kim, Dae-Hyoung;Jeong, Su-Hyeon;Seo, Il-Bok;Kim, Soon-Joong
    • Journal of Korean Medicine Rehabilitation
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    • v.21 no.1
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    • pp.57-77
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    • 2011
  • Objectives : This study was to investigate the suppression effects of Sopunghwalhyeol-tang(Shufenghuoxie-tang) on the monosodium iodoacetate-induced osteoarthritis in rats. Methods : Arthritis was induced by injection of monosodium iodoacetate(0.5 mg) into the both knee joints of rats. Arthritic rats were divided into control(n=8) and treated(n=8) group. Control group was taken distilled water for 20 days. Treated group was taken extracts of Sopunghwalhyeol-tang(Shufenghuoxie-tang) by oraly for same duration. Normal group(n=8) was injected with normal saline and was taken distilled water for 20 days. Macroscopic examination and histopathological study on articular cartilage of knee joint were operated at 20 days after injection. Proteoglycan(PG) content of articular cartilages of knee joint was represented by safranine O staining, was measured at 20 days after injection. Tumor necrosis $factor-{\alpha}(TNF-{\alpha})$, $interleukin-1{\beta}(IL-1{\beta})$, in synovial fluid were measured with enzyme-linked immuno sorbent assay(ELISA) kit at 20 days after injection. Immunohistochemical staining of cyclo-oxygenase-2(COX-2), inducible nitric oxide synthase(iNOS) in knee joints were observed at 20 days after injection. Results : 1. Lymphocytes in peripheral blood the treated group was significantly decreased compared with the control group. 2. PG content in articular cartilage of the treated group was significantly increased compared with the control group. 3. Histopathologically, osteoarthritic scores of the treated group was significantly decreased compared with the control group. 4. $TNF-{\alpha}$ content in synovial fluid of the treated group was significantly decreased compared with the control group. 5. COX-2 revelation index in chondrocytes and synovial membrane of the treated group was significantly decreased compared with the control group. 6. Matrix metalloproteinase-3(MMP-3) revelation index in chondrocytes and synovial membrane of the treated group was significantly decreased compared with the control group. Conclusions : On the basis of these results, we concluded that Sopunghwalhyeol-tang(Shufenghuoxie-tang) has inhibiting effects on the $TNF-{\alpha}$, COX-2 and MMP-3 secretion of chondrocytes and synovial membrane in Monosodium Iodoacetate-induced osteoarthritis model of rats.

Detection for Methicillin Resistant Staphylococcus aureus in Using Bio-Chip Based Loop Mediated Isothermal Amplification Assay (칩 기반 등온증폭법을 이용한 약제 내성 포도상구균의 검출)

  • Cho, Min-Ho;Jang, Won-Cheoul;Choi, Jae-Gu
    • Journal of the Korean Chemical Society
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    • v.57 no.1
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    • pp.81-87
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    • 2013
  • Staphylococcus aureus is the most important pathogen in nosocomial infections, including bloodstream infections. Prompt identification of S. aureus from blood cultures and detection of methicillin resistance are essential in cases of suspected sepsis. We have studied a new method for the sequence-specific visual detection of minute amounts of nucleic acids using intercalating reaction by addition of SYBR Green to amplicons of LAMP, and it's a unique gene amplification method in which DNA can be isothermally amplified using only one enzyme. Staphylococcus-LAMP, which targets the spa gene, encoding S.aureus-specific protein A, and the mecA gene, encoding penicillin-binding protein-2' for methicillin resistance, detected MRSA and MRSE. In this study, by using LAMP assay, I detected for Staphylococcus aureus and Staphylococcus epidermidis concentration in the clinical sample. The detection of Staphylococcus aureus and Staphylococcus epidermidis was tested by using serial 10-fold dilutions standard solution. I have accurate detected the limit of detection, sensitity, specificity and reproducibility of the assay. The Bio-chip based LAMP assay allowed easy, rapid, accurate and sensitive detection of infection with Staphylococcus and especially applicable in a resource-limited situation.

Effect of CD14, Toll-like receptors, cytoskeletal inhibitors and $NF-{\kappa}B$ inhibitor on MMP-8 release from human neutrophils induced by E. coli lipopolysaccharides. (E. coli lipopolysaccharides로 유도된 사람 호중구에서 CD14, Toll-like receptors, cytoskeletal inhibitors 그리고 $NF-{\kappa}B$ inhibitor가 MMP-8 분비에 미치는 영향)

