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Biomedical subjects

A M Jansen

Publications and source records attributed to A M Jansen.

At least 19 recordsLinked to original sources

Stable infection of primates with Trypanosoma cruzi I and II.

In order to better comprehend the putative association between genotype Trypanosoma cruzi II and primates, an evaluation of the infection in free ranging primates and specimens born in captivity from different geographical areas, the Amazon and the Atlantic forest, was carried out. Seroprevalences of the T. cruzi infection among the primates was similar in both biomes (45.5% and 46%). The parasites were isolated from 8 and 4 different species of primates, respectively from the Amazon and Atlantic forest. Multi-locus enzyme electrophoresis (MLEE) typed the isolates from Amazon as zymodeme 1. Mini-exon gene analysis characterized all these isolates as T. cruzi I, the main genotype circulating in the region. In the Atlantic forest, primates infected with TCI and TCII, as well as a mixed infection (TCI and TCII), were detected. These findings prove that primates may maintain stable infections by both genotypes. Moreover, data show that T. cruzi can occur in a wide range of primate genera, independent of their social behaviour, niches or habitats. Considering the high seroprevalence and stability of T. cruzi infection among the primates, these animals play an important role in the maintenance of the parasite in nature.

Animals↗

Sequence characterization and expression patterns of defensin and lysozyme encoding genes from the gut of the reduviid bug Triatoma brasiliensis.

The cDNAs encoding an intestinal defensin (def1) and lysozyme (lys1) of the reduviid bug Triatoma brasiliensis have been amplified by PCR using specific oligonucleotide primers and 5'- and 3'-RACE, cloned and sequenced. The 576 bp clone has an open reading frame of 282 bp and encodes a pre-prodefensin with 94 amino acid residues, containing a putative signal and activation peptide cleavage site at Ser19 and Arg51, respectively. The genomic DNA contains a second defensin gene with similar characteristics, 88.3% identity and also one intron of 107 nucleotides. The 538 bp clone has an open reading frame of 417 bp, encoding a pre-lysozyme with 139 amino acid residues. The putative signal peptide is cleaved at alanine 18. Using whole mount in situ hybridization, high expression of both genes has been found, distributed uniformly throughout the entire cardia and the blood-storing stomach and to a much lower extent in the digesting small intestine. Using quantitative real-time PCR, the expression level of def1 was also shown to be very low in small intestine, rectum and salivary glands; in the stomach, expression was 500-2500 times higher than in the cardia and fat body. No expression of lys1 could be detected in the salivary glands and rarely a very low expression in the small intestine, rectum and fat body. Lys1 expression in the stomach was 60-300 times higher than in the cardia. Comparing the levels in unfed fifth instars and up to 15 days after feeding, a strong def1 induction was evident in the fat body at 15 days after feeding and in the stomach a maximum level of def1 and lys1 at 5 days after feeding.

Amino Acid Sequence↗

Trypanosoma cruzi infection in wild mammals of the National Park 'Serra da Capivara' and its surroundings (Piaui, Brazil), an area endemic for Chagas disease.

We studied the prevalence of Trypanosoma cruzi infection among eight species of wild small mammals (n=289) in an area where human cases of infection/disease have occurred. Dogs (n=52) and goats (n=56) were also surveyed. The study was carried out inside a biological reserve, the National Park 'Serra da Capivara' and its surroundings in Piaui State, Brazil. The marsupial Didelphis albiventris and the caviomorph rodent Trichomys apereoides were found to be the most important reservoirs in the study area. Trichomys apereoides was the most abundant species (80%) and D. albiventris the most frequently infected (61%). Both T. cruzi I and T. cruzi II genotypes were isolated from these species. One specimen of Tr. apereoides displayed a mixed T. cruzi I/zymodeme 3 infection. Serum prevalence among dogs suggests that they may be involved in the maintenance of the parasite in the peridomestic environment, in contrast to goats, which are not apparently of any epidemiological importance. The distinct distribution and patterns of infection observed in the study areas suggest that even in the same biome, epidemiological studies or determination of control measures must take into account ecological peculiarities.

