Chagas disease is included in the neglected tropical diseases list and is endemic to 21 Latin American countries. The two drugs currently available for treating Chagas disease are nifurtimox and benznidazole and both result in many significant side effects. The study describes the synthesis and biological evaluation of 3,5-disubstituted isoxazoles. Isoxazoles were obtained by reaction of flavones and hydroxylamine and either alkylated at the free hydroxyl group and/or nitrated at the isoxazole ring. These compounds were evaluated for their in vitro anti-Trypanosoma cruzi activity against trypomastigote and amastigote forms of the parasite in T. cruzi-infected cell lineages. Benznidazole was used as a reference compound for the in vitro assay and mammalian L929 cells were employed to evaluate cytotoxicity. A majority of the compounds tested were very active and the most active isoxazole against amastigote and trypomastigotes of T. cruzi was slightly more potent than the current medicine benznidazole.

Annona cornifolia A. St. -Hil. é uma pequena árvore anual perene encontrada no cerrado brasileiro; seu fruto verde é usado popularmente no tratamento de úlceras. As acetogeninas isoladas a partir das sementes de Annona cornifolia anteriormente apresentaram atividade antioxidante. Continuando as nossas investigações sobre as atividades biológicas das acetogeninas, quatro misturas binárias e dez acetogeninas bis-tetra-hidrofurânicas adjacentes puras foram avaliadas quanto às atividades: citotóxica (sobre três linhagens de células tumorais humanas), antifúngica (sobre Paracoccidioides brasiliensis), tripanocida (sobre Trypanosoma cruzi) e leishmanicida (sobre Leishmania amazonensis). As acetogeninas apresentaram atividade citotóxica, confirmando seu uso potencial na terapia anticâncer. Quanto às atividades leishmanicida e tripanocida, foi observada uma inibição de 87% das formas amastigotas de L. amazonensis e 100% de formas amastigotas e tripomastigotas de T. cruzi, quando testadas a uma concentração de 20 µg mL–1. Além disso, seis acetogeninas mostraram maior atividade sobre todas as três cepas testadas de P. brasiliensis que o trimetoprim-sulfametoxazol, um medicamento usado no tratamento da paracoccidioidomicose. Assim, as acetogeninas podem ser uma alternativa no tratamento de uma série de doenças que têm um enorme impacto em milhões de pessoas no mundo. Este trabalho relata pela primeira as atividades antifúngica, leishmanicida e tripanocida para estas acetogeninas.

Annona cornifolia A. St. -Hil. is a small annual perennial tree found in the Brazilian savannah; their green fruit is popularly used in the treatment of ulcers. The acetogenins isolated from the seeds of Annona cornifolia previously showed to possess antioxidant activity. In continuation of our investigations on the biological activities of acetogenins, four binary mixtures and ten pure adjacent bis-tetrahydrofuran annonaceous acetogenins were evaluated: the cytotoxic (against three human tumor cell lines), antifungal (against Paracoccidioides brasiliensis), trypanocidal (against Trypanosoma cruzi) and leishmanicidal (against Leishmania amazonensis) activities. Acetogenins presented cytotoxic activity confirming their potential use in anti-cancer therapy. Regarding leishmanicidal and trypanocidal activities, an inhibition of 87% of L. amazonensis amastigotes and 100% of T. cruzi amastigotes and trypomastigotes was observed, when tested at the concentration of 20 µg mL–1. Moreover, six acetogenins showed more activity against all the three tested isolates of P. brasiliensis than trimethoprim-sulfamethoxazole, a drug used for treating paracoccidioidomycosis. Thus, acetogenins may be an alternative in treating a number of diseases that have a huge impact on millions of people worldwide. This paper reports for the first time the antifungal, leishmanicidal and trypanocidal activities for these acetogenins.

