• Title/Summary/Keyword: cell enzyme-linked immunosorbent assay

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Improvement of anti-oxidant and anti-inflammatory activities of aleurone layer extracts of black rice (Oryza sativa L.) by enzyme treatment (효소 처리에 의한 흑미 호분 추출물의 산화방지와 항염증 활성 증진)

  • Lee, Mi Kyoung;Ryu, Soo In;Lee, Min Ho
    • Korean Journal of Food Science and Technology
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    • v.50 no.5
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    • pp.528-534
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    • 2018
  • The current study investigated the effects of enzyme treatment on black rice (Oryza sativa L.) aleurone layer extracts. Different enzymes (lipase, lecitase ultra and lipopan 50BG) were used to test anti-oxidant and anti-inflammatory activities in vitro. The antioxidant activities of enzyme treated or non-enzyme treated extracts of black rice bran were evaluated via 1,1-diphenyl-2-picrylhydrazyl radical scavenging activity and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) radical scavenging activity. Lipase treated extracts of black rice bran showed higher antioxidant activity compared to that of non-enzyme treated extracts. Anti-inflammatory activities of enzyme treated black rice bran extracts on nitrite production and tumor necrosis $factor-{\alpha}$ ($TNF-{\alpha}$) secretion, were tested using a nitric oxide (NO) colorimetric assay kit and an enzyme-linked immunosorbent assay (ELISA) kit. The ethanolic extract of enzyme treated black rice bran decreased the levels of nitric oxide production and pro-inflammatory cytokines, such as tumor necrosis $factor-{\alpha}$, in a lipopolysaccharidestimulated RAW 264.7 cell culture. These findings indicate that enhanced anti-oxidant and anti-inflammation activities of the ethanolic extracts of enzyme treated black rice (Oryza sativa L.) aleurone layers, may be attributed to molecular conversion of ingredients in enzyme catalyzed reactions.

Characteristics of outbreak for the classical swine fever (CSF) at Incheon metropolitan area in 2002 (인천지역에서 발생한 돼지콜레라의 역학적 특성)

  • 권효정;변재원;이정구;김경호;박은정;이성모;황현순
    • Korean Journal of Veterinary Service
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    • v.26 no.1
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    • pp.1-9
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    • 2003
  • The aim of this survey was to investigate the characteristics of outbreak farm determined as the classical swine fever(CSF) at Gangwha-gun and Seo-gu, Incheon metropolitan area from October 7 to November 25 in 2002. Sixty pigs in six different farms were confirmed to the CSF and a total of 9,106 pigs containing 3,194 related epidemiologically was slaughtered to stop spreading of the disease. Clinical signs of pigs diagnosed with the CSF were high fever, anorexia, depression, paralysis of hindlimbs, cyanosis, etc and gross lesions were typically represented with hemorrhage of submandibular and superficial lymph node, infarction of spleen, and petechial (ecchymotic) hemorrhage of kidney and skin. But some outbreak farms had not shown remarkable symptoms, so they were confused with other bacterial diseases. White blood cell (WBC) counts, the classical swine fever virus(CSFV) antigen and antibody enzyme linked immunosorbent assay(ELISA) and reverse transcriptase polymerase chain reaction (RT-PCR) results about six farms indicated that total 60 pigs were infected with the CSFV. Although the origin and infection route of the CSFV were not clear, but the transmissions between farms were mainly through indirect contact such as the movement of farm personal and vehicles from outbreak farm.