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Enzootiology of Trypanosoma evansi in Pantanal, Brazil.

In order to better understand the enzootiology of trypanosomiasis caused by Trypanosoma evansi in the Brazilian Pantanal we examined domestic and wild mammals by microhematocrit centrifuge technique (MHCT), immunofluorescence antibody test (IFAT) and polymerase chain reaction (PCR). T. evansi infection was detected in all species sampled with exception of the sheep and the feral pig. High parasitemias were observed in capybaras (5/24), coatis (18/115), horses (31/321) and dogs (3/112). Among these species, only the capybaras did not develop anemia. Low parasitemias, only detected by PCR, were found in buffaloes (18/43), bovines (29/331), marsupials (1/4), small rodents (14/67), bats (7/18), and one armadillo (1/8). The highest prevalence of T. evansi infection was recorded in horses (73%), although no neurological signs in infected horses were observed. Diagnosis through standard parasitological tests and IFAT should be used with caution since they may overlook comprovedly infected horses. The relationship between ranch management and T. evansi infection in horse was investigated. The importance of other transmission mechanisms apart from the tabanids and reservoir hosts are discussed.

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Trypanosoma cruzi transmission in a captive primate unit, Rio de Janeiro, Brazil.

A breeding in captivity program of neotropical primates for subsequent reintroduction in nature is in progress at the Primatology Center of Rio de Janeiro (CPRJ). Almost 200 animals of 20 species that include both wild captured animals and specimens born in captivity are maintained in CPRJ. Here, we examined 198 primates of CPRJ for infection with the protozoan parasite Trypanosoma cruzi. The animals included 18 species of eight genera. We also performed an "ad lib" search for triatomines that could be incriminated as putative transmitters of the protozoan in this scenario. Anti-T. cruzi antibodies were observed (by indirect immunofluorescence assay-IFA) in 40 monkeys (26.5%). Four Panstrongylus megistus were collected in the monkey's food storage room near the cages and in human dwellings in the proximity to CPRJ. T. cruzi were isolated from nine primates of two genera (Leontopithecus and Saguinus) and from two individuals of the vector P. megistus. The transmission inside the cages could be attested by the isolation of the T. cruzi from primates born in captivity. Multi-locus enzyme electrophoresis (MLEE) demonstrated that the two isolates from Saguinus bicolor bicolor displayed a zymodeme 1 profile in four out of five tested enzymes, while all isolates derived from Leontopithecus showed zymodeme 2 for four out of the five tested enzymes. Mini-exon gene analysis genotyped all isolates as T. cruzi II, which is associated with human disease in Brazil. A wild primate unit such as CPRJ, located inside the forest and near to human dwellings and with T. cruzi II infected animals, deserves a careful surveillance in order to prevent expansion of the infection.

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Distinct patterns of Trypanosoma cruzi infection in Leontopithecus rosalia in distinct Atlantic coastal rainforest fragments in Rio de Janeiro--Brazil.

Previous studies on infection of Trypanosoma cruzi in the Poço das Antas Biological Reserve population of wild free-ranging Leontopithecus rosalia have shown the presence of genotype T. cruzi II, associated in Brazil with human disease. Herein, this study has been extended, the infection being evaluated in L. rosalia of 3 different tamarin populations, inhabiting distinct forest areas located in the same Atlantic Coastal Rainforest. Edentata, Marsupialia, Rodentia and Chiroptera were examined exclusively in the Poço das Antas Biological Reserve. Excluding Chiroptera, T. cruzi infection was found in all orders. Biochemical and molecular characterization demonstrated that golden lion tamarins maintained stable infections by T. cruzi II. The isolates from the other mammals corresponded to T. cruzi I, suggesting independent transmission cycles occurring among the sylvatic mammals inside Poço das Antas Biological Reserve. Significant differences in the infection patterns presented by the 3 populations of wild and captive-born golden lion tamarins were noticed. In Poço das Antas a considerably higher number of positive haemocultures from tamarins with positive serological titres was observed in comparison to those obtained from other areas. The implications for conservation and public health of an active sylvatic cycle in the Atlantic Coastal Rainforest of Rio de Janeiro are discussed.

