• Title/Summary/Keyword: Blood Assay

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Serologic survey of the ruminant bacterial infectious diseases in farmed deer and wild water deer in Jeonbuk province (사육사슴 및 야생고라니에서 소 세균성 전염병에 대한 혈청학적 연구)

  • Jo, Young-Suk;Chung, Yun-Shin;So, Seung-Young;Seol, Min-Suk;Cho, Ho-Seong;Kim, Bum-Seok;Lim, Chae-Woong
    • Korean Journal of Veterinary Service
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    • v.33 no.3
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    • pp.249-254
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    • 2010
  • Deer can be one of the susceptible animals to bovine infectious diseases, and thus, may play a role either as a reservoir or amplifier host for spreading the diseases to other species such as cattle and goat. This study was conducted to determine the serum antibodies to bacterial infectious diseases for brucellosis, tuberculosis (TB), paratuberculosis (Johne's disease) in deer. Serum samples were randomly collected from 78 deer from 31 farms at Jeonbuk province, and 7 wild water deer from Jeonbuk wild animal treatment center during 2005 to 2007, respectively. Four farm deer (5.1%) showed antibodies to tuberculosis using Antigen Rapid Bovine TB Ab Test Kit. One elk (1.3%) and one wild water deer had antibodies for paratuberculosis. Antibody against Brucellosis was not detected in tube agglutination test (TAT) and enzyme-linked immunosorbent assay (ELISA). These data suggest that caution should be applied to inspection of velvet, deer blood and meat for human consumption from deer because of zoonotic bacterial diseases in deer. In addition, farmed deer can be a transmissible host for zoonotic disease to diary or raising farm.

Purification and Characterization of Anticoagulant Protein from the Tabanus, Tabanus bivittatus

  • Ahn Mi-Young;Hahn Bum-Soo;Lee Pyeong-Jae;Wu Song-Ji;Kim Yeong-Shik
    • Archives of Pharmacal Research
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    • v.29 no.5
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    • pp.418-423
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    • 2006
  • Tabanus anticoagulant protein (TAP) was isolated from the whole body of the tabanus, Tabanus bivittatus, using three purification steps (ammonium sulfate fractionation, gel filtration on Bio-Gel P-60, and ion exchange chromatography on DEAE Sephadex gel). The purified TAP, with a molecular weight of 65 kDa, was assessed to be homogeneous by SDS-polyacrylamide gel electrophoresis, and an isoelectric point of 7.9 was determined by isoelectric focusing. The internal amino acid sequence of the purified protein was composed of Ser-Leu-Asn-Asn-Gln-Phe-Ala-Ser-Phe-lle-Asp-Lys-Val-Arg. The protein was activated by $Cu^{2+}\;and\;Zn^{2+}$, and the optimal conditions were found to be at pH $3\sim6\;and\;40\sim70^{\circ}C$. Standard coagulation screen assays were used to determine thrombin time and activated partial thromboplastin time. Chromogenic substrate assays were performed for thrombin and factor Xa activity. TAP considerably prolonged human plasma clotting time, especially activated partial thromboplastin time in a dose-dependent manner; it showed potent and specific antithrombin activity in the chromogenic substrate assay. Specific anti-factor Xa activity in TAP was not detected. Overall, this result suggested that TAP has significant anticoagulant activity on blood coagulation system.

Biological Activity of Human Dimeric Hyperglycosylated Erythropoietin (dHGEPO) Fusion Proteins

  • Naidansuren, Purevjargal;Min, Kwan-Sik
    • Reproductive and Developmental Biology
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    • v.34 no.4
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    • pp.289-297
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    • 2010
  • Erythropoietin (EPO) is a glycoprotein hormone secreted from primarily cells of the peritubular capillary endothelium of the kidney, and is responsible for the regulation of red blood cell production. We constructed and expressed dimeric cDNAs in Chinease hamster ovary (CHO) cells encoding a fusion protein consisting of 2 complete human EPO domains linked by a 2-amino acid linker (Ile-Asp). We described the activity of dimeric hyperglycosylated EPO (dHGEPO) mutants containing additional oligosaccharide chains and characterized the function of glycosylation. No dimeric proteins with mutation at the $105^{th}$ amino acid were found in the cell medium. Growth and differentiation of the human EPO-dependent leukemiae cell line (F36E) were used to measure cytokine dependency and in vitro bioactivity of dHGEPO proteins. MIT assay at 24 h increased due to the survival of F36E cells. The dHGEPO protein migrated as a broad band with an average molecular mass of 75 kDa. The mutant, dHGEPO, was slightly higher than the wild-type (WT) dimeri-EPO band. Enzymatic N-deglycosylation resulted in the formation of a narrow band with a molecular mass twice of that of of monomeric EPO digested with an N-glycosylation enzyme. Hematocrit values were remarkably increased in all treatment groups. Pharmacokinetic analysis was also affected when 2.5 IU of dHGEPO were intravenously injected into the tails of the mice. The biological activity and half-life of dHGEPO mutants were enhanced as compared to the corresponding items associated the WT dimeric EPO. These results suggest that recombinant dHGEPO may be attractive biological and therapeutic targets.

