• Title/Summary/Keyword: cELISA

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Comparison of Serum Osteopontin Levels in Patients with Stable and Chronic Obstructive Pulmonary Disease and Exacerbation (안정된 만성 폐쇄성 폐질환환자와 급성 악화상태의 혈중 Osteopontin 농도 비교)

  • Ma, Jeong-Eun;Lee, Seung-Hun;Kim, Yu-Eun;Lim, Su-Jin;Lee, Seung-Jun;Jeong, Yi-Yeong;Kim, Ho-Cheol;Lee, Jong-Deog;Hwang, Young-Sil;Cho, Yu-Ji
    • Tuberculosis and Respiratory Diseases
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    • v.71 no.3
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    • pp.195-201
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    • 2011
  • Background: Osteopontin (Opn) is recognized as an important adhesive bone matrix protein and a key cytokine involved in immune cell recruitment and tissue repair and remolding. However, serum levels of osteopontin have not been evaluated in patients with chronic obstructive pulmonary disease (COPD). Thus, the aim of this study was to evaluate and compare the serum levels of osteopontin in patients experiencing COPD exacerbations and in patients with stable COPD. Methods: Serum samples were obtained from 22 healthy control subjects, 18 stable COPD patients, and 15 COPD with exacerbation patients. Serum concentrations of osteopontin were measured by the ELISA method. Results: Serum levels of osteopontin were higher in patients with acute exacerbation than with stable COPD and in healthy control subjects ($62.4{\pm}51.9ng/mL$, $36.9{\pm}11.1ng/mL$, $30{\pm}11ng/mL$, test for trend p=0.003). In the patients with COPD exacerbation, the osteopontin levels when the patient was discharged from the hospital tended to decrease compared to those at admission ($45{\pm}52.1ng/mL$, $62.4{\pm}51.9ng/mL$, p=0.160). Osteopontin levels significantly increased according to patient factors, including never-smoker, ex-smoker and current smoker ($23{\pm}5.7ng/mL$, $35.5{\pm}17.6ng/mL$, $58.6{\pm}47.8ng/mL$, test for trend p=0.006). Also, osteopontin levels showed a significantly negative correlation with forced expiratory volume in one second ($FEV_1$%) predicted in healthy controls and stable COPD patients (r=-0.389; p=0.013). C-reactive protein (CRP) was positively correlated with osteopontin levels in patients with COPD exacerbation (r=0.775; p=0.002). Conclusion: The serum levels of osteopontin increased in patients with COPD exacerbation and tended to decrease after clinical improvement. These results suggest the possible role of osteopontin as a biomarker of acute exacerbation of COPD.

Effects of $TGF-{\beta}1$ on Cellular Activity of Minocycline-Pretreated Human Periodontal Ligament Cells (($TGF-{\beta}$)이 Minocycline을 전처리한 사람 치주인대세포의 활성에 미치는 영향)

