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Leishmania (Viannia) braziliensis is one of the main causes of cutaneous leishmaniasis in the Americas. This species presents genetic polymorphism that can cause destructive lesions in oral, nasal, and oropharyngeal tracts. In a previous study, the parasite caused several histopathological changes to hamster ileums. Our study evaluates immune response components, morphological changes, and effects on neurons in the ileums of hamsters infected by three different strains of L. (V.) braziliensis in two infection periods. For the experiment, we separated hamsters into four groups: a control group and three infected groups. Infected hamsters were euthanized 90- or 120-days post infection. We used three strains of L. (V.) braziliensis: the reference MHOM/BR/1975/M2903 and two strains isolated from patients who had different responses to Glucantime® treatment (MHOM/BR/2003/2314 and MHOM/BR/2000/1655). After laparotomy, ileums were collected for histological processing, biochemical analysis, and evaluation of neurons in the myenteric and submucosal plexuses of the enteric nervous system (ENS). The results demonstrated the increase of blood leukocytes after the infection. Optical microscopy analysis showed histopathological changes with inflammatory infiltrates, edemas, ganglionitis, and Leishmania amastigotes in the ileums of infected hamsters. We observed changes in the organ histoarchitecture of infected hamsters when compared to control groups, such as thicker muscular and submucosa layers, deeper and wider crypts, and taller and broader villi. The number of intraepithelial lymphocytes and TGF-ß-immunoreactive cells increased in all infected groups when compared to the control groups. Mast cells increased with longer infection periods. The infection also caused remodeling of intestinal collagen and morphometry of myenteric and submucosal plexus neurons; but this effect was dependent on infection duration. Our results show that L. (V.) braziliensis infection caused time-dependent alterations in hamster ileums. This was demonstrated by the reduction of inflammatory cells and the increase of tissue regeneration factors at 120 days of infection. The infected groups demonstrated different profiles in organ histoarchitecture, migration of immune cells, and morphometry of ENS neurons. These findings suggest that the small intestine (or at least the ileum) is a target organ for L. (V.) braziliensis infection, as the infection caused changes that were dependent on duration and strain.
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Íleo/parasitologia , Leishmania braziliensis , Leishmaniose/patologia , Animais , Cricetinae , HumanosRESUMO
Leishmaniasis is a worldwide problem and has been neglected in a wide range of fields, from diagnosis to treatment. This report describes a case of mucosal leishmaniasis, which may developped after seven decades of an inadequately treated cutaneous lesion. A female patient, 79 years old, from the non-endemic area for leishmaniasis in northern Paraná, presenting mucosal lesion in the nose and throat, reported an "angry ulcer" treated inappropriately as a child when she lived in an endemic region of the state of São Paulo. Indirect immunofluorescence and direct parasite screening were positive. Polymerase chain reaction detected a parasite belonging to the subgenus Leishmania (Viannia) sp. Due to patients limitations, such as low weight and advanced age, the therapeutic model adopted was the combined small doses of Glucantime™ to pentoxifylline, which ensured treatment success.
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Leishmaniose Mucocutânea/prevenção & controle , Antimoniato de Meglumina/uso terapêutico , Pentoxifilina/uso terapêutico , Tripanossomicidas/uso terapêutico , Idoso , Brasil , Quimioterapia Combinada , Feminino , Humanos , Leishmania/efeitos dos fármacosRESUMO
This study is aimed to investigate the in vitro anti-leishmanial activity of ethanolic, aqueous or dichloromethane extracts of leaves, flowers, fruits or roots, of six medicinal plant species, namely, Nectandra megapotamica, Brunfelsia uniflora, Myrcianthes pungens, Anona muricata, Hymenaea stigonocarpa and Piper corcovandesis. After isolation and analysis of chemical components by ultra-high performance liquid chromatography-high-resolution tandem mass spectrometry (UHPLC-HRMS/MS), the extracts were also tested for toxicity in J774.A1 macrophages and human erythrocytes. Phenolic acids, flavonoids, acetogenins, alkaloids and lignans were identified in these extracts. Grow inhibition of promastigotes forms of Leishmania amazonensis and Leishmania braziliensis and the cytotoxicity in J774.A1 macrophages were estimated by the XTT method. The most promising results for L. amazonensis and L. braziliensis were shown by the ethanolic extract of the fruits of Hymenaea stigonocarpa and dichloromethane extract of the roots of Piper corcovadensis, with IC 50 of 160 and 150 µg mL-1, resp. Ethanolic extracts of A. muricata (leaf), B. uniflora (flower and leaf), M. pungens (fruit and leaf), N. megapotamica (leaf), and aqueous extract of H. stigonocarpa (fruit) showed IC 50 > 170 µg mL-1 for L. amazonensis and > 200 µg mL-1 for L. braziliensis. The extracts exhibited low cytotoxicity towards J774.A1 macrophages with CC 50 > 1000 µg mL-1 and hemolytic activity from 0 to 46.1 %.
