• Title/Summary/Keyword: Mouse(Balb/c.)

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Cytotoxicity of lymphokine activated peritoneal macrophages against Trichomonas vaginalis (질트리코모나스에 대한 림포카인황성대식세포의 세포독성능)

  • Yoon, Kyong;Ryu, Jae-Sook;Min, Duk-Young
    • Parasites, Hosts and Diseases
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    • v.29 no.4
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    • pp.381-388
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    • 1991
  • Trichomonas vaginalis is a parasitic nagellate in the urogenital tract of human. Innate cytotonicity of macrophages against T. vaginalis has been recognized, but any report on the cytotoxicity of Iymphokine-activated macrophages to T vaginalis is not yet available. The present study aimed to elucidate the Iymphokine-activated cell mediated cytotoxic effect against T. vaginalis by mouse peritoneal macrophages. Cytotoxicity was measured by counting the release of $^3H-thymidine$ from prelabeled protozoa, and tested in U-bottom microtiter plates. Nitrite concentration in culture supernatants was measured by standard Griess reaction. The results obtained are as follows: 1, The cytotoxicity of macrophages was increased by addition of rIL-2 or $rIFN-{\gamma}$$. 2, Cytotoxicity of macrophages was reduced by addition of rIL-4 to rOM-CSV, rIL-2 or $rIFN-{\gamma}$. 3. Crude Iymphokine mixed with anti-lL-2 decreased the cytotoxity of macrophages. 4. In case of macrophages cultured with $rIFN-{\gamma}$ or rIL-4, the concentration of nitrite was related with cytotokity of macrophages against T. vaginalis, but the cytotoxicity of macrophages cultured with rIL-2 and $rIFN-{\gamma}$ was decreased in spite of its high production of llitrite. From the results obtained, it is assumed that rIL-2 and $rIFN-{\gamma}$ enhance the cytotoxicity of macrophages while rIL-4 inhibits the cytotoxicity against T. vaginalis, and that the production of nitrite does not relate with the cytotoxicity of macrophages, but nitric oxide may play a role as an inhibitory factor on the proliferation of T. vaginalis.

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DEU-7 Derived from Ulmus macrocarpa Improved Immune Functions in Cyclophosphamide-treated Mice (면역억제 마우스 모델에서 왕느릅나무 유래 DEU-7의 면역기능 증강)

  • Kang, Kyung-Hwa;Go, Ji Su;Lee, Inhwan;Lee, Sang Ho;Lee, Sung Do;Kim, Deok Won;Lee, Jong-Hwan;Hwang, HyeJin;Hyun, Sook Kyung;KIM, Byoung Woo;Kim, Chul Min;Chung, Kyung Tae
    • Journal of Life Science
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    • v.25 no.10
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    • pp.1156-1163
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    • 2015
  • The present study investigated the immunomodulatory properties of four different medicinal plants in a cyclophosphamide-treated Balb/c mouse model. One of the four plants, Ulmus macrocarpa, showed partial resistance against immune suppression induced by cyclophosphamide. The bark of U. macrocarpa, commonly known as the Chinese elm, has been used as a pharmaceutical material in Korean traditional medicine to treat bacterial inflammation and induce wound healing. In this study, water extract of U. macrocarpa, named DEU-7, was used for its immunomodulating functional activity. DEU-7 increased the weight of the spleen and the number of splenocytes but did not significantly affect the liver, kidney, and thymus in vivo. A splenocyte viability assay confirmed that DEU-7 influenced ex vivo splenocyte survival. DEU-7 also increased the levels of cytokines, such as IL-2 and IL-4, and immunoglobulins, such as IgM, IgG, and IgA. These results indicated that DEU-7 is involved in the activation of T and B lymphocytes. In addition, DEU-7 was able to maintain the production of cytokines, such as TNF-α, IL-12, and IFN-γ, in the condition of cyclophosphamide-induced immune suppression, suggesting that DEU-7 activated innate immune cells, even under immune suppression. We concluded that DEU-7 aids immunological homeostasis, thereby preventing immune suppression, and aids both innate and adaptive immune response by maintaining the levels of various cytokines and immunoglobulins. Consequently, it is worth investigating the potential of DEU-7 as a supplemental source for immune-enhancing agents.