Protective effect of Poncirus trifoliata and Citrus aurantium extract on acute pancreatitis in mice model (급성췌장염 마우스 모델에서 지실과 지각 추출물의 보호효과)

  • Park, Kyoung-Chel;Bae, Gi-Sang;Choi, Sun-Bok;Jo, Il-Joo;Gwak, Tae-Sin;Lee, Guem-San;Park, Sung-Joo;Song, Ho-Joon
    • The Korea Journal of Herbology
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    • v.27 no.5
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    • pp.9-14
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    • 2012
  • Objective : We investigated the effect of Poncirus trifoliata and Citrus aurantium extract in mice with cerulein-induced acute pancreatitis (AP) model. Methods : AP was induced via intraperitoneal injection of cerulein (50 ${\mu}g/kg$) given every hour for 6h. Poncirus trifoliata (PT: 200 or 400 mg/kg) and Citrus aurantium (CA: 200 or 400 mg/kg) extract were injected 1 h before in mice with cerulein-induced AP. Mice were sacrificed at 6 h after last injection of cerulein. Blood samples were taken to determine serum amylase and lipase levels. The pancreas and lungs were rapidly removed for morphological examination, myeloperoxidase assay. Results : PT pre-treatment significantly protected the pancreas and lung damages and reduced the MPO activity and serum amylase in cerulein-induced AP. However, CA pre-treatment did not significantly protected the pancreas and lung inflammation in cerulein-induced AP. Conclusion : These results suggest that PT but not CA could protect the cerulein-induced AP. Conclusion : These results suggest that PT but not CA could protect the cerulein-induced AP.

Study on Antiangiogenic Effect of Black Ginseng Radix (흑삼의 신생혈관 억제활성에 대한 연구)

  • Song, Gyu-Yong;Chung, Kyu-Jin;Shin, Young-Jin;Lee, Gye-Won;Lee, Sook-Young;Seo, Young-Bae
    • The Korea Journal of Herbology
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    • v.26 no.3
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    • pp.83-90
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    • 2011
  • Objectives : This study was performed to investigate the influence of black ginseng radix extracts (BG) and ginsenoside Rg3, Rg5 on basic fibroblast growth factor (bFGF) induced proliferation, migration and capillary tubule-like formation of human umbilical vein endothelial cells (HUVECs). Methods : HUVECs were cultured with BG and ginsenoside Rg3, Rg5 at different concentrations (60, 125, 250, 500, $1,000{\mu}g/m\ell$) for 2 day In the presence of bFGF, respectively. XTT was used to detect the proliferation. Migration and tube formations were examined to detect the antiangiogenesis. Also, the chick embryo chorioallantoic membrane (CAM) assay was performed to detect the antiangiogenesis. Results : BG and ginsenoside Rg3, Rg5 significantly inhibited bFGF-induced endothelial cell proliferation and migration in a dose-dependent manner. Tube formation in bFGF-induced HUVECs were suppressed by BG and ginsenoside Rg3, Rg5. Moreover, BG and ginsenoside Rg3, Rg5 (30-$50{\mu}g$/egg) inhibited new blood vessel formation on the growing CAM. Conclusions:Based on the present results, it can be suggested that BG has a potential chemopreventive agent via antiangiogenesis.