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BACKGROUND: We aimed to determine the cellular recruitment (leukocyte rolling and adhesion) by which the Leishmania (Viannia) braziliensis, L. (Leishmania) amazonensis, and L. (Leishmania) major species in the mesenteric microcirculation of BALB/c mice. METHODS: Five experimental groups were considered: group 1 (L. braziliensis); group 2 (L. amazonensis); group 3 (L. major); group 4 (control group with PBS); group 5 (negative control group), analyzed 3, 6, 12, and 24 h after parasite inoculation. RESULTS: Infections by the different Leishmania species caused an increase in the number of rolling leukocytes: L. braziliensis a peak at 6 h; L. amazonensis and L. major a peak at 3 h. The Leishmania infections induced leukocyte adhesion: L. major and L. amazonensis showed an increase after 3 and 6 h, respectively. CONCLUSION: The kinetics of cellular recruitment in Leishmania infections, leading to infection susceptibility or resistance, indicates that distinct mechanisms regulate the initial response to Leishmania infection and determine its course.
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Leishmaniasis is a neglected disease that affects millions of people worldwide. This study aimed to analyze antileishmanial activity of Campomanesia xanthocarpa leaf essential oil (EO) on promastigote and amastigote forms of Leishmania amazonensis, cytotoxicity in murine macrophages and sheep erythrocytes. The essential oil (EO) was analyzed by gas chromatography/mass spectrophotometry. The main and most abundant compounds were sesquiterpene hydrocarbons (71.22%) such as trans-caryophyllene (7.87%), bicyclogermacrene (11.28%), and δ-cadinene (8.34%). The IC50 for promastigote and amastigote forms of L. amazonensis was 70 µg mL-1 and 6 µg mL-1, respectively. C. xanthocarpa EO was not cytotoxic for murine macrophages (CC50 1860 µg mL-1) and sheep erythrocytes (1.5%), presenting high selectivity index for protozoan (310). C. xanthocarpa EO induced effects on the morphology and ultrastructure of this parasite. The high activity for intracellular amastigote forms, low toxicity to murine macrophages, and erythrocytes, suggest that C. xanthocarpa EO is promising for the treatment of leishmaniasis.
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Antiprotozoários , Leishmania mexicana , Óleos Voláteis , Animais , Antiprotozoários/farmacologia , Citoplasma , Camundongos , Camundongos Endogâmicos BALB C , Óleos Voláteis/farmacologia , Organelas , OvinosRESUMO
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.
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Humanos , Leishmania braziliensis/genética , Leishmaniose Cutânea/diagnóstico , Reação em Cadeia da Polimerase , DNA de Protozoário/genética , Sensibilidade e Especificidade , MicroscopiaRESUMO
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.
