• Title/Summary/Keyword: ELISA-inhibition test

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The Characteristics, Detection and Control of Bacteriophage in Fermented Dairy Products (발효유제품에서 박테리오파지의 특성, 검출과 제어)

  • Ahn, Sung-Il;Azzouny, Rehab A.;Huyen, Tran Thi Thanh;Kwak, Hae-Soo
    • Food Science of Animal Resources
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    • v.29 no.1
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    • pp.1-14
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    • 2009
  • This study was to review the classification, detection and control of bacteriophage in fermented dairy products. Bacteriophage has lytic and/or lysogenic life cycles. Epidemiologically speaking, detected major phages are c2, 936 and p335. Among them p335 has been the largest concern in dairy industry. Traditionally, various analytical technologies, such as spot, starter activity, indicator test, ATP measurement and conductimetric analysis, have been used for the phage detection. In recent years, advanced methods such as flow cytometric method, petrifilm, enzyme linked immunosorbent assay (ELISA) and multiflex PCR diagnostic kit have been deveoloped. The phage contamination has been controlled by using heat, high-pressure treatment, and the combinations of heat and pressure, and/or chemical. Also some starter cultures with phage-resistant character have been developed to minimize the concentration of phages in dairy product. Bacteriophage inhibition media such as calcium medium was also mentioned. To prevent the contamination of bacteriophage in dairy industry, further researches on the detection and control of phage, and phage resistant starters are necessary in the future.

Immunomodulatory Effects of Callophyllis japonica Ethanol Extract on Dendritic Cells (수지상세포에 대한 Callophyllis japonica 추출물의 면역조절효과)

  • Kim, Mi-Hyoung;Joo, Hong-Gu
    • IMMUNE NETWORK
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    • v.7 no.2
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    • pp.95-100
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    • 2007
  • Background: A red seaweed, Callophyllis japonica has been traditionally eaten in the oriental area. In a recent study, it has been demonstrated that the ethanol extract of C. japonica have antioxidant activity. However, there are few studies about the effects of C. japonica on the function of immune cells. We investigated the immunomodulatory effects of C. japonica on the function of dendritic cells, the potent antigen-presenting cells. Methods: Bone marrow-derived dendritic cells (DCs) were used and the viability was measured by 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide assay and trypan blue exclusion test. Cytokine and nitric oxide (NO) levels were determined by using ELISA and Griess reagent, respectively. The expression levels of DC surface markers were measured by flow cytometric analysis. Results: C. japonica ethanol extract did not significantly affect the DCs viability and the IL-12 production from DCs, irrespective of the presence of lipopolysaccharide (LPS). In addition, it did not significantly change the expression of DC surface markers. However, C. japonica ethanol extract significantly inhibited the LPS-induced NO production and also increased the proliferation of allogeneic lymphocytes activated by DCs. Conclusion: Our data suggests that C. japonica ethanol extract enhances the proliferation of allogeneic lymphocytes activated by DCs which is associated with inhibition of NO production from DCs induced by LPS.

Vp28 of Shrimp White Spot Syndrome Virus Is Involved in the Attachment and Penetration into Shrimp Cells

  • Yi, Guohua;Wang, Zhimin;Qi, Yipeng;Yao, Lunguang;Qian, Juan;Hu, Longbo
    • BMB Reports
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    • v.37 no.6
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    • pp.726-734
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    • 2004
  • White spot disease (WSD) is caused by the white spot syndrome virus (WSSV), which results in devastating losses to the shrimp farming industry around the world. However, the mechanism of virus entry and spread into the shrimp cells is unknown. A binding assay in vitro demonstrated VP28-EGFP (envelope protein VP28 fused with enhanced green fluorescence protein) binding to shrimp cells. This provides direct evidence that VP28-EGFP can bind to shrimp cells at pH 6.0 within 0.5 h. However, the protein was observed to enter the cytoplasm 3 h post-adsorption. Meanwhile, the plaque inhibition test showed that the polyclonal antibody against VP28 (a major envelope protein of WSSV) could neutralize the WSSV and block an infection with the virus. The result of competition ELISA further confirmed that the envelope protein VP28 could compete with WSSV to bind to shrimp cells. Overall, VP28 of the WSSV can bind to shrimp cells as an attachment protein, and can help the virus enter the cytoplasm.

