• Title/Summary/Keyword: mouse assay

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Evaluation and Optimization of a Serum-based Minimum Inhibitory Concentration Assay to Caspofungin in Candida albicans Clinical Isolates

  • Yoo, Young Bin;Kim, Sung-Soon;Kim, Young Kwon;Kim, Sunghyun
    • Biomedical Science Letters
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    • v.22 no.4
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    • pp.174-183
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    • 2016
  • In the present study, a serum-based minimum inhibitory concentration (MIC) testing to caspofungin was optimized and evaluated to solve the limitations of the conventional Clinical and Laboratory Standards Institute (CLSI) guideline-based antifungal agent MIC test and the usefulness of this testing for clinical application was determined. A total of 105 Candida albicans clinical isolates were used for measuring MIC to caspofungin. Results showed that growth characteristics were different according to types of serum and the mouse serum was the most suitable for this assay. In order to measure the optimal concentration of mouse serum, 0 to 100% mouse serum were added to the media during fungal culture. The optimal concentration of serum was 50% when consideration of antifungal agent administration and inoculum size, serum components and ease of hyphae separated, and the consideration of the degree of growth. In comparison of the usefulness between the conventional Alamar-modified broth microdilution MIC assay and 50% mouse serum-based MIC testing, the range of $MIC_{80}$ of the Alamar-modified broth microdilution MIC assay was $0.13{\sim}2.0{\mu}g/mL$ (SD ${\pm}0.42{\mu}g/mL$) and that of the 50% mouse serum-based MIC assay was $2.0{\sim}32.0{\mu}g/mL$ (SD ${\pm}9.01{\mu}g/mL$). The range of $MIC_{50}$ of the Alamar-modified broth microdilution MIC assay was $0.13{\sim}2.0{\mu}g/mL$ (SD ${\pm}0.40{\mu}g/mL$) and that of the 50% mouse serum-based MIC assay was $1.0{\sim}16.0{\mu}g/mL$ (SD ${\pm}2.36{\mu}g/mL$). The MICs of 50% mouse serum-based MIC testing was increased by up to 4 to 64 times than Alamar-modified broth microdilution MIC assay. In conclusion, a 50% mouse serum-based MIC assay was more useful for measuring MIC in Candida albicans clinical isolates than conventional colorimetric broth microdilution MIC testing.

Study on Mutagenicity of DehydroevodiamineㆍHCl(DHED) (치료제 DehydroevodiamineㆍHCl(DHED)의 변이원성 연구)

  • 성이숙;정성윤;정주연;채규영;진미령;최봉웅;장병모;김대경
    • YAKHAK HOEJI
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    • v.46 no.3
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    • pp.208-212
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    • 2002
  • Dehydroevodiamine HCl (DHED), which is a component separated from Evodia rutaecarpa Bentham, has novel anticholinesterase and antiamnesic activities in the scopolamine-induced amnesia model. Several studies suggest that DHED might be an effective drug for the Alzheimer's disease and the vascular type of dementia. In order to evaluate the mutagenic potential of DHED, Salmonella typhimurium reversion assay, chromosomal aberration test on Chinese hamster lung cells, in vivo micronucleus assay using mouse bone marrow cells, and comet assay were performed. DHED did not increase the number of revertant in the reverse mutation test using Salmonella typhimurium TA1535, TA1537, TA98, TA100. DHED HCl, at concentration of 5 and 10 $\mu\textrm{g}$/mι, increased the number of chromosome aberrated Chinese hamster lung cells with 5 and 10%, respectively. In mouse micronucleus test, no significant increase in the occurrence of micronucleated polychromatic erythrocyte was observed in ICR mice orally administered with DHED. DHED was tested for ability to induce genotoxic effect in L5178Y cells (mouse lymphoma cells) using the single cell gel electrophoresis assay (comet assay). In comet assay, tail moment did not increase in L5178Y cells treated with 10, 100, 300 $\mu$M DHED.

The effects of Chungpesagan-Tang and herbs on Mouse neuroblastoma 2a cells damaged by hypoxia-reoxygenation (청폐사간탕(淸肺瀉肝湯)과 단미(單味)들이 Hypoxia-Reoxygenation에 의해 손상받은 Mouse Neuroblastoma 2a Cells에 미치는 영향(影響))

