• Title/Summary/Keyword: ELISA.

Search Result 2,992, Processing Time 0.035 seconds

The Value of ICAM-1 Expression and the Soluble ICAM-1(sICAM-1) Level as a Marker of Activity in Sarcoidosis: The Relationship Between the ICAM-1 Level and the Clinical Course of the Disease (유육종증의 활동성 지표로서의 ICAM-1)

  • Kim, Dong-Soon;Paik, Sang-Hoon;Shim, Tae-Sun;Lim, Chae-Man;Lee, Sang-Do;Koh, Youn-Suck;Kim, Woo-Sung;Kim, Won-Dong
    • Tuberculosis and Respiratory Diseases
    • /
    • v.45 no.1
    • /
    • pp.116-127
    • /
    • 1998
  • Background: The natural course of sarcoidosis is variable from spontaneous remission to significant morbidity or death. So the assessment of disease activity is important but no single parameter was generally accepted as a good marker. Recently several studies suggested that adhesion molecules, especially ICAM-1 can be a marker, but there are some controversies. And only few data are available about the relationship of ICAM-1 with clinical follow-up course. Methods: We measured the expression of adhesion molecules on BAL cells by flow cytometry and the level of soluble ICAM-1(sICAM-1) in serum and BALF at the time of diagnosis in 12 patients with active disease and 7 inactive sarcoidosis(5 male, 14 female, mean age: $39.4{\pm}10.7$ years, mean follow-up : $20{\pm}15$ months). Follow-up clinical course were compared with the changes in serum sICAMA-1 level and the adhesion molecule on BAL cells. Results: In the patients with active disease, the ICAM-1 on AM(RMFI: $3.68{\pm}1.71$) and sICAM-1 level in serum($582{\pm}193$ng/ml) and BAL fluid($47.8{\pm}16.5$ng/ml) were all higher than those of 7 inactive disease(RMFI: $1.89{\pm}0.75$, p=0.0298, serum: $294{\pm}117$ ng/ml, p=0.0049, BALF: $20.9{\pm}8.3$ ng/ml). In the active sarcoidosis, ICAM-1 on AM(RMFI : $1.51{\pm}0.84$) and serum sICAM-1 were decreased after the therapy($250{\pm}147$ ng/ml) but no significant change was noted in inactive disease. Also we found the initial ICAM-1 on AM and serum sICAM-1 had a significant correlation with the degree of improvement in PFT after the therapy. During the follow-up, the disease relapsed in 4 patients after the discontinuation of steroid and the serum sICAM-1 level went-up again at the time of relapse. Conclusion: Our data suggest that the serum sICAM-1 level and the ICAM-1 expression on AM can be a good marker of disease activity and also a predictor of outcome in sarcoidosis.

  • PDF

Activity of Cytokines and Expression of CD62L in Patients with Bronchial Asthma (기관지 천식환자에서 CD62L의 발현 및 싸이토카인의 변화)

  • Song, Kwang-Seon;Lee, Won-Yeon;Hong, Ae-Ra;Kim, Hee-Sun;Yong, Suk-Joong;Shin, Kye-Chul
    • Tuberculosis and Respiratory Diseases
    • /
    • v.45 no.1
    • /
    • pp.90-98
    • /
    • 1998
  • Background : The CD4+ T-helper cells comprise functionally distinct subsets of Th1 and Th2 cells that are distinguished on the basis of differential cytokines production Th1 cells secrete interferon-$\gamma$, lymphotoxin, interleukin-2. Th2 cells produce interleukin-4, interleukin-5, interleukin-10. A previous study shown that Th2 cells and their cytokines increased in patients with atopic asthma. We compared cytokines(IL-4, IFN-$\gamma$) activity and subpopulation of T-lymphocytes in peripheral blood from atopic asthmatics versus non-asthmatics. Method: Fifteen patients with atopic asthma(nine men, six women), twelve patients with chronic bronchitis(six men, six women), five healthy persons(three men, two women) were studied. Activity of IL-4, IFN-$\gamma$ and T-cell subpopulation in peripheral blood were estimated. Results: Patients had a median age of 55yr. The mean activity of IL-4 of asthmatics was significantly increased(control $0.75{\pm}1.1pmol/L$, atopic asthmatics $3.50{\pm}0.75pmol/L$, chronic bronchitis $2.01{\pm}1.2pmol/L$), but IFN-$\gamma$ was not significantly increased. In the T lymphocyte sunsets the percent of CD62L+ T-lymphoeytes of asthmatics was not significantly increased (control $16.7{\pm}16.4%$, atopic asthmatics $24.8{\pm}23.6%$, chronic bronchitis $17.0{\pm}16.9%$). Conclusion: In this study elevated production of IL-4 was observed in atopic asthmatics. CD62L+T-lymphoeytes was not increased in atopic asthma.

