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F Guhl

Publications and source records attributed to F Guhl.

At least 37 records · Page 2Linked to original sources

Clinical Trypanosoma rangeli infection as a complication of Chagas' disease.

Laboratory studies on a group of 20 patients from the Rio Negro Valley, Colombia selected for detailed study showed that 14 gave antibody reactions on immunoassay consistent with Trypanosoma cruzi or T. rangeli infections. Four were diagnosed as having T. rangeli infection, 4 had mixed infections and 6 were infected with T. cruzi alone. Immunoprecipitation analysis showed that sera from T. cruzi-infected patients recognized a similar range of trypomastigote-derived polypeptides as sera from patients in Brazil, and all of the Colombian sera reacted with the 160 kiloDalton (kDa) polypeptide associated with active infection. Although sera from patients with T. rangeli infection alone gave a positive immunofluorescence or ELISA reaction with T. rangeli, they failed to bind to parasite polypeptides by either immunoprecipitation or Western blotting. Intriguingly, sera from patients with mixed infections consistently gave a stronger, but qualitatively similar, binding reaction in immunoprecipitation and Western blotting compared to sera from patients infected with T. cruzi alone.

Antibodies↗

Use of monoclonal antibodies for the differential detection of Trypanosoma cruzi and T. rangeli in epidemiological studies and xenodiagnosis.

T. cruzi and T. rangeli have the same insect and mammalian hosts, including man, and in addition share approximately half the antigenic determinants recognised by the humoral response. Thus serodiagnosis of T. cruzi infection in areas where T. rangeli is endemic may include an unknown rate of false positives due to this antigenic cross-reactivity. Similarly, the results of xenodiagnostic procedures and epidemiological surveys of insect vectors are prone to distortion because of the close morphological resemblance of the epimastigote stages. The description of a T. cruzi epimastigote specific monoclonal antibody, 2A2, which reacts with both culture and insect derived epimastigotes provides a more reliable basis for differential diagnosis of these two parasites.

Animals↗

The affinity of the lectins Ricinus communis and Glycine maxima to carbohydrates on the cell surface of various forms of Trypanosoma cruzi and Trypanosoma rangeli, and the application of these lectins for the identification of T. cruzi in the feces of Rhodnius prolixus.

Flagellates of Trypanosoma cruzi (stock Molino 1), obtained from the intestine of experimentally infected Rhodnius prolixus, grown in cellular or acellular culture, as well as from the blood of infected mice, were examined by a direct fluorescence test using the lectins RCA (Ricinus communis-120) and SBA (soy bean agglutinin; Glycine maxima), conjugated with fluorescein isothiocyanate, for the detection of beta-D-galactose and alpha,beta-N-acetyl-D-galactosamine on the membranes of the flagellates. The same reactions were carried out using Trypanosoma rangeli (stock San Agustin), obtained from the intestine, hemo-lymph or salivary glands of experimentally infected R. prolixus, as well as from cultures and from the blood of experimentally infected CFW mice. The results indicate that the membrane of T. rangeli in the salivary glands of the vector contains beta-D-galactose, but that this sugar is absent from all other developmental stages of this trypanosome. All stages of intestinal and cultured. T. cruzi presented positive reactions with RCA-FITC and SBA-FITC. The high specificity of this technique makes it useful for the examination of R. prolixus, previously used in xenodiagnosis of Chagas' disease and for the examination of intradomiciliary or sylvatic vectors in epidemiological surveys in areas where T. cruzi and T. rangeli coexist. Formaldehyde fixed samples can be examined months later and false reports due to T. rangeli can be avoided.

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[Laboratory maintenance of Trypanosoma (Herpetosoma) rangeli Tejera, 1920].

Two laboratory maintenance systems of Trypanosoma rangeli were compared. The maintenance by weekly subinoculations in Tobie's culture medium and the intrafemoral inoculation of Rhodnius prolixus with cultured flagellates, resulted in loss of infectivity of the metacyclic salivarian trypomastigotes for mice, ten months after maintenance in culture. With the system of cyclical passes through culture-Rhodnius-mouse-culture-Rhodnius, the infectivity of the metacyclic trypomastigotes for mice, was maintained during the three years of the experiment. The number and percentage of metacyclic trypomastigotes formed in the salivary glands of R. prolixus, previously inoculated intrafemorally or intracoelomically with culture forms of T. rangeli, did not show correlation with the inoculated dose, however the inoculated quantity demonstrated a direct relation with the mortality rate of the insects. The results indicate that T. rangeli requires an adequate maintenance system, so that under experimental condition the biological characteristics, normally expressed under natural conditions, are conserved.

Animals↗

Antigen-specific lymphocyte transformation in congenital toxoplasmosis.

Secreted (TSA) and water lysed antigens (WLA) derived from cell culture of the RH strain of Toxoplasma gondii have been used to induce antigen-specific mitogenesis of lymphocytes from children with congenital toxoplasmosis. Responsiveness to WLA was variable, with all patients giving false negative results. Responses to TSA, however, were highly specific, with no false negatives. This increased specificity was not due to an increased response to TSA (P less than 0.001), but a lower TSA response by the lymphocytes of the uninfected subjects (P greater than 0.1), compared with WLA in both cases. Preliminary studies have shown that while WLA induces high levels of non-specific immunity, such as natural killing, TSA is able to induce specific killing of Toxoplasma in vitro through the action of activated macrophages; this may partially explain the increased specificity of TSA in the lymphocyte transformation test.

Adult↗

Isozyme profiles of Trypanosoma cruzi stocks from Colombia and Ecuador.