The phytochemical investigation on the aereal parts of Lychnophora pinaster Mart., Asteraceae, was carried to isolation of triterpenes. 3-O-Acetyl-lupeol (1), 3-O-acetyl-pseudotaraxasterol (2), and 3-O-acetyl-α-amyrin (3) were isolated from hexanic extract and 4,4-dimethyl-cholesta-22,24-dien-5-ol (4), α-amyrin (5), and lupeol (6) were isolated from hexanic/dichlorometanic extract of the leaves. Compounds Δ7-bauerenyl acetate (7), friedelin (8), stigmasterol (9), and sitosterol (10) were isolated from the hexanic/dichlorometanic extract of the stems. The steroids 9 and 10 were also isolated from the hexanic/dichlorometanic extract of the flowers. Triterpenes 1, 3, 4, and 7 are described for the first time in the genus Lychnophora. The apolar fractions of the leaf and stem extracts and some isolated triterpenes showed low trypanocidal activity. Moreover, apolar fractions of the leaf and stem extracts and 5 showed antibacterial action against Staphylococcus aureus.

One hundred and twenty-one isolates of endophytic fungi were recovered from leaves of the bioactive Brazilian plant species Ageratum myriadenia, Palicourea tetraphylla, Piptadenia adiantoides, and Trixis vauthieri. All fungal isolates were cultivated in liquid media and crude extracts were obtained with ethyl acetate. The crude extracts were tested in bioassay panels using Leishmania amazonensis, Trypanosoma cruzi, the enzyme trypanothione reductase (TryR) from Trypanosoma cruzi, and three human cancer cell lines. Thirty-three extracts (27.2%) exhibited at least one biological activity. Seventeen extracts (14%) were cytotoxic against one or more human cancer cell line with the IC50 values ranged of >0.2 to 25 µg/mL. Twenty-four extracts (19.8%) inhibited the activity of TryR, and three showed ability to inhibit the growth of T. cruzi above 60% and their IC50 values ranged among 1 to 10 µg/mL. Eleven extracts (9%) were able to inhibit the growth of L. amazonensis and showed with IC50 values ranged among 4.6 to 24.4 µg/mL. The endophytic fungi were identified as belonging to the genera Alternaria, Arthrinium, Cochliobolus, Colletotrichum, Penicillium, Fusarium, and Gibberella. An interesting result was obtained for the bioactive isolates UFMGCB 508, 537, 899 and 903, which were related to fungi associated with medicinal plants native to Asia, Australia, Africa, and Polynesia. These results indicate that bioactive plants living in Brazilian ecosystems are a potential host of endophytic fungi able to produce bioactive prototype molecules for drug development against neglected tropical diseases.

Trypanosoma cruzi is a protozoan parasite that causes a severe disease (Chagas'disease) in Central and South America. The currently available chemotherapeutic agents against this disease are still inadequate. The enzyme trypanothione reductase (TR) is considered a validated molecular target for the development of new drugs against this parasite. In this regard, a series of arylfurans based on 2,5-bis-(4-acetamidophenyl)furan was synthesized and tested for their in vitro inhibitory activity against TR. Molecular modeling studies of putative enzyme-inhibitor complexes revealed a possible mechanism of interaction. From synthesized compounds, a benzylaminofuran derivative was found to be more active than the lead compound.

The natural lignans veraguensin and grandisin have been reported to be active against Trypanosoma cruzi bloodstream forms. Aiming at the total synthesis of these and related compounds, we prepared three 2-arylfurans and eight 2,5-diarylfurans. They were evaluated for their potential as T. cruzi trypanothione reductase (TR) inhibitors as well against the parasite's intracellular (amastigote) and bloodstream (trypomastigote) forms. Compound 12 was the most effective against TR with an IC50 of 48.5 µM while 7 and 14 were active against amastigotes, inhibiting the parasite development by 60% at 20 µg/ml (59 and 90 µM, respectively). On the other hand, none of the compounds was significantly active against the parasite bloodstream forms even at 250 µg/ml (0.6-1.5 mM).

Fourteen compounds were evaluated for their activity against Trypanosoma cruzi blood stream forms at the concentration of 500 µg/ml. Six compounds were active and re-tested at lower concentrations.

