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An. bras. dermatol ; 97(1): 89-92, Jan.-Feb. 2022. graf
Article in English | LILACS | ID: biblio-1360083


Abstract Cutaneous leishmaniasis represents a public health problem that affects 85 countries. It is an endemic disease in Brazil, having an important socioeconomic impact. An exuberant case of cutaneous leishmaniasis is reported herein. A 28-year-old male patient with Down syndrome had had verrucous plaques on the back for over a year, with progressive growth. PCR of a lesion sample was positive for Leishmania braziliensis. The patient's condition was classified as atypical cutaneous leishmaniasis. He was successfully treated with amphotericin B and miltefosine. The treatment remains a challenge, given the toxicity and low cure rate of the currently recommended drugs.

Humans , Male , Adult , Leishmania braziliensis , Leishmaniasis, Cutaneous/drug therapy , Leishmaniasis, Cutaneous/epidemiology , Antiprotozoal Agents/therapeutic use , Amphotericin B/therapeutic use , Endemic Diseases
An. bras. dermatol ; 96(3): 352-354, May-June 2021. graf
Article in English | LILACS | ID: biblio-1285072


Abstract Cutaneous leishmaniasis is characterized by ulcers with raised edges and a granular bottom, mainly on the lower limbs. This is a case report of a male patient with an ulcer on the left plantar region. The diagnosis was confirmed by positive PCR for L. braziliensis and the presence of amastigotes of Leishmania sp. in the histopathological examination. After treatment with Glucantime, the patient showed full healing of the ulcer. The unusual location of the ulceration calls attention to atypical presentations of leishmaniasis, and the importance of histopathological examination and PCR, leading to the appropriate diagnosis and treatment.

Humans , Male , Leishmania braziliensis , Leishmaniasis, Cutaneous/diagnosis , Leishmaniasis, Cutaneous/drug therapy , Foot Ulcer , Leishmania , Ulcer , Meglumine Antimoniate
Rev. Soc. Bras. Med. Trop ; 54: e00472020, 2021. tab, graf
Article in English | ColecionaSUS, LILACS, ColecionaSUS, SES-SP | ID: biblio-1143886


Abstract INTRODUCTION: The objective of this study was to evaluate the performance of filter paper (FP) for lesion scraping collection in a polymerase chain reaction (PCR) for cutaneous leishmaniasis (CL) diagnosis. METHODS: Lesion scrapings from 48 patients were collected and analyzed for PCR. RESULTS: PCR with FP detected up to three Leishmania braziliensis promastigotes. Considering the direct search by microscopy or PCR of samples collected in STE buffer as standards, the sensitivity of PCR with FP was 100%. CONCLUSIONS: FP can be useful for CL diagnosis in remote regions, allowing high sensitivity in the detection of the parasite by PCR.

Humans , Leishmania braziliensis/genetics , Leishmaniasis, Cutaneous/diagnosis , Polymerase Chain Reaction , DNA, Protozoan/genetics , Sensitivity and Specificity , Microscopy
An. bras. dermatol ; 95(5): 641-644, Sept.-Oct. 2020. tab, graf
Article in English | ColecionaSUS, LILACS, ColecionaSUS | ID: biblio-1130931


Abstract Genital lesions are an unusual presentation of American cutaneous leishmaniasis. Conditions such as disseminated cutaneous leishmaniasis and HIV infection may be associated with genital involvement. The authors present five cases of American cutaneous leishmaniasis with genital lesions and discuss the clinical and epidemiological aspects observed in this case series.

Humans , Leishmania braziliensis , Leishmaniasis, Mucocutaneous , HIV Infections , Leishmaniasis, Cutaneous/diagnosis , Leishmaniasis, Cutaneous/epidemiology , United States , Genitalia
Hist. ciênc. saúde-Manguinhos ; 27(3): 741-761, set. 2020. tab, graf
Article in Portuguese | LILACS | ID: biblio-1134073


Resumo O artigo faz análise histórica da emergência da leishmaniose tegumentar americana como objeto do conhecimento e desafio médico-sanitário no Amazonas desde a década de 1970. Fornece visão geral dessa época, as medidas sanitárias e os estudos científicos realizados no contexto de implantação dos principais projetos de desenvolvimento regionais executados em nome da política de integração nacional do governo federal. Utiliza como metodologia a análise documental de leis, produção científica, relatórios de pesquisa, boletins epidemiológicos e jornais. Os resultados da pesquisa mostram que a doença surgiu no Amazonas associando o grande problema de saúde com mudanças político-econômicas e alterações socioambientais.

