• 제목/요약/키워드: in vitro activity test

검색결과 529건 처리시간 0.027초

Effects of CpG Oligodeoxynucleotides on Immune Responses and Expression of Cytokine Genes in Cultured Olive Flounder Paralichthys olivaceus

  • Ahn, kyoung-Jin;Nam, Bo-Hye;Kim, Young-Ok;Kang, Jung-Ha;Kim, Bong-Seok;Jee, Young-Ju;Lee, Sang-Jun
    • Fisheries and Aquatic Sciences
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    • 제10권1호
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    • pp.1-7
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    • 2007
  • The induction of cellular and humoral immunity and cytokine gene expression by synthetic CpG oligodexoynucleotides (CpG-ODNs) has not been investigated systematically in olive flounder Paralichthys olivaceus in vivo. We optimized the proper concentration of CpG-ODNs using an in vitro assay for the superoxide anion $(O_2^-)$. CpG-ODNs induced $O_2^-$ and nitric oxide (NO) production, lysozyme activity, and the proinflammatory cytokine gene expression of $IL-1{\beta}$ and $TNF-{\alpha}$ in olive flounder significantly in vivo, whereas non-CpG-ODNs did not produce these effects or produced them to a lesser extent. This implied that CpG-ODNs could stimulate cellular and humoral immunity and cytokine gene expression in olive flounder. This is the first evidence of NO production and the first study on the mRNA expression of the proinflammatory cytokine genes $IL-1{\beta}$ and $TNF-{\alpha}$ in olive flounder in response to CpG-ODNs. Comparison of the variation in NO production and lysozyme activity to that of other studies led us to postulate that a group-specific difference exists in the immune responses of olive flounder against CpG-ODNs. Furthermore, the detailed immunostimulatory spectrum of CpG-ODNs in olive flounder could be a useful index with which to analyze the effect of CpG-ODNs against the challenge test prior to field applications.

Acmthmoeba culbertsoni 감염에 대한 silica 투여의 영향 - 대식세포의 역할을 중심으로 - (The effect of silica on the development of experimental Acanthamoeba meningoencephalitis with reference to the macrophage role in mice)

  • 이홍수;신호준
    • Parasites, Hosts and Diseases
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    • 제32권4호
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    • pp.259-266
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    • 1994
  • CSH/HeJ 마우스에 Acanaamoeba culbertsoni영양형 $3{\;}{\times}{\;}10^5$개를 비강으로 접종하 여 실험적 아메바성 수막뇌염을 일으킬 때 숙주의 방어기작에 대식세포가 미치는 영향을 알아 보았다. 대식세포 억제제인 silica를 마우스 복강 내로 투여한 경우의 사망율이 60.6%로 아메바만을 접종한 마우스의 사망율이 10.0%와 비하여 큰 차이를 보였으며 열처리하여 죽인 Toxoplasma gondii tachyzoites의 in vitro탐식능 관찰에서 기능을 억제시킨 대식세포의 탐식능이 3% 이하로써 아메바에 대한 복강대식세포의 기능의 저하로 인한 수막뇌염의 증가를 관찰하였다. A. culbertsoni에 대한 대식세포의 살해활동의 in vitro 실험에서 silica투여한 실험군의 복강대식세포의 뚜렷한 기능 저하를 관찰하였고, 효소표지 면역 검사법(ELISU)을 이용한 마우스 혈청 내의 $interleukin-1{\beta}(IL-1{\beta})$의 흡광도는 아메바 접종군과 생리식염수 투여군보다 silica투여를 수반한 실험군이 현저하게 낮게 나타나 대식세포가 억제되었을 때 실험적 수막뇌염이 증가함을 알 수 있었다. 결과적으로 대식세포가 A. culbertsoni 감염에 대한 숙주의 방어작용에 중요한 역할을 하는 것으로 생각된다.