Studies on Anti-Wrinkle and Whitening Effects of Liposomes Containing Acerola Extract Mixture (아세로라 추출물 혼합 리포좀의 주름, 미백 효과에 대한 연구)

  • Kim, Su Jin;Oh, Won Jun;Kwon, Sung Pil;Nam, Gaewon
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.47 no.4
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    • pp.341-352
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    • 2021
  • Acerola is an excellent ingredient because of its high natural vitamin C content, but it is difficult to stabilize and has hardly been studied as a cosmetic material. Therefore, this study developed a mixed liposome preparation for stabilizing acerola extract. As a safety test, the skin irritation test was evaluated by BCOP assay and HET-CAM assay. We evaluated the inhibition of tyrosinase activity, the whitening effect of melanin production, and the wrinkle effect of prochloragentype-I C-peptide production, and confirmed the possibility of functional cosmetics. In addition, a cream of liposomes containing acerola extract mixture was developed to evaluate the clinical studies of skin wrinkles and whitening. BCOP assay, HET-CAM assay and human skin primary irritation test results of liposomes containing acerola extract mixture showed no irritation and were safe from skin and eye. The result of tyrosinase activity by 75.8% at 1,000 ㎍/mL. As a result of the melanogenesis inhibition test, liposome with acerola extract showed the melanin content by 46.2% at 1,000 ㎍/mL that does not effect the viability of the B16F10 cell line. The result of collagen production test using ELISA kit, liposomes containing acerola extract mixture showed collagen synthesis ability by 152.1% at 1,000 ㎍/mL that does not affect the viability of the HS68 cell line. But it did not showed any inhibition of collagenase (MMP-1) activity at all concentrations in the MMP-1 activity inhibition test in the HS68 cell line. We performed clinical studies for the whitening and skin-wrinkle activity of cream containing acerola extract mixes liposome, was showed that the melanin contents and wrinkle was statistically significant reduction. These results suggest that liposomes containing acerola extract mixture have safe natural material, and skin wrinkle, whitening effects allowing their application in cosmetics as a natural product.

Fusion Peptide Improves Stability and Bioactivity of Single Chain Antibody against Rabies Virus

  • Xi, Hualong;Zhang, Kaixin;Yin, Yanchun;Gu, Tiejun;Sun, Qing;Shi, Linqing;Zhang, Renxia;Jiang, Chunlai;Kong, Wei;Wu, Yongge
    • Journal of Microbiology and Biotechnology
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    • v.27 no.4
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    • pp.718-724
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    • 2017
  • The combination of rabies immunoglobulin (RIG) with a vaccine is currently effective against rabies infections, but improvements are needed. Genetic engineering antibody technology is an attractive approach for developing novel antibodies to replace RIG. In our previous study, a single-chain variable fragment, scFv57R, against rabies virus glycoprotein was constructed. However, its inherent weak stability and short half-life compared with the parent RIG may limit its diagnostic and therapeutic application. Therefore, an acidic tail of synuclein (ATS) derived from the C-terminal acidic tail of human alpha-synuclein protein was fused to the C-terminus of scFv57R in order to help it resist adverse stress and improve the stability and half-life. The tail showed no apparent effect on the preparation procedure and affinity of the protein, nor did it change the neutralizing potency in vitro. In the ELISA test of molecular stability, the ATS fusion form of the protein, scFv57R-ATS, showed an increase in thermal stability and longer half-life in serum than scFv57R. The protection against fatal rabies virus challenge improved after fusing the tail to the scFv, which may be attributed to the improved stability. Thus, the ATS fusion approach presented here is easily implemented and can be used as a new strategy to improve the stability and half-life of engineered antibody proteins for practical applications.

Inhibition of Leptin and Leptin Receptor Gene Expression by Silibinin-Curcumin Combination

  • Nejati-Koshki, Kazem;Akbarzadeh, Abolfazl;Pourhasan-Moghaddam, Mohammad;Abhari, Alireza;Dariushnejad, Hassan
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.11
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    • pp.6595-6599
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    • 2013
  • Leptin and its receptor are involved in breast carcinogenesis as mitogenic factors. Therefore, they could be considered as targets for breast cancer therapy. Expression of the leptin receptor gene could be modulated by leptin secretion. Silibinin and curcumin are herbal compounds with anti-cancer activity against breast cancer. The aim of this study was to assess their potential to inhibit of expression of the leptin gene and its receptor and leptin secretion. Cytotoxic effects of the two agents on combination on T47D breast cancer cells was investigated by MTT assay test after 24h treatment. With different concentrations the levels of leptin, leptin receptor genes expression were measured by reverse-transcription real-time PCR. Amount of secreted leptin in the culture medium was determined by ELISA. Data were statistically analyzed by one-way ANOVA test. The silibinin and curcumin combination inhibited growth of T47D cells in a dose dependent manner. There were also significant difference between control and treated cells in leptin expression and the quantity of secreted leptin with a relative decrease in leptin receptor expression. In conclusion, these herbal compounds inhibit the expression and secretion of leptin and it could probably be used as drug candidates for breast cancer therapy through leptin targeting in the future.

