Márcia Hueb

Development and validation of PCR-based assays for diagnosis of American cutaneous leishmaniasis and identification of the parasite species

In this study, PCR assays targeting different Leishmania heat-shock protein 70 gene (hsp70) regions, producing fragments ranging in size from 230-390 bp were developed and evaluated to determine their potential as a tool for the specific molecular diagnosis of cutaneous leishmaniasis (CL). A total of 70 Leishmania strains were analysed, including seven reference strains (RS) and 63 previously typed strains. Analysis of the RS indicated a specific region of 234 bp in the hsp70 gene as a valid target that was highly sensitive for detection of Leishmania species DNA with capacity of distinguishing all analyzed species, after polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). This PCR assay was compared with other PCR targets used for the molecular diagnosis of leishmaniasis: hsp70 (1400-bp region), internal transcribed spacer (ITS)1 and glucose-6-phosphate dehydrogenase (G6pd). A good agreement among the methods was observed concerning the Leishmania species identification. Moreover, to evaluate the potential for molecular diagnosis, we compared the PCR targets hsp70-234 bp, ITS1, G6pd and mkDNA using a panel of 99 DNA samples from tissue fragments collected from patients with confirmed CL. Both PCR-hsp70-234 bp and PCR-ITS1 detected Leishmania DNA in more than 70% of the samples. However, using hsp70-234 bp PCR-RFLP, identification of all of the Leishmania species associated with CL in Brazil can be achieved employing a simpler and cheaper electrophoresis protocol.

Fatores associados ao insucesso do tratamento da leishmaniose cutânea com antimoniato de meglumina

We investigated factors associated with treatment failure in the treatment of cutaneous leishmaniasis with meglumine antimony in a reference service in Mato Grosso State. A retrospective cohort of 151 patients with cutaneous leishmaniasis was built using medical records. The incidence of therapeutic failure was 47% (IC95%=39.2%-55%). Antimoniate doses below 10mg/kg/day (RR=1.8; IC95:1.1-3.0), previous leishmaniasis treatment (RR=1.7; IC95:1.3-2.4), 3 or more skin lesions (RR=1.9; IC95:1.4-2.5), incomplete treatment (RR=1.9; IC95:1.3-2.6) and body weight above 68kg (RR=1.7; IC95:1.1-2.5) were associated with therapeutic failure. After adjustment, therapeutic failure was associated with having 3 or more cutaneous lesions (OR=4.6; IC95%=1.2-17.4), reports of previous leishmaniasis treatment (OR=4.5; IC95%=1.1-17.5), body weight above 68kg (OR=4.3; IC95=1.5-11.9) and incomplete treatment schedule (OR=12.5; IC95%=2.1-75.4), although body weight is possibly associated with treatment failure due to the limitation of the maximum daily dose. These results help to identify patients at risk of treatment failure of cutaneous leishmaniasis with antimony therapy.

Entomoftoromicose (zigomicose) causada por Conidiobolus coronatus em Mato Grosso (Brasil): relato de caso

Zygomycosis is a subcutaneous mycosis caused by soil fungi, such as Conidiobolus coronatus. In general, the main clinical manifestation is a chronic rhinofacial tumor. We report the first case of zygomycosis (entomophthoramycosis) caused by Conidiobolus coronatus, occurring in Mato Grosso, West Brazil.

Ocorrência de transmissão autóctone de leishmaniose visceral em Mato Grosso

O Estado de Mato Grosso, antes de sua divisao em 1979, apresentava transmissao de LV na regiao de Corumba, que se mantem ate hoje. Essa regiao, apos a separacao dos estados, ficou localizada no entao recem-criado Mato Grosso do Sul. O novo Estado de Mato Grosso, apesar de endemico para muitas doencas tropicais como malaria, paracoccidioidomicose, leishmaniose tegumentar, entre outras, nao tem registro da ocorrencia de casos autoctones de calazar9. No periodo de 1992 a 1994, quatro casos de LV foram diagnosticados e tratados no Hospital da Universidade Federal de Mato Grosso, em Cuiaba. Apos investigacao sobre procedencia e historia migracional dos pacientes, concluiu-se por tratar-se de transmissao autoctone da doenca no Estado. O primeiro caso foi de um paciente masculino de 49 anos, natural de Sao Paulo, e residente em Mato Grosso ha 12 anos, nao se tendo ausentado do Estado nesse periodo. Apresentava quadro de febre, emagrecimento, hepatoesplenomegalia, adenomegalia, lesoes cutâneas, pancitopenia e hipergamaglobulinemia. O diagnostico foi feito pelo achado do protozoario em medula ossea, linfonodos e pele ao exame direto. O paciente evoluiu bem com o tratamento convencional. O segundo paciente era uma mulher de 39 anos, natural e procedente de Santo Antonio

Expression of annexin A1 in Leishmania-infected skin and its correlation with histopathological features

INTRODUCTION The aim of this study was quantify annexin A1 expression in macrophages and cluster of differentiation 4 (CD4) + and cluster of differentiation 8 (CD8)+ T cells from the skin of patients with cutaneous leishmaniasis (n=55) and correlate with histopathological aspects. METHODS Infecting species were identified by polymerase chain reaction-restriction fragment length polymorphism, and expression of annexin A1 was analyzed by immunofluorescence. RESULTS All patients (n = 55) were infected with Leishmania braziliensis . Annexin A1 was expressed more abundantly in CD163 + macrophages in infected skin (p < 0.0001) than in uninfected skin. In addition, macrophages in necrotic exudative reaction lesions expressed annexin A1 at higher levels than those observed in granulomatous (p < 0.01) and cellular lesions p < 0.05). This difference might be due to the need to clear both parasites and necrotic tissue from necrotic lesions. CD4 + cells in cellular lesions expressed annexin A1 more abundantly than did those in necrotic (p < 0.05) and granulomatous lesions (p < 0.01). Expression in CD8 + T cells followed the same trend. These differences might be due to the pervasiveness of lymphohistiocytic and plasmacytic infiltrate in cellular lesions. CONCLUSIONS Annexin A1 is differentially expressed in CD163 + macrophages and T cells depending on the histopathological features of Leishmania -infected skin, which might affect cell activation.

Varicella zoster virus reactivation during or immediately following treatment of tegumentary leishmaniasis with antimony compounds

Antimony compounds are the cornerstone treatments for tegumentary leishmaniasis. The reactivation of herpes virus is a side effect described in few reports. We conducted an observational study to describe the incidence of herpes zoster reactivation during treatment with antimony compounds. The global incidence of herpes zoster is approximately 2.5 cases per 1,000 persons per month (or 30 cases per 1,000 persons per year). The estimated incidence of herpes zoster in patients undergoing antimony therapy is higher than previously reported.