  • PDF

Optimization of Human Thrombopoietin Production in Insert Cells Using Baculovirus Expression System (베큘로 바이러스 발현 시스템에 의한 곤충세포에서의 인간 트롬보포이에틴 생산 최적화)

  • 고여욱;손미영;박상규;안혜경;박승국;박명환;양재명
    • KSBB Journal
    • /
    • v.13 no.2
    • /
    • pp.181-186
    • /
    • 1998
  • In order to obtain high-level production of recombinant human thrombopoietin (rhTPO) in insect cell line, HTI-TN-5B1-4 (TN5), conditions for optimal rhTPO expression such as multiplicity of infection (MOI), the cell density at infection, harvesting time and type of culture method as well as growth media were determined. When TN5 cells were cultured as anchorage-dependent state in 60-mm dish, cell density $2\times^6$ cells,MOI of 10 and Garvesting the culture media at 72 hr post-infection wrere the cinditions for highest rh TPO production. High production of rhTPO was also achieved by using EXPRESS FIVE serum free media rather than SF900II serum free media-1. Anchorage-dependent TN5 cells were adapted as a suspension culture when they were grown in the presence of heparin. TN5 cells were successfully cultured at 0.2 L scale in suspension culture without having aggregation. When TN5 cells were cultured as suspension state, cell density of $0.6\times10^6$ cells/mL, MOI of 1 and harvesting the culture media at 72 hr post-infection, gave the highest yield of rhTPO.

  • PDF

Selection and Mechanism of Anti-Obesity Agents from Natural Products Based on Anti-Angiogenesis (신생혈관형성억제작용을 기반으로 한 항비만제제의 선별 및 작용기전)

  • Shin, Jin-Hyuk;Lee, Jin-Hee;Kang, Kyeong-Wan;Hwang, Jae-Ho;Han, Kyeong-Ho;Shin, Tai-Sun;Kim, Min-Yong;Kim, Jong-Deog
    • KSBB Journal
    • /
    • v.24 no.2
    • /
    • pp.122-130
    • /
    • 2009
  • Anti-angiogenic mechanism was examined for anti-obesity agents with the extract of P.radix, P.semen, S.hebra and C.furctus through anti-cell adhesion effect and western blot. Cell adhesion molecules, VCAM-1 was supressed with the order of P.radix (0.2 ppm, 125%) > P.semen (0.5 ppm, 100%) > S.hebra (5.0 ppm, 114%) > C. furctus (5.0 ppm, 111.8%), ICAM-1 was inhibited by P.radix (0.25 ppm, 130%) > P.semen (0.5 ppm, 100%) > S.hebra (5.0 ppm, 138%) > C. furctus (5.0 ppm, 66.7%), E-Selectin was also supressed P.radix (0.25 ppm, 100%) > P.semen (1.0 ppm, 128%) > S.hebra (5.0 ppm, 120%) > C. furctus (5.0 ppm, 100.7%). And signal molecules, VE-cadherin was supressed by P.radix and S.hebra, ${\beta}$-catenin was inhibited by P.radix, and Akt was supressed all these 4 kinds of natural products. These P.radix, P.semen, S.hebra and C.furctus were showed the possibility of anti-obesity agents based on anti-angiogenesis.

