Identification of causative Leishmania species in Giemsa-stained smears prepared from patients with cutaneous leishmaniasis in Peru using PCR-RFLP

Background: Dezful and its suburbs, as the second city in Khuzestan Province, southwest of Iran, has been an endemic area of cutaneous leishmaniasis (CL) with a low incidence rate since the last decades. However, the disease incidence has rapidly increased, and now is considered as a re-emerging parasitic disease in the area. Objectives: The aim of this study was to identify the most prevalent CL species in Dezful Region. Methods: A total of 196 microscopically confirmed slides from CL patients referred to Dezful Health Center were randomly collected in the period of 2015 - 2016. After DNA extraction from microscopically positive slides, polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) was carried out on 61 eligible specimens using ribosomal internal transcribed spacer 1 (ITS1) gene. The HaeIII restriction enzyme was used for the identification of species. Results: Samples were randomly collected from 196 acute CL cases, including 110 (56.2%) males and 86 (43.8%) females. Most infections were seen in the age range of 2 - 7 years (65/196, 33.1%). Totally, 60.1% of the cases had only one lesion, and half of the lesions appeared on hands. Furthermore, 162 (82.7%) cases were referred to Dezful Health Center in the cold seasons (autumn and winter). Results of PCR-RFLP on 61 eligible isolates showed that 60 (98.4%) isolates were Leishmania major, and only 1 (1.6%) isolate was Leishmania tropica. Conclusions: Our findings indicated that L. major is the main agent of re-emerged CL in Dezful and its suburbs, and the disease is a zoonosis.

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The Investigation of the Association of Cutaneous Leishmaniasis in Biopsy Specimens of the Patients with Granulomatous Disease and Skin Cancer Using the Molecular Method

Iranian Journal of Parasitology ◽

10.18502/ijpa.v15i3.4194 ◽

2020 ◽

Author(s):

Gülnaz ÇULHA ◽

Asena Çiğdem DOĞRAMACI ◽

Sibel HAKVERDİ ◽

İlke Evrim SEÇİNTİ ◽

Özkan ASLANTAŞ ◽

...

Keyword(s):

Differential Diagnosis ◽

Cutaneous Leishmaniasis ◽

Leishmania Species ◽

Pcr Analysis ◽

Biopsy Specimens ◽

Skin Cancers ◽

Pcr Rflp ◽

Granulomatous Dermatitis ◽

Tissue Specimens ◽

Granulomatous Diseases

Background: Clinically, cutaneous leishmaniasis (CL) can be confused with granulomatous diseases and skin cancers, and it may lead to erroneous diagnosis and treatment. Diagnosis based and histopathology can have some difficulties due to low number of parasites, especially in chronic CL cases. We aimed to emphasize the necessity of considering CL in the differential diagnosis for cases of granulomatous diseases and basal cell carcinoma, particularly in areas where CL is endemic. Methods: One hundred and seven paraffin-embedded tissue biopsy specimens were selected from the archive, as of 2002, of Pathology Department, School of Medicine, University of Hatay Mustafa Kemal in Hatay, Turkey. After DNA isolation, performed with the samples were used for PCR analysis with specific 13A, 13B primers targeting kinetoplastid DNA (kDNA) found in all Leishmania species. Another PCR was performed with LITSR and L5.8S primers targeting ITS-1 internal-transcribed-spacer-1 (ITS-1) region to subtype positive samples. Then these samples were further analyzed for subtyping with PCR-RFLP using HaeIII enzyme (BsuRI). Results: Ten out of 107 tissue specimens were positive via kDNA-PCR. Lupus vulgaris, sarcoidosis, skin lymphoma and Leishmania cutis appeared in 9 out of 10 positive specimens. One of the cases presented with a mass on the cheek and was pre-diagnosed with hemangioma, but leishmaniasis did not appear. All of 10 specimens were diagnosed as granulomatous dermatitis. Two out of 10 samples, found positive with kDNA-PCR, were analyzed with ITS-1-PCR and identified as L. infantum/donovani after RFLP. Conclusion: Molecular methods should be utilized in the differential diagnosis of CL to eliminate false diagnoses of granulomatous diseases and skin cancers.