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Trypanosoma cruzi I and Trypanosoma cruzi II: recognition of sugar structures by Arachis hypogaea (peanut agglutinin) lectin.

Epimastigote culture forms of different isolates of Trypanosoma cruzi from different mammal hosts, humans, and vectors were tested with FITC-conjugated peanut agglutinin lectin (PNA-FITC). The parasites maintained in axenic medium, liver infusion tryptose. were evaluated by flow cytometric analyses; whereas T. cruzi I (Tcl), which is associated with the sylvatic transmission cycle, was labeled in high percentages with PNA (88-99.2%), T. cruzi II (TcII) (parasites associated with domiciliar cycle) and T. cruzi, zymodeme 3 (Tc/Z3) (also associated with the sylvatic cycle) were labeled in low percentages (TcII, 0-26% and Tc/Z3, 0-12.6%). It was demonstrated that it is possible to differentiate the 2 main T. cruzi subpopulations, TcI and TcII, using Arachis hypogaea. These results also showed a higher variability in TcII in terms of PNA binding.

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Trypanosoma cruzi in the scent glands of Didelphis marsupialis: the kinetics of colonization.

This study examined the dynamics of colonization of Trypanosoma cruzi in the scent glands of the opossum Didelphis marsupialis following direct inoculation with 10(5) epimastigotes of isolate G-49 (an opossum-derived strain). One, three, and five days, 1 month, and 1 year after inoculation, scent glands were fixed for analysis using brightfield and electron microscopies. One day after inoculation the parasites, mainly as epimastigotes, were randomly distributed into the lumen. From the third day on, the parasites still in the form of epimastigotes tended to concentrate closer to the epithelium. The flagellates reached the definitive distribution pattern on the fifth day, when they formed huge clusters deep into the foveae. In samples collected 1 month and 1 year after inoculation, the ratio of epimastigotes:trypomastigotes was 1:1, with epimastigotes predominating near the epithelium and trypomastigotes far from it. Our observations suggest that T. cruzi grows continuously in the scent glands and does not depend on adhesion to promote metacyclogenesis. Metacyclogenesis far from the epithelium seems to be an important selective advantage to both host and parasite, since it assures the elimination of the infective forms of the parasite when the host expels the glands' contents, which occurs in frightening situations or at times of stress. The morphological characteristics of infected and noninfected scent glands using transmission and scanning electron microscopies were also described.

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Specific antibody levels and antigenic recognition of Wistar rats inoculated with distinct isolates of Trypanosoma evansi.

"Mal de Cadeiras", an enzootic disease caused by Trypanosoma evansi, is one of the most important trypanosomiases in the Brazilian Pantanal region. The disease affects mainly horses, which are widely used in extensive cattle production, an activity of greatest economical significance for the region. The parasite also infects sylvan (coatis and capybaras) and domestic (dogs) animals, respectively considered wild and domestic reservoirs of T. evansi. For a better understanding of the interaction of T. evansi with its rodent host, we evaluated the differences in the specific antibody level patterns and in the parasitic peptides recognition patterns of experimentally infected Wistar rats. The rats experimentally infected with T. evansi isolates obtained from coatis, dogs and horses were submitted to indirect immunofluorescence test (IgM e IgG) and Western blotting. The serological titers for IgM and IgG ranged between 1:40 and 1:160. The most recognized polypeptide profiles were in a range of 17 and 74 kDa. Our data suggest that the humoral immune response in Wistar rats is not sufficient for granting an effective control of T. evansi infections.

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Biological and biochemical characterization of isolates of Trypanosoma evansi from Pantanal of Matogrosso--Brazil.

