• Title/Summary/Keyword: BALB/C mouse

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Proteinase activity in the isolates of Trichomonas vaginalis according to their pathogenicity (질트리코모나스의 병원성과 단백 분해 효소와의 상관성)

  • 심영기;박경희
    • Parasites, Hosts and Diseases
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    • v.31 no.2
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    • pp.117-128
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    • 1993
  • Ten axonic isolates of Trichomonns uaginolis were subcutaneously injected to the BALB/c mice in order to assess their pathogenicity by means of so-called "mouse assay" method. All the isolates revealed neutral and acid proteinase activities both in their Iysates and in culture media, but the specific activities of both proteinases in the severely pathogenic group were significantly higher than the mildly pathogenic group (p < 0.05). In the SDS-PAGE system in which the electrophoretic gels contained 0.4% gelatin as the substrate, five different handing patterns of trichomonal proteinases were detected, and the patterns were closely related with the pathogenicity of the isolates of T. vosinalis. All five bands might be regarded as cysteine proteinases group in the inhibitor assays. The cytotoxicity of the Iysates of T. vaginalis to the target Chinese hamster ovarian (CHO) cell line was also significantly different according to the pathogenicity of the isolates, and generally lower in the Iysates treated with cysteine proteinase inhibitors than in the control Iysates. In summarizing the results, it might be considered that the proteinases of T.vaginalis showing five electrophoretic banding patterns are closely related with the pathogenicity and cytotoxicity of the isolates of T. voginolis.

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Production of Monoclonal Antibodies against Vibrio parahaemolyticus and Development of High Sensitive Immuno-Selective Filtration Method (Vibrio parahaemolyticus에 대한 단클론성 항체 개발과 고감도 면역선택여과법의 개발)

  • Kim, Jeong-Sook;Choi, Young-Dong;Shim, Won-Bo
    • Journal of Food Hygiene and Safety
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    • v.31 no.1
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    • pp.21-27
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    • 2016
  • The objectives of this study are to produce monoclonal antibodies (MAbs) against Vibrio parahaemolyticus and to develop an immuno-selective filtration (ISF) method for the rapid and sensitive detection of V. parahaemolyticus. The characterization of the MAb produced from HKVP 4H9-9 hybridoma cell was validated by enzyme-linked immunosorbent assay (ELISA) and western blot. The produced MAb was specific to V. parahaemolyticus and showed weak cross-reaction to V. alginolyticus, V. vulnificus and Staphylococcus aureus. After optimization of the method, $5{\times}10^1cell/mL$ of V. parahaemolyticus in a pure culture could be detectable. Although weak cross-reactivity to V. vulnificus, V. alginolyticus and Staphylococcus aureus was observed, the ISF was confirmed to be highly specific to V. parahaemolyticus. Especially, the ISF showed the most sensitivity compared to the immunoassays currently reported is easier to perform and quicker than ID-ELISA.

Immuno-stimulating Activities of Skipjack Tuna Katsuwonus pelamis Cooking Juice Concentrates on Mouse Macrophages and Spleen Cells (참치(Katsuwonus pelamis) 자숙액 농축물의 마우스 대식세포 및 비장세포에 대한 면역증강활성)

  • Kang, Bo-Kyeong;Kim, Koth-Bong-Woo-Ri;Ahn, Na-Kyung;Choi, Yeon-Uk;Kim, Min-Ji;Bark, Si-Woo;Pak, Won-Min;Kim, Bo-Ram;Park, Ji-Hye;Bae, Nan-Young;Ahn, Dong-Hyun
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.47 no.6
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    • pp.776-784
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    • 2014
  • Tuna cooking juice concentrate (TCJC) is by-produced during the canning processing of skipjack tuna Katsuwonus pelamis and it is well known that TCJC contains various nutritional components. Therefore, the immuno-stimulating activity of TCJC was investigated using macrophage RAW 264.7 cell line and the spleen cell isolated from BALB/c mice. The TCJC increased the production of IL-6, TNF-${\alpha}$, and IL-$1{\beta}$ in a dose-dependent manner compared to the control in RAW 264.7 cells without any toxicity. In particular, the production of TNF-${\alpha}$ was increased over 300-fold. The production of both Th1 cytokine (such as IFN-${\gamma}$, TNF-${\alpha}$, IL-2, and IL-12) and Th2 cytokine (IL-4, IL-6, IL-10) was also increased by TCJC treatment in splenocytes. Moreover, the TCJC increased the splenocyte proliferation in a concentration-dependent manner compared to control. These results indicate that TCJC may enhance immune function by promoting various cytokine production.