  • Yang, Seung-Min;Kim, Tae-Il;Seol, Yang-Jo;Lee, Yong-Moo;Ku, Young;Chung, Chong-Pyoung;Han, Soo-Boo;Rhyu, In-Chul
    • Journal of Periodontal and Implant Science
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    • v.35 no.2
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    • pp.427-436
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    • 2005
  • Objective: MMP-8 is a neutrophil enzyme and its level increases in some inflammatory diseases, including periodontal disease. We knew that the lipopolysaccharide of E.coli(E-LPS) induced MMP-8 release from human neutrophils. E-LPS is known to induce the production and release of inflammatory cytokines through CD14, Toll-like receptor(TLR). In the present study, we investigated whether MMP-8 release by E-LPS is induced via CD14-TLR pathway and the cellular mechanism of MMP-8 release in human neutrophils. Material and methods: Human neutrophils were isolated from the peripheral blood of healthy donors and pre-incubated in medium containing antibodies against CD14, anti-TLR2 and anti-TLR4 or several inhibitors of microtubules and microfilaments and then incubated with E-LPS. The cells were treated TPCK and E-LPS simultaneously. The MMP-8amount in the culture medium was determined using ELISA. Results: E-LPS increased MMP-8release from neutrophils and its induction was inhibited by anti-CD14 and anti-TLR4 but not by anti-TLR2 antibodies. The inhibitors of microtubule and microfilament polymerization significantly decreased E-LPS-induced MMP-8release. TPCK inhibited E-LPS-induced MMP-8 release. Conclusion: These results suggest that MMP-8 release is induced by E-LPS via the CD14-TLR4 signal pathway in human neutrophils and may be depedent on microtubule and microfilament systems and $NF-{\kappa}B$ pathway.

Modulation of G-CSF Secretion by Mutations of Non Alpha-Helical Region in N-Terminus (G-CSF 단백질 N-말단의 비 알파-Helix 영역의 돌연변이에 의한 분비 조절)

  • Park, Jeong-Hae;Park, Jung-Ae;Kang, Seok-Woo;Goo, Tae-Won;Chung, Kyung-Tae
    • Journal of Life Science
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    • v.21 no.12
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    • pp.1778-1783
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    • 2011
  • Hematopoietic cytokines regulate production of blood cells by stimulating proliferation and differentiation of bone marrow cells. Among these hematopoietic cytokines, called hematopoitic growth factors, glranulocyte-colony stimulating Factor (G-CSF), which regulates growth of neutrophils, is one of important therapeutic factors because cancer patients suffer with neutropenia which is severe reduction of neutrophils after chemotherapy. Two groups of recombinant G-CSF have approved and used for therapeutic purposes and many researches are still on-going to produce recombinant G-CSF by different techniques. We engineered human G-CSF with Bombyx specific endoplasmic reticulum (ER) signal sequence, therefore, secretion of human G-CSF protein was improved in Bombyx mori-origined cell line, Bm5. The Bombyx ER signal sequence and human G-CSF matured protein region chimera was further remodeled at the N-terminus of matured G-CSF protein to understand roles of N-terminus on outer cellular secretion and/or production. Three different mutants were generated deleting three amino acids in non alpha-helical region in N-terminus in order to scan important amino acids for G-CSF secretion. One of 3 different N-terminal deletion mutants showed dramatically reduction of secreted amount of G-CSF indicating its important role on secretion. The data suggest that remodeling in non alpha-helical region of N-terminus is also important for recombinant G-CSF production.

Seroprevalence of Brucellosis among Risk Population in Gyeongsangbuk-do, 2006 (경상북도 거주 고위험군에서 브루셀라 감염에 관한 혈청 역학적 연구)

  • Lee, Kwan;Park, Woo-Won;Kim, Sung-Hwan;Lee, Do-Young;Park, Mi-Yeoun;Hur, Young-Ju;Lim, Hyun-Sul
    • Journal of Preventive Medicine and Public Health
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    • v.40 no.4
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    • pp.285-290
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    • 2007
  • Objectives : Cases of human brucellosis in Korea have recently increased due to the increasing incidence of bovine brucellosis. The authors conducted this study to elucidate the status of brucellosis through seroepidemiologic study. Methods : We selected our study population from a high risk group. We conducted a questionnaire survey and obtained blood samples to determine the seroprevalence of brucellosis antibodies for 10 days in February, 2005. The titers of brucellosis were measured by the combination of standard tube agglutination test (STA) and enzyme-linked immunosorbent assay (ELISA) test. Results : Our study subjects comprised 1,075 cases: 971 livestock workers, 51 veterinarians, and 53 artificial inseminators. In the STA test, 27 cases (2.5%) had titers of greater than or equal to 1:20. Of 1,068 cases (7 cases were excluded due to previous brucellosis), 7 cases of brucellosis were diagnosed with titers of 1:160, giving a seroprevalence of brucellosis of 0.66%. The seroprevalence in the male group was 0.95%, and that of livestock workers, veterinarians, and artificial inseminators was 0.52%, 4.17%, and 0.00%, respectively. The Spearman's correlation coefficient between the positive rate of bovine brucellosis per capita and household and human brucellosis was 0.806 and 0.744, respectively. The concordance rate between the Korea National Institute of Health and the Gyeongsangbuk-do Institute of Health and Environment by the STA and ELISA tests was 94.7% and 100.0%, respectively. Conclusions : The study results indicated in higher seroprevalence rate among veterinarians than among livestock workers and artificial inseminators. Because veterinarians may be exposed to this high risk, effective working guidelines for veterinarians to guard against brucellosis must be developed. Moreover, more extensive epidemiologic research for laboratory workers and meat handlers is needed.