Protective Effect of Cortex Fraxini on Heart Injury in a Rat Model of Myocardial Infarction (흰쥐를 이용한 심근경색모델에서 진피(秦皮)의 심장손상 보호효과)

  • Lim, Sun-Ha;Lee, Jong-Won
    • The Korea Journal of Herbology
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    • v.26 no.4
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    • pp.149-154
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    • 2011
  • Objectives : Myocardial infarction is caused by heart cell death in a region where coronary arteries supplying blood to the region are occluded. In the present study, we determined whether ethanol extract of Cortex fraxini (HY5053) could attenuate heart injury by inhibiting apoptosis. Methods : Improvement of survival of HepG2 cells, a human hepatocellular carcinoma cell line, and reduction of apoptosis under hypoxic conditions (3% $O_2$) were assessed by trypan blue staining and DNA fragmentation assay, respectively. To assess the impact of HY5053 on the heart injury, Sprague-Dawley rats underwent 1 day of the left anterior descending coronary artery occlusion. HY5053 was given by intraperitoneal injection (200 mg/kg) 1 hr prior to the occlusion. Subsequently, the heart were harvested, excised into 4 slices, and the slices were stained with 2,3,5-triphenyl tetrazolium chloride. Finally, the extent of heart injury represented as ischemic index (%) was assessed. Results : Addition of HY5053 (400 ${\mu}g$/mL) into the culture medium for 1 day under ischemic conditions improved the cell survival by 50%, compared with control (0 ${\mu}g$/mL), consequently delayed apoptosis in 6 hr difference. Also, HY5053 (200 mg/kg) reduced the ischemic index by 44%, compared with control (0 mg/kg). Conclusions : These findings suggested that HY5053 attenuated myocardial infarction by inhibiting apoptosis. Thus, Cortex fraxini could be developed as a novel cardioprotectant to complement a currently available treatment, coronary angioplasty.

Electrospray Tandem Mass Spectrometry for the Quantification and Bioavailability Test of Gliquidone in Human Plasma (Electrospray Tandem Mass를 이용한 혈중 글리퀴돈의 정량법 개발 및 생체이용률시험)

  • Moon Chul-Jin;Lee Eun-Hee;Yang Song-Hyun;Moon Hae-Ran
    • YAKHAK HOEJI
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    • v.49 no.3
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    • pp.212-216
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    • 2005
  • A rapid, sensitive and selective electrospray tandem mass spectrometric (ESI-LC/MS/MS) method for the quantitation of gliquidone in human plasma was developed. A bioavailability study of gliquidone tablet (30 mg gliquidone, Boehringer Ingelheim Korea Co.) was performed using the validated ESI-LC/MS/MS method. The dose of 30 mg of gliquidone (1 tablet) was orally administered to 9 healthy Korean subjects. After administration, blood was taken at 0.5, 1, 1.5, 2, 2.5, 3, 3.5, 4, 5, 7, 9, 12, 24, and 33 hour. The validation data were as follows; the standard curve was linear ($r^2$=0.999) over the concentration range of $10\~1000 ng/ml$. The coefficient of variation for intra- and inter-day assay were $8.30\~18.86$, and $2.19\~12.92\%$, respectively. The lower limit of quantification for gliquidone was 10 ng/ml. The pharmacokinetic parameters obtained were as follows; $AUC_t$ was 3861.17$\pm$1328.61 ng-hr/ml, $C_{max}$ was 831.02$\pm$227.99 ng/ml, $T_{max}$ was $2.94{\pm}0.77 hr,\;K_e$, was 0.19$\pm$0.06 1/hr, and $t_{l/2}$ was 4.47$\pm$3.52 hr. Based on the validated analytical method and pharmacokinetic parameters, a standard guideline of the bioavailability test of gliquidone dosage forms was prepared successfully and could be used for the bioequivalence test of gliquidone preparation.

Tandem Mass Spectrometry for the Quantification and Bioavailability Test of Nicorandil in Human Plasma (Tandem Mass를 이용한 혈중 니코란딜의 정량법 개발 및 생체이용률시험)

  • Moon Chul-Jin;Lee Eun-Hee;Yang Song-Hyun;Moon Hae-Ran
    • YAKHAK HOEJI
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    • v.49 no.3
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    • pp.225-229
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    • 2005
  • A rapid, sensitive and selective tandem mass spectrometric method (LC-MS/MS) for the quantitation of nicorandil in human plasma was developed. A bioavailability study of Sigmat tablet (5 mg nicorandil, Choongwae Co.) was per-formed using the validated LC-MS/MS method. The dose of 5 fig of nicorandil (1 tablet) was orally administered to 9 healthy Korean subjects. After administration, blood was taken at 0.25, 0.5, 1, 2, 3, 4, 5, 6, 9, 12, and 24 hour. The validation data were as follows; the standard curve was linear ($r^2$=0.999) over the concentration range of $0.5\~200.0 ng/ml$. The coefficient of variation for intra- and inter-day assay were $3.55\~7.44$, and $2.17\~9.102\%$, respectively. The lower limit of quantification for nicorandil was 0.5 ng/ml. The pharmacokinetic parameters obtained were as follows; $AUC_t$ was 145.9$\pm$83.0 ng-hr/ml, Cmax was 83.8$\pm$32.2 ng/ml, $C_{max}$ was 0.42$\pm$0.13 hr, $K_e$ was 0.56$\pm$0.23 l/hr, and $t_{l/2}$ was 1.42$\pm$0.52 hr. Based on the validated analytical method and pharmacokinetic parameters, a standard guideline of the bioavailability test of nicorandil dos-age forms was prepared successfully and could be used for the bioequivalence test of nicorandil preparation.

