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Biomedical subjects

M A Basombrío

Publications and source records attributed to M A Basombrío.

At least 19 recordsLinked to original sources

Furoxan derivatives as cytotoxic agents: preliminary in vivo antitumoral activity studies.

Furoxan derivatives with in vitro cytotoxic activity were investigated as antitumoral agents in vivo. The compounds were tested in murine models of both CCRFS-180 II sarcoma and mammary adenocarcinoma. Two of the furoxan derivatives considered here, 3-formyl-4-phenyl-1,2,5-oxadiazole N2-oxide and 3-carbonitrile-4-phenyl-1,2,5-oxadiazole N2-oxide, present in vivo antitumoral activity. They were able to produce more than 90% of tumoral necrosis under the experimental protocol of administration and posology employed. NO-releasing capacity of furoxans may explain the anti-neoplastic activity of these compounds.

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Canine infection and the possible role of dogs in the transmission of American tegumentary leishmaniosis in Salta, Argentina.

Some Leishmania species affect humans in two principal forms: visceral and cutaneous leishmaniosis (CL). Several studies have identified dogs as the main reservoirs of the visceral leishmaniosis (VL) caused by Leishmania infantum. The purpose of this work was to carry out a survey of the canine population associated with human cases of American tegumentary leishmaniosis (ATL), in order to establish the clinical, parasitological, serological and immunological characteristics of the canine disease, in an endemic region for both ATL and Chagas' disease in the province of Salta, in northwestern Argentina. Two hundred and eight dogs from the endemic area were examined and 41 (19.7%) of them presented lesions compatible with leishmaniosis. In order to investigate the presence of antibodies against Leishmania spp. and Trypanosoma cruzi, sera were screened by ELISA using two complex antigens from these parasites and, because of cross-reactions between them, a specific antigen for diagnosis of T. cruzi infection. Sixty-two (29.8%) of 208 dogs were positive for the complex antigen F45 from Leishmania and 50 (24%) were positive for the complex antigen F105 from T. cruzi. Nine dogs (4.3%) were positive for the specific Ag163B6-cruzipain suggesting that these dogs were truly infected with T. cruzi. Furthermore, three of these nine dogs presented Leishmania sp. in their skin lesions and therefore were considered as infected by both, T. cruzi and Leishmania parasites. The prevalence of Leishmania infection detected by lesions and/or positive serology was 27.4% (57/208). On the basis of previous observations regarding the clustered appearance of human ATL, the dog population was divided into two groups: zone A, dogs living within a 100 m radius from houses with human cases, and zone B, dogs living beyond this limit. The prevalence of ATL in dogs was significantly higher in zone A (34.6%) than in zone B (7.3%), suggesting a strong correlation between canine and human cases. The average time required for a parasitological diagnosis by microscopy was six times longer for dog samples than human ones, and the average number of parasites per 100 microscopic fields was 14-fold lower in canine samples. The high prevalence of Leishmania infection and the close association with human cases, demonstrated that dogs are a very susceptible host for Leishmania infection, but the scarcity of parasites in their lesions suggests that they may not be the main reservoir of the parasite in this endemic area.

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Force of infection and evolution of lesions of canine tegumentary leishmaniasis in northwestern Argentina.

A clinical-serological follow-up was carried out in a canine population in endemic foci of Leishmania braziliensis spread in northwestern Argentina. Each dog was studied in at least two visits, 309+/-15 days (X+/-SE) apart. Some initially healthy dogs (n=52) developed seroconversion or lesions. The clinical evolution of the disease in dogs resembles in many aspects the human disease. Similarities include the long duration of most ulcers with occasional healing or appearance of new ones and the late appearance of erosive snout lesions in some animals. Yearly incidence rates of 22.7% for seroconversion and of 13.5% for disease were calculated as indicators of the force of infection by this parasite upon the canine population.

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Synthesis and antitrypanosomal evaluation of E-isomers of 5-nitro-2-furaldehyde and 5-nitrothiophene-2-carboxaldehyde semicarbazone derivatives. structure-activity relationships.