  • Yang, Seung-Oh;You, Hyung-Keun;Shin, Hyung-Shik
    • Journal of Periodontal and Implant Science
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    • v.26 no.2
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    • pp.469-490
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    • 1996
  • The initial events required for periodontal regeneration is the attachment, spreading, and proliferation of appropriated cells at the healing sites. These have been reported that minocycline stimulates the attachment of periodontal ligament cells, and also $TGF-{\beta}1$ enhances the proliferation of periodontal ligament cells. The purpose of the present study was to evaluate the effects of $TGF-{\beta}1$ on the cellular activity of minocycline treated human periodontal ligament cells. Periodontal ligament cells were obtained from the explants of healthy periodontal ligaments of extracted 3rd molars or premolar teeth extracted from the patients for orthodontic treatment. The cells were cultured in minimal essential medium(${\alpha}-MEM$) supplemented with 10.000units/ml penicillin, $10,000{\mu}g/ml$ streptomycin and 10% FBS(fetal bovine serum) at $37^{\circ}C$ in a humidified atmosphere of 5% carbon dioxide and the 5th to the 8th passages of the cells were used. To evaluate the effect of minocycline on cell attachment, the cells were seeded at a cell density of $1.5{\times}10^4$ cells/well in 24-well culture plates and treated with $20{\mu}g/ml$ and $100{\mu}g/ml$ of minocycline for 1.5 h. After trypsinization, the cells were counted with hemocytometer and were taken photographs for observation of cellular morphology. To evaluate the effect of $TGF-{\beta}1$ on minocycline-pretreated periodontal ligament cells, the cells were seeded at a cell density of $1{\times}10^4$ cells/ well in 24-well culture plates and treated with $20{\mu}g/ml$ and $100{\mu}g/ml$ of minocycline for 1.5 h. After incubation, 1 and 10ng/ml of $rh-TGF-{\beta}1$ were also added to the each well and incubated for 1 and 2 days, respectively. Then, MTT assay, DNA synthesis($^3H-thymidine\;assay$), and protein and collagen assay(3H-proline assay) were carried out. In the MIT assay, after 200ul MTT solutionlconeentration of 5mg/ml) were added to the each well of the 24-well plates and incubated for 3 hours, and 200 ul DMSO were added so as to dissolve insoluble blue formazan crystals which was formed in incubated period. Then it read plates on a ELISA reader. For mitogenic assay, 1 uCi/ml $^3H-thymidine$ was added to each well for the final 2 hours of the incubation periods. After labeling, the wells were washed 3 times with ice cold PBS and 4 times with 5% TCA to remove unincorporated label and precipitate the cellular DNA. DNA, with the incorporated $^3H-thymidine$, was solubilized with 500 ul of 0.1% NaOH/0.1% SDS. A 250 ul aliquot was removed from each well and placed in a scintillation vial with 4ml of scintillation cocktail. Using an liguid scintillation counter, counts per minute(CPM) were determined for each samples. 3 uCi/ml $^3H-proline$ was added to each well for the final 4 hours of the incubation periods and total protein and percent collagen synthesis were carried out. The results indicate that minocycline treated group with $100{\mu}g/ml$ concentration for 1.5 hours significantly increased than that of control in cell attachment, and cell process is also evident compared with that of control in cell morphology, and the cellular activity and DNA synthesis rate of cells treated minocycline and $TGF-{\beta}1$ significantly increased than that of control values, but were below to values of the $TGF-{\beta}1$ only treated group in MIT assay and $^3H-thymidine\;assay$, and the total protein synthesis of minocycline and $TGF-{\beta}1$ treated group also significantly increased than that of control values, but the percent collagen synthesis of tested group significantly decreased to compared with control. On the above the findings, the tested group of minocycline and $TGF-{\beta}1$ did not increase the effect on the cell activity than $TGF-{\beta}1$ only tested group and the tested group of minocycline inhibited cell activity. This results indicate that minocycline was effective on cell attachment in early stage, but it is harmful to cell activity, that inhibitory effect of minocycline was compensated with stimulatory effect of $TGF-{\beta}1$.

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Coagulation and Fibrinolysis in Exudative Pleural Effusions (삼출성 흉막액에서 응고 및 섬유소 용해계에 관한 연구)

  • Ryu, Jeong-Seon;Lee, Hong-Lyeol
    • Tuberculosis and Respiratory Diseases
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    • v.45 no.6
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    • pp.1214-1222
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    • 1998
  • Background : The intrapleural hypofibrinolysis is caused by mainly excessive concentration of pleural plasminogen activator inhibitor-1 antigen(PAI-1 Ag), which binds tissue type plasminogen activator. In pleural inflammation induced by sclerosing agents for pleurodesis, levels of pleural PAI-1 antigen increase in relation to decreasing D-dimer levels. It has been known that the pleural mesothelial cells have the capability of secreting PAI-1 Ag in response to inflammation in vivo. Therefore, we estimated whether pleural inflammation changes the balance between fibrinolytic and coagulative properties in exudative pleural effusions. Method : The thirty cases was included in our study. We determined the pleural levels of glucose, lactic dehydrogenase(LDH), pH and the counts of white blood cell(WBC), polymorpho leukocyte(PMN), lymphocyte as the parameters of pleural inflammation and cellular components of pleural fluid. The plasma level of fibrinogen in fluid and the neutrophil count in blood were determined. The levels of D-dimer, PAI-1 Ag and thrombinantithrombin III complex(TAT) were determined by ELISA(Behring, Marburg, Germany). Result : The causes of pleural effusion were as following : tuberculous in 14 cases, malignant in 10 cases and parapneumonic in 6 cases. The levels of pleural D-dimer, PAI-1 Ag and TAT was significantly higher than that of plasma(p<0..001). The severity of pleural inflammation did not correlated with pleural D-dimer, PAI-1 Ag, TAT and their plasma levels. But the level of pleural TAT correlated with pleural WBC and lymphocyte count. Conclusion : We found that the severity of pleural inflammations did not correlated with pleural D-dimer, PAI-1 Ag, TAT and the possibility of local production of PAI-1 antigen is present.