Photoprotection and Anti-inflammatory Effects of Chinese Medical Plants (약용식물추출물의 광보호 효과와 항염증 효과 연구)

  • Jin-Hwa, Kim;Sung-Min, Park;Gwan-Sub, Sim;Bum-Chun , Lee;Hyeong-Bae, Pyo
    • Journal of the Society of Cosmetic Scientists of Korea
    • /
    • v.30 no.2
    • /
    • pp.227-233
    • /
    • 2004
  • Chronic exposure to solar radiation, particularly ultraviolet (UV) light, causes a variety of adverse reactions on human skin, such as sunburn, photoaging and photocarcinogenesis. Free radicals and reactive oxygen species (ROS) caused by UV exposure or other environmental facts play critical roles in cellular damage. And, repeated-UV irradiation activated the expression of the matrix metalloproteinase (MMP) and induced skin irritation. Therefore, the development of effective and safe photoprotectants that can reduce and improve the skin damage has been required. The purpose of this study was to investigate the photo-protective effect of several chinese medical plants (Juniperus chinensis) on the UV -induced skin cell damages. We tested free radical and superoxide scavenging effect in vitro. Fluorometric assays of the proteolytic activities of MMP-1 (collagenase) were performed using fluorescent collagen substrates. UVA induced MMP-1 synthesis and activity were analyzed by enzyme-linked immunosorbent assay (ELISA) and gelatin-based zymography in skin fibroblasts. We also examined anti-inflammatory effects by the determination test of proinflammatory cytokine, interleukin 6 in HaCaT keratinocytes. Expression of prostaglandin E$_2$ (PGE$_2$) after UVB irradiation was measured by competitive enzyme immunoassay(EIA) using PGE$_2$ monoclonal antibody. In the human skin we tested anti-irritation effect on the SLS-induced damage skin after appling the extract containing emulsion. We found that Juniperus chinensis extract had potent radical scavenging effect by 98% at 100$\mu\textrm{g}$/mL. The extract of Juniperus chinensis showed strong inhibitory effect on MMP-1 activities by 97% at 100 $\mu\textrm{g}$/mL and suppressed the UVA induced expression of MMP-1 by 79% at 25$\mu\textrm{g}$/mL. This extract also showed strong inhibition on MMP-2 activity in UVA irradiated fibroblast by zymography. In the test of proinflammatory cytokines of human keratinocytes Juniperus chinensis extract decreased expression of interleukin 6 about 30%. The amount of PGE$_2$ by HaCaT keratinocytes was significantly increased at the doses of above 10 mJ/$\textrm{cm}^2$ of UVB (p < 0.05). At the concentrations of 3.2-25$\mu\textrm{g}$/mL of this extract, the production of PGE$_2$ by HaCaT keratinocytes (24 h after 10mJ/$\textrm{cm}^2$ UVB irradiation) was significantly inhibited in culture supernatants (p < 0.05). In SLS-induced skin irritation model in vivo, we found to reduce skin erythema and improve barrier recovery after appling Juniperus chinensis extract containing emulsion when compared to irritated non-treated and placebo-treated skin. Our results suggest that Juniperus chinensis extract can be effectively used for the prevention of UV and SLS-induced adverse skin reactions and applied as anti-aging and anti-irritation cosmetics.

Clinical Significance of Plasma TGF-${\beta}_1$ in Coal Workers' Pneumoconiosis (탄광부 진폐증에서 혈장 Transforming Growth Factor-${\beta}_1$의 의의)

  • Kim, Chong-Ju;Lee, Won-Yeon;Hong, Ae-Ra;Shin, Pyo-Jin;Yong, Suk-Joong;Shin, Kye-Chul
    • Tuberculosis and Respiratory Diseases
    • /
    • v.50 no.1
    • /
    • pp.76-83
    • /
    • 2001
  • Background : Coal workers' pneumoconiosis is a fibrotic lung disease resulting from chronic inhalation of coal dust. The precise mechanism of lung fibrosis in coal workers' pneumoconiosis is uncertain. However, a relationship between the stimulation of fibroblast proliferation and collagen production by mediators released from in flammatory and resident lung cells is thought to be a major factor. The transforming growth factor-$\beta$(TGF-$\beta$), a multifunctional cytokine and growth factor, plays a key role in the scarring and fibrotic processes due to its ability to induce extracellular matrix proteins and modulate the growth and immune function of many cell types. To determine the involvement of TGF-$\beta$ in the development of lung fibrosis in coal workers' pneumoconiosis, the TGF-${\beta}_1$ level in plasma was measured in patients with coal workers' pneumoconiosis. Methods : Plasma was collected from 40 patients with coal workers' pneumoconiosis (20 with simple coal workers' pneumoconiosis and 20 with complicated coal workers' pneumoconiosis) and from 10 normal controls. The ELISA method was used to measure the plasma TGF-${\beta}_1$ concentration. Results : Compared to the control group ($0.63{\pm}01.8$ ng/mL), there was no significant difference in the plasma TGF-${\beta}_1$ level in patients with simple coal workers' pneumoconiosis ($0.64{\pm}0.17$ ng/mL) (p>0.05). However, in patients with complicated coal workers' pneumoconiosis the plasma TGF-${\beta}_1$ level ($0.79{\pm}0.18$ ng/mL) was significantly higher than in patients with simple coal workers' pneumoconiosis and the control group (p<0.05). Conclusion : The data suggests that TGF-${\beta}_1$ has some influence in the development of lung fibrosis in coal workers' pneumoconiosis.

