ABSTRACT Leishmania infantum is a protozoan that causes visceral leishmaniasis (VL) in the Americas and some regions of Europe. The disease is mainly characterized by hepatosplenomegaly and fever, and can be fatal. Factors related to the host and parasite can contribute to the transmission of Leishmania and the clinical outcome. The intraspecific genetic variability of L. infantum strains may be one of these factors. In this study, we evaluated the genetic variability of L. infantum obtained from bone marrow smear slides from patients in the Sao Paulo State, Brazil. For this, the minicircle of the kDNA hypervariable region was used as target by Sanger sequencing. By analyzing the similarity of the nucleotides and the maximum likelihood tree (Fasttree), we observed a high similarity (98%) among samples. Moreover, we identified four different profiles of L. infantum. In conclusion, L. infantum strains from Sao Paulo State, Brazil, showed low diversity measured by minicircle of the kDNA hypervariable region.

RESUMO Introdução: Em função da urgência e demanda de uma resposta à pandemia do novo coronavírus (Covid-19), vários testes de detecção de anticorpos para a síndrome respiratória aguda grave do coronavírus 2 (Sars-CoV-2) têm sido desenvolvidos. Objetivo: Este estudo teve como objetivo avaliar o desempenho do teste rápido utilizado em um inquérito epidemiológico para Sars-CoV-2 em comparação com outros ensaios sorológicos. Métodos: Foram avaliadas 86 amostras de soro em três ensaios sorológicos: um imunoensaio de fluxo lateral - Wondfo Sars-CoV-2 Antibody Test (TRW) - e dois imunoensaios de quimioluminescência: Elecsys anti-Sars-CoV-2 (ECLIA) e Sars-CoV-2 IgG (CMIA-IgG). Resultados: As sensibilidades diagnósticas estimadas dos testes sorológicos na avaliação dessas amostras foram: TRW 59% [95% intervalo de confiança (IC) 43,4%-72,9%], ECLIA 66,7% (51%-79,4%) e CMIA-IgG 61,5% (47,1%-73%). Enquanto isso, a especificidade diagnóstica estimada para TRW foi 78,7% (95% CI 65,1%-88%), ECLIA 72,3% (58,2%-83,1%) e CMIA-IgG 76,6% (74%-95,5%). Os valores de sensibilidade e especificidade foram inferiores aos afirmados pelos fabricantes. Embora 16,2% (14/86) dos resultados tenham sido discordantes entre os três ensaios serológicos para Sars-CoV-2, o grau de concordância pelo índice Kappa foi adequado: TRW/CMIA-IgG [0,757 (95% IC 0,615-0,899)], TRW/ECLIA [0,715 (0,565-0,864)] e ECLIA/CMIA-IgG [0,858 (0,748-0,968)]. Conclusão: O teste sorológico pode ser uma ferramenta diagnóstica útil, o que reforça sua avaliação criteriosa, bem como o momento correto de sua utilização.

ABSTRACT Introduction: Due to urgency and demand of a response to the Covid-19 pandemic, numerous Sars-CoV-2 immunoassays have been rapidly developed. Objective: This study aimed at assessing the performance of rapid Sars-CoV-2 antibody test in comparison to high-throughput serological assays. Methods: A total of 86 serum samples were evaluated in the three assays: a lateral flow immunoassay - Wondfo Sars-CoV-2 Antibody Test (WRT) - and two chemiluminescence immunoassays: Elecsys Anti-Sars-CoV-2 (ECLIA), and Sars-CoV-2 IgG (CMIA-IgG). Results: The estimated diagnostic sensitivities of serological tests in the evaluation of serum samples from the epidemiological survey were: WRT 59% [95% confidence interval (CI) 43.4%-72.9%], ECLIA 66.7% (51%-79.4%), and CMIA-IgG 61.5% (47.1%-73%). Meanwhile, the estimated diagnostic specificity was for WRT 78.7% (95% CI 65.1%-88%), ECLIA 72.3% (58.2%-83.1%), and CMIA-IgG 76.6% (74%-95.5%). The sensitivity and specificity values were lower than manufacturers’ claimed. Although 16.2% (14/86) of serological results were discordant among the three Sars-CoV-2 serological assays, the degree of agreement by the kappa index was adequate: WRT/CMIA-IgG [0.757 (95% CI 0.615-0.899)], WRT/ECLIA [0.715 (0.565-0.864)], and ECLIA/CMIA-IgG [0.858 (0.748-0.968)]. Conclusion: The serological testing may be a useful diagnostic tool, which reinforces its careful evaluation, and, as well as the correct time to use it.