Antiinflammatory Effects of Heparin in Hemorrhage or LPS Induced Acute Lung Injury (출혈성 및 내독소 투여로 유발된 급성폐손상에서 heparin의 항염증효과)

  • Kim, Jae Yeol;Choi, Jae Chul;Lee, Young Woo;Jung, Jae Woo;Shin, Jong Wook;Park, In Won;Choi, Byoung Whui
    • Tuberculosis and Respiratory Diseases
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    • v.60 no.1
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    • pp.49-56
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    • 2006
  • 배경 : 급성 폐손상은 폐내, 외의 원인질환들에 의해 폐포-모세혈관의 투과성이 증가하며, 폐부종에 의해 급성 저산소성 호흡곤란이 유발되는 증후군이다. 헤파린은 항응고작용 외에 자체적으로 항염증효과를 가지고 있으나, 염증성질환에 헤파린을 투여하면 출혈성 합병증이 발생하기 때문에 실제로 임상에서 이용하는데 제약이 있다. 하지만 헤파린에서 2-O와 3-O sulfate를 제거하면, 항응고 효과가 제거되고 항염증효과는 지니고 있는 비항응고성 헤파린 (nonanticoagulant heparin)으로 변화한다. 본 연구에서는 흰쥐에게 내독소 (LPS)를 투여하거나, 출혈성 쇼크를 일으켜서 유발된 급성폐손상에서 비항응고성 헤파린의 치료효과를 살펴보았다. 방법 : 각 군당 5 마리 이상의 흰쥐 (Balb/c mouse)를 이용하였다. 미정맥 (tail vein)을 통해 생리식염수 또는 비항응고성 헤파린 (50 mg/kg)을 투여한 직후에 내독소를 복강으로 투여하거나 (1 mg/kg), 심장천자를 통해 총 혈액의 1/3 정도로 제거하여 출혈성 쇼크를 유도하여 급성폐손상을 유발하였다. 내독소 투여 또는 출혈성 쇼크 유발 1 시간 후에 흰쥐를 희생시키고 폐를 적출하였고, 폐의 염증성 변화는 사이토카인 ($TNF-{\alpha}$, MIP-2, $IL-1{\beta}$)을 측정하여 살펴보았고, 폐손상의 정도는 myeloperoxidase (MPO) assay와 wet-to-dry weight ratio를 측정하여 알아보았다. 결 과 : 내독소를 투여한 흰쥐의 폐에서 대조군의 폐에 비해 사이토카인의 발현이 증가하고 ($TNF-{\alpha}$; $196.1{\pm}10.8$ vs $83.7{\pm}18.4pg/ml$, MIP-2; $3,000{\pm}725$ vs $187{\pm}26pg/ml$, $IL-1{\beta}$; $6,500{\pm}1167$ vs $266{\pm}25pg/ml$, p<0.05, respectively), 폐의 MPO 활성이 증가하였다 ($27.9{\pm}6.2$ vs $10.5{\pm}2.3U/g$ of lung protein, p<0.05). 출혈성 쇼크를 일으킨 흰쥐의 폐에서 대조군의 폐에 비해 사이토카인의 발현은 증가되지 않았으나, MPO 발현은 증가되었다 ($16.5{\pm}3.2$ vs $10.5{\pm}2.3U/g$ of lung protein, p<0.05). 내독소 투여 또는 출혈성 쇼크에 의해 급성폐손상이 유발된 흰쥐에서 생리적 식염수를 투여하거나 비항응고성 헤파린을 투여한 군 사이에 사이토카인의 발현이나 MPO 활성에 의미있는 차이는 관찰되지 않았다. 결론 : 이상의 결과로 비항응고성 헤파린은 내독소를 투여하거나 출혈성 쇼크를 일으키고 한 시간 뒤에 측정한 흰쥐의 급성폐손상에서 의미있는 치료효과를 보이지 않았다.