Leptin potentiates Prevotella intermedia lipopolysaccharide-induced production of TNF-$\alpha$ in monocyte-derived macrophages

  • Kim, Sung-Jo
    • Journal of Periodontal and Implant Science
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    • v.40 no.3
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    • pp.119-124
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    • 2010
  • Purpose: In addition to regulating body weight, leptin is also recognized for its role in the regulation of immune function and inflammation. The purpose of this study was to investigate the effect of leptin on Prevotella (P.) intermedia lipopolysaccharide (LPS)-induced tumor necrosis factor (TNF)-$\alpha$ production in differentiated THP-1 cells, a human monocytic cell line. Methods: LPS from P. intermedia ATCC 25611 was prepared by the standard hot phenol-water method. THP-1 cells were incubated in the medium supplemented with phorbol myristate acetate to induce differentiation into macrophage-like cells. The amount of TNF-$\alpha$ and interleukin-8 secreted into the culture medium was determined by enzyme-linked immunosorbent assay (ELISA). TNF-$\alpha$ and Ob-R mRNA expression levels were determined by semi-quantitative reverse transcription-polymerase chain reaction analysis. Results: Leptin enhanced P. intermedia LPS-induced TNF-$\alpha$ production in a dose-dependent manner. Leptin modulated P. intermedia LPS-induced TNF-$\alpha$ expression predominantly at the transcriptional level. Effect of leptin on P. intermedia LPS-induced TNF-$\alpha$ production was not mediated by the leptin receptor. Conclusions: The ability of leptin to enhance P. intermedia LPS-induced TNF-$\alpha$ production may be important in the establishment of chronic lesion accompanied by osseous tissue destruction observed in inflammatory periodontal disease.

In Vitro Expression of the Recombinant hFSH Gene using Retrovirus Vector System (In Vitro에서 Retrovirus Vector System을 이용한 재조합 hFSH 유전자의 발현)

  • Min, Gyeong-Heon;Kwon, Mo-Sun;Kim, Teoan;Koo, Bon-Chul
    • Reproductive and Developmental Biology
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    • v.35 no.1
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    • pp.115-121
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    • 2011
  • hFSH is a glycoprotein secreted from anterior pituitary and consists of ${\alpha}$ and ${\beta}$ subunits. Because of its major biological functions including sperm formation in the male and for follicular growth, FSH is used to cure woman's sterility. In this study we tried to produce recombinant hFSH in vitro using a retrovirus expression vector. Two major components of the vector we constructed are: ( i ) a DNA fragment containing ${\alpha}$ and ${\beta}$ genes fused by a DNA sequence coding carboxyl terminal peptide (CTP) of human chorionic gonadotropin, (ii) a DNA fragment corresponding woodchuck hepatitis virus posttranscriptional regulatory element (WPRE). Evaluation of expression profile of the recombinant FSH using reverse transcription PCR and enzyme-linked immunosorbent assay (ELISA). Among three cell lines tested, HeLa cells were the best for hFSH expression (5,395 mIU/ml), then followed by chicken embryonic fibroblast (CEF) cells and Chinese hamster ovary (CHO) cells in the order of hFSH production. In addition to the amount, the FSH produced from HeLa cells was highest in terms of biological activity which was determined by measuring cAMP.

Citrus unshiu Water Extract Inhibits Trypsin-induced $TNF-{\alpha}$ and Tryptase Productions by Blocking the ERK Phosphorylation and Trypsin Activity

  • Kang, Ok-Hwa;Kim, Dae-Ki;Lee, Young-Mi
    • Natural Product Sciences
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    • v.10 no.5
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    • pp.211-216
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    • 2004
  • Citrus unshiu (Rutaceae) has long been known as an anti-inflammatory and anti-allergic agent. In the present study, the inhibitory effect of CUWE (Citus unshiu water extract) on the production of $TNF-{\alpha}$ and tryptase was examined. In addition, a possible mechanism for the inhibition of trypsin-stimulated human leukemic mast cell-1 (HMC- 1 ) activation was determined. To do so, $TNF-{\alpha}$ production from the HMC-1 cells that were stimulated by trypsin (100 nM) in the presence or absence of CUWE $(10,\;100,\;and\;100\;{\mu}g/ml)$ was measured by enzyme-linked immunosorbent assay (ELISA) and reverse transcription-PCR. The tryptase production was evaluated by reverse transcription-PCR. Extracellular signal-regulated kinase (ERK) activation was analyzed by Western blot. Trypsin activity was measured by using Bz-DL-Arg-p-nitroanilide (BAPNA) as substrate. Results showed that the CUWE inhibited production of both $TNF-{\alpha}$ and tryptase from the trypsin-stimulated HMC-1 in a dose-dependent manner. The CUWE a1so inhibited the ERK phosphorylation and trysin activity. These results indicate that the CUWE had an inhibitory effect on $TNF-{\alpha}$ and the tryptase productions by blocking the ERK phosphorylation and trypsin activity.

