• Title/Summary/Keyword: ELISA system

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Activation of the renin-angiotensin system in high fructose-induced metabolic syndrome

  • Kim, Mina;Do, Ga Young;Kim, Inkyeom
    • The Korean Journal of Physiology and Pharmacology
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    • 제24권4호
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    • pp.319-328
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    • 2020
  • High fructose intake induces hyperglycemia and hypertension. However, the mechanism by which fructose induces metabolic syndrome is largely unknown. We hypothesized that high fructose intake induces activation of the renin-angiotensin system (RAS), resulting in hypertension and metabolic syndrome. We provided 11-week-old Sprague-Dawley rats with drinking water, with or without 20% fructose, for two weeks. We measured serum renin, angiotensin II (Ang II), and aldosterone (Aldo) using ELISA kits. The expression of RAS genes was determined by quantitative reverse transcription polymerase chain reaction. High fructose intake increased body weight and water retention, regardless of food intake or urine volume. After two weeks, fructose intake induced glucose intolerance and hypertension. High fructose intake increased serum renin, Ang II, triglyceride, and cholesterol levels, but not Aldo levels. High fructose intake increased the expression of angiotensinogen in the liver; angiotensin-converting enzyme in the lungs; and renin, angiotensin II type 1a receptor (AT1aR), and angiotensin II type 1b receptor (AT1bR) in the kidneys. However, expression of AT1aR and AT1bR in the adrenal glands did not increase in rats given fructose. Taken together, these results indicate that high fructose intake induces activation of RAS, resulting in hypertension and metabolic syndrome.

상아질 결합제의 L929 세포 독성에 관한 연구 (CYTOTOXICITY OF DENTIN BONDING SYSTEM ON L929 CELLS)

  • 임미경;지정호;김상섭
    • Restorative Dentistry and Endodontics
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    • 제22권1호
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    • pp.209-219
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    • 1997
  • The research of the dentin bonding system was mainly on the chemistry and bonding strength. And in vitro assessement of biocompatibility of dentin bonding system was not completely developed. The purpose of this study was to evaluate the cytotoxic effect of several dentin primers. Scotchbond Multi-Purpose (3M Dental Products. USA). Gluma (BayerDental. Germany). All-Bond (Bisco. USA). ProBond (CaulkDensply, USA) and VeridonFil (Dongyang Nylon. Korea) were included. Cytotoxicity was tested using MTT cell viability test. 0.5 ul. 1 ul. 2 ul and 10 ul of each primer were added to the 96 well plate of incubated L929 cell lines. After 30-minute. 1. 4. 24 and 72-hour exposures. absorbance of L929 cells was observed with ELISA reader. All data were analyzed using t-test. All primers showed cytotoxicity on L929 cells under every conditions used in this study. Absorbance of L929 cells was decreased by time. Scotch bond group exhibited the lowest absorbance value in all exposure time and value.

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천연물 유래 물질이 감염성 질환에 미치는 영향과 효능 (Effects and Efficacy of Natural Product on Infectious Diseases of pseudomonas aeruginosa)

  • 박지원
    • 한국자원식물학회:학술대회논문집
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    • 한국자원식물학회 2020년도 추계국제학술대회
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    • pp.3-13
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    • 2020
  • Pseudomonas aeruginosa is a ubiquitous gram-negative bacterium causing serious infections. The P. aeruginosa T3SS is a syringe-like apparatus on the bacterial surface, with 4 effector toxins: ExoS, ExoT, ExoY, and ExoU. Here, we investigated the effect of ExoS and ExoT of the T3SS of P. aeruginosa K strain (PAK). The type three secretion system (T3SS) is a major virulence system of Pseudomonas aeruginosa (P. aeruginosa). The effector protein Exotoxin S (ExoS) produced by P. aeruginosa is secreted into the host cells via the T3SS. For the purpose of screening the inhibitors with regard to ExoS secretion, we developed the sandwich-type enzyme-linked immunosorbent assay (ELISA) system. PAK clinical strains induce proinflammatory cytokine production through the T3SS, and this involves NF-κB activation in pneumonia mouse models. We tried to confirm the role of the NF-κB transcription factor in ExoS- and ExoT-induced pneumonia mouse models. pro-inflammatory cytokines induction in response to ExoS and ExoT infection relied on NF-κB activation. Our findings highlight the roles of natural poduct in inhibiting proinflammatory cytokine expression during ExoS and ExoT exposure in PAK infections, paving the way for a novel therapeutic approach for the treatment of pulmonary infections.