  • PDF

GRO-${\alpha}$, IL-8 and ENA-78 : Expressed by Stimulated Endothelial Cells and Increased PMN Adhesion (활성화된 내피세포에서 GRO-${\alpha}$, IL-8 및 ENA-78의 발현양상과 호중구 부착에 미치는 영향)

  • Ryu, Ki-Chan;Kim, Yun-Seong;Kim, Yong-Ki;Kim, In-Ju;Kim, Young-Dae;Lee, Chang-Hun;Park, Do-Youn;Kim, Ji-Yeon;Ha, Tae-Jeong;Lee, Min-Ki;Park, Soon-Kew
    • Tuberculosis and Respiratory Diseases
    • /
    • v.52 no.2
    • /
    • pp.145-155
    • /
    • 2002
  • Background: Inflammation, where vascular endothelial cells are activated by cytokines, recruits circulating leukocytes such as neutrophils into the tissues. Mononuclear phagocytes as well as tissue cells activated by these stimuli produce these chemokines. In this study, thr effects of IL-1 and LPS on the expression of CXC chemokines such as GRO-${\alpha}$, IL-8 and ENA-78 in vascular endothelial cells and the neutrophil adhesion effects of ENA-78 and GRO-${\alpha}$ was investigated. Methods: Human umbilical vein endothelial cells were cultured and stimulated with various concentrations of IL-1 and LPS. The concentrations of the GRO-${\alpha}$, IL-8 and ENA-78 secreted were measured using enzymelinked immunosorbent assay. The effects of ENA-78 and GRO-${\alpha}$ on neutrophil adhesion to the endothelial cells were also investigated. Results: The addition of IL-1 and LPS to the vascular endothelial cells induced GRO-${\alpha}$ IL-8 and ENA-78 secretion in a time- and dose-dependent manner. The neutrophil adhesion was also increased by induction of ENA-78 and GRO-${\alpha}$ to the vascular endothelial cells in a dose-dependent manner. Conclusion: CXC chemokines such as GRO-${\alpha}$, IL-8 and ENA-78 secreted by the vascular endothelial cells play an important role in the acute inflammatory responses by stimulating neutrophil adhesion to the vascular endothelial cells, raising the possibility that the CXC chemokines are one of the targets in the clinical application of acute inflammation.

Interleukin Levels in the Bronchoalveolar Lavage Fluid of Patients with Pulmonary Sarcoidosis (유육종증환자의 기관지폐포세척액내 Interleukin에 대한 연구)

  • Song, Jeong-Sup;Ahn, Joong-Hyun;Kim, Chi-Hong;Kim, Kwan-Hyoung;Moon, Hwa-Sik;Park, Sung-Hak
    • Tuberculosis and Respiratory Diseases
    • /
    • v.45 no.5
    • /
    • pp.1047-1057
    • /
    • 1998
  • Background : Sarcoidosis is a systemic granulomatous disorder of unknown origin and characterized by accumulation of T cells and macrophages. Various cytokines may play crucial roles in the activation of T cells and macrophages, and thereby in the formation of granulomas. However, little is known about the balance between proinflammatory cytokines and antiinflammatory cytokines in the development of sarcoid granulomas and disease activities. In the present study, we measured IL-6, IL-8 and IL-10 in the bronchoalveolar lavage fluid(BALF) from patients with pulmonary sarcoidosis to find out whether there is an imbalance between proinflammatory cytokines and antiinflammatory cytokines in the lung. Methods: Fourteen subjects with the diagnosis of sarcoidosis and six healthy volunteers were included. BALF was concentrated ten-fold by pressure ultrafiltration and each cytokine levels were measured by EUSA method. Active sarcoidosis was defined by major organ involvement or clinically progressive diseases. Results: The mean IL-6 levels in the BALF of the active sarcoidosis group were significantly increased than in controls or inactive sarcoidosis group(p<0.05). Meanwhile, the IL-8 levels were increased and IL-10 levels were decreased in the active sarcoidosis group than in controls or inactive sarcoidosis group without significance(p>0.05). In active pulmonary sarcoidosis patients, the IL-6 levels in BALF correlated with the BALF CD4/CD8 ratio(r=0.768, p<0.05) and IL-8 levels(r=0.564, p<0.05). Conclusions : The data presented showed that pro-inflammatory cytokine IL-6 is important in the pathogenesis of sarcoidosis and decreased tendency of anti-inflammatory cytokine IL-10 might also be involved in the development of granulomatous inflammation in sarcoidosis.