Effect of Rheum undulatum Linne extract and Glycyrriza uralensis Fischer extract against arachidonic acid and iron-induced oxidative stress in HepG2 cell and CCl4-induced liver injury in mice (대황과 감초 병용의 항산화 및 간보호효과)

  • Lee, Eun Hye;Baek, Su Youn;Kim, Kwang-Youn;Lee, Seul-Gi;Kim, Sang Chan;Lee, Hyeong Sik;Kim, Young Woo
    • Herbal Formula Science
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    • v.24 no.3
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    • pp.163-174
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    • 2016
  • Objectives : Rheum undulatum Linne and Glycyrriza uralensis Fischer are widely used herbal medicine. In this study, anti-oxidant and liver protective effects of R. undunlatum extract (RUE) and G. uralensis extract (GUE) were investigated in HepG2 cells, respectively. Oxidative stress and liver fibrosis were induced by arachidonic acid (AA) and iron, and CCl4.Methods : MTT assay was assessed for cell viability, and immunoblotting analysis was performed to detect expression of apoptosis related proteins. In addition, reactive oxygen species (ROS) and mitochondrial dysfunction were measured. In vivo, BALB/c mouse were orally administrated with the aqueous extract of 10 mg/kg RUE and 100 mg/kg GUE for 3 days and then, injected with CCl4 0.5 ml/kg body weight to induce acute liver damage. Serum ALT level was measured, and histological change was observed in Harris's hematoxylin and eosin stainResults : RUE and GUE pre-treatment increased relative cell viability in concentration dependent manner and altered the expression levels of apoptosis-related proteins such as procaspase 3, PARP and Bcl-xL. RUE and GUE also inhibited the mitochondrial dysfunction and excessive reactive oxygen species (ROS) production induced by AA and iron. In addition, RUE and GUE activated liver kinase B1 (LKB1), by increasing phosphorylation. Moreover, RUE and GUE treatment decreased liver injuries induced by CCl4, as evidenced by decreases in histological liver damage as well as serum alanine amino transferase (ALT) level.Conclusions : These data suggest that RUE and GUE has anti-oxidant and liver protective effects against AA and iron-induced oxidative stress and CCl4-induced liver injury.

Antibacterial Effect of Immunoglobulin alone and in Combination with Ciprofloxacin against Pseudomonas aeruginosa (면역 글로불린 단독 및 Ciprofloxacin 병용에 의한 Pseudomonas aeruginosa에 대한 항균 효과)

  • Sung, Yeul-Oh;Kim, Hee-Sun;Jeon, Tae-Il;Kim, Sung-Kwang
    • Journal of Yeungnam Medical Science
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    • v.8 no.1
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    • pp.53-62
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    • 1991
  • Experiments were performed in mice(Balb/C) to support the basic efficacy of the human immunoglobulin (IgG) preparation. The antibacterial activity of IgG purified from human sera was examined with or without the quinolone agent, ciprofloxacin(CPFX), against Pseudomonas aeruginosa isolated from clinical specimens. Results were as follows: Antibacterial activities in terms of the percentage of survivors, after administration of Ps. aeruginosa into mouse intraperitoneal cavity were in the following order, single IgG group, CPFX administration after IgG pretreatment group, IgG and CPFX combined administration group and CPFX alone group. The number of living bacteria was monitored in blood and liver tissue of mice infected with Ps. aeriginosa and treated by IgG administration. The increase of living bacteria in liver was more drastic than that in blood. Leukocytosis was observed in mice injected with IgG, excluding those only with ciprofloxacin, after 8 hours of administration to see a decrease to normal number of bacteria after 18 hours. No significant difference was noticed between pretreatment group and post treatment group. In vitro susceptibility test of IgG against Ps. aeruginosa, minimal inhibitory concentration(MIC) was $250{\mu}g/ml$, resistant to IgG, regardless of a combined administration with CPFX. In vitro test revealed that the IgG itself did not have anti-Ps. aeruginosa activity.