Several novel semicarbazone derivatives were prepared from 5-nitro-2-furaldehyde or 5-nitrothiophene-2-carboxaldehyde and semicarbazides bearing a spermidine-mimetic moiety. All derivatives presented the E-configuration, as determined by NMR-NOE experiments. These compounds were tested in vitro as potential antitrypanosomal agents, and some of them, together with the parent compounds, 5-nitro-2-furaldehyde and 5-nitrothiophene-2-carboxaldehyde semicarbazone derivatives, were also evaluated in vivo using infected mice. Structure-activity relationship studies were carried out using voltammetric response and lipophilic-hydrophilic balance as parameters. Two of the compounds (1 and 3) displayed the highest in vivo activity. A correlation was found between lipophilic-hydrophilic properties and trypanocidal activity, high R(M) values being associated with low in vivo effects.

Aldehydes↗

Studies on the virulence and attenuation of Trypanosoma cruzi using immunodeficient animals.

Tissue invasion and pathology by Trypanosoma cruzi result from an interaction between parasite virulence and host immunity. Successive in vivo generations of the parasite select populations with increasing ability to invade the host. Conversely, prolonged in vitro selection of the parasite produces attenuated sublines with low infectivity for mammals. One such subline (TCC clone) has been extensively used in our laboratory as experimental vaccine and tested in comparative experiments with its virulent ancestor (TUL). The experiments here reviewed aimed at the use of immunodeficient mice for testing the infectivity of TCC parasites. It has not been possible to obtain virulent, revertant sublines by prolonged passaged in such mice.

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[The transmission de Chagas disease in Salta and the detection of congenital cases].

Data on the prevalence of Trypanosoma cruzi infection is presented for the province of Salta, Argentina. Special emphasis is given to the detection of congenital transmission and to the economic benefits of preventing Chagas' disease. Seroepidemiological data obtained from 20 year old army draftees revealed a reduction, from 22.7 to 11.11% between 1964 and 1985. In university students, a rate of 0.96% was found in 1998. Surveys carried out during 1996 showed that more than 15% of the pregnant women analyzed carried T. cruzi infection, particularly in the north of the province. This situation brings about a high risk of appearance of congenital cases and represents an opportunity to test the most adequate strategies for detection. By applying systematically microhematocrit, hemoculture and PCR methods, to umbilical chord blood, an increase in the early detection of congenitally infected babies is being achieved. In 1992-94, very high seroprevalence rates of infection were found among indians of the Chaco region of Salta. The overall rate was 37%, but there were 5 localities where more than 54% of the population was infected. These numbers indicate that, in vast areas of the provincial territory, fight against vector bugs must not merely consist of surveilance activities, but rather of renewed spraying attacks. The fight must include control of pregnant women and blood banks. An economic analysis of the economic return, calculated only for spraying activities and for the Department of Anta (Salta), indicated a net present value of over 7 million dollars and an internal rate of return exceeding 60%.

Argentina↗

Immunogenicity of the recombinant SAPA protein of Trypanosoma cruzi for mice.

The humoral and cellular immune responses induced by the recombinant SAPA (shed acute phase antigen) of Trypanosoma cruzi were studied in mice and correlated with the immunologic control of parasitemia. The immunizing schedule used consisted of 2 weekly injections of 50 micrograms glutathione-S-transferase (GST)-SAPA in Freund's adjuvant. Specific alpha GST-SAPA antibodies were detected by enzyme-linked immunosorbent assay 1 wk after each antigen dose, the concentration of antibodies after the second injection being 30-fold higher than after the first. Immediate- (ITH) and delayed-type hypersensitivity (DTH) reactions were observed as footpad swelling after injecting 50 micrograms GST-SAPA in preimmunized mice as compared to naive controls. Adoptive transfer experiments indicated that these cutaneous reactions were mediated by lymphoid cells and not by serum. Both humoral and cellular responses were specific for the GST-SAPA antigen and did not cross-react with either the GST or the recombinant GST-1 T. cruzi antigen. Immunized mice that had developed high levels of antibody and DTH reaction to GST-SAPA were able to control the level of parasitemia after challenge with 10(3) blood trypomastigotes. The levels of parasitemia obtained were lowered to about 1/3 (P < 0.05) and mortality at day 60 was reduced from 67 to 25% (P = 0.085). Comparison of this immunizing method with other schedules involving more injections or higher antigen doses indicates that control of parasitemia can be obtained with low amounts of antigen and seems to be associated with the development of DTH.