Panax ginseng Meyer prevents radiation-induced liver injury via modulation of oxidative stress and apoptosis

  • Kim, Hyeong-Geug;Jang, Seong-Soon;Lee, Jin-Seok;Kim, Hyo-Seon;Son, Chang-Gue
    • Journal of Ginseng Research
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    • v.41 no.2
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    • pp.159-168
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    • 2017
  • Background: Radiotherapy is one of the most important modalities in cancer treatment; however, normal tissue damage is a serious concern. Drug development for the protection or reduction of normal tissue damage is therefore a clinical issue. Herein, we evaluated the protective properties of Panax ginseng Meyer and its corresponding mechanisms. Methods: C56BL/6 mice were orally pretreated with P. ginseng water extract (PGE; 25 mg/kg, 50 mg/kg, or 100 mg/kg) or intraperitoneally injected melatonin (20 mg/kg) for 4 d consecutively, then exposed to 15-Gy X-ray radiation 1 h after the last administration. After 10 d of irradiation, the biological properties of hematoxicity, fat accumulation, histopathology, oxidative stress, antioxidant activity, pro-inflammatory cytokines, and apoptosis signals were examined in the hepatic tissue. Results: The irradiation markedly induced myelosuppression as determined by hematological analysis of the peripheral blood. Steatohepatitis was induced by X-ray irradiations, whereas pretreatment with PGE significantly attenuated it. Oxidative stress was drastically increased, whereas antioxidant components were depleted by irradiation. Irradiation also notably increased serum liver enzymes and hepatic protein levels of pro-inflammatory cytokines. Those alterations were markedly normalized by pretreatment with PGE. The degree of irradiation-induced hepatic tissue apoptosis was also attenuated by pretreatment with PGE, which was evidenced by a terminal deoxynucleotidyl transferase 2'-deoxyuridine 5'-triphosphate nick-end labeling assay, western blotting, and gene expressions analysis, particularly of apoptotic molecules. Conclusion: We suggest that PGE could be applicable for use against radiation-induced liver injury, and its corresponding mechanisms involve the modulation of oxidative stress, inflammatory reactions, and apoptosis.

Isolation of Brucella spp from sere-positive native bulls and calves below twelve months old (브루셀라병 혈청검사 양성 수소와 12개월령 이하 소에서의 균 분리 및 동정)

  • Ryu, Jae-Yun;Bun, Jung-Hyun;Lee, Hee-Young;Lee, Yong-Chang;Lee, Jong-Jin;Song, Young-Gak;Nam, Hyang-Mi
    • Korean Journal of Veterinary Service
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    • v.30 no.3
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    • pp.375-384
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    • 2007
  • This study was conducted to investigate the characteristics of brucellosis in Korean native cattle in a farm where bovine brucellosis was confirmed 3 times from September 2006 to March 2007. Of 74 bulls serum samples examined, 21 (28.4%) were positive by Rose-Bengal test (RBT) and Standard tube agglutination test (STAT). In the isolation test from seropositive bulls, B abortus was isolated and identified from 2 specimens (testis, intestinal lymph node) among 6 kinds of specimens including blood, urine, feces and soil. Isolation rate of intestinal lymph node and testis was 25% (3/12 cases) and 16.7% (2/12), respectively. B abortus was also isolated from calves below 12 months old, i.e., 1 isolate (25.0%) was confirmed from testis, 4 (40.0%) from supra-mammary lymph nodes and 1 (25.0%) from intestinal lymph node. All isolates had Brucella specific 16s r-RNA with 905-bp band detected by PCR assay. For the more effective control of bovine brucellosis in korea, this paper would like to suggest that all of bulls and calves should be included in the screening tests.