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Ginsenoside Rh2 attenuates microglial activation against toxoplasmic encephalitis via TLR4/NF-κB signaling pathway

  • Xu, Xiang;Jin, Lan;Jiang, Tong;Lu, Ying;Aosai, Fumie;Piao, Hu-Nan;Xu, Guang-Hua;Jin, Cheng-Hua;Jin, Xue-Jun;Ma, Juan;Piao, Lian-Xun
    • Journal of Ginseng Research
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    • v.44 no.5
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    • pp.704-716
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    • 2020
  • Background: Ginsenoside Rh2 (GRh2) is a characterized component in red ginseng widely used in Korea and China. GRh2 exhibits a wide range of pharmacological activities, such as anti-inflammatory, antioxidant, and anticancer properties. However, its effects on Toxoplasma gondii (T. gondii) infection have not been clarified yet. Methods: The effect of GRh2 against T. gondii was assessed under in vitro and in vivo experiments. The BV2 cells were infected with tachyzoites of T. gondii RH strain, and the effects of GRh2 were evaluated by MTT assay, morphological observations, immunofluorescence staining, a trypan blue exclusion assay, reverse transcription PCR, and Western blot analyses. The in vivo experiment was conducted with BALB/c mice inoculated with lethal amounts of tachyzoites with or without GRh2 treatment. Results and conclusion: The GRh2 treatment significantly inhibited the proliferation of T. gondii under in vitro and in vivo studies. Furthermore, GRh2 blocked the activation of microglia and specifically decreased the release of inflammatory mediators in response to T. gondii infection through TLR4/NF-κB signaling pathway. In mice, GRh2 conferred modest protection from a lethal dose of T. gondii. After the treatment, the proliferation of tachyzoites in the peritoneal cavity of infected mice markedly decreased. Moreover, GRh2 also significantly decreased the T. gondii burden in mouse brain tissues. These findings indicate that GRh2 exhibits an antieT. gondii effect and inhibits the microglial activation through TLR4/NF-κB signaling pathway, providing the basic pharmacological basis for the development of new drugs to treat toxoplasmic encephalitis.

Evaluation of cytotoxicity of electroplated stainless steel orthodontic wire (전기도금한 교정용 스테인레스스틸 선재의 세포독성에 관한 연구)

  • Lee, Gye-Hyeong;Cho, Jin-Hyoung;Lee, Ki-Heon;Hwang, Hyeon-Shik
    • The korean journal of orthodontics
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    • v.35 no.2 s.109
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    • pp.127-136
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    • 2005
  • The purpose of this study was to examine the cytotoxicity of orthodontic wire which had an increased diameter through electroplating, and to evaluate its possible clinical applications, First. nickel plating was carried out on the commercially available stainless steel wire using an electroplating technique For the comparison of the electroplated wire with ready made stainless steel wire and titanium or copper. each wire was incubated for 72 hours in a medium. The release of the metal ion was measured using ICP-AES (Inductively Coupled Plasma Atomic Emission Spectrophotometer). Balb/c 3T3 mouse fibroblast was put on a microplate and placed in an incubated medium of 75%, 50%, and 20% dilation. An MTT analysis was used to compare with the medium only. The change in absorbency value of each wire group and the difference of absorbency value according to the change of dilution was measured The results of ICP-AES analysis showed that great amount nickel iou was isolated from electroplated orthodontic wires and great amount copper ion was isolated from copper. The results of the MTT analysis showed that there was no difference in the absorbency value of titanium at any dilution. However, the electroplated wires (p<0.001) the stainless steel wires (p<0.05) and the copper (P<0.001) were statistically significantly lower than those of medium only at all dilutions. Assessment as per ISO 10993, part 5, showed that electroplated wire was alloted to 'moderate cytotoxic' the titanium and stainless steel wire were 'non-cytotoxic' The results of this study indicate that the electroplated orthodontic wires need additional efforts to decrease cytotoxicity for their clinical applications.