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The Role of CD1a expression in the diagnosis of cutaneous leishmaniasis, its relationship with Leishmania species and clinicopathological features

Dermatologic Therapy ◽

10.1111/dth.14977 ◽

2021 ◽

Author(s):

Yasemin Yuyucu Karabulut ◽

Funda KuŞ Bozkurt ◽

Ümit Türsen ◽

Gül Bayram ◽

Gülhan Örekeci Temel ◽

...

Keyword(s):

Cutaneous Leishmaniasis ◽

Leishmania Species ◽

Clinicopathological Features

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Film-Forming Systems for the Delivery of DNDI-0690 to Treat Cutaneous Leishmaniasis

Pharmaceutics ◽

10.3390/pharmaceutics13040516 ◽

2021 ◽

Vol 13 (4) ◽

pp. 516

Author(s):

Katrien Van Bocxlaer ◽

Kerri-Nicola McArthur ◽

Andy Harris ◽

Mo Alavijeh ◽

Stéphanie Braillard ◽

...

Keyword(s):

Drug Delivery ◽

Cutaneous Leishmaniasis ◽

Mouse Skin ◽

Parasite Load ◽

Leishmania Species ◽

Lesion Size ◽

Drying Time ◽

Sustained Drug Delivery ◽

Film Forming ◽

The Impact

In cutaneous leishmaniasis (CL), parasites reside in the dermis, creating an opportunity for local drug administration potentially reducing adverse effects and improving treatment adherence compared to current therapies. Polymeric film-forming systems (FFSs) are directly applied to the skin and form a thin film as the solvent evaporates. In contrast to conventional topical dosage forms, FFSs strongly adhere to the skin, favouring sustained drug delivery to the affected site, reducing the need for frequent applications, and enhancing patient compliance. This study reports the first investigation of the use of film-forming systems for the delivery of DNDI-0690, a nitroimidazole compound with potent activity against CL-causing Leishmania species. A total of seven polymers with or without plasticiser were evaluated for drying time, stickiness, film-flexibility, and cosmetic attributes; three FFSs yielded a positive evaluation for all test parameters. The impact of each of these FFSs on the permeation of the model skin permeant hydrocortisone (hydrocortisone, 1% (w/v) across the Strat-M membrane was evaluated, and the formulations resulting in the highest and lowest permeation flux (Klucel LF with triethyl citrate and Eudragit RS with dibutyl sebacate, respectively) were selected as the FFS vehicle for DNDI-0690. The release and skin distribution of the drug upon application to Leishmania-infected and uninfected BALB/c mouse skin were examined using Franz diffusion cells followed by an evaluation of the efficacy of both DNDI-0690 FFSs (1% (w/v)) in an experimental CL model. Whereas the Eudragit film resulted in a higher permeation of DNDI-0690, the Klucel film was able to deposit four times more drug into the skin, where the parasite resides. Of the FFSs formulations, only the Eudragit system resulted in a reduced parasite load, but not reduced lesion size, when compared to the vehicle only control. Whereas drug delivery into the skin was successfully modulated using different FFS systems, the FFS systems selected were not effective for the topical application of DNDI-0690. The convenience and aesthetic of FFS systems alongside their ability to modulate drug delivery to and into the skin merit further investigation using other promising antileishmanial drugs.

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Macrophage Polarization in the Skin Lesion Caused by Neotropical Species of Leishmania sp

Journal of Immunology Research ◽

10.1155/2021/5596876 ◽

2021 ◽

Vol 2021 ◽

pp. 1-8

Author(s):

Carmen M. Sandoval Pacheco ◽

Gabriela V. Araujo Flores ◽

Kadir Gonzalez ◽

Claudia M. de Castro Gomes ◽

Luiz F. D. Passero ◽

...

