Xenodiagnostico con Lutzomyia youngi en casos venezolanos de leishmaniasis cutaned por Leishmania braziliensis

Xenodiagnósticos con Lutzomya yungi aplicados en los bordes de las úlceras de pacientes infectados con Leishmania braziliensis antes y después del tratamiento con 10 dosis de antimonial pentavalente y un aminoglicósido, evidencian la condición reservoria de leishmanias del enfermo, para flebótomos endofágicos y la utilidad de un tratamiento específico-temprano que no solamente conduce a la curación clínica, sino a la eliminación del riesgo de una eventual transmisión intradomiciliar por insectos que pican dentro del domicilio durante la noche.

Evaluation of antibody responses in american visceral leishmaniasis by ELISA and immunoblot

American visceral leishmaniasis (AVL) is an important disease among children of northeast Brazil. In order to characterize antibody responses during AVL, sera of hospitalized patients were analyzed by ELISA and Western blot using a Leishmania chagasi antigen preparation. The ELISA was positive (asorbance [greater than or equals to] 0.196) at a serum dilution of 1:1024 in all patients at presentation, and fell to ward control levels over the following year. Only one of 72 control subjects tested positive, and that donor had a sibling with AVL. Immunoblots of the patients' sera recognized multiple bands, the most frequent of which were at approximately 116 kDa, 70 kDa, and 26 kDa. Less frenquently observed were bands at approximately 93 kDa, 74 kDa, 62 kDa, 46 kDa and 32 kDa. The ELISA responses and patterns of banding were distinctive for AVL, and could be used to differentiate patients with AVL from those with Chagas' disease of cutaneous leishmaniasis. Sera from six AVL patients followed for up to six weeks after treatment identified no new bands. Sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) of surface iodinated parasite proteins showed one major band and four minor bands, whereas SDS-PAGE of biotinylated prarasite proteins revealed a banding pattern similar to those of patient sera. AVL appears to produce characteristic immunoblot patterns which can be used along with a sensitive screening ELISA to diagnose AVL.

The genus lobatostoma (Trematoda: Aspidocotylea) in the pacific coast of South America, with description of Lobatostoma veranoi new species, parasite of Menticirrhus ophycephalus (Teleostei: Sciaenidae)

The presence of three aspidocotyleans trematodes in marine fishes from Perú and Chile is reported. One of them, Lobatostoma veranoi from the intestine of Menticirrhus ophicephalu (Sciaenidae) is considered a new species. Distinct characteristcs of the new species are:a cirrus sac smaller than the pharynx; tail overlapping posteriorly the ventral disk; testis in the last third of the body and the presence of 64-66 marginal alveoli. The two other species are Lobatostoma pacificum Manter, 1940 found in Trachinotus paitensis Cuvier, 1830 from Perú and Chile and Lobatostoma Anisotremum Oliva & Carvajal, 1984 from the intestine of Anisotremus scapularis (Tschudi, 1844) from Perú.

Evaluación de la respuesta de isotipos de inmunoglobulina especifica a Leishmania en leishmaniasis tegumentaria Americana

Con el fin de determinar las clases de anticuerpo producido contra el parásito y la cinética de los mismos en relación a la evolución de la infección, se estudiaron los sueros de 133 pacientes infectados con Leishmania del complejo braziliensis. Se utilizó la prueba de inmunofluorescencia indirecta y amastigotas de L. mexicana amazonensis como antígeno. En los sueros obtenidos al momento de consultar para el diagnóstico se encontró IgM en 54 de los sueros absorbidos con Straphylococcus aureus Cowan I y en 5 de los no absorbidos. La IgM sólo se encontro en los sueros de pacientes con tiempo devolución de las lesiones < o = de 2 meses. la IgG se detectó en todos los sueros no absorbidos. Los sueros tomados durante recurrencia y después de cicatrización sólo presentaron IgG. El uso combinado de la prueba de Montenegro y/o título de IgM específico, aumentó el porcentage de pacientes con un diagnóstico inmunológico positivo en aquéllos cuyas lesiones tenían un tiempo de evolución menor de 2 meses. En los sueros de los 10 individuos sanos no se detectó inmunoglobulina específica a Leishmania y ninguno presentó reacción positiva a la prueba de Montenegro. Entre los 16 pacientes con otra etiología, 3 con esporotricosis, mostraron en su suero IgG reactiva con Leishmania pero ninguno incluyendo 2 con menos de dos meses de evolución de las lesiones presentó IgM. concluimos que en pacientes infectados con L. braziliensis la presencia de IgM e IgG específica a Leishmania esta asociado con el tiempo de evolución de las lesiones y el estado primario recurrente de la infección; demás la detección de IgM anti-Leishmania combinada con la respuesta de Mn sería de potencial utilidad en el diagnóstico clínico de la leishmaniasis tegumentaria temprana.

MARCKS-related protein (MRP) is a substrate for the Leishmania major surface protease leishmanolysin (gp63).

Myristoylated alanine-rich C kinase substrate (MARCKS) and MARCKS-related protein (MRP; MacMARCKS) are protein kinase C substrates in diverse cell types. Activation of murine macrophages by cytokines increases MRP expression, but infection with Leishmania promastigotes during activation results in MRP depletion. We therefore examined the effect of Leishmania major LV39 on recombinant MRP. Both live promastigotes and a soluble fraction of LV39 lysates degraded MRP to yield lower molecular weight fragments. Degradation was independent of MRP myristoylation and was inhibited by protein kinase C-dependent phosphorylation of MRP. MRP was similarly degraded by purified leishmanolysin (gp63), a Leishmania surface metalloprotease. Degradation was evident at low enzyme/substrate ratios, over a broad pH range, and was inhibited by 1,10-phenanthroline and by a hydroxamate dipeptide inhibitor of leishmanolysin. Using mass spectrometric analysis, cleavage was shown to occur within the effector domain of MRP between Ser(92) and Phe(93), in accordance with the substrate specificity of leishmanolysin. Moreover, an MRP construct in which the effector domain had been deleted was resistant to cleavage. Thus, Leishmania infection may result in leishmanolysin-dependent hydrolysis of MRP, a major protein kinase C substrate in macrophages.

Distinct ultrastructural aspects in different biopsies of a single patient with diffuse cutaneous leishmaniasis

The authors investigated the relation between parasites and host-cells in active and regressed lesions of a patient with diffuse cutaneous leishmaniasis, evaluating the frequency of different cell types, and the location and integrity of amastigotes. No correlation was found between parasite integrity and size of parasitophorous vacuoles. They observed ultrastructural findings characterizing a cell mediated immune response: macrophages lysis, parasitic destruction inside macrophages, close contact between parasitized macrophages and lymphocytes and between parasites and lymphocytes, lymphocytic infiltration and fibrosis. They suggest that in DCL there is a limited cellular immune response, although insufficient to control infection.