Cytochemical comparison of distribution of proteins and lipids in axenically vs. monoxenically grown Entamoeba histolytica |
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| Authors: | N N Sharma R A Albach J G Shaffer |
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| Affiliation: | 1. Department of General Internal Medicine, Medical College of Wisconsin, Milwaukee, WI, USA;2. Aurora Cardiovascular Services, Aurora Sinai/Aurora St. Luke''s Medical Centers, Milwaukee, WI, USA;3. Division of Cardiology, Medical College of Wisconsin, Milwaukee, WI, USA;4. Kalamoon Medical School, Damascus, Syria;5. Johns Hopkins Bloomberg School of Public Health, Baltimore, MD, USA;6. University of California, Los Angeles, CA, USA;1. Department of Chemistry and Chemical Engineering, Royal Military College, Kingston, Ontario, Canada;2. Department of Anatomy and Cell Biology, McGill University, Montreal, Quebec, Canada |
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| Abstract: | Ninhydrin-Schiff (NS) method revealed a higher level of protein-bound amino acids in diffuse and particulate form in cytoplasm of 72-hr, axenically grown amebae as compared to 72-hr cells from monoxenic cultures. Younger (24-hr) monoxenically grown amebae had more such cytoplasmic NS-staining material than older cells. Nuclear NS-positive material in both axenic and monoxenic cells, which was restricted to the peripheral chromatin and endosome, was more intense than that in the cytoplasm.Tyrosine-containing proteins, as determined by Millon's reaction, were present in small amounts in the cytoplasm of both axenic and monoxenic amebae. As was true of the NS method, more intense staining was observed in the nuclei of both cell types. In contrast to the NS technic, however, the nuclear stain was evenly dispersed.Axenic 72-hr amebae had more intensely stained granular and diffuse sudanophilic material (stained with Sudan black B) in the cytoplasm than 72-hr monoxenic cells. Twenty-four-hour monoxenic cells had more sudanophilic inclusions than 48- and 72-hr amebae, primarily associated with large bodies. In some amebae (axenic and monoxenic) intense sudanophilic material was in close association with the nuclear membrane, rarely with the endosome. Cold acetone extracted all sudanophilic material from axenic amebae but left some lightly stained cytoplasmic inclusions in monoxenic cells, especially younger (24-hr) cells. Hot chloroform:methanol extracted all lipids from all cells. The phosphine 3R secondary fluorescence technic used to demonstrate neutral fats gave negative results with both cell types except for a faint reaction in young (24-hr) monoxenic cells.The amount of neutral fat detected was very low. Oil red O demonstrated only a faint diffuse and particulate reaction in 72-hr axenic and 72-hr monoxenic amebae. However, younger (24-, 48-hr) monoxenic amebae had more neutral fat (= oil red Ostaining material) than 72-hr cells. In some young cells (24-, 48-hr) oil red O-positive material was often in association with the nuclear membrane. Phospholipids could not be demonstrated in either cell type with Nile blue sulfate. |
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