Keyword(s):

Cutaneous Leishmaniasis ◽

Macrophage Polarization ◽

Leishmania Species ◽

Skin Lesions ◽

Host Cells ◽

M2 Macrophage ◽

M2 Macrophages ◽

Intracellular Parasites ◽

Skin Biopsies ◽

Acquired Immune Responses

Macrophages play important roles in the innate and acquired immune responses against Leishmania parasites. Depending on the subset and activation status, macrophages may eliminate intracellular parasites; however, these host cells also can offer a safe environment for Leishmania replication. In this sense, the fate of the parasite may be influenced by the phenotype of the infected macrophage, linked to the subtype of classically activated (M1) or alternatively activated (M2) macrophages. In the present study, M1 and M2 macrophage subsets were analyzed by double-staining immunohistochemistry in skin biopsies from patients with American cutaneous leishmaniasis (ACL) caused by L. (L.) amazonensis, L. (V.) braziliensis, L. (V.) panamensis ,and L. (L.) infantum chagasi. High number of M1 macrophages was detected in nonulcerated cutaneous leishmaniasis (NUCL) caused by L. (L.) infantum chagasi ( M 1 = 112 ± 12 , M 2 = 43 ± 12 cells/mm2). On the other side, high density of M2 macrophages was observed in the skin lesions of patients with anergic diffuse cutaneous leishmaniasis (ADCL) ( M 1 = 195 ± 25 , M 2 = 616 ± 114 ), followed by cases of localized cutaneous leishmaniasis (LCL) caused by L. (L.) amazonensis ( M 1 = 97 ± 24 , M 2 = 219 ± 29 ), L. (V.) panamensis ( M 1 = 71 ± 14 , M 2 = 164 ± 14 ), and L. (V.) braziliensis ( M 1 = 50 ± 13 , M 2 = 53 ± 10 ); however, low density of M2 macrophages was observed in NUCL. The data presented herein show the polarization of macrophages in skin lesions caused by different Leishmania species that may be related with the outcome of the disease.

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Label-free quantitative proteomic analysis reveals potential biomarkers for early healing in cutaneous leishmaniasis

PeerJ ◽

10.7717/peerj.6228 ◽

2019 ◽

Vol 6 ◽

pp. e6228 ◽

Cited By ~ 4

Author(s):

Andrés Montoya ◽

Manuel Carlos López ◽

Ivan D. Vélez ◽

Sara M. Robledo

Keyword(s):

Cutaneous Leishmaniasis ◽

Differential Expression ◽

Calcium Binding ◽

Therapeutic Response ◽

Label Free ◽

Early Biomarkers ◽

Parasitological Diagnosis ◽

Pcr Rflp ◽

Before And After ◽

Globally Distributed

Background Leishmaniasis is a parasitic disease caused by more than 20 species of the Leishmania genus. The disease is globally distributed and is endemic in 97 countries and three territories in the tropical and subtropical regions. The efficacy of the current treatments is becoming increasingly low either due to incomplete treatment or resistant parasites. Failure of treatment is frequent, and therefore, the search for early biomarkers of therapeutic response in cutaneous leishmaniasis (CL) is urgently needed. Objective The aim of this study was to compare the proteomic profiles in patients with CL before and after 7 days of treatment and identify early biomarkers of curative response. Methods Four patients with a parasitological diagnosis of leishmaniasis with confirmation of species by PCR-RFLP were recruited. All patients had a single lesion, and a protein from the middle of the ulcer was quantified by liquid chromatography and mass spectrometry. Results A total of 12 proteins showed differential expression in the comparative LC-electrospray ionization MS/MS (LC-ESI-MS/MS) triplicate analysis. Seven of them were up-regulated and five of them were down-regulated. Calcium binding proteins A2, A8, and A9 and hemoglobin subunits alpha-2 and delta showed high correlation with epidermis development and immune response. Conclusion We identified changes in the profiles of proteins that had a positive therapeutic response to the treatment. The proteins identified with differential expression are related to the reduction of inflammation and increased tissue repair. These proteins can be useful as biomarkers for early monitoring of therapeutic response in CL.

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