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청열윤부탕(淸熱潤膚湯)이 DNFB로 유발된 알레르기 피부염에 미치는 효과 (Effects of Cheongyeolyunbu-tang on DNFB-induced Allergic Dermatitis)

  • 이경기;김진주;정희재;정승기
    • 대한한방내과학회지
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    • 제29권3호
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    • pp.730-741
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    • 2008
  • Objective : To examine the efficacy of CY, the improvement of atopy dermatitis(AD)-like lesions on the rostral back of the NC/Nga mice, which took CY orally and were treated with a chemical substance called DNFB, the inhibition of ear swelling, and the inhibition of inflammatory response of the ear tissue of the mice were observed and compared, and the inhibition of the serum IgE count and the IL-4 and IFN-${\gamma}$ count produced by CD4+ T cells of the lymph node were observed and compared. Materials and Methods : For measurement of ear thicknesses, 0.15% DNFB $25{\mu}l$ mixed with acetone/olive oil(3:1) was applied to the right ear and dorsal skin of the NC/Nga mice 5 times at an interval of 7 days. Ear thickness was measured every day over a five-week period after the first application of DNFB. The total serum IgE count was measured with ELISA by taking blood samples 24 hours after the fifth application of DNFB. To measure the IL-4 and IFN-${\gamma}$ count, the lymph node was cut off, and the CD4+ T cells were purified and stimulated to activate the T cells, and then the IL-4 and IFN-${\gamma}$ count was measured with ELISA. Results : Continuous oral administration of CY improved the AD-like skin lesions on the rostral back and inhibited the ear swelling in NC/Nga mice treated with DNFB and inhibited the inflammatory response in the NC/Nga mice treated with DNFB NC/Nga mice. Continuous oral administration of CY did not significantly inhibit the serum IgE count and failed to significantly reduce the IL-4 count generated after TCR stimulation in the CD4+ T cells of the NC/Nga mice treated with DNFB. Continuous oral administration of CY significantly reduced the IFN-${\gamma}$ count generated after TCR stimulation in the CD4+ T cells of the NC/Nga mice treated with DNFB.

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NC/Nga 마우스 전혈을 이용한 항 아토피 피부염 물질 탐색 (Screening of Anti-Atopic Dermatitis Material by Using NC/Nga Mouse Whole Blood System)

  • 박동훈;김연욱
    • IMMUNE NETWORK
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    • 제8권3호
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    • pp.98-105
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    • 2008
  • Background: Allergic inflammation was induced by activated Th2 lymphocytes, leading to IgE production and eosinophil activation. A Th2 disproportion was shown in atopic children soon after birth. During specific allergen stimulation, an increase of Th2 cells was observed in most cases. In this study, we prepared new screening "whole blood" system for searching the anti-atopic materials. Cytokine production and IgE secretion from whole blood system were assessed and we confirmed the results by using animal system. Methods: Pathological features in NC/Nga mice are similar to those observed in human atopic dermatitis. Whole blood from NC/Nga mouse was stimulated by using TNCB (Th2 activator) or candidate materials of anti-atopic dermatitis, and the production of cytokines (IL-4, IL-12, and IFN-${\gamma}$) were measured by ELISA. In order to confirm the results of whole blood system, in vivo test was done by using NC/Nga mice. Results: In whole blood system, LPS and extracts of green tea, hardy orange and onion induced the production of IL-12 and IFN-${\gamma}$ while they reduced the production of IL-4. Also, LPS and extracts of onion reduced IgE production. Though atopic dermatitis was observed from a mouse stimulated with TNCB, it was not when a mouse was co-stimulated in LPS or extracts of onion. The results are same as those observed in whole blood system. Conclusion: Whole blood system was simple and speedy methods for searching a materials compared with the conventional high-cost animal system. And the results using whole blood system was proved to be reliable in our experiments for screening anti-atopic material. We expect that the system can be applied to other experiments for searching similar materials.

The Effects of Phragmitis Rhizoma Herbal-acupuncture Solution on Inflammation in Human Mast Cells and Human Alveolar Epithelial Cell Lines - Phragmitis Rhizoma's Effects -

  • Kim, Byung-Soo
    • 대한한의학회지
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    • 제35권4호
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    • pp.1-9
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    • 2014
  • Objectives: This study was designed to find the effect of Phragmitis Rhizoma (PR) herbal-acupuncture solution on the inflammatory cytokine and chemokine secretion in human mast cell (HMC) and human alveolar epithelial cell 549 (A549) lines. Methods: Histamine levels in HMC after PR herbal-acupuncture solution treatment were measured with ELISA. Other cytokines and chemokines levels such as interleukin 8 (IL-8), monocyte chemoattractant protein-1 (MCP-1), and chemokine (C-C motif) ligand 5 (Ccl5, RANTES) in A549 were measured with flow cytometry CBA system. Results: In the PR herbal-acupuncture solution treatment group, the expression of histamine, IL-8, MPC-1, Ccl5, and RANTES decreased significantly. Conclusions: The results support that PR herbal-acupuncture solution had a suppressive effect on cytokine-induced inflammation.