Abstract The history of the emergence of American cutaneous leishmaniasis in the Brazilian state of Amazonas since the 1970s is analyzed as an object of knowledge and a medical and public health challenge. An overview of the period is provided, including the public health measures and scientific studies undertaken in the context of the execution of large-scale regional developments pursued in the name of national integration by the federal government. The methodology uses documental analysis of laws, the scientific literature, research reports, epidemiological bulletins, and newspapers. The results show that American cutaneous leishmaniasis emerged as a major health problem in Amazonas in close association with the political, economic, and socioenvironmental changes seen in the period.

Humans , Animals , Public Health/history , Leishmaniasis, Cutaneous/history , Conservation of Natural Resources , Leishmania/isolation & purification , Psychodidae/parasitology , Urbanization/history , Leishmania braziliensis/isolation & purification , Brazil/epidemiology , Insect Control/history , Leishmaniasis, Cutaneous/transmission , Leishmaniasis, Cutaneous/epidemiology , Leishmania guyanensis/isolation & purification , Industrial Development/history , Insect Vectors
An. Fac. Cienc. Méd. (Asunción) ; 53(2): 145-152, 20200800.
Article in Spanish | LILACS | ID: biblio-1119624


La leishmaniasis cutánea o mucocutánea presenta variedad de formas clínicas, siendo la más frecuente la úlcera moldurada. Una forma poco frecuente es la llamada leishmaniasis cutánea difusa caracterizada por presentar pápulas, tubérculos, nódulos, placas e infiltración. Inicialmente localizadas, pero con tendencia a la progresión, pudiendo llegar a ser diseminadas. Es una forma aún no comunicada en Paraguay. Presentamos el caso de una mujer adulta mayor, con placas y nódulos en ambos miembros inferiores, y cuyo frotis y anatomía patológica confirmaron el diagnóstico de leishmaniasis. Clínicamente clasificada como leishmaniasis cutánea difusa, la PCR y HRM demostraron ser producida por Leishmania (Viannia) brasiliensis.

Cutaneous or mucocutaneous leishmaniasis presents a variety of clinical forms, the most common being a molded ulcer. A rare form is the so-called diffuse cutaneous leishmaniasis characterized by presenting papules, tubers, nodules, plaques and infiltration; initially located but with a tendency to progression, and may become widespread. It is a form not reported in Paraguay. We present the case of an older adult woman, with plaques and nodules on both lower limbs, and whose smear and pathological anatomy confirmed the diagnosis of leishmaniasis. Clinically classified as diffuse cutaneous leishmaniasis, PCR and HRM were shown to be produced by Leishmania (Viannia) brasiliensis.

Leishmania braziliensis , Leishmaniasis, Diffuse Cutaneous , Leishmaniasis, Diffuse Cutaneous/epidemiology
Mem. Inst. Oswaldo Cruz ; 115: e200113, 2020. tab, graf
Article in English | SES-SP, LILACS, SES-SP | ID: biblio-1135235


BACKGROUND Lutzomyia longipalpis-derived cell line (Lulo) has been suggested as a model for studies of interaction between sandflies and Leishmania. OBJECTIVES Here, we present data of proteomic and gene expression analyses of Lulo cell related to interactions with Leishmania (Viannia) braziliensis. METHODS Lulo cell protein extracts were analysed through a combination of two-dimensional gel electrophoresis and mass spectrometry and resulting spots were further investigated in silico to identify proteins using Mascot search and, afterwards, resulting sequences were applied for analysis with VectorBase. RESULTS Sixty-four spots were identified showing similarities to other proteins registered in the databases and could be classified according to their biological function, such as ion-binding proteins (23%), proteases (14%), cytoskeletal proteins (11%) and interactive membrane proteins (9.5%). Effects of interaction with L. (V.) braziliensis with the expression of three genes (enolase, tubulin and vacuolar transport protein) were observed after an eight-hour timeframe and compared to culture without parasites, and demonstrated the impact of parasite interaction with the expression of the following genes: LLOJ000219 (1.69-fold), LLOJ000326 (1.43-fold) and LLOJ006663 (2.41-fold). CONCLUSIONS This set of results adds relevant information regarding the usefulness of the Lulo cell line for studies with Leishmania parasites that indicate variations of protein expression.