RESUMEN Introducción: Debido a la urgencia y la demanda de una respuesta a la pandemia de Covid-19, se han desarrollado rápidamente numerosos inmunoensayos del Sars-CoV-2. Objetivo: Este estudio tuvo como objetivo evaluar el rendimiento de la prueba rápida de anticuerpos contra el Sars-CoV-2 en comparación con los ensayos serológicos de alto rendimiento. Métodos: Se evaluaron un total de 86 muestras de suero en los tres ensayos: un inmunoensayo de flujo lateral - Wondfo Sars-CoV-2 Antibody Test (TRW) - y dos inmunoensayos de quimioluminiscencia: Elecsys Anti-Sars-CoV-2 (ECLIA) y Sars-CoV-2 IgG (CMIA-IgG). Resultados: Las sensibilidades diagnósticas estimadas de las pruebas serológicas en la evaluación de muestras de suero de la encuesta epidemiológica fueron: WRT 59% [intervalo de confianza (IC) del 95%: 43,4%-72,9%], ECLIA 66,7% (51%-79,4%) y CMIA-IgG 61,5% (47,1%-73%). Mientras tanto, la especificidad diagnóstica estimada fue para WRT 78,7% (95% CI 65,1%-88%), ECLIA 72,3% (58,2%-83,1%) y CMIA-IgG 76,6% (74%-95,5%). Los valores de sensibilidad y especificidad fueron más bajos que los declarados por los fabricantes. Aunque el 16,2% (14/86) de los resultados fueron discordantes entre los tres ensayos serológicos del Sars-CoV-2, el grado de concordancia del índice kappa fue adecuado: WRT/CMIA-IgG [0,757 (IC del 95%: 0,615-0,899)], WRT/ECLIA [0,715 (0,565-0,864)] y ECLIA/CMIA-IgG [0,858 (0,748-0,968)]. Conclusión: La prueba serológica puede ser una herramienta diagnóstica útil, lo que refuerza su evaluación cuidadosa, así como el momento adecuado para usarla.

ABSTRACT Leishmaniasis is one of the most neglected tropical diseases worldwide caused by the parasite Leishmania. In Paraguay the species responsible for cutaneous leishmaniasis (CL) is L. (Viannia) braziliensis. Here we report a case diagnosed with Leishmaniasis, and molecular analyses using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) demonstrate that the case was caused by L. (V.) lainsoni. This is the first record of this species for Paraguay, with which we extend the distribution range of the parasite 1,430 km southwards from the southernmost previous known locality. More studies are needed to know the actual incidence of this species in cases of CL in Paraguay, and to identify natural reservoirs in the wild.

RESUMEN La leishmaniasis es una de las enfermedades tropicales más desatendidas, causada por el parásito Leishmania. En Paraguay, la especie responsable de la leishmaniasis cutánea es (LC) es L. (Viannia) braziliensis. Aquí se reporta un caso diagnosticado de Leishmaniasis, y análisis moleculares empleando reacción en cadena de la polimerasa - polimorfismos de restricción de fragmentos de restricción (PCR-RFLP) demostraron que el caso fue causado por L. (V.) lainsoni. Este es el primer registro de esta especie para Paraguay, con lo cual se extiende el rango de distribución conocido de la especie, unos 1.430 km al sur de localidades previamente conocidas. Se necesitan más estudios para conocer la incidencia real de esta especie en casos de LC en Paraguay, y para identificar reservorios naturales del parásito en la naturaleza.