Autometallography for Zinc Detection in the Central Nervous System (중추신경계통내 분포하는 Zinc의 조직화학적 동정)

  • Jo, Seung-Mook;Gorm, Danscher;Kim, Sung-Jun;Park, Seung-Kook;Kang, Tae-Cheon;Won, Moo-Ho
    • Applied Microscopy
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    • v.30 no.4
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    • pp.347-355
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    • 2000
  • Zinc is one of the most abundant oligoelements in the living cell. It appears tightly bound to some metalloproteins and nucleic acids, loosely bound to some metallothioneins or even as free ion. Small amounts of zinc ions (in the nanomolar range) regulate a plentitude of enzymatic proteins, receptors and transcription factors, thus rolls need accurate homeostasis of zinc ions. Zinc is an essential catalytic or structural element of many proteins, and a signaling messenger that is released by neural activity at many central excitatory synapses. Growing evidences suggest that zinc may also be a key mediator and modulator of the neuronal death associated with transient global ischemia and sustained seizures, as well as perhaps other neurological disease stoles. Some neurons have developed mechanisms to accumulate zinc in specific membrane compartment ('vesicular zinc') which can be evidenced using histochemical techniques. Substances giving a bright colour or emitting fluorescence when in contact with divalent metal ions are currently used to detect them inside cells; their use leads to the so called 'direct' methods. The fixation and precipitation of metal ions as insoluble salt precipitates, their maintenance along the histological process and, finally, their demonstration after autometallographic development are essential steps for other methods, the so called 'indirect methods'. This study is a short report on the autometallograhical approaches for zinc detection in the central nervous system (CNS) by means of a modified selenium method.

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Effects of Plant Water Extract Mixture Ixeris Sonchifolia Hance, Oenanthevjavanica, Fagopyrum Esculentum Moench, Hizikia Fusiforme, Zingiber Officinale Roscoe on Mouse Immune Cell Activation Ex vivo (5가지 (고들빼기, 돌미나리, 메밀, 톳, 생강)혼합식품 물 추출물의 마우스 면역세포 활성화 효과)

  • Ryu, Hye-Sook;Kim, Jung-Hee;Kim, Hyun-Sook
    • Journal of Nutrition and Health
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    • v.41 no.2
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    • pp.141-146
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    • 2008
  • Ixeris sonchifolia Hance (Godulbaegi), Oenanthe javanica (Dolminari), Fagopyrum esculentum Moench (Buckwheat>, Hizikia fusiforme (Seaweed Fusiforme) and Zingiber ojficinale Roscoe (Ginger) have been used respectively as one of folk remedies as well as food materials. However, reportedly few studies on their immunomodulating effects have been made, although it has been known from other preceding studies that the ex vivo supplementation of each Ish, OJ, Fem, Hf, Zor water extracts tends to enhance the proliferation of splenocyte in comparison to the control group. This study on the combined immunomodulative effect of water extract mixture of these five food materials (Ish + Oj + Fem +Hf+Zor) lasted covering seven or eight weeks. The old mice (balb/c) was fed ad libitum on chow diet, and the water extract of plant mixture was orally administrated every other day for four weeks at two different concentrations (50 and 500 mg/kg B.W) . After preparing the single cell suspension, the proliferation of splenocyte was determined by MTT (3-[4,5-dimethylthiazol-2-y]-2,5-diphenyl terazolium bromide) assay. The production of cytokine ($IL-{\beta}$, IL-6, and $TNF-{\alpha}$) which was secreted by macrophages stimulated with LPS or not was detected by ELISA assay using the cytokine kit. After the 48 hours of incubation with the mitogen (ConA or LPS) stimulation, the proliferation of the mice splenocyt in the experimental group statisticaly increased at both of two different concentrations in comparison to the control group. The cytokines production was more significantly enhanced at the lower supplementation (50 mg/kg B.W.) group than at the higher concentration (500 mg/kg B.W.). The result of this study may suggest that the supplementation of water extract of plant mixture can regulate and enhance the immune function by increasing the splenocyte proliferation and regulating the cytokine production capacity by the activated macrophages in mice.