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Immune Activity of Mosidae and Quality Characteristics of Brown Rice Dasik Using Mosidae Powder (모시대의 면역 활성 탐색 및 모시대 분말 첨가 현미다식의 품질 특성)

  • Kim, Ae-Jung;Han, Myung-Ryun;Kim, Myung-Hwan;Tae, Ki-Hwan;Lee, Soo-Jung
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.38 no.5
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    • pp.548-554
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    • 2009
  • The principal objective of this study was to evaluate the immune activity of Mosidae and the physiochemical characteristics of brown rice Dasik prepared with Mosidae (Adenophora remotiflora) powder. We assessed the effects of Mosidae ethanol extract (MEE) on the production of IL-6T, IL-12 and TNF-$\alpha$ by peritoneal exudate macrophages (PEMs) using ELISA. We also determined general compositions, and conducted Hunter's color values, sensory evaluation, and the mechanical characteristics of Mosidae Dasik stored at room temperature ($20^{\circ}C$). With MEE treatment, ILI-6 (75% of LPS: positive control), IL-12 (35.7% of LPS) and TNF-$\alpha$ (27.32% of LPS) were proliferated at a dose of $1000{\mu}g/mL$. In the general compositions of the samples, fat contents of Mosidae Dasik significantly decreased (p<0.05). The more Mosidae powder was added to the samples, the more was the luminance, and Hunter's a and b were significantly decreased (p<0.05). As more Mosidae powder was added to the samples, springiness score was significantly decreased, but the score of hardness, gumminess and chewiness were increased (p<0.05). The results of sensory evaluation showed that there were significant differences in the color, taste and overall quality of the samples (p<0.05), but there was no significant difference in texture. We note that, among the samples evaluated herein, Mosidae stimulates some kinds of cytokines from machrophage and 1% Mosidae Dasik (MPD1) for the best commercial value.

Detection of TNF-alpha in Serum as the Effect of Corticosteroid to the Myocardial Protection in Cardiopulmonary Bypass (체외순환시 스테로이드의 심근보호효과에 관한 혈청내 TNF-alpha 측정의 의의)

  • 최영호;김욱진;김태식;조원민;김학제
    • Journal of Chest Surgery
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    • v.31 no.5
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    • pp.502-508
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    • 1998
  • Proinflammatory cytokines such as tumor necrosis factor-$\alpha$(TNF-$\alpha$) have been implicated in myocardial and organ dysfunction associated with postperfusion syndrome. We tested the hypothesis that cytokine productions are depressed by preoperative cortiosteroid injection for cardiopulmonary bypass(CPB) and the postoperative courses will be better than without steriod pretreated cases. Cardiac surgery was performed in randomized blind fashion for 20 patients from June 1996 to September 1996. In the steroid group(n=10), corticosteroid(dexamethasone 1 mg/kg) was injected 1 hour before anesthetic induction, but in the control group(n=10), nothing was injected. Each of groups were sampled 11 times as scheduled for TNF-$\alpha$ bioassays. We have checked EKG, cardiac enzymes(CPK, LDH with isoenzyme), WBC count preoperative day, one day and three days after operation. Viatal signs were continuously monitored for three postoperaive days. In the postoperative period three patients in the control group had elevated body temperature and four patients had hypotension that required considerable intravenous fluid administration. But steroid injected patients showed normal body temperture and acceptable blood pressures without supportive treatment. CPK enzymes rose in control group higher than steroid group at postoperative 1st and 3rd day(CPK; 1122$\pm$465 vs 567$\pm$271, 864$\pm$42 vs 325$\pm$87), and CPK-MB enzymes rose in control group higher than steroid group at postoperative 1st day(106.4$\pm$115.1 vs 29.5$\pm$22.4)(P=0.02). Arterial tumor necrosis factor-$\alpha$ rose during cardiopulmonary bypass, peaking at 5 minutes before the end of aortic cross clamping(ACC-5min) in steroid group(11.9$\pm$4.7 pg/ml), and 5 minutes before the end of cardiopulmonary bypass(CPB-5min) in control group(22.3$\pm$6.8 pg/ml). The steroid pretreated patients had a shorter period of time in respirator suport time, ICU stay day, hospital admission day. We conclude that corticosteroid suppress cytokine production during and after cardiopulmonary bypass, and may improve the postoperative course through inhibition of reperfusion injury such as myocardial stunning and hemodynamic instability.