Effect of Persicae Semen for Atopic Dermatitis Skin Tissue and Regulate to Inflammation Mediator in Serum (도인(桃仁)의 아토피 피부염 모델 피부조직 및 혈청 내 염증매개물질 조절 효과)

  • Kim, Sangwoo;Hong, SooYeon;Kwon, Boguen;Kim, Myunghyun;Kim, Sang-bae;Jin, Dae-hwan;Choi, Woochan;Sohn, Youngjoo;Jung, Hyuk-Sang
    • The Korea Journal of Herbology
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    • v.35 no.4
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    • pp.51-60
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    • 2020
  • Objective : The objective of this study was to demonstrate the effect of Persicae Semen (PS) in DNCB-induced atopic dermatitis mouse and HaCaT cell. Methods : The BALB/c mice were divided into four groups. To develop atopic dermatitis, 200 ㎕ of 1 and 0.5% DNCB solution was put on the back of mice in the Control group, the PS-Low group and the PS-High group once a day. After application of DNCB, 200 ㎕ of the PS extract was also treated. The Normal group was given PBS. The mice dorsal skin was stained with Masson's trichrome, H&E, and toluidine blue to evaluate the thickness of the epidermis and dermis, infiltration of eosinophils and mast cells respectively. ELISA was applied to measure the serum level of IgE and IL-6. Toxicity of PS was measured by MTS assay in HaCaT cell. To investigate the effects of PS on HaCaT cells, cells were pre-treated with PS for 1h, and then stimulated with TNF-α and IFN-γ. After 24 hours, the expression of TARC was analyzed using RT-PCR. Results : PS not only significantly diminished the thickness of the epidermis and dermis, but also reduced the infiltration of eosinophil and mast cell in skin lesion. PS also reduced the serum IgE and IL-6 level which plated important roles in the atopic dermatitis. The expression of TARC was decreased significantly in TNF-α/IFN-γ stimulated HaCaT cell. Conclusion : These results suggest that PS may be effective in alleviating the atopic dermatitis induced by DNCB and inflammation by TNF-α/IFN-γ.

A Study on the Protective Effect and Its Mechanism of Zinc against Immuno-cytotoxicity of Methylmercury (유기수은의 세포면역독성과 이에 대한 아연의 방어효과 및 기전)

  • 고대하;염정호;오경재
    • Journal of Environmental Health Sciences
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    • v.27 no.2
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    • pp.82-91
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    • 2001
  • This study was carried out to elucidate the protective effect of zinc chloride(ZnCl$_2$) and its mechanism against the immuno-cytotoxicity of methylmercury chloide($CH_3$HgCl). This study was observed in the culture of EMT-6 cells which are originated from mammary adenocarcinoma of Balb/c mouse. Cytotoxicity of metals was measured by cell viability and NO$_2$$^{[-10]}$ , and mitochondrial function was evaluated by adenosine triphosohate (ATP) production. $CH_3$HgCl significantly decreased the sythesis of nitric oxide(NO), ATP and glutathione(GSH) in a dose-dependent manner. ZnCl$_2$ significantly increased the synthesis of GSH in a dose-dependent manner, but synthesis of NO and ATP were not changed. The immuno-cytotoxicity of $CH_3$HgCl was not fully protected when combined addition of ZnCl$_2$, whereas ZnCl$_2$ prior to addition of $CH_3$HgCl completly protected the Hg-induced immuno-cytotoxicity. Similarly, intracellular accumulation of mercury significantly decreased by ZnCl$_2$. Degree of diminution of intracellular mercury was larger in ZnCl$_2$ prior to addition of $CH_3$HgCl than in combined addition of ZnCl$_2$ and $CH_3$HgCl.. Dithiothreitol(DTT) or buthionine sulfoximine(BSO) addition at 50$\mu$M or less, which was not toxic to the cells, did not affect synthesis of NO and ATP. DTT increased intracellular GSH level and DTT pretreatment protected toxicity induced by $CH_3$HgCl as shown complete recover in the NO and ATP values. BSO decreased intracellular GSH level and BSO pretreatment exaggerated toxicity induced by $CH_3$HgCl as shown synergistic reduction in the NO and ATP values. These results indicated that the protective effects of zinc against immuno-cytotoxicity of methylmercury associated with increasing cellular level of GSH. Increased intracellular GSH transports methylmercury to out of cells. In accordance with intracellular level of mercury decreased, immuno-cytotoxicity of methylmercury decreased. These result also suggest that the protective mechanism of zinc against the mercury toxicity would be exerted in the immune system in vivo.

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