Foram investigados aspectos clínicos, laboratoriais, terapêuticos e evolutivos da leishmaniose tegumentar americana em Belo Horizonte. O estudo incluiu 358 pacientes com leishmaniose cutânea (LC) e 25 com leishmaniose mucosa (LM). Comparados aos pacientes com LC, aqueles com LM apresentaram maior tempo de doença e relato de outras doenças concomitantes, sugerindo que a debilitação pela leishmaniose e/ou outras doenças podem contribuir para a ativação e/ou disseminação mucosa do parasito. As sensibilidades das reações intradérmica, de imunofluorescência indireta e da pesquisa direta do parasito foram de 78,4, 79,3 e 68,3%, respectivamente. O tratamento com antimoniato de meglumina foi 100% eficaz, com 59% de efeitos colaterais ao longo do tratamento. A recidiva após tratamento ocorreu em 32 (10,1%) dos 318 casos seguidos por até dois anos. A maioria das recidivas (31 dos 32 casos) ocorreu em pacientes com LC tratados com 15mg Sb5+/kg/dia. Na investigação de critérios de cura, a reação intradérmica negativa foi o único fator associado a um risco três vezes maior de recidiva. Um aumento da dose ou do tempo de tratamento talvez melhore o prognóstico nestes pacientes.

This study investigated clinical, laboratorial, therapeutic and prognostic aspects of American cutaneous leishmaniasis in Belo Horizonte in 358 patients with cutaneous leishmaniasis (CL) and 25 with mucocutaneous leishmaniasis (MCL). Compared to CL patients, the MCL patients reported longer duration of disease and higher frequency of other diseases, suggesting that debilitation caused by leishmaniasis or other conditions might contribute to activation and/or mucous dissemination of the parasite. The sensitivity of skin test, indirect immunofluorescence reactions and direct detection of parasites was 78.4, 79.3 and 68.3%, respectively. The treatment with meglumine antimoniate presented 100% efficacy, but 59% patients had side-effects. During two years of follow-up, there were 32/318 relapses after successful treatment. Most relapses (31/32) were of CL patients treated with 15 mg Sb5+/kg/day. The negative response to skin test was the only factor associated with a significant threefold increased risk of relapse. Higher dose or longer duration of treatment might improve the prognosis in these patients.

Due to the overlapping distribution of Trypanosoma rangeli and T. cruzi in Central and South America, sharing several reservoirs and triatomine vectors, we herein describe a simple method to collect triatomine feces and hemolymph in filter paper for further detection and specific characterization of these two trypanosomes. Experimentally infected triatomines feces and hemolymph were collected in filter paper and specific detection of T. rangeli or T. cruzi DNA by polymerase chain reaction was achieved. This simple DNA collection method allows sample collection in the field and further specific trypanosome detection and characterization in the laboratory.

While relapses following clinical cure of American cutaneous leishmaniasis are frequent, no test has been described until now to predict such relapses. A cohort of 318 American cutaneous leishmaniasis patients was followed up for two years after treatment with meglumine antimoniate, during which time 32 relapses occurred, 30 in the first year and two in the second (accumulated risk: 10.5%). No association was found between these relapses and the parasite-specific antibody response before and after treatment, or between the relapses and stratification by sociodemographic and clinical characteristics. However when Leishmania was used as antigen, patients with a negative skin test at the time of diagnosis presented a 3.4-fold higher risk (hazard risk = 3.4; 95% confidence interval, 1.7-7.0) of American cutaneous leishmaniasis relapse, compared with patients with a positive response. This result shows that the skin test can be a predictor of American cutaneous leishmaniasis relapse after treatment.