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Leishmania braziliensis , Leishmaniose Cutânea , DNA de Protozoário/genética , Humanos , Leishmania braziliensis/genética , Leishmaniose Cutânea/diagnóstico , Microscopia , Reação em Cadeia da Polimerase , Sensibilidade e EspecificidadeRESUMO
INTRODUCTION: Aberrant Crypt (AC) and Aberrant Crypt Focus (ACF) are considered pre-neoplasic lesions, ranging from hyperplasia to different degrees of dysplasia in the colon. This work aimed to evaluate and quantify the chemopreventive activity of Zingiber officinale essential oil in the colorectal region of Wistar rats. MATERIALS AND METHODS: We extracted the essential oil from ginger rhizomes and carried out ACF induction, in rats, with 1.2 Dimethylhydrazine (DMH) at a 20 mg/kg dose. The experimental groups were GI (negative control); GII (positive induction control); GIII (DMH + essential oil); GIV (DMH +5-Florouracil) and GV (essential oil). The histological techniques used were methylene blue, hematoxylin-eosin (HE) dyeing, and immunohistochemistry (IHQ). RESULTS: The major essential oil compounds were citral (17.25%), δ-citral (10.25%), camphene (9.55%), α-zingiberene (7.57%), nerol (6.37%) and plelandrene (6.83%). For the presence of AC or ACF, we did not observe them in GI and GV, while in GII and GIII, they were observed, in high values, in both regions, but only in the distal region, there was a significant difference between them. For GIV, for both regions, there were significant lower numbers of AC when compared to GIII. As observed, with HE, there were hyperplastic and dysplastic ACF in the proximal and distal portions of the colon. For IHQ analyses, there were positively PCNA antibody marked cells in all experimental groups. Yet, there was no significant correlation of mitotic index among them. Moreover, the results of GIII compared to GIV were very similar. CONCLUSION: In this sense, the Zingiber officinale essential oil has good antioxidant potential because it presents a mixture of monoterpene and sesquiterpene compounds. Thus, it is able to develop a chemoprotective effect, as it presented similar results to the standard drug, showing cell proliferation control.
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1,2-Dimetilidrazina/toxicidade , Focos de Criptas Aberrantes/tratamento farmacológico , Antineoplásicos/farmacologia , Neoplasias Colorretais/tratamento farmacológico , Óleos Voláteis/farmacologia , Zingiber officinale/química , Focos de Criptas Aberrantes/induzido quimicamente , Focos de Criptas Aberrantes/patologia , Animais , Neoplasias Colorretais/induzido quimicamente , Neoplasias Colorretais/patologia , Masculino , Ratos , Ratos WistarRESUMO
Studies of the primers that were designed to detect New World Leishmania were systematically reviewed to report the characteristics of each target, detection limit, specificity of the primers designed and diagnostic sensibility. The papers identified in the databases PubMed and Web of Science involved 50 studies. Minicircle is the most applied target in molecular research for diagnosis, due to its high sensitivity in detecting Leishmania in different clinical samples, a characteristic that can be partially attributed to the higher number of copies of the minicircle per cell. The other molecular targets shown in this review were less sensitive to diagnostic use because of the lower number of copies of the target gene per cell, but more specific for identification of the subgenus and/or species. The choice of the best target is an important step towards the result of the research. The target allows the design of primers that are specific to the genus, subgenus or a particular species and also imparts sensitivity to the method for diagnosis. The findings of this systematic review provide the advantages and disadvantages of the main molecular targets and primers designed for New World Leishmania, offering information so that the researcher can choose the PCR system best suited to their research need. This is a timely and extremely thorough review of the primers designed for New World Leishmania.
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Primers do DNA/análise , DNA de Protozoário/análise , Leishmania/genética , Leishmaniose Cutânea/parasitologia , Reação em Cadeia da Polimerase/métodos , Humanos , Leishmania/isolamento & purificação , Limite de Detecção , Sensibilidade e EspecificidadeRESUMO
INTRODUCTION: Genetic polymorphisms define the cytokine production leading to susceptibility or resistance to diseases. We studied the cytokine polymorphism in the development of tegumentary leishmaniasis (TL). METHODS: Genotyping of TNF-α, TGF-ß1, IFN-γ, IL-6, and IL-10 were performed by polymerase chain reaction assay. RESULTS: G and C alleles of TGF- ß1 (codon 25) were the most common in controls and patients, respectively. G/G was the most frequent genotype in controls, and G/C and C/C in patients. CONCLUSIONS: G/G genotype of codon 25 in TGF-ß1 appeared to confer resistance, and G/C and C/C genotypes, susceptibility to TL in this population.