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Taurine Possesses In vitro Antimutagenic Activity Comparable to Major Antioxidants

  • Sung, Mi-Kyung;Jeon, Hye-Seung;Park, Taesun
    • Preventive Nutrition and Food Science
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    • 제4권1호
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    • pp.43-46
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    • 1999
  • Taurine is known to suppress oxidant-induced tissue injury by stabilizing biomembrane and scavanging free radicals. The purpose of this study was to determne the antioxidative and antimutabenic acitvities of taurine, ad to compare those acitivities with major antioxidants. For the measurement of antioxidative capacity, 0.05 , 0.1,0.5 and 1.0mg/ml of taurine, L-Ascorbic acid, alpha-tocopherol, and BHT (dibuty hydroxiy toluene)were prepared and tested for their ability to donate electrons to DPPH (1,1-diphenyl-2-picryl-hydrazyl). Antimutagenic acitivity was examined using the Ames salmonela test system at concentrations of 600, 900 and 1200ug/ml. Results indicated that taurine possesses electron-donating capacity, however, the degree of donation was very weak compared to the major antioxidants tested. However, taurine was evaluated as a potent mutation suppressor. Antimutagenic capacity was in increasing order BHT>taurine>L-ascorbic acid>alpha-tocopherol at concentrations of 600 and 900ug/ml. There was a dose-dependent increase in antimutabenicity of these compounds , however, antimutagenity of the 900ug taurie/plate was not significantly differently from that of 1200ug taurine/plate. These results indicate that taurine effectively suppresses the mutagenicity of AFB1 without noticeable elelctron donating ability.

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골수기질세포와 진피섬유모세포의 이식이 교원질 합성에 미치는 영향 (Effect of Transplantation of Bone Marrow Stromal Cells and Dermal Fibroblasts on Collagen Synthesis)

  • 최원일;한승규;이병일;김우경
    • Archives of Plastic Surgery
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    • 제34권2호
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    • pp.156-162
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    • 2007
  • Purpose: In the previous in vitro studies the bone marrow stromal cells(BSCs) have shown the superior effect for wound healing activity than fibroblasts, which includes cell proliferation, type I collagen synthesis, and the production of bFGF, VEGF and TGF-${\beta}$ in chronic wound healing. The aim of this study is to compare the effects of BSCs and fibroblasts on wound healing activity in vivo, especially on collagen synthesis. Methods: The fibroblasts and BSCs were harvested from patients and cultured. The cultured cells were infiltrated into the pores of polyethylene discs. These discs were divided into three groups according to the mixed cells. In groups I, II and III the discs were loaded with no cells, fibroblasts and BSCs, respectively. Twelve discs per group(total 36 discs) were made for this study. After creating 6 pockets in the back of each rats, each discs was implanted into each pockets. At three time intervals from 1 to 3 weeks, the implanted discs were harvested for the histological and quantitative analysis. The amount of collagen produced was evaluated using ELISA. Statistical comparisons were made using the Mann-Whitney U-test. Results: There was great difference in the collagen synthesis among the three groups by the 1st and 2nd weeks. The BSC group showed highest collagen level, followed by fibroblast group and no cell group(p<0.05). The 3rd week specimens also showed greater collagen amount in BSC and fibroblast groups compared to those of no cell group(p<0.05). However, there was little difference between BSC and fibroblast groups. Conclusion: This result demonstrates that BSC has superior effect on stimulating wound healing than fibroblast, which is currently used for wound healing.

B형 간염 바이러스 X 항원을 면역한 A2Kb Transgenic Mice에서 CD8+ T Cell의 활성화에 의한 X 항원 표현 재조합 Vaccinia Virus에 대한 방어 효과; in vitro 배양을 통한 XEP-3 특이적인 CTL의 반응 (Protective Effects on A2Kb Transgenic Mice That Were Immunized with Hepatitis B Virus X Antigen Peptides by the Activation of CD8+ T Cells; XEP-3 Specific CTL Responses in the in vitro Culture)