La leishmaniasis cutánea americana, causada por Leishmania (Viannia) brazifensis, da lugar a recaídas en aproximadamente un 10% de los pacientes tratados en el Brasil. A falta de un criterio fiable, las curaciones postratamiento de la leishmaniasis cutánea americana se han definido principalmente atendiendo a los datos clínicos. En este artículo se presentan los resultados de un estudio de seguimiento de dos años de las recaídas sufridas por una cohorte de 318 pacientes aquejados de leishmaniasis cutánea americana después de ser tratados con antimoniato de meglumina. Se obtuvo el consentimiento informado por escrito y los pacientes fueron entrevistados y examinados por el mismo médico, con arreglo a un protocolo predefinido con información sobre factores sociodemográficos (sexo, edad, color de la piel, lugar de nacimiento, educación y nivel de ingresos, ocupación y lugar de residencia) y factores clínicos (cuadro clínico y momento de aparición de la leishmaniasis cutánea americana, número de lesiones y lugar de las mismas, historia familiar de leishmaniasis cutánea americana, concurrencia de otras afecciones y tratamiento anterior). Se evaluó la respuesta de hipersensibilidad retardada mediante una prueba cutánea en la que se empleó antígeno preparado a partir de suspensiones de promastigotes muertos. Se realizaron pruebas serológicas antes del tratamiento y 10 días después del mismo.Todos los pacientes fueron tratados con antimoniato de meglumina intramuscular (Glucantimet): dosis de 15 mg de Sb5+ * kg-1 * día-1 en los pacientes con leishmaniasis cutánea, y de 20 mg de Sb5+ * kg-1 * día-1 en los pacientes con leishmaniasis mucocutánea, durante al menos 20 y 30 días, respectivamente (7). Hubo remisión completa de las lesiones en todos los pacientes. Se realizaron evaluaciones clínicas antes e inmediatamente después del tratamiento, y cada seis meses durante dos años después del tratamiento. Se definió como recaída postratamiento toda: (a) aparición de una lesión nueva dentro de los límites de una cicatriz anterior, (b) aparición de una nueva lesión mucosa en pacientes con una cicatriz cutánea, o (c) reactivación de una cicatriz en el lugar de una lesión mucosa previa. Se trazaron las curvas de Kaplan-Meier, que muestran la probabilidad de que no se produzca ninguna recaída durante el seguimiento, para el conjunto de la cohorte y para cada factor específico investigado, y se utilizó el modelo de riesgo instantáneo proporcional de Cox para determinar el efecto independiente de los diferentes factores de recidiva.La cohorte estaba constituida por 207 hombres y 111 mujeres, con edades de entre 6 meses y 80 años (media = 35,4 + 18,9 años); 300 (94,3%) pacientes sufrían leishmaniasis cutánea, y 18 (5,7%) leishmaniasis mucocutánea. El nivel de educación y de ingresos de la mayoría de los pacientes era bajo. Sólo 71 (22,3%) realizaban tareas agrícolas o silvícolas. Se registraron 32 recaídas, 26 dentro de los 6 primeros meses de tratamiento, 4 al cabo de entre 7 y 12 meses, y 2 durante el segundo año tras el tratamiento. Veintinueve de las 32 recaídas se manifestaron en el mismo lugar afectado por la lesión cutánea previa en los pacientes con leishmaniasis cutánea. Otros dos pacientes con leishmaniasis cutánea presentaron tanto una recaída cutánea como nuevas lesiones mucosas. Sólo en un paciente con leishmaniasis mucocutánea se observó una reactivación de una lesión mucosa cicatrizada. La mayoría de las recaídas (30/32) ocurrieron en el término de un año, lo que corrobora la recomendación de la OMS (1990) de que se debe seguir a los pacientes durante al menos un año después del tratamiento. El riesgo acumulado de recaída fue del 10,5%. No se observó ninguna relación entre la recaída y la respuesta de producción de anticuerpos específicos contra el parásito antes y después del tratamiento, o entre la recaída y la estratificación por características sociodemográficas y clínicas.La prueba cutánea provoca una reacción de hipersensibilidad retardada y es un buen marcador de la respuesta inmunitaria celular en la leishmaniasis. Los pacientes con una prueba cutánea negativa en el momento del diagnóstico presentaron un riesgo 3,4 veces mayor de recaída que los que habían dado positivo. Se prevé que el uso de la prueba cutánea como factor predictivo de recidiva de leishmaniasis cutánea americana tendrá gran repercusión en la salud pública, ya que la prueba es segura y económica, amén de ampliamente usada para el diagnóstico de la leishmaniasis cutánea americana. La prueba parece indicar también que a los pacientes en los que no se observa ninguna reacción de hipersensibilidad retardada en la prueba cutánea en el momento del diagnóstico se les debe tratar con una dosis mayor o durante más tiempo para prevenir las recaídas. Es necesario realizar ahora nuevas investigaciones en las zonas endémicas en que coexisten otras especies de Leishmania a fin de evaluar la coherencia de estos resultados.