Animals , Psychodidae/parasitology , Leishmania braziliensis/genetics , Proteomics , Leishmania/genetics , Cell Line , Transcriptome
Mem. Inst. Oswaldo Cruz ; 115: e200272, 2020. graf
Article in English | SES-SP, LILACS, SES-SP | ID: biblio-1135255


BACKGROUND Metformin (MET) is a hypoglycemic drug used for the treatment of diabetes, despite interference in host immunity against microorganisms. Cutaneous infection caused by pathogens such as Leishmania braziliensis (Lb), the agent responsible for cutaneous leishmaniasis (CL) in Brazil, represents an interesting model in which to evaluate the effects associated with MET. OBJECTIVE To evaluate the modulatory effect of MET in Lb infection. MATERIAL AND METHODS Experimental study of Lb infection and MET treatment in BALB/c mice and Raw 264.7 macrophages. FINDINGS MET treatment interfered with lesion kinetics, increased parasite load and reduced macrophage proliferation. Low concentrations of MET in Lb culture allow for the maintenance of stationary parasite growth phase. Lb-infected cells treated with MET exhibited increased parasite load. While both MET and Lb infection alone promoted the production of intracellular reactive oxygen species (ROS), reduced levels of ROS were seen in MET-treated Lb-infected macrophages. MAIN CONCLUSION Experimental treatment with MET interfered with the kinetics of cutaneous ulceration, increased Lb parasite load, altered ROS production and modulated cellular proliferation. Our experimental results indicate that MET interfere with the evolution of CL.

Animals , Mice , Leishmaniasis, Cutaneous/drug therapy , Leishmania/drug effects , Metformin/pharmacology , Leishmania braziliensis , Brazil , Mice, Inbred BALB C
Mem. Inst. Oswaldo Cruz ; 115: e190469, 2020. graf
Article in English | SES-SP, LILACS, SES-SP | ID: biblio-1135243


BACKGROUND Oxidative stress is responsible for generating DNA lesions and the 8-oxoguanine (8-oxoG) is the most commonly lesion found in DNA damage. When this base is incorporated during DNA replication, it could generate double-strand DNA breaks and cellular death. MutT enzyme hydrolyzes the 8-oxoG from the nucleotide pool, preventing its incorporation during DNA replication. OBJECTIVES To investigate the importance of 8-oxoG in Leishmania infantum and L. braziliensis, in this study we analysed the impact of heterologous expression of Escherichia coli MutT (EcMutT) enzyme in drug-resistance phenotype and defense against oxidative stress. METHODS Comparative analysis of L. braziliensis and L. infantum H2O2 tolerance and cell cycle profile were performed. Lines of L. braziliensis and L. infantum expressing EcMutT were generated and evaluated using susceptibility tests to H2O2 and SbIII, cell cycle analysis, γH2A western blotting, and BrdU native detection assay. FINDINGS Comparative analysis of tolerance to oxidative stress generated by H2O2 showed that L. infantum is more tolerant to exogenous H2O2 than L. braziliensis. In addition, cell cycle analysis showed that L. infantum, after treatment with H2O2, remains in G1 phase, returning to its normal growth rate after 72 h. In contrast, after treatment with H2O2, L. braziliensis parasites continue to move to the next stages of the cell cycle. Expression of the E. coli MutT gene in L. braziliensis and L. infantum does not interfere in parasite growth or in susceptibility to SbIII. Interestingly, we observed that L. braziliensis EcMutT-expressing clones were more tolerant to H2O2 treatment, presented lower activation of γH2A, a biomarker of genotoxic stress, and lower replication stress than its parental non-transfected parasites. In contrast, the EcMutT is not involved in protection against oxidative stress generated by H2O2 in L. infantum. MAIN CONCLUSIONS Our results showed that 8-oxoG clearance in L. braziliensis is important to avoid misincorporation during DNA replication after oxidative stress generated by H2O2.

Humans , Animals , Mice , Rats , Pyrophosphatases/genetics , Pyrophosphatases/metabolism , Superoxide Dismutase/metabolism , Leishmania braziliensis/drug effects , Leishmania infantum/drug effects , Escherichia coli Proteins/genetics , Escherichia coli , Guanine/analogs & derivatives , Antimony/toxicity , Rabbits , Superoxide Dismutase/genetics , Leishmania braziliensis/enzymology , Leishmania infantum/enzymology , Oxidative Stress/drug effects , Oxidative Stress/physiology , Escherichia coli Proteins/metabolism , Guanine/pharmacology , Hydrogen Peroxide/toxicity , Antiprotozoal Agents/pharmacology
Mem. Inst. Oswaldo Cruz ; 115: e190413, 2020. tab, graf
Article in English | LILACS | ID: biblio-1101274


BACKGROUND The leishmaniases are complex neglected diseases caused by protozoan parasites of the genus Leishmania. Leishmania braziliensis is the main etiological agent of cutaneous leishmaniasis in the New World. In recent studies, genomic changes such as chromosome and gene copy number variations (CNVs), as well as transcriptomic changes have been highlighted as mechanisms used by Leishmania species to adapt to stress situations. OBJECTIVES The aim of this study was to determine the effect of short-term minor temperature shifts in the genomic and transcriptomic responses of L. braziliensis promastigotes in vitro. METHODS Growth curves, genome and transcriptome sequencing of L. braziliensis promastigotes were conducted from cultures exposed to three different temperatures (24ºC, 28ºC and 30ºC) compared with the control temperature (26ºC). FINDINGS Our results showed a decrease in L. braziliensis proliferation at 30ºC, with around 3% of the genes showing CNVs at each temperature, and transcriptomic changes in genes encoding amastin surface-like proteins, heat shock proteins and transport proteins, which may indicate a direct response to temperature stress. MAIN CONCLUSIONS This study provides evidence that L. braziliensis promastigotes exhibit a decrease in cell density, and noticeable changes in the transcriptomic profiles. However, there were not perceptible changes at chromosome CNVs and only ~3% of the genes changed their copies in each treatment.