  • 황유경;김형일;김남경;박정민;정홍석
    • IMMUNE NETWORK
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    • 제2권1호
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    • pp.41-48
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    • 2002
  • Background: Viral antigens presented on the cell surface in association with MHC class I molecules are recognized by CD8+ T cells. MHC restricted peptides are important in eliciting cellular immune responses. As peptide antigens have a weak immunigenicity, pH-sensitive liposomes were used for peptide delivery to induce effective cytotoxic T lymphocyte (CTL) responses. In the previous study, as the HBx peptides could induce specific CTLs in vitro, we tested whether the HLA-A2/$K^b$ transgenic mice that were immunized by HBx-derived peptides could be protected from a viral challenge. Methods: HBx-peptides encapsulated by pH-sensitive liposomes were prepared. $A2K^b$ transgenic mice were immunized i.m. on days one and seven with the indicated concentrations of liposome-encapsulated peptides. Three weeks later, mice were infected with $1{\times}10^7pfu$/head of recombinant vaccinia virus (rVV)-HBx via i.p. administration. The ovaries were extracted from the mice, and the presence of rVV-HBx in the ovaries was analyzed using human TK-143B cells. IFN-${\gamma}$ secretion by these cells was directly assessed using a peptide-pulsed target cell stimulation assay with either peptide-pulsed antigen presenting cells (APCs), concanavalin A ($2{\mu}g/ml$), or a vehicle. To generate peptide-specific CTLs, splenocytes obtained from the immunized mice were stimulated with $20{\mu}g/ml$ of each peptide and restimulated with peptide-pulsed APC four times. The cytotoxic activity of the CTLs was assessed by standard $^{51}Cr$-release assay and intracellular IFN-${\gamma}$ assay. Results: Immunization of these peptides as a mixture in pH-sensitive liposomes to transgenic mice induced a good protective effect from a viral challenge by inducing the peptide-specific CD8+ T cells. Mice immunized with $50{\mu}g/head$ were much better protected against viral challenge compared to those immunized with $5{\mu}g$/head, whereas the mice immunized with empty liposomes were not protected at all. After in vitro CTL culture by peptide stimulation, however, specific cytotoxicity was much higher in the CTLs from mice immunized with $5{\mu}g/head$ than $50{\mu}g/head$ group. Increase of the number of cells that intracellular IFN-${\gamma}$ secreting cell among CD8+ T cells showed similar result. Conclusion: Mice immunized with XEPs within pH-sensitive liposome were protected against viral challenge. The protective effect depended on the amount of antigen used during immunization. XEP-3-specific CTLs could be induced by peptide stimulation in vitro from splenocytes obtained from immunized mice. The cytotoxic effect of CTLs was measured by $^{51}Cr$-release assay and the percentage of accumulated intracellular IFN-${\gamma}$ secreting cells after in vitro restimulation was measured by flow cytometric analysis. The result of $^{51}Cr$-release cytotoxicity test was well correlated with that of the flow cytometric analysis. Viral protection was effective in immunized group of $50{\mu}g/head$, while in the in vitro restimulation, it showed more spectific response in $5{\mu}g$/head group.

길항미생물에 의한 Rhizoctonia spp.의 억제 및 길항미생물의 농약 혼용시 생존율 (Suppression of Rhizoctonia spp. by Antagonistic Microorganisms and Their Compatibility with Fungicides)

  • 이상재;심경구;김영권;허근영
    • 아시안잔디학회지
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    • 제12권1호
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    • pp.23-30
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    • 1998
  • 174 isolates of soil microorganisms were isolated from E-golf club from Apr.1997 through Oct. 1997. And 27 strains of them were selected through the inhihition test of mycelial growth. In the same period, soil-borne diesease pathogens, "Rhizoctonia", causing Large patch, Brown patch, Spring dead spot, and Yellow patch were isolated from the diseased areas in E-golf and S-golf club. The antagonistic activity of the strains against the pathogens was tested to select the excel-lent antagonists. In contact with the fungicides, the survivability of the antagonists was tested to assess the compatibility of the antagonists with the pesticides. The results were as follows: 1.Suppression of Rhizoctonia by Antagonists. Antagonistic activity of 27 strains against the pathogens was: tested in vitro. In the result, 3 isolates(B-7, B-15, B-41) of bacteria and 2 isolates(F-5, F-47) of fungi were superior to the rest. 2.Compatibility of the antagonists: with the fungicides: With 13 kinds of pesticides widely using Golf Club, Compatibility of 5 antagonists: were finally tested to select the strains: that mostly survived in contact with pesticides. In the results:, two of five strains: were selected : one strain was bacteria B-15, the other strain was fungi F-47. 24h after the mixing with pesticides:, these two strains were shown to survive at 90% level and these were identified as Bacillus and Trichoderma, respectively. And the most compatible pesticides: with the antagonists were shown to Polytoxin-D thirarn(s:urvivability 99.4%) and Validamycin-A (survivability 98.6%). Keywords:Antagonist, Large Patch, Trichoderma, Compatibility, Fungicide.Fungicide.