Au Brésil, la leishmaniose cutanée américaine, due à Leishmania (Viannia) braziliensis, donne des rechutes chez environ 10% des malades traités. En l’absence de critère fiable, la guérison après traitement de la leishmaniose cutanée américaine est principalement définie d’après les données cliniques. Cet article présente les résultats d’une étude de suivi de deux ans portant sur les rechutes chez 318 malades traités par l’antimoniate de méglumine. Le consentement éclairé a été recueilli par écrit et les malades ont été interrogés et examinés par le même médecin, suivant un protocole préétabli comportant la collecte d’informations sur les facteurs socio-démographiques (sexe, âge, couleur de la peau, lieu de naissance, niveau d’études et de revenus, profession, lieu de résidence) et cliniques (tableau clinique et durée de la leishmaniose cutanée américaine, nombre et localisation des lésions, antécédents familiaux de leishmaniose cutanée américaine, présence d’autres affections et traitements antérieurs). Une réaction d’hypersensibilité retardée a été recherchée à l’aide d’un test cutané utilisant un antigène préparé à partir de suspensions de promastigotes morts. Des tests sérologiques ont été effectués avant le traitement et 10 jours après celui-ci.Tous les malades ont été traités par l’antimoniate de méglumine par voie intramusculaire (Glucantime à la dose de 15 mg Sb5+ · kg-1 · jour-1 pour la leishmaniose cutanée et 20 mg Sb5+ · kg-1 · jour-1 pour la leishmaniose cutanéo-muqueuse pendant au moins 20 et 30 jours respectivement. La rémission des lésions a été complète chez tous les patients. Un examen clinique a été réalisé avant le traitement, immédiatement après puis tous les 6 mois pendant 2 ans. Une rechute après traitement se définissait comme suit : a) survenue d’une nouvelle lésion à l’intérieur d’une ancienne cicatrice, b) survenue d’une nouvelle lésion muqueuse chez des patients porteurs d’une cicatrice cutanée, ou c) réactivation d’une cicatrice à l’emplacement d’une ancienne lésion muqueuse. Des courbes de Kaplan-Meier montrant la probabilité de l’absence de rechute pendant la période de suivi ont été établies pour l’ensemble de la cohorte et pour chacun des facteurs étudiés. Le modèle de régression de Cox à risques proportionnels a été utilisé pour déterminer l’effet indépendant des différents facteurs sur la survenue d’une rechute.La cohorte se composait de 207 malades de sexe masculin et 111 de sexe féminin, âgés de 6 mois à 80 ans (moyenne 35,4 + 18,9 ans), dont 300 (94,3 %) étaient atteints de leishmaniose cutanée et 18 (5,7%) de leishmaniose cutanéo-muqueuse. Le niveau d’études et de revenus de la plupart des malades était faible. Seuls 71 d’entre eux (22,3 %) travaillaient dans l’agriculture ou la sylviculture. On a observé 32 rechutes, dont 26 pendant les 6 mois suivant le traitement, 4 entre 7 et 12 mois après le traitement et 2 au cours de la deuxième année de suivi. Vingt-neuf rechutes ont eu lieu à l’emplacement d’une lésion cutanée antérieure chez les malades atteints de la forme cutanée. Deux autresmalades atteints de cette forme ont présenté à la fois une rechute cutanée et de nouvelles lésions muqueuses. Seul un malade atteint de leishmaniose cutanéo-muqueuse a présenté une réactivation d’une ancienne lésion muqueuse cicatrisée. La plupart des rechutes (30/32) sont survenues dans l’année qui a suivi le traitement, ce qui confirme la recommandation de l’OMS (1990) selon laquelle les patients doivent être suivis pendant au moins un an après le traitement. Le risque cumulatif de rechute était de 10,5 %. Il n’y avait pas d’association entre les rechutes et la réponse en anticorps spécifiques du parasite avant et après traitement, ni entre les rechutes et la stratification selon les caractéristiques socio-démographiques et cliniques des malades.Le test cutané est basé sur une réaction d’hypersensibilité retardée et constitue un bon marqueur de la réponse immunitaire cellulaire dans la leishmaniose. Chez les malades dont le test cutané était négatif au moment du diagnostic, le risque de rechute de leishmaniose cutanée américaine était 3,4 fois plus éleve que chez ceux dont le test était positif. L’utilisation du test cutané comme indicateur prédictif des rechutes de leishmaniose cutanée américaine devrait avoir un impact important en santé publique car ce test est sans danger, peu coûteux et largement utilisé pour le diagnostic de la maladie. Il peut également signaler la nécessité, pour empêcher les rechutes, de traiter par des doses plus élevées ou pendant plus longtemps les malades n’ayant pas présenté de réaction d’hypersensibilité retardée au moment du diagnostic. De nouvelles investigations dans des zones d’endémie où coexistent d’autres espèces de Leishmania sont maintenant nécessaires pour confirmer nos observations.