Animals , Temperature , Leishmania braziliensis/genetics , Adaptation, Physiological/genetics , DNA Copy Number Variations/genetics , Transcriptome/genetics , Adaptation, Physiological/physiology , Gene Expression Profiling , Genetic Profile
Mem. Inst. Oswaldo Cruz ; 115: e190408, 2020. graf
Article in English | LILACS | ID: biblio-1101276


BACKGROUND The mechanism of resistance to SbIII in Leishmania is complex, multifactorial and involves not only biochemical mechanisms, but also other elements, such as the immune system of the host. OBJECTIVES In this study, putative changes in the immunological profile of human monocytes infected with wild-type (WT) and antimony (SbIII)-resistant Leishmania (Viannia) braziliensis and Leishmania (Leishmania) infantum lines were evaluated. METHODS Susceptibility assays WT and SbIII-resistant L. braziliensis and L. infantum were performed using lines THP-1 human monocytic lineage. Phagocytic capacity, cytokine profile, intracellular nitric oxide (NO) production and surface carbohydrate residues profile were performed in peripheral blood monocytes by flow cytometry. FINDINGS The phagocytic capacity and intracellular NO production by classical (CD14++CD16-) and proinflammatory (CD14++CD16+) monocytes were higher in the presence of L. infantum lines compared to L. braziliensis lines. The results also highlight proinflammatory monocytes as the cellular subpopulation of major relevance in a phagocytosis event and NO expression. It is important to note that L. infantum induced a proinflammatory cytokine profile characterised by higher levels of TNF-α in culture supernatant than L. braziliensis. Conversely, both Leishmania lines induce high levels of IL-6 in culture supernatant. Analysis of the expression profile of surface carbohydrates showed that L. braziliensis presents 4.3-fold higher expression of galactose(β1,4)N-acetylglucosamine than L. infantum line. Interestingly, the expression level of α-N-acetylgalactosamine residues was 2-fold lower in the SbIII-resistant L. braziliensis line than its counterpart WT line, indicating differences in surface glycoconjugates between these lines. MAIN CONCLUSIONS Our results showed that L. braziliensis and L. infantum induce different innate immune responses and a highly inflammatory profile, which is characteristic of infection by L. infantum, the species associated with visceral disease.

Humans , Male , Female , Adult , Young Adult , Phagocytosis/immunology , Leishmania braziliensis/immunology , Monocytes/parasitology , Leishmania infantum/immunology , Antimony/pharmacology , Nitric Oxide/biosynthesis , Antiprotozoal Agents/pharmacology , Leishmania braziliensis/drug effects , Drug Resistance , Monocytes/immunology , Leishmania infantum/drug effects , Flow Cytometry , Immunity, Innate
Rev. bras. parasitol. vet ; 29(2): e003520, 2020. tab, graf
Article in English | LILACS | ID: biblio-1138090


Abstract Blood samples and swabs from ocular conjunctiva and mouth were obtained from 64 cats. Of 64 serum samples, 19 were positive for Leishmania antibodies by ELISA (29.80%). Eight cats were positive by PCR (12.5%) in swab samples from mouth and/or ocular mucosa. Poor kappa agreement between serological and molecular results (k = 0.16) was obtained. From five positive PCR samples one was L. braziliensis and four were L. infantum. Phylogenetic analysis performed with the five isolates of Leishmania, showed that samples of L. infantum isolated from the cats were phylogenetically close to those isolated from domestic dogs in Brazil, while the L. braziliensis is very similar to the one described in humans in Venezuela. The study demonstrated that, despite high seropositivity for Leishmania in cats living in the study region, poor agreement between serological and molecular results indicate that positive serology is not indicative of Leishmania infection in cats. Parasite DNA can be detected in ocular conjunctiva and oral swabs from cats, indicating that such samples could be used for diagnosis. Results of phylogenetic analyzes show that L. infantum circulating in Brazil is capable of infecting different hosts, demonstrating the parasite's ability to overcome the interspecies barrier.