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Trypanosoma cruzi: effect of immunization on the risk of vector-delivered infection in guinea pigs.

The protective effect of experimental immunization was studied in guinea pigs exposed to vectorial infection by Trypanosoma cruzi. Immunized animals received an inoculum of live-attenuated T. cruzi epimastigotes into a granuloma previously induced by Freund's complete adjuvant in the hind footpad. Seven days later, a delayed-type hypersensitivity reaction was triggered by reinjection of the parasites in the front footpad. The animals were then placed in Triatoma infestans-colonized corrals and exposed to vectorial T. cruzi transmission of the parasite for up to 200 days. The effectiveness of this immunizing protocol was controlled in terms of the number of bites necessary for infection (NBNI) in immunized as compared with control animals. Periodic entomological census allowed for the determination of vector biting and infection rates and the calculation of NBNI. Although this measurement was quite variable between yards, an overall average of 4,973 bites was enough to infect a control guinea pig in 4 separate experiments. The corresponding figure for the experimental group was 21,307 bites, implying that immunized animals could resist a 4.28-fold increase (range: 1.99-8.32) in the number of vector bites before becoming infected.

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Evaluation of an attenuated Trypanosoma cruzi strain in rats. Analysis of survival, parasitemia and tissue damage.

Infection and tissue damage induced by parasites of an attenuated Trypanosoma cruzi culture strain (TCC) were studied in "I" line of inbred rats. Suckling rats (S), 3-5 day old were inoculated i.p. with 10(6) TCC (S1), 10(7) TCC (S2) and 10(8) TCC (S3). Weaned rats (W), 21-25 day old were inoculated s.c. with 10(6) TCC (W1), 10(7) TCC (W2) and 10(8) TCC (W3). The cultures yielded up to 2% of trypomastigotes. Controls inoculated either i.p. or s.c. with 10(6) blood form trypomastigotes (SC and WC) as well as normal controls (NC) were included. Survival was 100% in S1, S2 and S3, and 0% in SC on day 13 post-infection (p.i.). The latter animals died with acute Chagas disease signs. Survival was 100% in the W groups. In the first 30 days p.i. parasites were detected in S1, S2 and S3 and W1, W2, W3 groups after exhaustive examination. Parasites were easily found in WC and SC until day 13. Xeno-diagnoses were positive (5/5) at 2 months p.i. and negative at 6 months p.i. (W1, W2, W3, 0/23; WC, 0/5). Only cardiac lesions were slightly increased. The frequency of focal chronic myocarditis seemed to be increased in a dose-independent manner (S1, S2, S3, 26%; W1, W2, W3, 46%) but was not significant in comparison with NC, and even was lower than usually found in WC (61.3%). The reduced virulence and pathogenicity suggest that the TCC strain suffered a remarkable attenuation after long term in vitro culture.

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Morphometry of skeletal muscle involvement in mice infected or preimmunized with live attenuated Trypanosoma cruzi.

A morphometric study was undertaken in the quadriceps muscle of Swiss mice in order to assess the effects of immunization with attenuated T. cruzi upon tissue lesions. interfascicular lymphocytic infiltration, presence of amastigote nests, vascular lesions, degeneration and fibrosis were evaluated independently. Each of these alterations was drastically prevented in preimmunized animals. These results indicate that immunity against T. cruzi not only reduces circulating parasites but also clears most of the organic damage caused by infection.

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Myocardial involvement in Cavia porcellus naturally infected with Trypanosoma cruzi.

This paper describes the myocardial involvement analyzed by histopathological examination in rural Cavia porcellus during natural T. cruzi infection. Four Cavia porcellus of both sexes were bred in a house free of Triatoma infestans. In contrast, four animals were born and lived in a rural yard colonized by T. infestans. Autopsies were performed at 6-9 months of age, in animals weighing 550 to 750 grams. The naturally infected Cavia porcellus presented moderate and severe lymphocyte and plasmocyte infiltrates, focally or diffusedly distributed. Replacement of myocytes both in atria and ventricles was often found and consisted of loose or dense connective tissue infiltration. Regarding the conducting system, polymorphonuclear cell infiltrates were observed in the A-V node and in the left bundle branch. Uninfected Cavia porcellus did not show these lesions. Typical chagasic cysts were not found in the naturally infected Cavia porcellus hearts. Parasitism was not observed in the skeletal muscles. It is concluded that naturally infected Cavia porcellus develop consistent lesions similar to those described in human chronic chagasic myocardiopathy. The high susceptibility of naturally infected Cavia porcellus must be taken into account when these animals are used in studies regarding chronic chagasic myocardiopathy.