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The effect of tumor necrosis factor (TNF)-α to induce matrix metalloproteinase (MMPs) from the human dental pulp, gingival, and periodontal ligament cells (사람의 치수, 치은, 치주인대 세포에 tumor necrosis factor (TNF)-α로 자극 시 matrix metalloproteinase (MMPs)의 분비에 관한 연구)

  • Rhim, Eun-Mi;Park, Sang-Hyuk;Kim, Duck-Su;Kim, Sun-Young;Choi, Kyoung-Kyu;Choi, Gi-Woon
    • Restorative Dentistry and Endodontics
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    • v.36 no.1
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    • pp.26-36
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    • 2011
  • Objectives: In the present study, three kinds of tissues cells (pulp, gingiva, and periodontal ligament) were investigated if those cells express MMP and TIMP when they were stimulated with neuropeptides (substance P, CGRP) or proinflammatory cytokine, TNF-$\alpha$. Materials and Methods: The cells cultured from human dental pulp (PF), gingiva (GF) and periodontal ligament were (PDLF) stimulated with Mock, SP, TNF-$\alpha$, and CGRP for 24 hrs and 48 hrs. for an RNase protection assay and Enzyme Linked Immunosorbent Assay. Cells (PF, GF and PDLF) seeded in 100 mm culture dish were stimulated with SP ($10^{-5}$, $10^{-8}\;M$) or only with medium (Mock stimulation) for 4hrs and for 24 hrs for RNase Protection Assay, and they were stimulated with CGRP ($10^{-5}\;M$) and TNF-$\alpha$(2 ng/mL) for 24 hrs and with various concentraion of TNF-$\alpha$(2, 10, and 100 ng/mL) for Rnase Protection Assay with a human MMP-1 probe set including MMP 1, 2, 8, 7, 8, 9, 12, and TIMP 2, 3. In addition, cells (PF, GF and PDLF) were stimulated with Mock and various concentraion of TNF-$\alpha$(2, 10, and 100 ng/mL) for 24 hrs and with TNF-$\alpha$(10 ng/mL) for 48 hrs, and the supernatents from the cells were collected for Enzyme Linked Immunosorbent Assay (ELISA) for MMP-1 and MMP-13. Results: The expression of MMPs in PF, GF, PDLF after stimulation with SP and CGRP were not changed compared with Mock stimulation for 4 hrs and 24 hrs. The expression of MMP-1, -12, -13 24 hrs after stimulation with TNF-$\alpha$ were upregulated, however the expression of TIMP-3 in PF, GF, PDLF after stimulation with TNF-$\alpha$ were downregulated. TNF-$\alpha$(2 ng/mL, 10 ng/mL, 100 ng/mL) increased MMP-1 and MMP-12 expression in PF dose dependently for 24 hrs. Conclusions: TNF-$\alpha$ in the area of inflammation may play an important role in regulating the remodeling of dentin, cementum, and alveolar bone.

Community-based Helicobacter pylori Screening and its Effects on Eradication in Patients with Dyspepsia (지역사회에서 소화불량 환자의 Helicobacter pylori 감염에 대한 집단검진 및 치료효과)