Resumo Amostras de sangue e swabs da conjuntiva ocular e oral foram obtidas de 64 gatos. Das 64 amostras de soro, 19 foram positivas para anticorpos contra Leishmania por ELISA (29,80%). Oito gatos foram positivos por PCR (12,5%) em amostras de swab da boca e / ou mucosa ocular. Demonstrou-se baixa concordância kappa entre os resultados sorológicos e moleculares (k = 0,16). Das cinco amostras positivas para PCR, uma era L. braziliensis e quatro eram L infantum. A análise filogenética realizada com os cinco isolados de Leishmania, mostrou que amostras de L. infantum, isoladas dos gatos, eram filogeneticamente próximas às isoladas de cães domésticos do Brasil enquanto L. braziliensis era muito semelhante ao descrito em humanos na Venezuela. O estudo demonstrou que, apesar da alta soropositividade para Leishmania, em gatos que vivem na região do estudo, pouca concordância entre os resultados sorológicos e moleculares indica que a sorologia positiva não é indicativa de infecção por Leishmania em gatos. O DNA do parasita pode ser detectado na conjuntiva ocular e nas zaragatoas orais de gatos, indicando que essas amostras podem ser usadas para o diagnóstico. . Resultados de análises filogenéticas mostram que L. infantum, circulando no Brasil, é capaz de infectar diferentes hospedeiros, demonstrando a capacidade do parasita de superar a barreira interespécies.

Animals , Cats , Leishmania braziliensis/isolation & purification , Cat Diseases/parasitology , Leishmaniasis/parasitology , Leishmania infantum/isolation & purification , Phylogeny , Leishmania braziliensis/genetics , Leishmania braziliensis/immunology , Enzyme-Linked Immunosorbent Assay/veterinary , Antibodies, Protozoan/blood , Cat Diseases/diagnosis , Leishmaniasis/diagnosis , Polymerase Chain Reaction/veterinary , DNA, Protozoan/analysis , Leishmania infantum/genetics , Leishmania infantum/immunology
Rev. Soc. Bras. Med. Trop ; 53: e20200255, 2020. tab, graf
Article in English | ColecionaSUS, LILACS, ColecionaSUS, SES-SP | ID: biblio-1143862


Abstract INTRODUCTION: In the Belém Metropolitan Region (BMR), Pará State, Brazil, American cutaneous leishmaniasis (ACL) is endemic; however, very little is known regarding its causative agents. Therefore, we used our standard diagnostic approach combined with an RNA polymerase II largest subunit (RNAPOIILS)-polymerase chain reaction (PCR) followed by analysis of restriction fragment length polymorphism (PCR-RFLP) to identify Leishmania spp. ACL agents in this region. METHODS: Thirty-two Leishmania spp. isolates from patients with ACL in the BMR during 1995-2018 were analyzed. Leishmania spp. DNA samples were amplified using the primers RPOR2/RPOF2, and the 615-bp PCR products were subjected to enzymatic digestion using TspRI and HgaI endonucleases. RESULTS: ACL etiological agents in the BMR comprised Leishmania (Viannia) lindenbergi (43.7%) followed by Leishmania (Viannia) lainsoni (34.4%), Leishmania (Leishmania) amazonensis (12.5%), and Leishmania (Viannia) braziliensis (9.4%). CONCLUSIONS: To our knowledge, the results of the study revealed for the first time that L. (V.) lindenbergi and L. (V.) lainsoni are the main ACL agents in BMR.

Humans , Leishmania braziliensis/genetics , Leishmaniasis, Cutaneous/diagnosis , Leishmaniasis, Cutaneous/epidemiology , Leishmania/genetics , United States , Brazil/epidemiology , Leishmaniasis, Cutaneous
Rev. bras. anal. clin ; 51(4): 315-321, 2019/12/30. ilus ilus, tab
Article in Portuguese | LILACS | ID: biblio-1104009


Objetivo: Realizar uma análise histopatológica e molecular em biópsia de pele entre as lesões de dermatites de pacientes com suspeita de Leishmaniose Tegumentar Americana (LTA) no hospital de referência do estado de Pernambuco entre o período de 2016 e 2017. Métodos: Trata-se de um estudo descritivo observacional, no qual todos os pacientes com lesões clinicamente sugestivas para LTA incluídos no estudo foram submetidos à coleta de biópsia de pele das lesões, as quais foram analisadas pela técnica histopatológica e PCR (Reação em Cadeia de Polimerase). Resultados: Foram analisadas 24 amostras de biópsia de pele de pacientes com suspeita clínica de LTA, por testes histopatológicos e confirmação pela PCR. As amostras foram caracterizadas pela busca do DNA de Leishmania braziliensis através da PCR. Das 24 amostras estudadas, em nenhuma foi encontrado DNA de L. braziliensis. Apenas em um caso foi detectada presença de amastigotas de Leishmania pela técnica histopatológica. Outros achados microscópicos observados foram: dermatite granulomatosa (33,33%), úlcera crônica (20,83%), carcinoma basocelular (16,66%), Leishmaniose, dermatite plasmocitária e inflamação granulomatosa (8,33%) e Hanseníase (4,16%). Conclusão: O diagnóstico histopatológico detectou um caso de LTA, porém, a PCR não encontrou DNA do parasito. A análise histopatológica mostrou que as lesões dermatotrópicas dos pacientes são oriundas principalmente de úlceras, tumores de pele e hanseníase.