  • Moon, Ha-Kyoung;Kim, Jong-Woo;Kang, Chul-Hun;Whang, Wei-Wan
    • Journal of Oriental Neuropsychiatry
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    • v.16 no.2
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    • pp.89-112
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    • 2005
  • Object : This study was designed to assess effect of Chungpesagan-Tang and herbs on Mouse neuroblastoma 2a cells damaged by hypoxia-reoxygenation. Method : Mouse neuroblastoma 2a (N2a) cells were measured by MTT assay and LDH assay after 48h hypoxia and 6h reoxygenation. Mouse neuroblastoma 2a (N2a) cells were treated by Chungpesagan-Tang and herbs. Result : In MTT assay of hypoxia all of herbs were almost ineffective and Hubak was a little effective. 2. In MTT assay of reoxygenation most of herbs were not effective. But Hubak was some effective. 3. In LDH assay of hypoxia all of herbs were effective. Especially Chungpesagan-Tang were equally effective on all of concentration. 4. In LDH assay of reoxygenation all of herbs were generally effective. Especially Chungpesagan-Tang and Baekji were highly effective and Kilkyung was also effective on low concentration. 5, The herbs were generally effective on LDH assay of hypoxia and reoxygenation. Conclusion : The results suggest that Chungpesagan-Tang and all of herbs may have protective effect on condition of oxidative stress and can be applied on the development of a new medicine for neurodegenerative disease like dementia.

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Effects of Taeumin Chungsimyeunjatang on the Cerebral neurons injured by Hydrogen Peroxide (태음인(太陰人) 청심연자탕(淸心蓮子湯)이 Hydrogen Peroxide에 손상(損傷)된 백서(白鼠)의 대뇌신경세포(大腦神經細胞)에 미치는 영향(影響))

  • Ok, Yun-young;Ryu, Do-gon;Kim, Kyung-yo
    • Journal of Sasang Constitutional Medicine
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    • v.11 no.2
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    • pp.251-266
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    • 1999
  • 1. Purpose : The purpose of this study was to determine the effects of Chungsimyeunjatang on the cerebral neurons injured by hydrogen peroxide($H_2O_2$). 2. Methods : I observed cell viability in mouse cerebral neurons exposed to hydrogen peroxide by NR assay and MTT assay and determined lipid peroxidation and amounts of LDH release in mouse cerebral neurons exposed to hydrogen peroxide. After administration of Chungsimyeunjatang water extracts, I observed significant changes of cell viability, lipid peroxidation and amounts of LDH release in mouse cerebral neurons exposed to hydrogen peroxide. 3. Results : Hydrogen peroxide showed neurotoxicity. Cell viability in mouse cerebral neurons exposed to hydrogen peroxide decreased in NR assay and MTT assay. Lipid peroxidation and amounts of LDH release in mouse cerebral neurons exposed to hydrogen peroxide increased. Chungsimyeunjatang was very effective in blocking hydrogen peroxide-induced neurotoxicity.

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Cytotoxicity Evaluation of Cosmetic Materials to Mouse Fibroblast : by Tetrazolium salt, MTT Colorimetric Assay (Tetrazolium salt, MTT Colorimetric Assay를 이용한 Mouse Fibroblast에 대한 화장품원료 물질의 세포독성 평가)

  • Jo, Jae- Hoon
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.15 no.1
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    • pp.37-50
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    • 1989
  • The in Vitro chemosensitivity of fibroblast cell strains was determined using a semiautomated tetrazolium-based colorimetric assay(MTT assay) to 16 cosmetic materials. This assay is useful method to evaluate toxic effects of the chemicals. From assay results, we determined that the preservatives are more toxic than moisteurizers. The chemicals in the same group have a different toxicity. That is, in preservatives, Germall -115 is more toxic than Danisol -M, -p, and in surfactant sodium laurel sulfate than Myrj 52, and in moisteurizers, 1, 3-butylene glycol is more safe than the others. When the results from this assay for preservatives were compared with patch test results, good correlation was observed. Forthemore, this assay method can be used together with Patch test for the evaluation of the chemical toxicity, particularly in cosmetic field.

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Effects of Baepungtang water extract on Cultured Spinal Sensory neurons Damaged by Xanthine Oxidase/Hypoxanthine (배풍탕(排風湯) 전탕액(煎湯液)이 XO/HX에 의해 손상(損傷)된 배양(培養) 척수감각신경세포(脊髓感覺神經細胞)에 미치는 효과(效果))

  • Yu Jin-Deok;Yun Yong-Gap
    • Herbal Formula Science
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    • v.8 no.1
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    • pp.319-328
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    • 2000
  • To evaluate the effect of Baepungtang(BPT) water extract on cultured mouse spinal sensory neuron which was inhibited by xanthine oxidase(XO) and hypoxanthine(HX)-induced oxigen radicals, MTT assay, NR assay, Neurofilament enzymeimmuno assay and LDH activity assay were carried out after the cultured mouse spinal sensory neuron were preincubated with various concentrations of BPT water extract for 3 hours prior to exposure of XO/HX. The results obtained were as follows: 1. XO/HX, a oxigen radical, decreased the survival rate of the cultured mouse spinal sensory neuron cell on NR assay and MTT assay. 2. $MTT_{50}$ value and $NR_{50}$ value of XO/HX were 30 mU/ml XO/O.2 mM HX. 3. BPT water extract have efficacy of increasing neurofilament. 4. BPT water extract have efficacy of increasing LDH activity. From above the results, It is concluded that BPT has marked efficacy as a treatment for the damages caused in the XO/HX-mediated oxidative process.