Effect of Mixture extracted from Bupleuri Radix and Physalidis Herba on the LPS-induced depression in Rats (시호(柴胡)와 산장(酸漿) 복합제가 LPS로 유도된 우울증에 미치는 효과)

  • Lee, Won-Haeng;Kim, Do-Hoon;Lee, Tae Hee
    • The Korea Journal of Herbology
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    • v.30 no.3
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    • pp.69-76
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    • 2015
  • Objectives : The effect of mixture extracted from Bupleuri Radix and Physalidis Herba(BR+RH) on the LPS-induced Depression in rats was investigated. Methods : Rats were administered intragastrically BR+PH after injectio of LPS to induce deprssion. Immobility was examined using Tail Suspension Test(TST), Forced Swimming Test(FST). The level of plasma corticosterone was measured by an Enzyme-Linked Immunosorbent Assay(ELISA) method. The expressions of c-Fos, Corticotropin Releasing Factor(CRF), NADPH-d in the Paraventricular nucleus(PVN) and TH in the Locus coeruleus(LC) were measured by immunohistochemical method. Results : In the effect of BR + PH on TST, immobility was significantly decreased comparing with the LPS group. In FST, immobility was shown decrease tendency in the BR+PH group. The expression of c-Fos in the PVN was significantly decreased at BR + PH400 group, comparing with the LPS group. The expression of CRF in PVN was shown dto have the decrease tendency in the BR+PH group, comparing with the LPS group. The expression of NADPH-d in PVN was not significantly decreased at BR+PH groups, comparing with the LPS group. The expression of TH in the LC was shown to have the decrease tendency at BR + RH groups, but not significantly, comparing with the LPS group. Conclusions : Anti-depressant effect of mixture after extracted from Bupleuri Radix and Physalidis Herba was through the anti-inflammatory effect via inhibition of HPA axis. NO and catecholamine system is not involved.

Effect of Banggibongnyeongtang on LPS-induced Depression in rats (방기복령탕(防己茯苓湯)이 흰쥐에서 LPS로 유도된 우울증에 미치는 효과)

  • Park, Sung jun;Lee, Tae Hee
    • Herbal Formula Science
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    • v.27 no.2
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    • pp.137-149
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    • 2019
  • Objective : This study is conducted in order to investigate the effect of Banggibongnyeongtang(BBT) on Lipopolysaccharide (LPS)-induced depression. Method : LPS $5{\mu}g$ was injected to lateral ventricle. Experimental groups were administered BBT intraperitoneally. Depressive behavior was confirmed by weight change, sucrose preference, open field test(OFT), and forced swimming test(FST). The plasma concentration of $IL-1{\beta}$ and $TNF-{\alpha}$, Corticotropin-Releasing Factor(CRF), Adrenocorticotropin Hormone(ACTH) and Corticosterone(CORT) were measured by ELISA. Result : BBT did not change the body weight significantly than LPS group, but on sucrose preference, BBT increased significantly in LPS+BBT400 group compared to LPS group (P<0.05). In the OFT, BBT increased spending time in the central zone and decreased grooming number. LPS+BBT400 group increased central zone-spending time, and decreased grooming number than LPS group significantly (P<0.05). In the FST, LPS+BBT400 group decreased immobility time than LPS group significantly (P<0.05). BBT decreased $IL-1{\beta}$ concentration does-dependently, but only with significant decrease in LPS+BBT400 group than LPS group in plasma (P<0.05). But BBT did not decrease $TNF-{\alpha}$ concentration significantly in plasma. BBT decreased plasma CRF, ACTH, and CORT. And CRH and CORT of LPS+BBT400 group were shown significant decrease comparing with LPS group (P<0.05). Conclusion : It is postulated that the anti-depressant effect of BBT can be validated through inhibition of HPA axis abnormal activity by the anti-inflammatory effect.