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Predisposição Genética para Doença/genética , Leishmaniose Cutânea/genética , Polimorfismo Genético/genética , Fator de Crescimento Transformador beta1/genética , Adulto , Idoso , Alelos , Biomarcadores , Brasil , Estudos de Casos e Controles , Citocinas/genética , Feminino , Genótipo , Humanos , Interferon gama/genética , Interleucina-10/genética , Interleucina-6/sangue , Interleucina-6/genética , Masculino , Pessoa de Meia-Idade , Estudos Retrospectivos , Fator de Necrose Tumoral alfa/genéticaRESUMO
Leishmaniasis is a neglected disease and drugs approved for its treatment often lead to abandonment, failure of therapy and even death. Photodynamic therapy (PDT) has been shown to be a promising, non-invasive and selective for a target region without requiring high-cost technology. Usually, it is employed a photosensitizing agent (PS) incorporated into nanoparticles (NP). Pluronics® P-123 and F-127 micelles are very interesting aqueous NP promoting efficient and selective delivery and less adverse effects. This study aimed to detect the activity of Pluronics® P-123 and F-127 themselves since there is a scarcity of data on these NP activities without drugs incorporation. This study evaluated, in vitro, the activity of Pluronics® against promastigotes and amastigotes of Leishmania amazonensis and also their cytotoxicities. Additionally, the determination of the mitochondria membrane potential in promastigotes, internalization of these Pluronics® in the parasite membrane and macrophages and its stability in the culture medium was evaluated. Results showed that Pluronics® did not cause significant damage to human red cells and promastigotes. The P-123 and F-127 inhibited the survival rate of L. amazonensis amastigotes, and also presented loss of mitochondrial membrane potential on promastigotes. The Pluronics® showed low cytotoxic activity on J774A.1 macrophages, while only P-123 showed moderate cytotoxicity for BALB/c macrophages. The stability of P-123 and F-127 in culture medium was maintained for ten days. In conclusion, the NP studied can be used for incorporating potent leishmanicidal chemotherapy, due to their selectivity towards macrophages, being a promising system for the treatment of cutaneous leishmaniasis.
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Sistemas de Liberação de Medicamentos/métodos , Leishmania/efeitos dos fármacos , Nanopartículas/química , Fotoquimioterapia/métodos , Poloxâmero/farmacologia , Animais , Relação Dose-Resposta a Droga , Eritrócitos/efeitos dos fármacos , Feminino , Macrófagos/efeitos dos fármacos , Potencial da Membrana Mitocondrial , Camundongos , Camundongos Endogâmicos BALB CRESUMO
The treatment of cutaneous leishmaniasis in associated with several adverse effects and therapeutic failure, resulting in patients' abandonment of treatment. Research on new drugs with leishmanicidal potential from medicinal plants is essential. The anti-Leishmania activity of Tetradenia riparia essential oil (TrEO) and its derivatives, such as the diterpene 6,7-dehydroroyleanone (TrROY), and the immunomodulatory effects of TrEO have been reported. However, few studies have investigated the effects of TrROY. The present study evaluated the modulation of cytokine production by murine macrophages that were infected with Leishmania amazonensis (6 parasites/macrophage) and treated with TrROY (0.1, 1, and 100 µg/ml). Cytokine levels were measured by flow cytometry. The results were analyzed using Student's t test at a 95% confidence interval. Microscopic counting was performed to evaluate the inhibitory effects of TrROY on intracellular infection. TrROY modulated the production of cytokines that are essential for the immune defense response to Leishmania, with a decrease in interleukin-4 (IL-4) levels and an increase in IL-12 levels. A TrROY concentration of 0.1 µg/ml was chosen for the subsequent experiments. This dose was chosen because it modulated IL-4/IL-12 release by murine macrophages that were infected with Leishmania and because it presented no cytotoxic effects. TrROY (0.1 µg/ml) induced a 31% reduction of the rate of infection in murine macrophages compared with untreated cells. TrROY may be a promising leishmanicidal agent. Further in vitro and in vivo studies should be conducted to evaluate the anti-Leishmania and immunomodulatory activity of TrROY.