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The Antifungal Test: An Efficient Screening Tool for the Discovery of Microbial Metabolites with Respiratory Inhibitory Activity

  • Han, Jae Woo;Kim, Bomin;Oh, Mira;Choi, Jaehyuk;Choi, Gyung Ja;Kim, Hun
    • Mycobiology
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    • 제48권4호
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    • pp.326-329
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    • 2020
  • Valuable natural compounds produced by a variety of microorganisms can be used as lead molecules for development of new agrochemicals. Furthermore, high-throughput in vitro screening systems with specific modes of action can increase the probability of discovery of new fungicides. In the current study, a rapid assay tested with various microbes was developed to determine the degree of respiratory inhibition of Saccharomyces cerevisiae in two different liquid media, YG (containing a fermentable carbon source) and NFYG (containing a non-fermentable carbon source). Based on this system, we screened 100 fungal isolates that were classified into basidiomycetes, to find microbial secondary metabolites that act as respiratory inhibitors. Consequently, of the 100 fungal species tested, the culture broth of an IUM04881 isolate inhibited growth of S. cerevisiae in NFYG medium, but not in YG medium. The result is comparable to that from treatment with kresoxim-methyl used as a control, suggesting that the culture broth of IUM04881 isolate might contain active compounds showing the inhibition activity for respiratory chain. Based on the assay developed in this study and spectroscopic analysis, we isolated and identified an antifungal compound (-)-oudemansin A from culture broth of IUM04881 that is identified as Oudemansiella venosolamellata. This is the first report that (-)-oudemansin A is identified from O. venosolamellata in Korea. Taken together, the development of this assay will accelerate efforts to find and identify natural respiratory inhibitors from various microbes.

고추 탄저병 방제제 Burkholderia lata CAB13001 선발 및 포장방제 효과 (Selection of Antifungal Bacteria Burkholderia lata CAB13001 for Control on Red Pepper Anthracnose and Its Control Efficacy in Field)

  • 함수상;김병련;권미경;한광섭;박인희;서경원
    • 한국유기농업학회지
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    • 제26권4호
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    • pp.649-660
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    • 2018
  • Colletotrichum acutatum에 의해 발생하는 고추탄저병을 방제하기 위해 토양 근권으로부터 분리한 미생물 중 길항효과가 있는 미생물을 선발하여 주요 작물병원균에 대한 항균활성 효과와 포장에서의 방제효과를 검정하였다. 기내 대치배양법을 이용한 길항력을 검정하였을 때, CAB13001 균주는 고추 탄저병을 일으키는 Colletotrichum acutatum을 포함한 식물 병원균 Sclerotinia cepivorum, Sclerotinia sclerotium, Botrytis cinerea에 대한 뛰어난 항균활성 효과를 보였다. 또한 풋고추를 이용한 생체 검정에서 선발균주는 고추 탄저병의 병반진전 억제효과가 우수하였으며, 포장에서의 방제가 역시 무처리 대비 82.4%로 매우 우수하였다. 이러한 결과로 볼 때 CAB13001 균주는 공기 전염을 하는 고추 탄저병의 생물적 방제제로 매우 유용할 것으로 판단된다. 한편, 본 연구에 사용된 길항미생물 CAB13001 균주는 16s rRNA gene의 계통학적 위치를 분석한 결과 Burkholderia lata로 동정하였다.

Platycodi radix beverage ameliorates postprandial lipemia response through lipid clearance of triglyceride-rich lipoprotein: A randomized controlled study in healthy subjects with a high-fat load