Chemical and Immunobiological Characterization of Lipopolysaccharides from Prevotella intermedia and Prevotella nigrescens (Prevotella intermedia와 Prevotella nigrescens의 세균내독소에 대한 연구;화학적 분석 및 면역생물학적 활성 평가)

  • Kim, Sung-Jo
    • Journal of Periodontal and Implant Science
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    • v.34 no.2
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    • pp.461-474
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    • 2004
  • The purpose of this study was to assess some biological activities of lipopolysaccharides (LPSs) from P. intermedia and P. nigrescens. LPS was prepared by the standard hot phenol-water method. NO production was assayed by measuring the accumulation of nitrite in culture supernatants. $TNF-{\alpha}$ production was determined by enzyme-linked immunosorbent assay. Western blot analysis of iNOS and analysis of reverse transcription (RT)-PCR products were carried out. LPS from P. intermedia demonstrated higher KDO content than those from two stains of P. nigrescens. LPSs from P. intermedia and P. nigrescens were mitogenic for spleen cells of BALB/C mouse. The present study clearly shows that LPSs from P. intermedia and P. nigrescens fully induced iNOS expression and NO production in RAW264.7 cells in the absence of other stimuli. Moreover, LPSs from P. intermedia and P. nigrescens clearly induced $TNF-{\alpha}$ production in RAW264.7 cells. The biological activities of LPS from P. intermedia was found to be comparable to those of P. nigrescens LPS. The ability of LPSs from P. intermedia and P. nigrescens to promote the production of NO and $TNF-{\alpha}$ may be important in the pathogenesis of inflammatory periodontal disease.

Preparation of the Monoclonal Antibodies against the Zppspores of Allomyces macrogynus (Allomyces macrogynus의 유주자와 반응하는 단일클론항체의 준비)

  • Choi, So-Young;Hwang, Jung-Sook;Kim, Jung-Seoup;Park, Kyung-Hee;Cho, Chung-Won;Youn, Hyun-Joo
    • Journal of Life Science
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    • v.6 no.4
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    • pp.264-269
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    • 1996
  • Monoclonal antibodies against the zoospores of Allomyces macrogynus were prepared using standard hybridoma technique. Mice were immunized either with the fixed zoospores or the zoospore proteins, and the production of the antibodies from the resulting hybridomas were screened by enzyme-linked immunosorbent assay (ELISA). Thirty hybridomas were initially identified ans six hybridomas were purified to the single cell clones. Culture supernatants from the hybridomas were tested for the effects on the growth of the germ tubes, and some of the hybridoma culture supernatants studied showed growth stimulatory effects.

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Effects of Draconis Resina on the Collagenase Activities and the Procollagen Synthesis in Hs68 Human Fibroblasts, and Tyrosinase Activity