Medicinal Herb Extracts Attenuate 1-Chloro-2,4dinitrobenzene-induced Development of Atopic Dermatitis-like Skin Lesions (한약재 단일 추출물 및 복합 추출물을 이용한 아토피성 피부염 억제 효과)

  • Lee, Moon Hee;Han, Min Ho;Yoon, Jung Jeh;Song, Myung Kyu;Kim, Min Ju;Hong, Su Hyun;Choi, Byung Tae;Kim, Byung Woo;Hwang, Hye Jin;Choi, Yung Hyun
    • Journal of Life Science
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    • v.24 no.8
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    • pp.851-859
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    • 2014
  • The present study was designed to investigate whether ethanol extracts of Sophora flavescens (GS), Glycyrrhiza uralensis (GC), Dictamnus dasycarpus (BSP), and their mixtures (GGB-1, -2, -3, and -4) inhibit 1-chloro-2,4-dinitrobenzene (DNCB)-induced atopic dermatitis (AD) in a mouse model. DNCB was topically applied on the dorsal surface of Balb/c mice to induce AD-like skin lesions. The pathological phenotypes of AD, such as erythema, ear thickness, edema, scabs, and discharge, were significantly decreased in the GGB (DNCB + GS:GC:BSP = 3:1:1 mixture)-1-treated groups compared with the other treated groups. The weight of the spleen in immune organs was significantly decreased in the GGB-1-treated groups, whereas the weight of the liver in a control group was similar to that of the groups treated with the samples. Furthermore, toluidine blue staining analysis, a method used to specifically identify mast cells, showed that master cell infiltration into the dermis of the GGB-1-treated group was significantly decreased. The immunoglobulin E concentration was lower in the GGB-1-treated group. In addition, the levels of inflammatory cytokines (interferon-${\gamma}$, interleukin-1, 4, 5, 6, and 13, $1{\beta}$, and tumor necrosis factor-${\alpha}$) were also significantly reduced in the GGB-1-treated group. Taken together, these results suggest that a mixture of GS, GC, and BSP in a proportion of 3:1:1 (GGB-1) may contribute to the relief of AD symptoms and may be considered an excellent candidate for an AD therapeutic drug.

Effect of Solvent Fractions from Doenjang on Antimutagenicity, Growth of Tumor Cells and Production of Interleukin-2 (된장 분획물의 항돌연변이 및 암세포 증식 억제효과와 interleukin-2 생성에 미치는 영향)

  • Kim, Kwang-Hyuk;Park, Kun-Young;Lee, Sook-Hee;Lim, Sun-Young
    • Journal of Life Science
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    • v.17 no.6 s.86
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    • pp.791-797
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    • 2007
  • We studied the inhibitory effect of solvent fractions from doenjang on mutagenicity using Salmonella typhimurium TA100 in Ames test. We also investigated the effect of solvent fractions from doenjang on the growth of tumor cells and the production of interleukin-2 (IL-2). The treatment of dichlorormethane and ethylacetate fractions (2.5 mg/assay) from doenjang to Ames test system inhibited aflatoxin B$_1$ (AFB$_1$) induced mutagenicity by 96% and 97%, respectively, and showed a higher antimutagenic effect than other solvent fractions. In case of N-methyl-N'-nitro-N-nitrosoguamidine (MNNG) induced mutagenicity, the ethylacetate fraction showed the highest inhibitory effect (by 75%) among the other sol-vent fractions, although the inhibitory effect was not stronger compared to AFB$_1$ induced mutagenicity. The treatment of dichloromethane and ethylacetate fractions markedly inhibited the growth of Yac-1 (by 80% and 94%, respectively) and sacroma-180 cancer cells (by 60% and 96%, respectively) after 4 days of incubation at 37${\circ}$C. To elucidate the immunological mechanism of antitumor activity of doenjang, spleen cells of Balb/c mouse were exposed to the dichloromethane and ethyl-acetate fractions for 24 hours at 37${\circ}$C . The culture supernatants following the treatment of djchloromethane and ethylacetate factions to spleen cells increased the production of IL-2. These results indicated that the anticarcinogenic effect of doenjang was mediated by the production of IL-2.

