• Journal of Microbiology and Biotechnology
• /
• v.13 no.4
• /
• pp.509-516
• /
• 2003

A model system for the immnunochemical detection of Escherichia coli O157:H7 using a combined immunomagnetic separation (IMS) and test-strip liposome immunoassay (LIA) procedure was developed. Immunomagnetic beads coated with anti-E. coli O157 IgG antibodies were used to separate the E. coli O157 (including the H7 serotype) from culture. Immunoliposomes, whose surface was conjugated to goat anti-E. coli O157:H7 IgG and which encapsulated the marker dye, sulforhodamine B, were used as a detection label. The test strip, onto which antibodies to goat IgG were immobilized, was the immunosensor capturing immunoliposomes that did not bind to E. coli O157:H7 on the immunomagnetic bead-E. coli O157:H7 complexes. In experiments, pure cell culture suspensions of $10^5 E.$ coli O157:H7 organisms per ml produced a measurable signal inhibition, whereas a weak yet detectable signal inhibition occurred with $10^3CFU/ml$. The inhibition signals increased, when the incubation time for IMS was extended to 90 min and higher IgG-tag density (0.4mol%) was used on the liposomes. With 0.2 and 0.4mol% IgG-tagged liposomes, the IMS-LIA procedure showed more improved signal inhibitions than those of a direct (no IMS) LIA. The combined assay, which measures the instantaneous signal from immunoliposomes, can be completed within 90 min, making it significantly faster than conventional plating methods and enzyme-linked immunosorbent assay (ELISA). Accordingly, it is quite feasible to use the combined immunoassay format of IMS and dye-loaded immunoliposomes for the detection of E. coli O157:H7.

• Journal of Periodontal and Implant Science
• /
• v.24 no.1
• /
• pp.144-154
• /
• 1994

Final goal of periodontal treatment is to reconstruct the destructed periodontal tissue as well as to remove the necrotic pathologic elements. The purpose of this study is to investigate on the effect of Zizyphi extract to the inhibitory ability on collagenolytic activity of P gingivalis, biologic activity of gingival fibroblasts, and on the collagen and protein synthesis of gingival fibroblasts. Gingival fibroblast from giniva of first bicuspids from patient for orthodontic treatment were used and cultured. For the measurement of inhibitory ability of collagenolytic activity, crude enzyme was extracted and used on the basis of modified Ono's method. On the inhibition of collagenolytic enzyme from herbal extracts, collagenokit CLN-100 were used. The cellular activity of gingival fibroblast, were studied using MTT solution and measured optical density on 570mm by ELISA reader. To measure the effects on the ability of whole protein and collagen synthesis, cell membrane was destructed with ultrasonic grinder after culturing, centrifuged and counted by liquid scintilation counter. The inhibitory effects on producing of $IL-l{\beta}$ by monocyte, after promotion of producing $IL-l{\beta}$ by LPS, were compared with the mixture of herbal extracts and other drugs using thymocyte stimulation assay. About inhibitory effects of $PGF_2$. by gingival fibroblasts, herbal extract was compared with the addition of the other control groups using enzyme imunoassay. On the inhibition of collagenolytic activity by P. gingivalis, benzene extracts showed the most efficient inhibitory effects among the $19{\mu}g/ml$ of the compared extracts and 40.5% by Tetracycline. On the cellular activity promoting effects, compared extracts showed a bit of more effects than PDGF of $100{\mu}g/ml$ concentration and IGF of $20{\mu}g/ml$ concentration. All of the PDGF, IGF, Zizyphi Fructus extract should increase in collagen synthesis, but especially 70% ethylalcohol extracts of Zizyphi Fructus showed comparably high effects among the compared extracts. Effects on whole protein synthesis were slightly increased on every extract but especially 70% ethylalcohol extract showed significantly effective than any other estract. On the inhibitory effects of Zizyphi Fructus $IL-l{\beta}$ production by monocyte, compared extracts showed 70% of highly inhibitory effect than that of 60% inhibition effects on controlled group and each extracts showed no significant difference. In $PGF_2$ production inhibitroy effect of Zizyphi Fructus gingival fibroblasts, Herbal extracts showed 70% of inhibition comparing with tat of 90.2% of controlled group, but each extracts showed similar effects excluding the $H_2O$ extracts. These results suggested that Zizyphi Fructus might be useful medicine for inhibition of inflammatory mediator including $IL-l{\beta}$ and $PGF_2$.