Ten isolates of Trypanosoma evansi from the Pantanal region of Brazil, recently derived from coati (Nasua nasua, carnivora, Procyonidae), horses and dogs, were characterized on the basis of biological (experimental infections in Wistar rats) and biochemical (multilocus enzyme eletrophoresis) data. Biological data were analyzed by Nested analysis of variance and Kruskal-Wallis. Marked heterogeneity in virulence was observed in the isolates. Some of the isolates showed an undulating parasitaemia, typical for African trypanosomes. This biological heterogeneity did not correspond with the biochemical homogeneity observed in the T. evansi isolates. T. evansi has one of the widest distributions and greatest range of mammalian hosts and is widely recognized to have evolved from Trypanosoma brucei. Adaptability of T. evansi was not reflected in the variability of biochemical and molecular parameters studied to date. The variability in virulence was very significant, but not correlated with the host from which it was derived. These data suggested that, in the region studied, T. evansi is transmitted among both domestic and sylvatic animals in one single transmission cycle.

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Homogeneity of Trypanosoma evansi isolates from domestic and sylvatic mammals from the Pantanal of Mato Grosso.

'Mal de Cadeiras' is a disease which causes great mortality in horses in the Pantanal Matogrossense region, Brazil. The agent of this disease is Trypanosoma evansi, a kinetoplastid flagellate which belongs to the Trypanosomatidae family, classified into the Salivarian section. Transmission occurs mechanically by haematophagous Diptera, mainly by Stomoxys sp. and Tabanus sp. and vampire bats. Outbreaks of Mal de Cadeiras in horses result in economic losses, thus limiting their use in cattle raising. Ten isolates of T. evansi recently derived from coati (Nasua nasua, Carnivora, Procyonidae), horses and dogs were compared, using schizodeme analyses from DNA digested by the restriction enzyme Hin fl. The results showed similar electrophoretic profiles for all isolates from wherever the host came. Homogeneity of isolates from domestic and sylvatic animals suggested two hypotheses: (1) the parasites circulated in only one transmission cycle;, and (2) independent cycles were not established in sufficient time to modify the molecular profiles of the isolates.

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The complexity of the sylvatic cycle of Trypanosoma cruzi in Rio de Janeiro state (Brazil) revealed by the non-transcribed spacer of the mini-exon gene.

American trypanosamiasis occurs in nature as a sylvatic cycle, where Trypanosoma cruzi interacts with wild triatomines and mammalian reservoirs, such as marsupials, rodents, armadillos and other animals. Due to difficulties in trying to isolate T. cruzi stocks from the sylvatic cycle, very few studies have been performed in order to understand the parasite infection in natural environments. Traditionally T. cruzi has been considered to be composed of a highly heterogeneous population of parasites. In contrast, the mini-exon and the 24S alpha rRNA gene loci have shown that T. cruzi stocks can be clustered in 2 major phylogenetic groups: lineage 1 and lineage 2. In this report, 68 recently isolated T. cruzi samples from the sylvatic cycle belonging to different geographical areas in Rio de Janeiro, Brazil, have been typed based on a variable spot in the non-transcribed spacer of the mini-exon gene. Eight isolates were from triatomines, 26 stocks were from golden-lion tamarins, 31 from opossums, 2 from rodents and 1 from a three-toed sloth. Thirty (44%-30/68) isolates were typed as lineage 1, while 36 (53%-36/68) isolates were typed as lineage 2. Two opossums presented mixed infection. Therefore, 3% (2/68) of the isolates were typed as lineage 1 + lineage 2. Using these geographical regions as models of sylvatic environments, it was observed that 96% of the Didelphis marsupialis were infected by lineage 2 isolates, while all 26 golden-lion tamarins were infected by lineage 1. The results show preferential association of the 2 lineages of T. cruzi with different hosts, composing the complexity of the sylvatic cycle.

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Amidine derivatives are highly effective against Trypanosoma evansi trypomastigotes.