Synthesis and Preliminary Evaluation of $9-(4-[^{18}F]Fluoro-3-hydroxymethylbutyl)$ Guanine $([^{18}F]FHBG)$ in HSV1-tk Gene Transduced Hepatoma Cell (9-(4-$[^{18}F]Fluoro-3-hydroxymethylbutyl)$guanine $([^{18}F]FHBG)$의 합성과 헤르페스 단순 바이러스 티미딘 키나아제 이입 간암 세포주에서의 기초 연구)

  • Moon, Byung-Seok;Lee, Tae-Sup;Lee, Myoung-Keun;Lee, Kyo-Chul;An, Gwang-Il;Chun, Kwon-Soo;Awh, Ok-Doo;Chi, Dae-Yoon;Choi, Chang-Woon;Lim, Sang-Moo;Cheon, Gi-Jeong
    • Nuclear Medicine and Molecular Imaging
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    • v.40 no.4
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    • pp.218-227
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    • 2006
  • Purpose: The HSV1-tk reporter gene system is the most widely used system because of its advantage that direct monitoring is possible without the introduction of a separate reporter gene in case of HSV1-tk suicide gene therapy. In this study, we investigate the usefulness of the reporter probe (substrate), $9-(4-[^{18}F]Fluoro-3-hydroxymethylbutyl)$guanine ($[^{18}F]FHBG$) for non-invasive reporter gene imaging using PET in HSV1-tk expressing hepatoma model. Materials and Methods: Radiolabeled FHBG was prepared in 8 steps from a commercially available triester. The labeling reaction was carried out by NCA nucleophilic substitution with $K[^{18}F]/K2.2.2.$ in acetonitrile using N2-monomethoxytrityl-9-14-(tosyl)-3-monomethoxytritylmethylbutyl]guanine as a precursor, followed by deprotection with 1 N HCl. Preliminary biological properties of the probe were evaluated with MCA cells and MCA-tk cells transduced with HSV1-tk reporter gene. In vitro uptake and release-out studies of $[^{18}F]FHBG$ were performed, and was analyzed correlation between $[^{18}F]FHBG$ uptake ratio according to increasing numeric count of MCA-tk cells and degree of gene expression. MicroPET scan image was obtained with MCA and MCA-tk tumor bearing Balb/c-nude mouse model. Results: $[^{18}F]FHBG$ was purified by reverse phase semi-HPLC system and collected at around 16-18 min. Radiothemical yield was about 20-25%) (corrected for decay), radiochemical purity was >95% and specific activity was around >55.5 $GBq/{\mu}\;mol$. Specific accumulation of $[^{18}F]FHBG$ was observed in HSV1-tk gene transduced MCA-tk cells but not in MCA cells, and consecutive 1 hour release-out results showed more than 86% of uptaked $[^{18}F]FHBG$ was retained inside of cells. The uptake of $[^{18}F]FHBG$ was showed a highly significant linear correlation ($R^2=0.995$) with increasing percentage of MCA-tk numeric cell count. In microPET scan images, remarkable difference of accumulation was observed for the two type of tumors. Conclusion: $[^{18}F]FHBG$ appears to be a useful as non-invasive PET imaging substrate in HSV1-tk expressing hepatoma model.