  • PDF

Neuroprotective Effects of Modified Yuldahanso-tang (MYH) in a Parkinson's Disease Mouse Model (MPTP로 유도된 Parkinson's disease 동물 모델에서 열다한소탕 가감방 (MYH)의 신경 세포 보호 효과)

  • Go, Ga-Yeon;Kim, Yoon-Ha;Ahn, Taek-Won
    • Journal of Sasang Constitutional Medicine
    • /
    • v.27 no.2
    • /
    • pp.270-287
    • /
    • 2015
  • Objectives To evaluate the neuroprotective effects of modified Yuldahanso-tang (MYH) in a Parkinson's disease mouse model. Methods 1) Four groups (each of 8 rats per group) were used in this study. 2) The neuroprotective effect of MYH was examined in a Parkinson's disease mouse model. C57BL/6 mice treated with 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP, 30 mg/kg/day), intraperitoneal (i.p.) for 5 days. 3) The brains of 2 mice per group were removed and frozen at $-20^{\circ}C$, and the striatum-substantia nigra part was seperated. The protein volume was measured by Bradford method following Bio-Rad protein analyzing kit. Using mouse/Rat Dopamine ELISA Assay Kit. 4) The brains of 2 mice per group were separated and removed. TH-immunohistochemical was examined in the MPTP-induced Parkinson's disease mice to evaluate the neuroprotective effects of MYH on ST and SNpc. 5) Two mice out of each group were anesthetized and skulls were opened from occipital to frontal direction to take out the brains. The brains added TTC solution for 20 minutes for staining. 6) The water tank used for morris water maze test was filled with $28^{\circ}C$ water, and a round platform of 10cm in diameter was installed for mice to step on. The study was carried out once a day within 30 seconds, keep exercising to step on the platform in the pool. 7) The brains of two mice out of each group were fixed in 10% formaldehyde solution and paraphillin substance was infiltrated. They were fragmented by microtome, and observed under an optical microscope after Hematoxylin & Eosin staining. 8) A round acrylic cylinder with its upper side open was filled with clean water and depressive mouse models were forced to swim for 15 minutes. After 24 hours the animals were put in the same equipment for 5 minutes and were forced to swim. 9) The convenient, simple, and accurate high-performance liquid chromatography (HPLC) method was established for simultaneous determination of Neurotransmitters in MPTP-MYH group. Results 1) MYH possess Dopamine cell protective effect on MPTP-induced injury in striatum and substantia nigra pars compacta. 2) MYH inhibits the loss of tyrosine hydroxylase-immunoreacitive (TH-IR) cells in the striatum and substantia nigra pars compacta on MPTP-induced injury in C57BL/6 mice. 3) MYH possesses improvement effect on MPTP-induced memory deterioration in C57BL/6 mice through the reduction of prolongated Sort of lost time by MPTP injection using the Morris water maze test. 4) MYH possesses hippocampal neuron protective effect on MPTP-induced injury in C57BL/6 mice. 5) MYH possesses improvement effect on MPTP-induced motor behaviour deficits and depression in C57BL/6 mice through the reduction of prolongated losing motion by MPTP injection using the Forced swimming test. 6) MYH increases serotonin product amount on MPTP-induced injury in C57BL/6 mice. Conclusions This experiment suggests that the neuroprotective effect of MYH is mediated by the increase in Dopamin, TH-ir cell, Hippocampus and Serotonin. Furthermore, MYH essential oil may serve as a potential preventive or therapeutic agent regarding Parkinson's disease.

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
    • /
    • v.38 no.5
    • /
    • pp.548-554
    • /
    • 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.