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Abietanos/farmacologia , Antiprotozoários/farmacologia , Lamiaceae/química , Leishmania/efeitos dos fármacos , Leishmaniose Cutânea/imunologia , Macrófagos/imunologia , Óleos Voláteis/farmacologia , Abietanos/química , Animais , Antiprotozoários/química , Humanos , Interleucina-12/imunologia , Interleucina-4/imunologia , Leishmania/fisiologia , Leishmaniose Cutânea/tratamento farmacológico , Leishmaniose Cutânea/parasitologia , Macrófagos/parasitologia , Camundongos , Óleos Voláteis/química , Células RAW 264.7RESUMO
Abstract INTRODUCTION: Genetic polymorphisms define the cytokine production leading to susceptibility or resistance to diseases. We studied the cytokine polymorphism in the development of tegumentary leishmaniasis (TL). METHODS: Genotyping of TNF-α, TGF-β1, IFN-γ, IL-6, and IL-10 were performed by polymerase chain reaction assay. RESULTS: G and C alleles of TGF- β1 (codon 25) were the most common in controls and patients, respectively. G/G was the most frequent genotype in controls, and G/C and C/C in patients. CONCLUSIONS: G/G genotype of codon 25 in TGF-β1 appeared to confer resistance, and G/C and C/C genotypes, susceptibility to TL in this population.
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Humanos , Animais , Masculino , Feminino , Mordeduras de Serpentes/diagnóstico , Serpentes/anatomia & histologia , Mordeduras de Serpentes/epidemiologia , Serpentes/classificação , Serpentes/fisiologia , Brasil/epidemiologia , Colubridae , Erros de DiagnósticoRESUMO
AIM: The present study compared and evaluated morphological and quantitative alterations in the ileum of hamsters infected by two L. (V.) braziliensis strains isolated from patients with different lesion aspects and treatment responses. MAIN METHODS: Hamsters were infected in the left hindpaw with a suspension of promastigotes (2 × 107/100 µl) of two different strains of L. (V.) braziliensis. After 90 or 120 days, the animals were euthanized. Samples of the ileum and mesenteric lymph node were collected for histological examination and quantitative polymerase chain reaction. KEY FINDINGS: All infected animals developed similar profile of paw lesions. In peripheral blood there was an increase in the number of mononuclear cells which contributed to elevated global leukocytes count. Increases in the width and height of villi and width and depth of crypts were observed. The thickness of the muscular layers, submucosa, and intestinal wall also increased. Histopathological alterations were observed, including inflammatory infiltrate in crypts and a large number of immune cells in the lamina propria, submucosa, and muscular layer. Immune cells were found inside myenteric ganglia, with an increase in the number of intraepithelial lymphocytes. Leishmania DNA was detected in the ileum and mesenteric lymph node at both times of infection. The presence of amastigotes in the ileum was revealed by immunohistochemistry. SIGNIFICANCE: The infection with different strains of L. (V.) braziliensis causes morphological and quantitative alterations in the ileum of hamsters and the parasite can migrate to the mesenteric lymph node and intestine.
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Íleo/parasitologia , Leishmania braziliensis/patogenicidade , Leishmaniose Cutânea/parasitologia , Animais , DNA de Protozoário/genética , Modelos Animais de Doenças , Feminino , Interações Hospedeiro-Parasita , Íleo/imunologia , Íleo/patologia , Leishmania braziliensis/genética , Leishmania braziliensis/imunologia , Leishmaniose Cutânea/imunologia , Leishmaniose Cutânea/patologia , Linfonodos/parasitologia , Mesocricetus , Carga Parasitária , Fatores de TempoRESUMO
This clinical case presents a patient with a raised and ulcerative lesion with erythematous edges in the mouth, on the lower lip that was unsuccessfully treated as herpes labialis. Clinical data and laboratory tests (Montenegro skin test, indirect immunofluorescence, direct parasite search and polymerase chain reaction) led to the diagnosis of American tegumentary leishmaniasis caused by Leishmania (Viannia) sp. Treatment with pentavalent antimonial (Glucantime®) for 120 days was not effective and administration of amphotericin B for 30 days resulted in wound healing. Glucantime® treatment protocol was longer than the recommended by the Brazilian Ministry of Health in the handbook of mucosal leishmaniasis. This suggests that amphotericin B should have been administered earlier, preventing the psychological and social problems faced by the patient. This study reports a rare clinical case of primary mucosal leishmaniasis on the lip that had a delayed diagnosis, highlighting the precariousness in the management of disease and showing that, despite the importance of leishmaniasis in Brazil, it is still neglected by health professionals.