Para o estudo de variabilidade genética em Triatoma brasiliensis, o principal vetor da doença de Chagas no Nordeste brasileiro, espécimes de três diferentes populações intradomiciliares foram analisados. Regiões do DNA genômico foram amplificadas utilizando dois iniciadores randômicos através da técnica de RAPD (Random Amplified Polymorphic DNA), visualizados em géis de poliacrilamida corados pela prata. Os perfis originados se mostraram bastante homogêneos quando comparados intrapopulacionalmente. Populações capturadas em duas regiões diferentes do Estado do Ceará também apresentaram homogeneidade entre si, mas, quando comparadas com a população proveniente do Piauí, foi possível diferenciá-las. Esses resultados, preliminares, indicam que o RAPD pode ser usado com sucesso nos estudos de variabilidade em triatomíneos, bem como sugerem a existência de variabilidade entre diferentes populações de T. brasiliensis pertencentes a uma mesma subespécie.

We evaluated the genetic variability of Triatoma brasiliensis, the main vector of Chagas disease in Northeast Brazil, using specimens from three populations. Regions of genomic DNA were amplified by RAPD (Random Amplified Polimorphic DNA), using two primers. The products were visualized after polyacrylamide gel electrophoresis followed by silver staining. A dendrogram constructed through the Dice similarity coefficient allowed for separation of the tested specimens into three distinct groups. The populations captured in areas from Ceará State showed similar profiles, but different from that captured in Piauí State. Our results indicate that RAPD can be used successfully in triatomine studies and suggest the presence of genetic variability between different populations of T. brasiliensis.

Para o estudo de variabilidade genética em Triatoma brasiliensis, o principal vetor da doença de Chagas no Nordeste brasileiro, espécimes de três diferentes populações intradomiciliares foram analisados. Regiões do DNA genômico foram amplificadas utilizando dois iniciadores randômicos através da técnica de RAPD (Random Amplified Polymorphic DNA), visualizados em géis de poliacrilamida corados pela prata. Os perfis originados se mostraram bastante homogêneos quando comparados intrapopulacionalmente. Populações capturadas em duas regiões diferentes do Estado do Ceará também apresentaram homogeneidade entre si, mas, quando comparadas com a população proveniente do Piauí, foi possível diferenciá-las. Esses resultados, preliminares, indicam que o RAPD pode ser usado com sucesso nos estudos de variabilidade em triatomíneos, bem como sugerem a existência de variabilidade entre diferentes populações de T. brasiliensis pertencentes a uma mesma subespécie.