Abstract Background: Three drugs - pentavalent antimonials, amphotericin B and pentamidine - are currently used for leishmaniasis treatment. They are administered for long periods, only parenterally, and have high cardiac, renal and hepatic toxicities. Therefore, the investigation of new compounds is required. Nitro-heterocyclic derivatives have been used as possible drug candidates to treat diseases caused by trypanosomatids. Methods: Leishmania (L.) amazonensis promastigotes (MHO/BR/73/M2269), maintained in the Laboratório de Soroepidemiologia - Instituto de Medicina Tropical- USP, were exposed to five nitroheterocyclic derivatives, with differences at phenyl-ring position 4: BSF-C4H9, BSF-H, BSF-NO2, BSF-CH3 and BSF-Cl, for 48 hours. After analyzing viability (MTT assay), we evaluated cellular-morphology activity of compounds by transmission electron microscopy (TEM) and measurement of apoptosis (phosphatidylserine expression) by flow cytometry. Results: EC50 of amphotericin B and BSF-CH3 were 0.50 (M and 0.39 (M respective. Other nitro-heterocyclic compounds presented EC50 higher than amphotericin B. All compounds showed greater AV- and PI-positive expression than amphotericin B at 100 (M, except BSF-NO2. TEM showed complete nuclear disfigurement with 100 (M of BSF-NO2, 25 and 6.25 (M of BSF-H, and 6.25 (M BSF-Cl; presence of vesicles within the flagellar pocket with 25 (M BSF-H; alteration of the kinetoplast with 25 (M BSF-C4H9, 25 (M of BSF-H, 6.25 (M BSF-CH3 and 6.25 (M of BSF-Cl. Conclusions: Nitro-heterocyclic compounds have shown activity against promastigotes of L. amazonensis, at lower concentrations. However, improvement of compound scaffolds are needed to assist the elucidation of the mechanism of action and to achieve greater activity.

Abstract Visceral leishmaniasis is common in Brazil and is caused by Leishmania (Leishmania) infantum/chagasi. Post-kala-azar dermal leishmaniasis frequently follows visceral leishmaniasis caused by L. donovani, and para-kala-azar dermal leishmaniasis refers to an uncommon presentation wherein it occurs simultaneously along with visceral leishmaniasis. While post-kala-azar dermal leishmaniasis only occurs occasionally in L. infantum/chagasi infections, it frequently occurs in patients with concomitant immunosuppression (HIV co-infection). Here, we describe the first case of para-kala-azar dermal leishmaniasis in Brazil. It is important to raise awareness of post- and para-kala-azar dermal leishmaniasis in L. infantum endemic areas as these patients may contribute to visceral leishmaniasis transmission.

ABSTRACT Exoantigens (exo) from Leptomonas seymouri and Crithidia fasciculata were used in an enzyme linked immunosorbent assay (ELISA), showing 100% reactivity with sera from visceral leishmaniasis (VL) cases, and no reactivity with American tegumentary leishmaniasis (ATL) ones. Our results have indicated that these exoantigens can be applied in the discrimination of VL and ATL cases.

RESUMOEste estudo avaliou a aplicabilidade do kDNA-PCR como método de rotina para diagnóstico de leishmaniose tegumentar americana (ATL) no Instituto de Infectologia Emílio Ribas (IIER), São Paulo, SP, Brasil. O método kDNA-PCR detectou DNA de Leishmania em 87,5% (112/128) dos pacientes com suspeita de ter leishmaniose e, os métodos tradicionais apresentaram as seguintes porcentagens de positividade: 62,8% (49/78) para o teste de Montenegro, 61,8% (47/76) para a pesquisa direta e 19,3% (22/114) para cultura in vitro. O método molecular confirmou a doença em amostras negativas ou inconclusivas pelos métodos laboratoriais tradicionais e, mostrou-se capaz de auxiliar na identificação de infecções causadas pela espécie Leishmania (V.) braziliensis. Além disso, a revisão dos prontuários médicos confirmou a importância do método PCR-RFLP no diagnóstico final de ATL, prognóstico e escolha do tratamento. Assim, recomendamos a inclusão do PCR como método diagnóstico de ATL na rotina hospitalar, e sugerimos um fluxograma para solicitação de exames laboratoriais.