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Anfotericina B/uso terapêutico , Antiprotozoários/uso terapêutico , Leishmaniose Mucocutânea/diagnóstico , Doenças Labiais/diagnóstico , Adulto , Técnica Indireta de Fluorescência para Anticorpo , Humanos , Leishmania/imunologia , Leishmaniose Mucocutânea/tratamento farmacológico , Doenças Labiais/tratamento farmacológico , Doenças Labiais/parasitologia , MasculinoRESUMO
OBJECTIVE: We researched articles that used photodynamic therapy (PDT) for skin wound healing in humans. METHODS: The systematic review was conducted through scientific articles that investigated the action of PDT on wound healing in humans, published from July 2005 to March 2017, in the data bases PubMed and LILACS. RESULTS: The main types of wound described in selected articles in this review were chronic ulcer and non-melanoma skin cancer. For accomplishing the PDT, second generation of photosensitizing agents with laser or light emitting diode were used. The studies demonstrated that PDT contribute in several ways to the wound healing process: leading to cellular death; reducing or increasing inflammation; stimulating fibroblasts proliferation and, consequently, of collagen and elastin; raising transforming growth factor beta and metalloproteinases. Based on this, PDT provided good results in wound healing process, acting in several steps and accelerating tissue repair. CONCLUSIONS: PDT improved healing in many wound models in humans, revealing itself as a promising therapeutic modality for stimulating wound healing and remodelling.
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Fotoquimioterapia/métodos , Fármacos Fotossensibilizantes/uso terapêutico , Cicatrização/efeitos da radiação , Morte Celular/efeitos da radiação , Colágeno/biossíntese , Elastina/biossíntese , Fibroblastos/metabolismo , Humanos , Metaloproteases/biossíntese , Fotoquimioterapia/efeitos adversos , Fotoquimioterapia/instrumentação , Neoplasias Cutâneas/radioterapia , Úlcera Cutânea/radioterapia , Fatores de Crescimento Transformadores/biossínteseRESUMO
Introduction: American tegumentary leishmaniasis (ATL) is a zoonotic disease caused by protozoan parasites of the genus Leishmania that affects the skin and mucous membrane. Currently, the available drugs for the treatment are injectable, with side effects, long-term treatment regimen and there is the possibility of drug resistance. Thus, alternative therapies have been tested, including photodynamic therapy (PDT). We evaluated the efficacy of PDT on its own and associated with the prescribed ATL treatment. Methods: BALB/c mice were infected with Leishmania (Leishmania) amazonensis and divided into 6 groups: Gluc+PDT, treated with Glucantime® and photodynamic therapy (PDT) with methylene blue (MB)/red LED (light-emitting diode); Gluc, treated with Glucantime®; PDT, treated with PDT with MB/red LED; Ampho+PDT, treated with amphotericin and PDT with MB/red LED; Ampho, treated with amphotericin; and control, which were infected but not treated. Two treatment cycles were performed. After 165 days of infection, the parasite load was determined. Results: Statistical differences were not found (P>0.05) between measures of volume and thickness of the infected footpads in the treated groups when compared with the control group. However, there was a significant reduction (P<0.05) in the parasitic load of the popliteal lymph nodes of the Gluc+PDT, Gluc, PDT and Ampho groups when compared to the control group. In the histological analysis of the infected footpads, the Gluc+PDT group presented a smaller amount of amastigote nests and lower intensity of the mononuclear infiltrate when compared to the Gluc and PDT groups. Conclusion: The results showed that although there is no significant difference in the evaluations of footpad size (thickness and volume), there is a downward measurement tendency in the Gluc+PDT group, as it can be observed by volume data and corroborated by parasite negative load.