  • Kim, Seong-Ho;Hong, Dae-Yong;Lee, Kyeong-Soo;Kim, Seok-Beom;Kim, Sang-Kyu;Suh, Jeong-Ill;Kim, Mee-Kyung;Kang, Pock-Soo
    • Journal of Preventive Medicine and Public Health
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    • v.33 no.3
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    • pp.285-298
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    • 2000
  • Objectives : To investigate the positive rate of Helicobacter pylori in patients with dyspepsia; medical compliance and related factors; the eradication rate a year after screening and related factors; the relationship between the eradication of Helicobacter pylori and the improvement of symptoms; and the estimated cost of three alternative approaches to treat Helicobacter pylori in the community. Methods : A total of 510 subjects with dyspeptic symptoms were selected and given the serological test in March 1998. The subjects were all adults over 30 years of age residing in Kyongju city. Results : Of the 510 selected subjects, 375 (73.5%) subjects proved positive for Helicobacter pylori on serological testing. Of these 304 (81.1%) who consented to an endoscopic examination, underwent a Campylobacter-like organism (CLO) test. Of these 304 subjects, 204 (67.1%), who had positive CLO test results, were given the triple therapy - tripotassium dicitrato bismuthate, amoxicillin, and metronidazole. To determine the eradication rate of Helicobacter pylori, 181 (88.1%) out of the 204 subjects who were given the triple therapy completed a follow-up urea breath test one year later. Of these, the Helicobacter pylori of 87(48.1%) subjects was eradicated. Among the 122 subjects who were medication compliant, the Helicobacter pylori eradication rate was 57.4% (70 subjects), while the eradication rates was only 28.8% (17subjects) in the non-compliant group. The Helicobacter pylori eradication was significantly related to compliance (p<0.01), but not to other characteristics and habits. The symptom improvement rate tended to be higher 62.1%), in the Helicobacter pylori eradicated group than in the non-eradicated group (59.6%). Conclusions : When the advantages and disadvantages of each alternative treatment were considered in the light of cost, antibiotic tolerance and the number of patients to be treated, alternative II was favorable in terms of cost. Alternative III was favorable in terms of the number of patients to be treated, antibiotic tolerance and early detection of gastric cancer. Further long-term research analyzing the cost-benefit and cost-effectiveness of each treatment will be needed as supporting material in creating new policies.

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Evaluation of the Radioimmunotherapy Using I-131 labeled Vascular Endothelial Growth Factor Receptor2 Antibody in Melanoma Xenograft Murine Model (흑색종에서의 I-131표지 혈관내피세포성장인자 수용체2항체를 이용한 방사면역치료 평가)

  • Kim, Eun-Mi;Jeong, Hwan-Jeong;Park, Eun-Hye;Cheong, Su-Jin;Lee, Chang-Moon;Jang, Kyu-Yun;Kim, Dong-Wook;Lim, Seok-Tae;Sohn, Myung-Hee
    • Nuclear Medicine and Molecular Imaging
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    • v.42 no.4
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    • pp.307-313
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    • 2008
  • Purpose: Vascular endothelial growth factor (VEGF) and its receptor, fetal liver kinase 1 (Flk-1), play an important role in vascular permeability and tumor angiogenesis. The aim of this study is to evaluate the therapeutic efficacy of $^{131}I$ labeled anti-Flk-1 monoclonal antibody (DC101) on the growth of melanoma tumor, which is known to be very aggressive in vivo. Materials and Methods: Balb/c nude mice were injected subcutaneously with melanoma cells in the right flank. Tumors were allowed to grow up to $200-250\;mm^3$ in volume. Gamma camera imaging and biodistribution studies were performed to identify an uptake of $^{131}I$-DC101 in various organs. Mice with tumor were randomly divided into five groups (10 mice per group) and injected intravenously; control PBS (group 1), $^{131}I$-DC101 $50\;{\mu}g/mouse$ (group 2), non-labeled DC101 $50\;{\mu}g/mouse$ (group 3), $^{131}I$-DC101 $30\;{\mu}g/mouse$ (group 4) and $15\;{\mu}g/mouse$ (group 5) every 3 or 4 days for 20 days. Tumor volume was measured with caliper twice a week. Results: In gamma camera images, the uptake of $^{131}I$-DC101 into tumor and thyroid was increased with time. Biodistribution results showed that the radioactivity of blood and other major organ was gradually decreased with time whereas tumor uptake was increased up to 48 hr and then decreased. After 4th injection of $^{131}I$-DC101, tumor volume of group 2 and 4 was significantly smaller than that group 1. After 5th injection, the tumor volume of group 5 also significantly reduced. Conclusion: These results indicated that delivery of $^{131}I$ to tumor using FlK-1 antibody, DC101, effectively blocks tumor growth in aggressive melanoma xenograft model.