Objective: Accomplish a histopathological and molecular analysis in skin biopsy between the dermatitis lesions of patients with suspected American Cutaneous Leishmaniasis (ATL) at the Hospital of Reference of the State of Pernambuco between the period of 2016 and 2017. Methods: This is a descriptive, observational study in which all patients with clinically suggestive lesions for ATL included in the study were submitted to skin biopsy of the lesions and analyzed by the histopathological technique and PCR (Polymerase Chain Reaction). Results: Were analyzed 24 skin biopsy samples from patients with clinical suspicion of ATL, by histopathological tests and confirmation by PCR. Samples were characterized by the search of Leishmania braziliensis DNA through PCR. Of the 24 samples studied, no DNA of L. braziliensis was found. Only in one case was detected presence of Leishmania amastigotes by histopathological technique. Other microscopic findings were granulomatous dermatitis (33.33%), chronic ulcer (20.83%), basal cell carcinoma (16.66%), Leishmaniasis, plasmacytoma dermatitis and granulomatous inflammation (8.33%) and leprosy, 16%). Conclusion: The histopathological diagnosis detected a case of ATL, however, the PCR did not find DNA of the parasite. The histopathological analysis showed that the dermatotropic lesions of the patients come mainly from ulcers, skin tumors and leprosy.

Humans , Male , Female , Adult , Middle Aged , Aged , Leishmania braziliensis , Polymerase Chain Reaction , Leishmaniasis, Cutaneous , Dermatitis
Con-ciencia (La Paz) ; 7(2): 39-51, nov. 2019. ilus., tab.
Article in Spanish | LILACS, LIBOCS | ID: biblio-1178628


La leishmaniasis cutánea se caracteriza por la inducción de una respuesta inmune celular y humoral, siendo los niveles de anticuerpos IgG e IgE indicadores serológicos del diagnóstico, la progresión y resolución de la enfermedad. En el presente estudio se estandarizaron dos pruebas de ELISA indirecto para determinar los niveles séricos de anticuerpos IgG o IgE contra antígeno soluble de Leishmania braziliensis (ASL). Se trabajó a partir de la cepa de L. braziliensis (MHOM/BR/75/M2903). En el ELISA IgE se hizo un pretratamiento del suero con anticuerpos antiIgG humano y Polietilenglicol (PEG); la dilución óptima de la muestra para ambos ELISA fue 1/21, la dilución del conjugado fue 1/500 para el ELISA IgG y 1/100 para el IgE. Para el ELISA IgG estandarizado se encontró un índice de correlación de Kappa de 1 (p<0,05), frente al ELISA comercial rBiopharm®. Con una sensibilidad del 100%; 100% de especificidad, 100% valor predictivo positivo y 100% valor predictivo negativo. Sin embargo ambos test de ELISA presentaron reacciones cruzadas con anticuerpos anti-Trypanosoma. La validez de los resultados demuestran que los ELISAs estandarizados pueden ser utilizados como herramienta de diagnóstico y seguimiento al tratamiento leishmanicida.

Cutaneous leishmaniasis is characterized by the induction of a cellular and humoral immune responses, at levels of IgG and IgE indicators serological diagnosis, progression and resolution of disease. In this study two indirect ELISA tests were standardized to determine serum levels of IgG or IgE antibodies against soluble Leishmania braziliensis antigen (ASL). We worked from the strain of L. braziliensis (MHOM / BR /75 / M2903). In the IgE ELISA pretreatment serum was done with anti-human IgG and Polyethylene glycol (PEG) antibodies; the optimal dilution of the sample for both ELISA was 1/21, the conjugate dilution was for 1/500 and 1/100 IgG ELISA for IgE. For standardized ELISA IgG index Kappa correlation of 1 (p <0.05) compared to commercial ELISA r-Biopharm® found. With a sensitivity of 100%; 100% specificity, 100% positive predictive value and negative predictive value 100%. However both ELISA test showed cross-reactions anti-trypanosome antibodies. The results demonstrate that both ELISA can be used as diagnostic and monitoring tool. Being necessary to continue studies to validate these procedures and identify other possible cross-reactions.