• Journal of Life Science
• /
• v.12 no.4
• /
• pp.460-463
• /
• 2002

Twelve clones of monoclonal antibodies (mAbs) against thyroglobulin were produced and characterized. Among them, three mAbs (TN-1, TN-2 and TN-3) showed high binding affinity to thyroglobulin. from ELISA inhibition assay, TN-2 showed considerable reactivity with soluble thyroglobulin. TN-2 also reacted with phosphatidylserine which has a negative charge in aqueous condition. These results suggest that TN-2 has characteristics of autoantibody concerned with thyroiditis.

• Proceedings of the Korean Society of Applied Pharmacology
• /
• 1994.04a
• /
• pp.255-255
• /
• 1994

MIEF는 소의 초유중에 함유되어 있는 분자량 22,000 dalton의 peptide로서 말초혈액 림파구의 배양액에 첨가할 경우 자연살해세포를 활성화 시켜 암세포에 대한 자연살해능을 증가시키는 것으로 이미 보고된 바 있다. 본 연구에서는 수유기간별 MIEF의 함량 및 MIEF가 B세포의 증식 및 분화에 미치는 영향을 조사하였다. 수유기간별 MlEF의 함량을 competitive ELISA inhibition assay를 이용하여 조사한 결과 MIEF의 함량은 수유 첫째날의 초유에 109$\mu\textrm{g}$/ml로 가장 높았으며, 그 이후로 급격히 감소하여 3일째 이후에서는 3-4$\mu\textrm{g}$/ml이하로 존재하였다. MIEF가 B세포의 분화에 미치는 영향은 사람의 편도선에서 분리한 림파구의 배양액에 MIEF를 가하고 5-9일간 배양한후 항체를 생산하는 세포의 수를 ELISPOT assay로 측정하여 조사하였다. MIEF는 10-30$\mu\textrm{g}$/ml의 농도에서 B세포의 분화를 촉진시켰으며, 그 정도는 양성대조군으로 사용한 LPS와 유사하여 MIEF가 B세포의 분화를 유도하는 작용이 아주 강력함을 알수 있었다.

• Molecular & Cellular Toxicology
• /
• v.2 no.4
• /
• pp.223-228
• /
• 2006

Several plant-derived cannabinoids and endogenous ligands for cannabinoid receptors such as 2-arachidonyl-glycerol have been known to inhibit interleukin-2 (IL-2) expression. In the present study, we utilized arachidonylethanolamide (AEA), a putative endogenous ligand for cannabinoid receptors, to determine whether AEA modulated the expression of IL-2. AEA inhibited phorbol 12-myristate 13-acetate (PMA) plus ionomycin (Io)-induced IL-2 protein secretion and mRNA expression in EL-4 mouse T-cells as determined by ELISA and RT-PCR, respectively. To further characterize the inhibitory mechanism of AEA at the transcriptional level, we performed promoter study for IL-2 gene in PMA/Io-stimulated EL-4 cells. AEA decreased the transcriptional activity of the nuclear factor of activated T-cells (NF-AT) as well as the IL-2 promoter activity. These results suggest that AEA suppresses IL-2 expression and that the inhibition is mediated, at least in part, through the down-regulation of NF-AT.

• Journal of Environmental Health Sciences
• /
• v.16 no.1
• /
• pp.45-53
• /
• 1990

To investigate the effect of cultural condition on the aflatoxin production of Aspergillus flayus ATCC 15517, mixed culture with Aspergillus niger, better kind of media and size of Cultural vessels were examined. YES medium was better than SLS medium for this study. Small scale test tube culture was showed the possibility to simply examine the growth, total acidity, pH and aflatoxin production during cultivation, and also could reduce the second contamination of aflatoxin B1 from large scale broth cultured. Especially ELISA method is simple, sensitive and specific and therefore well suited to small scale of test tube culture. Mixed culture significantly reduced the aflatoxin production of Aspergillus fiavus ATCC 15517 and showed almost 95% inhibition of that level during the incubtation.

