| Project/Area Number |
06670269
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| Research Category |
Grant-in-Aid for General Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Research Field |
寄生虫学(含医用動物学)
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| Research Institution | KEIO UNIVERSITY |
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Principal Investigator |
KOBAYASHI Seiki RESEARCH ASSOCIATE,SCHOOL OF MEDICINE,KEIO UNIVERSITY, 医学部, 助手 (70112688)
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| Co-Investigator(Kenkyū-buntansha) |
SANUKI Jun-ichi RESEARCH ASSOCIATE,SCHOOL OF MEDICINE,KEIO UNIVERSITY, 医学部, 助手 (90255571)
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| Project Period (FY) |
1994 – 1995
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| Project Status |
Completed (Fiscal Year 1995)
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| Budget Amount *help |
¥2,100,000 (Direct Cost: ¥2,100,000)
Fiscal Year 1995: ¥700,000 (Direct Cost: ¥700,000)
Fiscal Year 1994: ¥1,400,000 (Direct Cost: ¥1,400,000)
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| Keywords | Entamoeba histolytica / Entamoeba invadens / DNA synthetic cycle / flow cytometry / mathematical modeling / Entamoeba dispar / axnic cultivation / DNA polymerase / Pseudomonas aeruginosa |
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| Research Abstract |
We developed a method to study the DNA synthetic cycles of Entamoeba histolytica and Entamoeba invadens by flow cytometry (FCM) based on a preparative procedure to reduce both high levels of natural fluorescence and non-specific adsorption of flurochromes. We modeled G_1, S,and G_2 phases as a series of overlapping Gaussian curves. Both E.histolytica and E,invadens displayd G_1, S,and G_2 proportions that are consistent with eukaryotic cell populations in exponential of stationary growth phase. Exponetial phase E.histolytica populations contained a hypodiploid subset with a mass of about 20% less than the diploid value which we estimate by FCM to be 24 x 10^<-14>gDNA/cell.
Exponential phase E.invadens populations contained a hypodiploid subset with a mass of about 6% less than the diploid value which estimate by FCM to be 30 x 10^<-14>gDNA/cell.
We developed also an axnic cultivation system for Entamoeba dispar which was nonpathogenic amoeba, by using newly developed BCSI-S medium in the presence of Crithidia fasciculata. sterilized by treating with 1% hydrogen peroxide for 24 hours at 4ﾟC.
DNA polymerase activity of the ameba which was grown in this culture system, was detected and characterized. DNA polymerase activity was high at pH 2 to pH 6, but at pH 8 and 10 the activity was very low in nuclear extracts from both trophozoites of E.histolytica and E.dispar which were grown axnically.
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