The Effects of Pulsatilla Koreana for Anti - Inflammatory and Cellular Activity of Periodontal Tissue (백두옹 추출물의 치주조직 세포에 활성도 및 항염 효과에 관한 연구)

  • Jung, Jin-Gwang;Chung, Chin-Hyung;Lim, Sung-Bin;Kim, Jung-Keun;So, Eun-Hee
    • Journal of Periodontal and Implant Science
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    • v.31 no.1
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    • pp.149-165
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    • 2001
  • This study was performed to define the cytotoxicity and the anti-inflammatory action of Pulsatilla koreana extracts. To analyze cytotoxic effects, gingival and periodontal ligament fibroblasts were used, and anti-inflammatory actions related to reduction of $IL-1{\beta}$ and $PGE_2$ production were performed in vitro, for the suggestion of efficacy and safety on periodontal therapeutic use of Pulsatilla koreana extracts. We extracted ethylacetate and butylalcohol from well-dried and ground Pulsatilla koreana throughout multiple processing, then used different concentration solution(0.1 %, 0.2 %, 0.4 %, 0.01 %, 0.02 %, 0.04 %, 1 %, 2 %) of ethylacetate and butylalcohol extracts to examine eytotoxic effects and anti-inflammatory actions Cytotoxic effects were examined by ELISA reader using MTT(Methyl Thiazol-2-YL-2, 5-diphenyl Tetrazolium bromide)solution following culture of human gingival and periodontal ligament fibroblasts. Synthesis of $IL-1{\beta}$was examined by $IL-1{\beta}$ enzyme-immunoassay(EIA)system after separation and culture of monocyte, and $PGE_2$ was examined by $PGE_2EIA$ system after culture of gingival fibroblasts. The results were as follows: 1. In the MTT test of gingival fibroblasts, the change of optical density was decreased significantly at 2 % of butylalcohol extracts and 0.04 %, 0.1 %, 0.2 %, 0.4 %, 1 %, 2 % of ethylacetate extracts.(p<0.05) 2. In the MTT test of periodontal ligament cells, the change of optical density were not differ significantly. but butylalcohol and ethylacetate extracts except from butylalcohol 0.01 % showed high cell cytotoxity. 3. Both ethylacetate and butylalcohol extracts from Pulsatilla koreana inhibited the synthesis of $IL-1{\beta}$and inhibition effect of ethylacetate extracts were higher than butylalcohol extracts. 4. Both ethylacetate and butylalcohol extracts from Pulsatilla koreana inhibited the synthesis of $PGE_2$, and ethylacetate extracts were higher than butylalcohol extracts. In conclusion, ethylacetate and butylalcohol extracts from Pulsatilla koreana showed little cell cytotoxity for gingival and periodontal ligament fibroblasts, and the inhibition of $IL-1{\beta}$ and $PGE_2$ sysnthesis, therefore it is considered that these extracts can be developed as the therapeutics of the periodontal disease.

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Xylitol Down-Regulates $1{\alpha},25$-Dihydroxy Vitamin D3-induced Osteoclastogenesis via in Part the Inhibition of RANKL Expression in Osteoblasts

  • Ohk, Seung-Ho;Jeong, Hyunjoo;Kim, Jong-Pill;Yoo, Yun-Jung;Seo, Jeong-Taeg;Shin, Dong-Min;Lee, Syng-Ill
    • International Journal of Oral Biology
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    • v.38 no.3
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    • pp.127-134
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    • 2013
  • Xylitol is a sugar alcohol with a variety of functions including bactericidal and anticariogenic effects. However, the cellular mechanisms underlying the role of xylitol in bone metabolism are not yet clarified. In our present study, we exploited the physiological role of xylitol on osteoclast differentiation in a co-culture system of osteoblastic and RAW 264.7 cells. Xylitol treatment of these co-cultures reduced the number of tartrate-resistant acid phosphatase (TRAP)-positive multinucleated cells induced by 10 nM $1{\alpha},25(OH)_2D_3$ in a dose-dependent manner. A cell viability test revealed no marked cellular damage by up to 100 mM of xylitol. Exposure of osteoblastic cells to xylitol decreased RANKL, but not OPG, mRNA expression in the presence of $10^{-8}M$ $1{\alpha},25(OH)_2D_3$ in a dose-dependent manner. Furthermore, bone resorption activity, assessed on bone slices in the coculture system, was found to be dramatically decreased with increasing xylitol concentrations. RANKL and OPG proteins were assayed by ELISA and the soluble RANKL (sRANKL) concentration was decreased with an increased xylitol concentration. In contrast, OPG was unaltered by any xylitol concentration in this assay. These results indicate that xylitol inhibits $1{\alpha},25(OH)_2D_3$-induced osteoclastogenesis by reducing the sRANKL/OPG expression ratio in osteoblastic cells.