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Inhibitory Effects of the Ethanol Extract of Ulmus davidiana on Apoptosis Induced by Glucose-glucose Oxidase and Cytokine Production in Cultured Mouse Primary Immune Cells

  • Lee, Jeong-Chae;Lim, Kye-Taek
    • BMB Reports
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    • v.34 no.5
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    • pp.463-471
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    • 2001
  • The bark of Ulmus darvidiana var. japonica Nakai (UDN) has been used for a long time to cure inflammation in oriental medicine. In the present study, two types of extracts, Ulmus water-eluted fraction (UWF) and Ulmus ethanol-eluted fraction (UEF), were prepared from the UDN stem bark, and employed to test the extracts to see if they had anti-oxidative properties against hydroxyl radicals that could alter immune reactivity in mouse immune cells. Deoxyribose assay, DNA nicking assay, and glucose/glucose oxidase assay showed that both fractions had scavenging activity against oxygen free radicals at 50 mg/ml. In addition, hydroxyl radical-mediated apoptosis in mouse thymocytes was not protected by UEF treatment, but the apoptosis was protected by UWF at the same concentration. DNA synthesis and cytokine production that were induced in splenocytes by mitogens (Concanavalin A and lipopolysaccharide) were reduced by the addition of both fractions. These results indicate that both extracts that were prepared from the UDN stem bark have anti-oxidative activities, anti-apoptotic effects, and inhibitory effects on DNA synthesis and cytokine production in mouse immune cell cultures.

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Hygienic Quality and Safety of Gamma Irradiated Angelicae Gigantis Radix and Aloe (감마선조사에 의한 당귀와 알로에의 위생화 및 안전성 평가)

  • 강일준;이수용;이상준;김광훈;이병훈
    • Toxicological Research
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    • v.13 no.1_2
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    • pp.55-60
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    • 1997
  • Gamma irradiation was applied to Angelicae gigantis radix and Aloe to improve their hygienic quality. The effective dose of irradiation was 7 kGy in Angelicae gigantis radlx and 5 kGy in Aloe for the sterilization of all contaminated microorganisms tested. After 8 months of storage at room temperature, no growth of microorganisms was observed in the irradiated products. The safety of these products were evaluated by Salmonella typhimurium reversion assay and in vivo micronucleus assay using mouse bone marrow cells. They were negative in the bacterial reversion assay with S. typhimurium TA 98, TA100, TA1535 and TA1537. In the in vivo mouse micronucleus assay, they did not show any clastogenic effect at all doses tested. These results indicate that the gamma irradiation of Angelicae gigantis radix at 12 kGy and of Aloe at 10 kGy have no genotoxic effects under these experimental conditions.

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Effects of Naetakcheonkeumsan and It’s Gamypang on the Lymphocytes and Cancer cells (內托千金散 및 그 加味方이 마우스의 免疫細胞 및 癌細胞에 미치는 效果)

  • Yang, Gi-ho;Jeong, Hyun-woo;Choi, Jung-hwa
    • The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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    • v.13 no.1
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    • pp.44-59
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    • 2000
  • Naetakcheonkeumsan(NCS) was a drug that treated carbuncle and cellulitis. So, the purpose of this Study was to investigate effect of NCS on the anti-cancer and proliferation of lymphocytes in normal mouse group, L1210 cells-transplanted mouse group and anti-cancer drug (vincristine) 0.005mg/kg were injected mouse(Ll210 cells-transplanted) group. We used NCS extract with freeze-dried, 8wks-old male mice, and Ll210 cell lines for this Study, The proliferation of cells was tested using a colorimetric tetrazoliun assay(MTT assay). The results of this Study were obtained as follow ; Group C(NCS plus Rehmanniae Radix Preparat administered group) inhibited proliferaion of lymphocytes in normal mouse group and Ll210 cells transplanted mouse group. Group A(NCS administered group) and Group B(NCS plus Cervi pantotrichum Cornu administered group) inhibited proliferation of Ll210 cells in Ll210 cells-transplanted mouse group and anti-cancer drug were injected mouse(Ll210 cells-transplanted) group. Group C incresed proliferation of L1210 cells in L1210 cells-transplanted mouse group, but inhibited in anti-cancer drug(vincristine) 0.005mg/kg were injected mouse(L1210 cells-transplanted) group.

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