  • Lee, Hansol;Lim, Yeni;Park, Soo-yeon;Cho, Soo-Muk;Choe, Jeong-Sook;Jeong, Sewon;Kwak, Jin Sook;Kwon, Oran
    • Nutrition Research and Practice
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    • 제12권5호
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    • pp.371-377
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    • 2018
  • BACKGROUND/OBJECTIVES: Elevation of postprandial lipemia characterized by a rise in triglyceride (TG)-rich lipoproteins can increase the risk of atherogenesis. The objective of this study was to investigate postprandial lipemia response to a single dietary fat/sugar load test and monitor beneficial changes induced by the consumption of Platycodi radix (AP) beverage in healthy subjects. SUBJECTS/METHODS: A total of 52 subjects were randomly assigned to either placebo or AP beverage group with a high-fat shake in a randomized controlled crossover trial. Postprandial blood was collected at 0, 1, 2, 4, and 6 h and analyzed for TG and lipoprotein lipase mass. Inhibition of pancreatic lipase was determined in vitro. RESULTS: AP inhibited pancreatic lipase activity in vitro ($IC_{50}=5mg/mL$). Compared to placebo beverage, AP beverage consumption with a high-fat shake induced significant increase of plasma lipoprotein lipase mass (P = 0.0111, ${\beta}$ estimate = 4.2948) with significant reduction in very low-density lipoprotein (VLDL) TG concentration (P = 0.038, ${\beta}$ estimate = -52.69) at 6 h. Based on significant correlation between high-fat dietary scores MEDFICTS and postprandial TG responses in VLDL (P = 0.0395, r = 0.2127), subgroup analysis revealed that 6 h-postprandial VLDL TG response was significantly decreased by AP consumption in subjects with MEDFICTS ${\geq}40$ (P = 0.0291, ${\beta}$ estimate = -7214). CONCLUSIONS: AP beverage might have potential to alleviate postprandial lipemia through inhibiting pancreatic lipase activity and elevating lipoprotein lipase mass. Subgroup analysis revealed that subjects with high-fat dietary pattern could be classified as responders to AP beverage among all subjects.

Vanadate 처리가 종양세포의 방사선 감수성에 미치는 영향 (Enhanced Radiosensitivity of Tumor Cells Treated with Vanadate in Vitro)

  • 이명자;이원영
    • Radiation Oncology Journal
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    • 제12권2호
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    • pp.129-141
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    • 1994
  • Intracellular ions which have a major role in cellular function have been reported to affect repair of radiation damage. Recently it has been reported that ouabain sensitizes A549 tumor cellls but not CCL-120 normal cells to radiation. Ouabain inhibits the $Na^+-K^+$-pump rapidly thus it increases intracellular Na concentration, Vanadate which is distributed extensively in almost all living organisms is known to be a $Na^+-K^+$-ATPase inhibitors, This study was performed to see any change in radiosensitivity of tumor cell by vanadate and any role of $Na^+-K^+$ATPase in radiosensitization. Experiments have been carried out by pretreatment with vanadate in human cell line(A549, JMG) and mouse cell line(L1210, spleen). For the cell survival MTT assay was performed for A549 and JMC cells and frypan blue dye exclusion test for L120, and spleen cells. Measurements of $Na^+-K^+$-ATPase activity in control, vanadate treated cell, radiation treated cell (9 Gy for A549 and JMG, 2 Gy for L1201, spleen), and combined $10^{-6}M$ vanadate and radiation treated cells were done. The results were summerized as fellows. 1. L1210 cell was most radiosensitive, and spleen cell and JMG cell were intermediate, and A549 cell was least radiosensitive. 2. Mininum or no cytotoxicity was seen with vanadate below concentration of $10^{-6}M$. 3. In A549 cells there was a little change in radiosensitivity with treatment of vanadate. However radiation sensitization was shown in low dose level of radiation i. e. 2- Gy. In JMG cells no change in radiosensitivity was noted. Both L1210 and spleen cell had radiosensitization but change was greater in tumor cell. 4. $Na^+-K^+$-ATPase activity was inhibited significantly in tumor cell by treatment of vanadate. 5. Radiaiton itself inhibited $Na^+-K^+$-ATPase activity of tumor cell with high $Na^+-K^+$-ATPase concention. Increase in radiosensitivity by vanadate was closely associated with orginal $Na^+-K^+$-ATPase contents. From the above results vanadate had little cytotoxicity and it sensitized tumor cells to radiation. Inhibitory effect of vanadate on $Na^+-K^+$-ATPase activity might be one of the contributing factors for radiosensitization to tumor cells which has greater enzyme activity than that of normal cell. It was suggested vanadate could be used as a potential radiosensitizer for tumor cells.

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