Humans , Trypanosoma , Leishmania braziliensis , Enzyme-Linked Immunosorbent Assay , Aftercare , Antibodies , Disease , Diagnosis , Indicators and Reagents
Arq. bras. med. vet. zootec. (Online) ; 71(3): 939-943, May-June 2019. tab
Article in English | LILACS, VETINDEX | ID: biblio-1011298


Alveolar macrophages (AMs) are an essential part of defense mechanisms within the lungs and their phagocytic activity is important for organ homeostasis. The phagocytic ability of AMs obtained from bronchoalveolar lavage from 17 mature mixed-breed pleasure horses (8 healthy and 9 diagnosed with mild equine asthma) was studied through assays with Leishmania (Viannia) braziliensis promastigotes, which enabled the calculation of a phagocytic index (PI) and a survival index (SI). Results indicate that phagocytic activity of AMs in asthma affected horses is similar to healthy horses, while leishmanicidal activity is significantly increased in horses with asthma.(AU)

Os macrófagos alveolares (MAs) são uma parte essencial dos mecanismos de defesa dentro dos pulmões e sua atividade fagocítica é importante para a homeostase desse órgão. A capacidade fagocitária dos MAs obtidos do lavado broncoalveolar de 17 equinos adultos, sem raça definida (oito saudáveis e nove com diagnóstico de asma equina leve), foi estudada por meio de ensaios com promastigotas de Leishmania (Viannia) braziliensis. Foi calculado o índice fagocítico e o índice de sobrevivência. Os resultados indicam que a atividade fagocítica de MAs em cavalos com asma é semelhante a cavalos saudáveis, enquanto a atividade leishmanicida está significativamente aumentada em cavalos com essa enfermidade.(AU)

Animals , Asthma/veterinary , Leishmania braziliensis , Macrophages, Alveolar/parasitology , Horses/parasitology , Phagocytosis
Rev. Univ. Ind. Santander, Salud ; 51(1): 33-42, ene.-mar. 2019. tab, graf
Article in Spanish | LILACS | ID: biblio-1003153


Resumen Introducción: La diversidad de las formas clínicas de la leishmaniasis del Nuevo Mundo (desde formas cutáneas localizadas a diseminadas o formas mucosas) causada por especies del subgénero Viannia podría inferir en la eficacia de los tratamientos tópicos. El objetivo del presente trabajo fue determinar las características de la leishmaniasis cutánea producida por infecciones con Leishmania (V.) braziliensis y L.(V.) panamensis en ratones BALB/c y la eficacia de un mismo tratamiento tópico. Materiales y métodos: Después de la infección con cada una de las especies se realizó seguimiento de las lesiones determinando su tamaño (mm ) y características macroscópicas, cada siete días por 150 días. Las características histopatológicas (en lesiones y órganos) fueron determinadas 70, 106 y 150 días post-infección y la eficacia de un tratamiento tópico (cura de lesión y parasitológica) fue determinada después del tratamiento con un gel de miltefosina aplicado una vez al día por 20 días sobre las lesiones. Resultados: Se observó un aumento del tamaño de las lesiones en ambos grupos de ratones, sin embargo, un mayor tamaño de las lesiones e intensidad de la respuesta inflamatoria con menos alteraciones epidérmicas fue encontrada en los ratones infectados con L. (V.) braziliensis. En ningún grupo se encontraron parásitos en órganos (nódulos, bazo e hígado) ni diferencias en la efectividad del tratamiento tópico utilizado. Conclusión: La eficacia del tratamiento tópico utilizado no fue afectada por las diferencias macro y microscópicas encontradas en la leishmaniasis producida por las dos especies de Leishmania evaluadas.

Abstract Introduction: The efficacy of topical treatments could be affected by the diversity of clinical forms (localized or disseminated cutaneous forms, mucosal forms) of New World-leishmaniasis caused by species of Leishmania from the subgenus Viannia. The aim of this study was to determine the cutaneous leishmaniasis features produced after infection with Leishmania (V.) braziliensis and L. (V.) panamensis in BALB/c mice and to determine the efficacy of one topical treatment. Materials and methods: Cutaneous leishmaniasis lesions were followed up after infection determining their lesion-size (mm2) and other macroscopic characteristics every 7 days for 150 days. Histopathological patterns (in lesions and organs) were determined 70, 106 and 150 days post-infection and the efficacy (lesion and parasitological cure) of miltefosine gel applied topical once a day for 20 days was determined. Results: An increase of size-lesions was observed in both groups of mice, however, a higher lesion- size and inflammatory response but lower epidermal changes were observed in L. (V.) braziliensis compared with L. (V.) panamensis infected ones. No parasites were observed in organs (nodules, spleen and liver) and no differences were observed in the effectiveness of the used topical treatment. Conclusion: The efficacy of the topical treatment used was not affected by the macro and microscopic differences produced after infection by the two Leishmania species evaluated.