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Field trial of vaccination against American trypanosomiasis (Chagas' disease) in domestic guinea pigs.

Domestically bred South American guinea pigs received 3 to 5 immunizing intradermal inocula of 28 X 10(6) live attenuated Trypanosoma cruzi epimastigotes (TCC strain) per kg. These inocula were unable to produce patent infections or to propagate through vectors. Groups of experimental and control guinea pigs were exposed to natural T. cruzi infection in a field yard for periods of up to 551 days. Xenodiagnoses were applied periodically to all animals. This showed that the incidence of natural T. cruzi infection was significantly lowered at various periods post-exposure. The final proportion of infected animals was 39% (20/51) among vaccinees vs. 63% (32/51) among controls (P less than 0.02). The protective effect was exerted particularly upon males and lasted for over a year in one experimental series (infection in 1/7 vaccinees vs. 6/7 controls, P = 0.014). Vaccination reduced vector transmission rates from 38% to 18% (P less than 0.001). These results agree with previous laboratory experiments in showing a partial resistance which does not eliminate residual T. cruzi infection. However, the field work indicates that even this kind of resistance may have epidemiological impact, reducing both the number of reservoirs spreading the disease and the rate of vector transmission.

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Side effects of immunization with liver attenuated Trypanosoma cruzi in mice and rabbits.

Immunity against lethal, bloodstream forms of Trypanosoma cruzi was achieved in mice by preinoculation of approximately equal to 10(5) culture epimastigotes of an attenuated T. cruzi strain (TCC). The risks of TCC inoculation in terms of pathogenicity or eventual increase in virulence of TCC progeny were evaluated. No pathogenic parasites could be selected from TCC progeny by either mouse, triatome, or culture passages. Immunizing doses of live TCC did not induce in adult mice alterations resembling chronic Chagas' disease, as judged by patterns of mortality, tissue damage, autoantibodies, or parasite recovery. On the basis of the same criteria, However, a remarkable similarity could be established between the disease caused in mice by inoculation of low numbers (10(2)) of pathogenic trypomastigotes and human chronic Chagas' disease. Although patent parasitemias were never revealed in fresh blood mounts obtained from TCC-inoculated mice, a few hemocultures and xenodiagnoses gave positive results, particularly soon after inoculations at birth. The parasites recovered by either method remained in the attenuated, epimastigote stage. In rabbits, no local lesions, fever, weight loss, or histopathological alterations were detected after subcutaneous inoculation of 10(7) TCC organisms, although one fifth of the animals yielded positive hemocultures of epimastigotes. The contrasting host response to cultured epimastigotes as compared with blood trypomastigotes indicates that, in experimental Chagas' disease, immunoprotection is not necessarily associated with immunopathology.

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Trypanosoma cruzi culture used as vaccine to prevent chronic Chagas' disease in mice.

The development of chronic pathology in mice at 2 to 10 months after inoculation of 10(2) T. cruzi trypomastigotes can be prevented by preimmunization with live, attenuated culture parasites (strain TCC). Swiss mice received one or three immunizing inoculations of 10(6) TCC organisms and were challenged with 10(2) Tulahuén blood trypomastigotes. Control groups received only the immunizing or the challenge inoculations. Immunized groups as compared with nonimmunized controls had lower mortality rates at 2 months postchallenge (9% versus 23%; P = 0.059), lower early peaks of parasitemia, lower percentages of positive xenodiagnoses at 5.5 months (40 versus 80%; P = 0.061), and lower incidences of tissue lesions in the skeletal muscle (P less than 0.005) at 2,6, and 10 months postchallenge. Tissue lesions in the heart and smooth muscle were also reduced, reaching statistical significance after 10 months (P less than 0.02). Chronic pathology parameters were never enhanced in preimmunized groups. In spite of the putative role that autoimmunity may play in the development of chronic chagasic lesions, the preventive effect of vaccination is readily exerted upon the chronic murine model of Chagas' disease.

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