The in vitro effect of N,N'-diphenyl-4-R-benzamidine (where R = H, CN, Br, Cl, CH3, OCH3 and NO2) in three isolates of Trypanosoma evansi was studied. The compounds were solubilized in dimethysulphoxide (DMSO) and tested in a concentration range of 5 to 160 micrograms/ml. The parasites were isolated from a horse, a dog and a coati. They were maintained in immunosuppressed rats, since they could not be cultured in vitro, and further purified through a diethylaminoethanol (DEAE) column. The trypomastigotes obtained were mixed with different concentrations of the drugs and after incubation at 26 degrees C for 24 h, the remaining parasites were counted in a Neubauer chamber. The percentage of inhibition was evaluated compared with the control, without the drugs. Most of the amidine derivatives showed high activity against the three T. evansi isolates, but different patterns of sensitivity to the tested compounds were observed. At least four compounds with Br, Cl, OCH3 and NO2 as substituents, were much more effective than Berenil [4,4'-(diazoamine)-dibenzamidine aceturate], the reference drug used, which is included in the same chemical class of amidines.

Animals↗

Hashimoto's disease in a papillary carcinoma of the thyroid originating in a teratoma of the ovary (malignant struma ovarii).

Adenocarcinomas represent a relatively rare complication of a cystic teratoma of the ovary. Those of thyroid origin have been reported in only a few cases. In this paper we report a case of papillary carcinoma of the thyroid arising from a cystic teratoma. The patient had no thyroid symptoms, but because of the presence of antimicrosomal and antithyroglobulin antibodies the diagnosis of Hashimoto's disease was made.

Autoantibodies↗

Trypanosoma cruzi in the opossum Didelphis marsupialis: a study of the correlations and kinetics of the systemic and scent gland infections in naturally and experimentally infected animals.

The genus Didelphis (Marsupialia, Didelphidae) has the unique capacity of supporting both multiplication cycles of Trypanosoma cruzi simultaneously; besides the intracellular forms, the epimastigotes can be found multiplying and differentiating abundantly in the lumen of the scent glands. The biological significance of the life cycle of T. cruzi within the scent glands of Didelphis marsupialis, as well as its contribution to the epidemiology of the disease, is presently unclear. In order to clarify the mechanisms involved in the colonization of this singular habitat by T. cruzi, as well as to understand its biological role, we have carried out a serological and parasitological follow-up of both natural and experimental infections of young and adult opossums. Although all natural infections were stable and long lasting, no infected scent glands were found, indicating that the stability of the systemic infections does not depend on the presence of flagellates in the scent gland. In 84% of the experimentally infected animals the colonization of the scent glands was preceded by a period of patent parasitemia. Parasitism of the scent glands was essentially permanent and bilateral, and its maintenance was independent of circulating parasites. Moreover, the course of the infection differed depending on the source (scent glands versus axenic culture-derived) of the metacyclic forms. Our results suggest that parasitism of the SG of D. marsupialis is most likely a secondary acquisition, a step toward independence from the insect vector, similarly to what is accepted for Trypanosoma equiperdum.

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The potential effects of new synthetic drugs against Leishmania amazonensis and Trypanosoma cruzi.

Amidine derivatives, never used before on trypanosomatids, were tested against Leishmania amazonensis and Trypanosoma cruzi. These drugs in doses up to 500 mg/kg inoculated into Swiss mice did not show any toxic effect (Santos, 1993). The in vitro effect of N,N'-diphenyl-4-R- benzamidine was evaluated. L. amazonensis promastigotes, epimastigotes and blood forms of T. cruzi, were assayed with/without the drugs in axenic media, using pentamidine isethionate and benznidazole, as reference drugs. The results were very promising for L. amazonensis, showing that the most active compounds were the metoxy and Br-derivatives, with LD50 of 20 microM and 22 microM, respectively. In general the amidines showed lower activity against T. cruzi than L. amazonensis. The most active compounds against blood trypomastigotes were the same metoxy and Br-derivatives, but in much higher concentrations, e.g. as LD50 of 59 nM and 251 nM, respectively. All amidines had a very low activity against epimastigotes, and the only active compounds were the halogen-derivatives with LD50 = 424 nM for the Br-derivative and LD50 of 474 nM for the C1-derivative.

Animals↗