SUMMARYThis study evaluated the applicability of kDNA-PCR as a prospective routine diagnosis method for American tegumentary leishmaniasis (ATL) in patients from the Instituto de Infectologia Emílio Ribas (IIER), a reference center for infectious diseases in São Paulo - SP, Brazil. The kDNA-PCR method detected Leishmania DNA in 87.5% (112/128) of the clinically suspected ATL patients, while the traditional methods demonstrated the following percentages of positivity: 62.8% (49/78) for the Montenegro skin test, 61.8% (47/76) for direct investigation, and 19.3% (22/114) for in vitro culture. The molecular method was able to confirm the disease in samples considered negative or inconclusive by traditional laboratory methods, contributing to the final clinical diagnosis and therapy of ATL in this hospital. Thus, we strongly recommend the inclusion of kDNA-PCR amplification as an alternative diagnostic method for ATL, suggesting a new algorithm routine to be followed to help the diagnosis and treatment of ATL in IIER.

A pobreza está intrinsicamente relacionada com a ocorrência de doenças tropicais negligenciadas (DTNs). Os principais países com os menores índices de desenvolvimento humano (IDH) e a maior carga de DTNs estão nas regiões tropicais e subtropicais do globo terrestre. O Brasil é o 70º país no ranking do IDH e concentra nove das 10 principais doenças tropicais consideradas negligenciadas pela OMS. Leishmanioses, tuberculose, dengue e hanseníase ocorrem em quase todo o território do Brasil. Mais de 90% dos casos de malária ocorrem na região norte e há surtos de filariose linfática e oncocercose. As regiões norte e nordeste apresentam o menor IDH e concentram o maior número das DTNs. Essas doenças são consideradas negligenciadas devido à falta de investimento no desenvolvimento de novas drogas e vacinas e também pela pouca eficácia dos programas de controle. Um problema preocupante em relação às DTNs é a co-infecção com HIV, que favorece manifestações clínicas graves e falência terapêutica. Neste artigo, a situação das principais DTNs no Brasil é descrita e correlacionada com o IDH e a pobreza.

Poverty is intrinsically related to the incidence of Neglected Tropical Diseases (NTDs). The main countries that have the lowest human development indices (HDI) and the highest burdens of NTDs are located in tropical and subtropical regions of the world. Among these countries is Brazil, which is ranked 70th in HDI. Nine out of the ten NTDs established by the World Health Organization (WHO) are present in Brazil. Leishmaniasis, tuberculosis, dengue fever and leprosy are present over almost the entire Brazilian territory. More than 90% of malaria cases occur in the Northern region of the country, and lymphatic filariasis and onchocerciasis occur in outbreaks in a particular region. The North and Northeast regions of Brazil have the lowest HDIs and the highest rates of NTDs. These diseases are considered neglected because there is not important investment in projects for the development of new drugs and vaccines and existing programs to control these diseases are not sufficient. Another problem related to NTDs is co-infection with HIV, which favors the occurrence of severe clinical manifestations and therapeutic failure. In this article, we describe the status of the main NTDs currently occurring in Brazil and relate them to the HDI and poverty.

O trabalho consiste na re-investigação dos frutos de Iryanthera juruensis e Virola pavonis e das folhas de V. michelii, bem como no estudo dos frutos de V. mollissima. A partir de I. juruensis foram isoladas uma neolignana ariltetralínica (1) e uma neolignana tetraidrofurânica (2), enquanto que de Virola pavonis foram isoladas neolignanas benzofurânicas (6-9), tetraidrofurânicas (2, 11-13, 17), bifenílica (10), diastereoisômeros de uma hidróxi-neolignana 8.O.4'(14-15) e outras. V. mollissima acumula a neolignana ariltetralônica 4 ou o seu derivado seco (5). As folhas de V. michelii apresentaram a ocorrência de lignanas furofurânicas (18-19). Os lignóides 1 e 2 obtidos dos arilos de I. juruensis apresentaram atividade leishmanicida contra a forma promastigota de Leishmania amazonensis.

This work revisits the fruits of Iryanthera juruensis and Virola pavonis and the leaves from V. michelii, as well as describing a study of the fruits of V. mollissima. In I. juruensis aryltetraline neolignan (1) and tetrahydrofuran neolignan (2), were found while from V. pavonis neolignans of benzofuran type (6-9), the tetrahydrofuran type (2, 11-13, 17) and the biphenyl type (10), in addition to diastereoisomers of the 8.O.4'-oxyneolignan type (14 and 15) and others were isolated. The V. mollissima accumulates the aryltetralone neolignan 4 and its seco derivative (5). The lignoids 1 and 2 obtained from I. juruensis arils possess antileishmanial activity against the promastigote form of Leishmania amazonensis.