We evaluated the genetic variability of Triatoma brasiliensis, the main vector of Chagas disease in Northeast Brazil, using specimens from three populations. Regions of genomic DNA were amplified by RAPD (Random Amplified Polimorphic DNA), using two primers. The products were visualized after polyacrylamide gel electrophoresis followed by silver staining. A dendrogram constructed through the Dice similarity coefficient allowed for separation of the tested specimens into three distinct groups. The populations captured in areas from Ceará State showed similar profiles, but different from that captured in Piauí State. Our results indicate that RAPD can be used successfully in triatomine studies and suggest the presence of genetic variability between different populations of T. brasiliensis.

Trypanosoma rangeli is a hemoflagelate parasite that infects domestic and sylvatic animals, as well as man, in Central and South America. T. rangeli has an overlapping distribution with T. cruzi, the etiological agent of Chagas disease, sharing several animal reservoirs and triatomine vectors. We have isolated T. rangeli strains in the State of Santa Catarina, in southern Brazil, which dramatically increased the distribution area of this parasite. This brief review summarizes several studies comparing T. rangeli strains isolated in Santa Catarina with others isolated in Colombia, Honduras and Venezuela. The different methods used include indirect immunofluorescence and western blot assays, lectin agglutination, isoenzyme electrophoresis and random amplified polymorphic DNA analysis, triatomine susceptibility, in vitro cell infection assays, and mini-exon gene analysis.

Four Trypanosoma cruzi strains from zymodermes A, B, C and D were successively clonedon BHI-LIT-agar-blood BLAB). Twenty clones from the first generation (F1), 10 from The second (F2) and 4 from the third (F3) from the strains A138, B147 and C23 were isolated. The D150 strain provied 29 F1 and F2 clones. The strains and clones had their isoenzyme and K-DNA patterns determined. The clones from A138, Bl47 and C231 strains presented isoemzyme and K-DNA patterns identical between thewmselves and their respective parental strains. Therefore showing the homogenety and stability of isoenzyme and K-DNA patterns after successive cloning. The Dl50 strain from zymodeme D (ZD) showed heterogeneity. Twenty-eight out of 29 clones of the first generation were of zymodeme A and only one was of zymodeme C, confirming previous reports that ZD strains consisted of ZA and ZC parasite populations. The only D150 strain clone of zymodeme C showed a K-DNA pattern identical to its parental strain. The remining clones although similar among themselves were different from the parental strain. Thus the T. cruzi strains had either homonogeneus or heterogeneous populations. The clones produced by successive cloning provided genetically homonogeous populations. Their experimental use will make future results more reliable and reproducible.

The author investigated the distribution of lectin receptors on Trypanosoma cruzi blood forms collected from mice inoculated with, respectively, the drug-resistant and drug-sensitive strains VL-10 and CL, and treated with the two standard active nitroheterocyclic compounds nifurtimox and benznidazole used for treatment of human Chagas' disease. Blood trypomastigotes purified in Fycoll-Hypaque were incubated with fluorescein-labelled lectins Con A, WGA, EE, WFA, TPA and PNA and then microscopically examined. Neither qualitative or quantitative differences in the fluorescence intensity could be detected between parasites from VL-10 and CL strains submitted or not to treatment. The results suggest that both strains do not differ in their surface membrane carbohydrate moieties. Moreover, the rapid clearance of blood forms the drug-sensitive strain in animals treated with singlo doses of both compounds is not likely to depend on membrane alterations expressed by changes in the carbohydrate components. furthermore, resistance or sensitivity to drugs is not apparently related to carbohydrate distribution on T. cruzi blood forms.