A total of 74 of 82 domestic Rhodnius prolixus from the same locality in eastern Colombia were found to be infected with Trypanosoma cruzi or T. rangeli. One of three domestic Triatoma dimidiata from Ecuador also showed T. cruzi infection. A total of 59 T. cruzi stocks from these and five other localities in Colombia were isolated from man, marsupials and triatomine bugs. Cellulose-acetate electrophoresis of nine or ten enzymes characterized all T. cruzi stocks as zymodeme 1 (reference clone Silvio X10/1). Differences in electrophoretic patterns between the newly isolated stocks and the zymodeme 1 standard were seen with the enzymes G6PD and HK. These results are in agreement with the previously described geographical distribution of T. cruzi zymodemes. Stocks were isolated from both low and high altitudes and there was no evidence of adaptative significance of T. cruzi enzyme polymorphism.

Animals↗

Antibody response to experimental Trypanosoma rangeli infection and its implications for immunodiagnosis of South American trypanosomiasis.

Differential immunodiagnosis of T. rangeli and T. cruzi infections in man poses a particular problem, not only because these parasites share antigenic determinants, as detected by immunofluorescence, but also because they have a similar geographical distribution, the same host range and often identical insect vectors. We show here that whereas mouse anti-T. rangeli sera have significant cross reactivity with T. cruzi by immunofluorescence, they are entirely specific when tested by ELISA, using apparently similar antigen preparations. Immunoprecipitation analysis detected relatively little cross-reactivity between heterologous antisera and parasite combinations. Intriguingly, immunization with T. rangeli epimastigotes was much more powerful than similar immunization with trypomastigotes, and the majority of the antibody was directed against a single polypeptide of apparent Mr 73kDa.

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Purified Trypanosoma cruzi specific glycoprotein for discriminative serological diagnosis of South American trypanosomiasis (Chagas' disease).

Chagas' disease, leishmaniasis, and Trypanosoma rangeli infection are endemic and their distributions overlap in vast regions of South and Central America. Serological cross-reactivities can confuse epidemiological studies of these infections, and their differential diagnosis has been assigned a high priority by the World Health Organisation. A lectin-affinity-purified, 90,000 molecular weight glycoprotein (GP90) is present in the known principal strains (zymodemes) of Trypanosoma cruzi and absent from Leishmania and T rangeli. Patients with T cruzi infection have antibody to GP90, whereas patients with leishmaniasis do not and the two infections can be distinguished in an ELISA system using this antigen. In a mouse model, the same test can differentiate between T cruzi and T rangeli infections. Antigens purified by affinity chromatography clearly provide a practical basis for very precise, even strain-specific, diagnostic tests.

Antibodies↗

Trypanosoma magdalenae sp. n (Protozoa: Kinetoplastida) from a freshwater teleost, Petenia kraussii, in Colombia.

A new, monomorphic trypanosome, Trypanosoma magdalenae sp. n. was found in five of 38 fish, Petenia kraussii, from the Río Magdalena in Colombia, South America. It is the first trypanosome species designated from freshwater teleosts in Colombia. The trypomastigotes measured in 42.4 micron +/- 2.05 SD (range, 39.8 - 45.6) by 2.1 micron +/- 0.20 (2.0 - 2.5). Their nuclear index was 1.6 micron +/- 0.22 (1.25 - 1.84) and their kinetoplastic index equaled 1.1 micron +/- 0.02 (1.08 - 1.15). Infection intensity was usually limited to two or three flagellates per 40 microliter of packed blood cells.

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A modification of Diamond's medium for the axenic culture of Entamoeba histolytica.

The use of casein hydrolysate in Diamond's axenic culture medium TPS-1 in replacement of trypticase allowed good growth of the trophozoites of Entamoeba histolytica. This modified medium also supported growth of trophozoites preserved for 16 months in liquid nitrogen. Considerable labour and cost of serum can be saved by using 5% instead of 10% bovine serum in combination with this modified medium.

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Chagas disease and human migration.

Human Chagas disease is a purely accidental occurrence. As humans came into contact with the natural foci of infection might then have become infected as a single addition to the already extensive host range of Trypanosoma cruzi that includes other primates. Thus began a process of adaptation and domiciliation to human habitations through which the vectors had direct access to abundant food as well as protection from climatic changes and predators. Our work deals with the extraction and specific amplification by polymerase chain reaction of T. cruzi DNA obtained from mummified human tissues and the positive diagnosis of Chagas disease in a series of 4, 000-year-old Pre-Hispanic human mummies from the northern coast of Chile. The area has been inhabited at least for 7,000 years, first by hunters, fishers and gatherers, and then gradually by more permanent settlements. The studied specimens belonged to the Chinchorro culture, a people inhabiting the area now occupied by the modern city of Arica. These were essentially fishers with a complex religious ideology, which accounts for the preservation of their dead in the way of mummified bodies, further enhanced by the extremely dry conditions of the desert. Chinchorro mummies are, perhaps, the oldest preserved bodies known to date.

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[Antigenic differentiation among 3 life-cycle stages of Toxoplasma gondii using fluorescent antibodies].

Antigenic differentiation between three stages of the life cycle of Toxoplasma gondii by fluorescent antibodies. Antisera were prepared in three groups of twenty mice each with three different antigens of Toxoplasma gondii: the first group was inoculated with tachyzoites of RH strain and received sulfadiazine treatment; the second with tissue cysts of T-100-cat-6751 strain and the third with oocysts of the same strain, both without treatment. In the indirect immunofluorescent antibody technique each antigen was tested with its homologous and heterologous antisera, determining qualitative and quantitative antigenic differences according to the fluorescence patterns. Some stages of Toxoplasma when reacting with their heterologous antibodies showed a central, partial posterior or total fluorescence during a certain period of development. The difference in fluorescence was sufficient to distinguish whether the origin of the infection was via cyst, oocysts and/or via tachyzoite when observation was made before day 65 post-infection.

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