성장중인 수토끼에서 혈청 IGF-I 수준과 Flow Cytometry 측정 정자형성의 변화

  • 김창근;이주형;방명걸;류재원;장유민;박민영;지달영;정영채
    • 한국동물번식학회:학술대회논문집
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    • 한국동물번식학회 2004년도 춘계학술발표대회
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    • pp.279-279
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    • 2004
  • 성장중인 수토끼에서 성성숙 전후의 혈청내 IGF-I 수준과 정자형성의 변화를 12주령부터 28주령까지 2주 간격으로 조사하였다. IGF-I 수준 측정은 non-extraction IGF-I ELISA kit(Biognostic System, Lab, USA)를 이용하였으며 정소내 정자형성은 biopsy needle(22G×3¹/₂ 72㎜×8.89㎝, Becton Dickinson, USA)로 채취한 정소조직을 fled ctyometric ploidy analysis로 DNA histogram을 4개 peak(peak Ⅰ과Ⅱ :1N, peak Ⅱ :2N, peak Ⅳ : 4N, peak Ⅲ과 Ⅳ사이 : S기)로 구분하였다. (중략)

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Immune reactions and allergy in experimental anisakiasis

  • Cho, Sung-Weon;Lee, Haneul-Nari
    • Parasites, Hosts and Diseases
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    • 제44권4호
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    • pp.271-283
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    • 2006
  • The third-stage larvae (L3) of the parasitic nematode, Anisakis simplex, have been implicated in the induction of hyperimmune allergic reactions in orally infected humans. In this work, we have conducted a review of an investigation into immune reactions occurring in animals experimentally infected with A. simplex L3. The patterns of serum antibody productions if the experimental animals against excretory-secretory products (ESP) of A. simplex L3 contributed to our current knowledge regarding specific humoral immune reactions in humans. In our review, we were able to determine that L3 infection of experimental animals may constitute a good model system for further exploration of immune mechanisms and allergy in anisakiasis of humans.

어류생체지표를 이용한 내분비계장애 연구 (Studies on the Endocrine Disruption in Wildlife Fish)

  • 구자민;류지성;정규혁;이철우;박응로;박광식
    • Environmental Analysis Health and Toxicology
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    • 제16권4호
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    • pp.197-204
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    • 2001
  • Endocrine disruption in crucian carp (Carassius auratus) living in the branch of Han River were examined. Vitellogenin level in plasma was measured using ELISA system and aromatase mRNA level in brain was observed using RT-PCR technique. In all female fish, vitellogenin levels were in the range of 20∼40 $\mu\textrm{g}$/ml and aromatase mRNA expression could be detected on the agarose gel after RT-PCR. However, in case of males, vitellogenin level was elevated in only one fish, while vitellogenin was hardly detected in others. Aromatase was expressed in all males although the levels were relatively lower than the level in female fish. Testis-ova and any other histological changes of reproductive organ were not shown in both sexes.

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대장균내에서 발현된 돼지 TGF-$\beta$1의 분리 및 면역학적 항원성 보유검증

  • 최은영;김현태;김평현;변우현
    • 한국미생물·생명공학회지
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    • 제25권2호
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    • pp.137-143
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    • 1997
  • Porcine transforming growth factor-$\beta$1 (TGF-$\beta$1) was expressed in Escherichia coli using cDNA of TGF-$\beta$1 and glutathione S-transferase (GST) fusion vector pGEX-1$\lambda$T. An ApoI-Tth111I fragment of cDNA which correspond to the amino acid residues from 123 to 390 of the precursor TGF-$\beta$1 was inserted into EcoRI-Tth111I digested pGEM#-l$\lambda$T and the recombined plasmid was named pGET-12. Gene products from the cloned regions of the recombinant plasmids pGET-12 was not detected in soluble fraction of cell free extract but detected in insoluble fraction. The solubilization of insoluble gene product was achieved by the treatment of N-laurylsarcosine. Molecular weight of partially purified proteins determined by electrophoresis was same as expected from cloned fragment. The ELISA test results of the purified proteins showed that immunologically detectable epitope was preserved in recombinant protein.

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