  • Kim, Tae Yeon;Leem, Kang-Hyun
    • The Korea Journal of Herbology
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    • v.30 no.6
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    • pp.1-6
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    • 2015
  • Objectives : Draconis Resina (DR), the resin of Daemonorops draco Bl., is used to circulate the blood and to stop bleeding. It also has been used to generate flesh including ulceration. The present study investigated the effects of DR extract on collagen metabolism in human fibroblasts and tyrosinase activity in mushroom tyrosinase.Methods : The effect of DR extract on type I procollagen production (collagen type I synthesis) and collagenase (matrix metalloproteinase-1, henceforth referred as MMP-1) activity in human normal fibroblasts cell line. Hs68 cells after ultraviolet B (UVB, 312 nm) irradiation was measured using the enzyme - linked immunosorbent assay (ELISA). The tyrosinase activity was also measured to find out the whitening effects in mushroom tyrosinase by ELISA method.Results : There was no cytotoxicity at DR extract at concentrations of 10 μg/ml, 30 μg/ml, and 100 μg/ml. DR extract significantly inhibited the increase of collagenase activity, whereas it did not show on the reduction of type I procollagen in UVB damaged Hs68 cells. DR extract did not reduce the L - DOPA oxidation. However, it significantly reduced the tyrosinase activity by DR extract at concentraions of 0.1 mg/ml, 1 mg/ml and 10 mg/ml.Conclusions : In conclusion, DR showed the anti-wrinkle and whitening effects via the inhibition of collagenase production and the tyrosinase activity. These results suggest that DR may have potential as an anti-aging ingredient in cosmetic herb markets.

The Neuroprotective Effects of Carnosine in Early Stage of Focal Ischemia Rodent Model

  • Park, Hui-Seung;Han, Kyung-Hoon;Shin, Jeoung-A;Park, Joo-Hyun;Song, Kwan-Young;Kim, Doh-Hee
    • Journal of Korean Neurosurgical Society
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    • v.55 no.3
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    • pp.125-130
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    • 2014
  • Objective : This study was conducted to elucidate neuroprotective effect of carnosine in early stage of stroke. Methods : Early stage of rodent stroke model and neuroblastoma chemical hypoxia model was established by middle cerebral artery occlusion and antimycin A. Neuroprotective effect of carnosine was investigated with 100, 250, and 500 mg of carnosine treatment. And antioxidant expression was analyzed by enzyme linked immunosorbent assay (ELISA) and western blot in brain and blood. Results : Intraperitoneal injection of 500 mg carnosine induced significant decrease of infarct volume and expansion of penumbra (p<0.05). The expression of superoxide dismutase (SOD) showed significant increase than in saline group in blood and brain (p<0.05). In the analysis of chemical hypoxia, carnosine induced increase of neuronal cell viability and decrease of reactive oxygen species (ROS) production. Conclusion : Carnosine has neuroprotective property which was related to antioxidant capacity in early stage of stroke. And, the oxidative stress should be considered one of major factor in early ischemic stroke.

Dioscorea japonica Thunb. Ethanolic Extract Attenuated Oxazolone-Induced Atopic Dermatitis-like Skin Lesions in BALB/c Mice

  • Jegal, Jonghwan;Park, No-June;Jo, Beom-Geun;Kim, Su-Nam;Yang, Min Hye
    • Natural Product Sciences
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    • v.25 no.3
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    • pp.261-267
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    • 2019
  • The rhizomes of Dioscorea japonica Thunb. are widely consumed as food and also used to treat diabetes and polyuria in Korea. This study was undertaken to study the anti-atopic dermatitis effects of a 95% ethanolic extract (DJE) of D. japonica in an oxazolone-stimulated murine model of atopic dermatitis (AD). The therapeutic effects of DJE on AD-like skin lesions were assessed on both ears. DJE (1%) or dexamethasone (0.5%; the positive control) were applied to skin lesions for three weeks. Serum levels of IgE and IL-4 were assessed by ELISA (enzyme-linked immunosorbent assay). Histopathological examinations were performed by hematoxylin and eosin (H&E) and toluidine blue staining and revealed DJE significantly reduced dermal thickness and inflammatory cell infiltration when applied to oxazolone-treated ear skin. DJE-treated AD mice also showed lower serum levels of IgE and IL-4 than oxazolone-stimulated controls. Our findings demonstrate DJE might be a useful safe, topical agent for the treatment of atopic diseases.