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Biomedical subjects

E Meerovitch

Publications and source records attributed to E Meerovitch.

At least 19 recordsLinked to original sources

Studies on a morphologically distinct colchicine-resistance variant of Entamoeba sp.

Colchicine has a temperature-dependent cytotoxic effect on Entamoeba sp. (Laredo isolate) that is most apparent when the drug is applied during the initiation of cultures at a concentration of 7.5 mM or higher. Continued transfer of cultures in medium containing progressively increasing concentrations of colchicine has resulted in a variant that grows prolifically in the presence of colchicine (7.5 mM) with a generation time comparable to that of the parent stock, Comparison of a number of parameters of the 2 variants revealed that colchicine resistance was accompanied by a change in cell shape, a reduced membrane permeability, which could partially be overcome by the addition of dimethyl sulfoxide (DMSO), and a reduced tolerance to osmotic stress. However, the parent strain and resistant variant were equally susceptible to cycloheximide and puromycin suggesting that the acquired colchicine resistance may not be explained on the basis of an entirely unspecific generalized reduced ability for drug uptake. Colchicine resistance and altered structure were found to be stable over a long period of time. The possible interdependence of these 2 parameters and their relation to cell motility in Entamoeba sp. are discussed.

Animals

Pathogenicity of axenically cultivated Entamoeba histolytica, strain 200:NIH, in the hamster.

Entamoeba histolytica strain 200:NIH cultured axenically for about 15 years produced lesions and abscesses in inbred adult hamsters following intrahepatic, intraperitoneal, and intradermal inoculation. These amebae, even in much higher doses, inoculated into the ceca of adult and young hamsters failed to produce lesions or to become established there. On the other hand, 200:NIH axenic amebae passaged through hamster liver (substrain 200:NIH-L) were able to become established and to produce lesions in the ceca of very young hamsters (1- and 3-week-old), the degree of pathogenicity appearing to be inversely proportional to age of the hamster.

Age Factors

Restoration of virulence of axenically cultivated Entamoeba histolytica by cholesterol.

The lost pathogenicity of two strains of Entamoeba histolytica, one isolated in 1924 and the other in 1967, grown in axenic culture for the past 5 and 6 years respectively, was restored by supplementing the culture medium with cholesterol through a number of transfers. The number of passages in the cholesterol-supplemented medium, necessary to restore a certain degree of pathogenicity of the two strains in hamsters, was proportional to the total time of in vitro cultivation of the strain, and not just the time of cultivation under axenic conditions. Pathogenicity, once restored, persisted for a long time after cholesterol treatment was stopped.

Animals

Behavior of axenic IP-106 strain of Entamoeba histolytica in the golden hamster.

Strain IP-106 of Entamoeba histolytica was isolated 12 yr ago from a case of amebic dysentery and has been maintained in axenic culture for the past 11 yr. Hamsters were inoculated intrahepatically, intraperitoneally, and intracecally with 6.5, 5.0, and 8.0 X 10(4) axenic trophozoites, respectively. With the first two routes of inoculation all the animals developed liver abscesses, and most developed amebic metastases to other sites. After intracecal inoculation only 3 out of 6 animals developed abscesses at the primary site and metastases to other sites. Most numerous metastases were obtained following intrahepatic inoculation. Intrahepatic inoculation with 1 and 2 X 10(4) axenic amebae resulted in an increase in the size of the liver abscess for a period of 2 wk and its apparent total resorption by day 17 post-inoculation, accompanied by a corresponding gain of body weight. With 4 and 6 X 10(4) amebae there was a steady increase of the size of the liver abscess and corresponding loss of body weight. High antiamebic indirect hemagglutination titers were observed only on day 17 post infection in animals inoculated with 2, 4, and 6 X 10(4) amebae. The extent of amebic metastases was roughly proportional to the size of the inoculum.

Animals

The presence of liver auto-antibodies induced by Entamoeba histolytica in the sera from both naturally infected humans and immunized rabbits.

Auto-antibodies against normal human liver have been detected in the sera of humans with highly positive indirect hemagglutination (IHA) amebiasis titers and with clinically-proven amebic liver abscess. Sera of amebiasis patients and rabbits immunized with killed Entamoeba histolytica were tested for anti-amebic antibodies by the IHA test and for auto-antibodies by the complement fixation test, using the antigens prepared from extracts of human liver and rabbit liver. A direct correlation was found to exist between high anti-Entamoeba antibody titers and the presence of anti-liver antibody in the serum. It is proposed that, in addition to direct parasite damage to host tissue, immunological damage could result from the attachment of circulating antigen to the cell surfaces of host tissues such as the liver.

Adolescent

Protective immunity and possible autoimmune regulation in amebiasis.

Hamsters were injected intradermally with axenic trophozoites of a pathogenic strain of Entamoeba histolytica, and controls similarly with culture medium. Localized lesions were produced in all animals injected with amebae. After the lesions healed all the animals were challenged intrahepatically with axenic amebae. Extensive liver abscesses were produced in all the controls, but a small abscess in only one of the vaccinated hamsters. In the protected animals the indirect hemagglutination antibody titers were high (I:512), while in the controls they ranged between 1:32 and 1:128. In the vaccinated hamster with the small abscess the IHA titer was 1:32. To investigate a possibility of autoimmune regulation in amebiasis, sera of hamsters with experimental amebic liver abscesses were acidified and tested by counter-current-immunoeletrophoresis against untreated autologous sera and amebic antigen. In most cases there was a direct correlation between the two titers. With human antisera in autologous systems there was reactivity against anti-amebic IgG antibodies. These experiments showed that autoimmune complexes may be present in cases of hepatic amebiasis. As in chronic hepatitis and some malignancies, it is possible that immune complexes have an immunoregulatory function in amebiasis.

Animals

Restoration of virulence of axenically-cultivated Entamoeba histolytica by cholesterol.

The lost pathogenicity of two strains of Entamoeba histolytica, one isolated in 1924 and the other in 1967, grown in axenic culture for the past five and six years respectively, was restored by supplementing the culture medium with cholesterol through a number of transfers. The number of passages in the cholesterol supplemented medium, necessary to restore a certain degree of pathogenicity of the two strains in hamsters, was proportional to the total time of in vitro cultivation of the strain, and not just the time of cultivation under axenic conditions. Pathogenicity, once restored, persisted for a long time after cholesterol treatment was stopped.

Animals

The subcellular distribution and properties of Crithidia sp. hydrolases with particular reference to pyrophosphate and orthophosphate monoester phosphohydrolases.

A cell fractionation scheme was developed for studying the distribution of certain hydrolases, especially phosphohydrolases in a Crithidia sp. (Trypanosomatidae). Whilst between 26-56% of the total cellular hydrolase activities were soluble (probably of flagellar pocket origin), a certain percentage, 5-40%, was sedimentable. A particulate fraction obtained after isopycnic density gradient centrifugation (p = 1.187-1.241), designated fraction FA/FB, was enriched in various acid hydrolases (relative specific activities 1.33-6.24) and displayed latent phosphohydrolase activities. The density gradient distributions of this hydrolytic enzymes were compared with reference to one another and malate dehydrogenase (mitochondrial marker). From the results obtained it appears that the sedimentable acid hydrolases of Crithidia are associated with a heterogeneous population of subcellular particles. Cytochemical observations on the FA/FB fraction supported this finding and revealed the association of acid phosphatase reaction product with subcellular elements resembling multivesicular bodies.

Animals