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Leishmaniasis is caused by protozoa of the Leishmania genus, which is divided into subgenus Viannia and Leishmania. In humans, the course of infection largely depends on the host-parasite relationship and primarily of the infective species. The objective of the present study was to design specific primers to the identification of Leishmania species using multiplex PCR. Four primers were designed, based on the GenBank sequences of the kDNA minicircle, amplifying 127 bp for subgenus Viannia, 100 bp for L. amazonensis, and 60 bp for Leishmania donovani complex and L. major. None of the primers amplified Trypanosoma cruzi or L. mexicana. The limit of detection of multiplex PCR was 2 × 10-5 parasites for L. braziliensis, 2 x 10-3 parasites for L. amazonensis, and 1.4 × 10-3 parasites for L. infantum. The high sensitivity of multiplex PCR was confirmed by the detection of parasites in different biological samples, including lesion scrapings, spleen imprinting of a hamster, sandflies, and blood. The multiplex PCR that was developed herein presented good performance with regard to detecting and identifying the parasite in different biological samples and may thus be useful for diagnosis, decision making with regard to the proper therapeutic approach, and determining the geographic distribution of Leishmania species.
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Primers do DNA/genética , DNA de Cinetoplasto/genética , Leishmania/classificação , Leishmania/genética , Leishmaniose/diagnóstico , Reação em Cadeia da Polimerase Multiplex/métodos , Animais , Sequência de Bases , Cricetinae , Interações Hospedeiro-Parasita , Humanos , Leishmania/isolamento & purificação , Leishmaniose/parasitologia , Psychodidae/parasitologia , Sensibilidade e Especificidade , Baço , Trypanosoma cruzi/genéticaRESUMO
Introduction: The treatment of cutaneous leishmaniasis (CL) is based primarily on the use of pentavalent antimonials, which may lead to many side effects limiting their use. Photodynamic therapy (PDT) is an alternative for the treatment of CL, and some xanthene dyes have the potential for use in PDT. Methods: The xanthenes rose bengal B (RB) and its derivatives rose bengal methyl ester (RBMET), and butyl ester (RBBUT) were analyzed for leishmanicidal activity against promastigotes and intracellular amastigotes of Leishmania amazonensis. Cytotoxicity was assessed in J774.A1 macrophages. Results: RB derivates RBMET (IC50 9.83 µM), and RBBUT (IC50 45.08 µM) showed leishmanicidal activity, however, were toxic to J774.A1 macrophages, resulting in low selectivity index. Conclusion: The RBMET and RBBUT showed to be effective against the L. amazonensis and the low selectivity index presented may not be a limitation for their use in PDT to CL treatment.
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The aim of this study was to better understand the dynamics of Leishmania sand flies and reservoirs in São Domingos ranch, Arapongas municipality, Paraná State, an anthropic environment in an endemic area of cutaneous leishmaniasis (CL). Sand flies were collected in wild animal burrows, residences and in the forest, with Falcão light trap (FA), Shannon trap (SH) and quadrangular pyramidal trap (QP). The search for Leishmania was made on sand flies, biological samples of wild rodents and dogs using PCR and culture; while parasite direct search (DS) was carried out on animal skin lesions; infection of gold hamsters; and indirect immunofluorescence (IIF) test in dog blood samples. Eighty eight (88) sand flies were collected with FA traps and 526 sand flies using the SH trap, with a predominance of Pintomyia fischeri. Six hundred and one (601) specimens of Brumptomyia brumpti were collected in armadillo burrows, with the QP trap. Seventeen (17) wild rodents were captured, six of them had skin lesions with characteristics of Leishmania infection. Even though no positive test was found for Leishmania, epidemiological surveillance should be maintained, remembering that the human buildings are situated only 50 m from the forest. Considering the species of wild animals and sandflies found in São Domingos, the negative test found do not exclude the existence of the Leishmania transmission cycle in this preservation area.