• Journal of the Korean Society of Food Science and Nutrition
• /
• v.29 no.3
• /
• pp.437-441
• /
• 2000

감마선 조사기술을 이용하여 새우 알러지를 감소시키기 위해 본 연구를 실시하였다. 감마선 조사구(A), 감마선 조사후 가열처리구(B), 가열처리 후 감마선 조사구(C)로 실험구를 설정하고 1,3,5,7,10 kGy의 감마선을 조사하여 실험에 사용하였다. A 처리구로부터 새우의 근혈장 단백질 용액과 근섬유 단백질 용액을 준비하였고, B와 C처리구에서도 각각의 단백질 용액을 준비하였다. 각 단백질 용엑에 대한 환자 IgE와 mAb 4.9.5의 결합력을 ELISA로 조사하였다. 두 항체 모두 감마선 조사구의 단백질을 용액과의 결합력이 감마선 조사선량이 증가할수록 낮게 나타났고, 감마선 조사후 가열처리가 항체의 결합력을 감소시키는데 더 효과적이었다. SDS-PAGE 결과 감마선 조사된 새우 단백질의 전기영동적 분리 pattern에는 변화가 없었다. 이 결과는 적절한 선량의 감마선 조사가 새우 알러지를 감소시킬 수 있다는 것을 시사한다.

• YAKHAK HOEJI
• /
• v.36 no.2
• /
• pp.188-190
• /
• 1992

Two clones of monconal antibodies(Co-1 and Co-2) against BSA-benzoylecgonine(BSABE) were produced. Both monoclonal antibodies showed high binding affinity to BSA-BE. Observing from ELISA inhibition assay, Co-1 reacted only weakly with soluble benzoylecgonine, while Co-2 showed considerable reactivity with soluble benzoylecgonine.

• The Korea Journal of Herbology
• /
• v.23 no.2
• /
• pp.99-109
• /
• 2008

Objectives : The present study was to investigate the effect of extract of Chelidonii herba (ECH) on the proliferation and activation of eosinophils which were prepared from lung cells of asthma-induced mouse by ovalbumin (OVA) treatment. Methods : C57BL/6 mouse was exposed to OVA three times a week for 6 weeks. The mouse lung tissues were dissected out, chopped and dossiciated with collagenase (1 ${\mu}g$/ml). Eosinophils were activated by rIL-3/rmIL-5 co-treatments. The lung cells were treated with ECH, incubated for 48 hr at $37^{\circ}C$, and analyzed by flow cytometery, ELISA, RT-PCR, and immuno-histochemical analysis. Results : In FACS analysis, number of granulocyte/lymphocyte, $CD3e^-$/$CCR3^+$, $CD3e^+$/$CD69^+$, $CD4^+$ and $CD23^+$/$B220^+$ in asthma-induced lung cells were significantly decreased by ECH treatment compared to the control group. And mRNA expression for IL-4, IL-5, IL-13, CCR3 and eotaxin in asthma-induced lung cells, which was induced by rIL-3 plus rmIL-5 treatments, was significantly decreased by ECH treatment. In ELISA analysis, production levels of IL-3, IL-5, IL-13 and histamine in asthma-induced lung cells, which were induced by rIL-3 plus rmIL-5 co-treatment, were significantly decreased by ECH treatment. ECH treatments significantly inhibited the proliferation of eosinohils prepared from asthma-induced mouse lung tissues compared to the non-ECH treated control cells. Immunohistochemical analysis revealed that ECH treatment significantly decreased the levels of eosipnphil activation compared to non-treated cells. Conclusions : The present data suggested that Chelidonium majus L. may have an effect on the inhibition of parameters associated with asthma responses in eosinpophils, and thus implicate the possibility for the clinical application of Chelidonium majus L.

• Journal of Digital Convergence
• /
• v.13 no.8
• /
• pp.503-514
• /
• 2015

The effects of Flavonol contents from Ginkgo biloba leaf on anti-atopy activity have not rarely been verified. This study is to investigate the effects of flavonol on Th2 cytokine production in MC/9 mast cells. For this, flavonol was analyzed by ELISA and Real-time PCR. Analysis results showed that flavonol significantly suppressed production of Th2 cytokines(IL-13, MIP-1a) in a dose dependent manner. The mRNA expression of IL-4, IL-5, IL-13, TNF-a were effectively restrained by Flavonol at the concentration 25,50,$100{\mu}g/m{\ell}$. And decrease of expression of NFAT-1, c-jun protein was confirmed by western blot analysis. These results indicate that flavonol has effects of decreasing the Th2 cytokine production in the MC/9 mast cell causing inhibition of transcription factors such as NFAT-1, c-jun. Thus, we would like to brief that flavonol may have the applicability as therapeutic agent for atopic dermatitis.