Animals , Leishmania braziliensis , Leishmania guyanensis , Mice, Inbred BALB C , Anti-Infective Agents, Local
An. bras. dermatol ; 94(1): 9-16, Jan.-Feb. 2019. tab, graf
Article in English | LILACS | ID: biblio-983744


Abstract: Disseminated leishmaniasis is a severe and emerging form of American tegumentary leishmaniasis. Disseminated leishmaniasis is defined by the presence of more than 10 polymorphic cutaneous lesions, distributed over more than two noncontiguous parts of the body. Nasal mucosal involvement is observed in almost half of cases. Disseminated leishmaniasis patients present with a decreased production of Th1 cytokines in the peripheral blood due to the attraction of leishmania- activated T cells to the multiple cutaneous lesions. Disseminated leishmaniasis development is poorly understood and is related to a complex network involving environmental, host immune response, and parasite factors, in which L. braziliensis polymorphism plays an important role. Disseminated leishmaniasis is a challenging disease to cure, presenting a high failure rate of 75% to pentavalent antimony therapy. Despite its importance and severity, this form of American tegumentary leishmaniasis has been poorly studied and documented, deserving greater attention from professionals working in endemic areas.

Humans , Leishmania braziliensis , Leishmaniasis, Cutaneous/pathology , Leishmaniasis, Cutaneous/drug therapy , Amphotericin B/therapeutic use , Treatment Outcome , Leishmaniasis, Cutaneous/immunology , Antiprotozoal Agents/therapeutic use
Mem. Inst. Oswaldo Cruz ; 114: e180438, 2019. tab, graf
Article in English | LILACS | ID: biblio-1040619


Leishmania braziliensis is the etiological agent of American mucosal leishmaniasis, one of the most severe clinical forms of leishmaniasis. Here, we report the assembly of the L. braziliensis (M2904) genome into 35 continuous chromosomes. Also, the annotation of 8395 genes is provided. The public availability of this information will contribute to a better knowledge of this pathogen and help in the search for vaccines and novel drug targets aimed to control the disease caused by this Leishmania species.

Leishmania braziliensis/genetics , DNA, Protozoan/genetics , Sequence Analysis, DNA
Mem. Inst. Oswaldo Cruz ; 114: e190147, 2019. tab, graf
Article in English | LILACS | ID: biblio-1040618


BACKGROUND Calpains are proteins belonging to the multi-gene family of calcium-dependent cysteine peptidases that undergo tight on/off regulation, and uncontrolled proteolysis of calpains is associated with severe human pathologies. Calpain orthologues are expanded and diversified in the trypanosomatids genome. OBJECTIVES Here, we characterised calpains in Leishmania braziliensis, the main causative agent of cutaneous leishmaniasis in Brazil. METHODS/FINDINGS In total, 34 predicted calpain-like genes were identified. After domain structure evaluation, reverse transcription-quantitative polymerase chain reaction (RT-qPCR) during in vitro metacyclogenesis revealed (i) five genes with enhanced expression in the procyclic stage, (ii) one augmented gene in the metacyclic stage, and (iii) one procyclic-exclusive transcript. Western blot analysis revealed that an antibody against a consensus-conserved peptide reacted with multiple calpain-like proteins, which is consistent with the multi-gene family characteristic. Flow cytometry and immunocytochemistry analyses revealed the presence of calpain-like molecules mainly in the cytoplasm, to a lesser extent in the plasma membrane, and negligible levels in the nucleus, which are all consistent with calpain localisation. Eventually, the calpain inhibitor MDL28170 was used for functional studies revealing (i) a leishmaniostatic effect, (ii) a reduction in the association index in mouse macrophages, (iii) ultra-structural alterations conceivable with autophagy, and (iv) an enhanced expression of the virulence factor GP63. CONCLUSION This report adds novel insights into the domain structure, expression, and localisation of L. braziliensis calpain-like molecules.

Animals , Mice , Leishmania braziliensis/chemistry , Calpain/genetics , Macrophages, Peritoneal/metabolism , Genome, Protozoan/genetics , Leishmania braziliensis/genetics , Leishmania braziliensis/metabolism , Leishmania braziliensis/ultrastructure , Immunohistochemistry , Calpain/drug effects , Calpain/metabolism , Calpain/ultrastructure , Cysteine Proteinase Inhibitors/pharmacology , Gene Expression Regulation , Blotting, Western , Reverse Transcriptase Polymerase Chain Reaction , Virulence Factors , Microscopy, Electron, Transmission , Dipeptides/pharmacology , Flow Cytometry , Mice, Inbred BALB C