Diagnosis Of Leishmaniasis Research Articles

Design and characterization of high-affinity synthetic peptides as bioreceptors for diagnosis of cutaneous leishmaniasis.

Cutaneous leishmaniasis (CL) is one of the illnesses caused by Leishmania parasite infection, which can be asymptomatic or severe according to the infecting Leishmania strain. CL is commonly diagnosed by directly detecting the parasites or their DNA in tissue samples. New diagnostic methodologies target specific proteins (biomarkers) secreted by the parasite during the infection process. However, specific bioreceptors for the in vivo or in vitro detection of these novel biomarkers are rather limited in terms of sensitivity and specificity. For this reason, we here introduce three novel peptides as bioreceptors for the highly sensitive and selective identification of acid phosphatase (sAP) and proteophosphoglycan (PPG), which have a crucial role in leishmaniasis infection. These high-affinity peptides have been designed from the conservative domains of the lectin family, holding the ability to interact with the biological target and produce the same effect than the original protein. The synthetic peptides have been characterized and the affinity and kinetic constants for their interaction with the targets (sAP and PPG) have been determined by a surface plasmon resonance biosensor. Values obtained for KD are in the nanomolar range, which is comparable to high-affinity antibodies, with the additional advantage of a high biochemical stability and simpler production. Pep2854 exhibited a high affinity for sAP (KD = 1.48 nM) while Pep2856 had a good affinity for PPG (KD 1.76 nM). This study evidences that these peptidomimetics represent a novel alternative tool to the use of high molecular weight proteins for biorecognition in the diagnostic test and biosensor devices for CL.Graphical abstract Supplementary InformationThe online version contains supplementary material available at 10.1007/s00216-021-03424-2.

Canine Visceral Leishmaniasis in an Area of Sporadic Transmission in Brazil.

Canine visceral leishmaniasis is a zoonotic disease caused by the protozoan Leishmania infantum in Latin America. Visceral leishmaniasis (VL) diagnosis in Brazil includes two serological tests according to the Ministry of Health (MH) protocol. Sensitivity and specificity of diagnostic tests, as well as clinical signs of VL, are usually reported in disease-endemic areas; however, it is known that local epidemiological factors can influence these results. This study aimed to evaluate the clinical features, sensitivity, and specificity of TR-DPP® and EIE-LVC in naturally infected dogs in a region of sporadic VL transmission to humans in Brazil. A total of 288 dogs were clinically evaluated and serological and parasitological (lymph node aspirates) samples were collected for VL diagnosis. TR-DPP and EIE-LVC showed poor sensitivity (0.62 and 0.44, respectively) to detect infected animals, compared with the direct parasitological examination, which is considered a gold standard method. Thus, the protocol of MH presented low sensitivity (0.42) to estimate prevalence and control measures in this region. TR-DPP presented a high negative predictive value (0.89), resulting in its indication as a confirmatory test in sporadic transmission areas. Classical clinical signs of VL were not frequently observed; therefore, clinical scoring systems might not be useful in this region. Veterinarians of nonendemic areas should be alert for asymptomatic dogs, especially those presenting lymph adenomegaly.

Assessment of a recombinant protein from Leishmania infantum as a novel tool for Visceral Leishmaniasis (VL) diagnosis in VL/HIV co-infection cases.

Visceral Leishmaniasis and HIV-AIDS coinfection (VL/HIV) is considered a life-threatening pathology when undiagnosed and untreated, due to the immunosuppression caused by both diseases. Serological tests largely used for the VL diagnosis include the direct agglutination test (DAT), ELISA and immunochromatographic (ICT) assays. For VL diagnosis in HIV infections, different studies have shown that the use of the DAT assay facilitates the VL diagnosis in co-infected patients, since the performance of the most widely used ELISA and ICT tests, based on the recombinant protein rK39, are much less efficient in HIV co-infections. In this scenario, alternative recombinant antigens may help the development of new serological diagnostic methods which may improve the VL diagnosis for the co-infection cases. This work aimed to evaluate the use of the recombinant Lci2 antigen, related to, but antigenically more diverse than rK39, for VL diagnosis in co-infected sera through ELISA assays. A direct comparison between recombinant Lci2 and rK39 was thus carried out. The two proteins were first tested using indirect ELISA with sera from VL afflicted individuals and healthy controls, with similar performances. They were then tested with two different sets of VL/HIV co-infected cases and a significant drop in performance, for one of these groups, was observed for rK39 (32% sensitivity), but not for Lci2 (98% sensitivity). In fact, an almost perfect agreement (Kappa: 0.93) between the Lci2 ELISA and DAT was observed for the coinfected VL/HIV patients. Lci2 then has the potential to be used as a new tool for the VL diagnosis of VL/HIV co-infections.

Immunoblot for the Diagnosis of Cutaneous Leishmaniasis in French Guiana.

.Cutaneous leishmaniasis (CL) is firmly established in South America. We aimed to assess the detection of IgG antibodies against 14 and/or 16 kDa antigens by immunoblot (IB) for CL serological diagnosis in French Guiana, an area where many endemic pathogens could interfere with it. This study was performed retrospectively on sera from 141 patients at the Cayenne tertiary hospital: 30 were patients with confirmed CL, 71 were diagnosed with various other endemic pathogens, 11 were diagnosed with an autoimmune disease, and 29 controls had no history of CL. Antibodies bound to the 14 and/or 16 kDa antigens in 27 of the 30 CL patients’ sera and in 39 of the 111 non-CL patients’ sera (26 from the infectious diseases group, four from the autoimmune diseases group, and nine from the dermatology department). The method tested showed a high sensitivity (90%) and a low specificity (66%), and a diagnosis odds ratio of 17.5 (95% CI [4.6–78.0]). This IB may be helpful to exclude the diagnosis of CL, prompting physicians to look for another diagnosis in the case of a negative IB.

Presumptive Diagnosis of Cutaneous Leishmaniasis

Introduction: Cutaneous Leishmaniasis is a neglected tropical disease caused by a parasite. The most common presumptive diagnostic tool for this disease is the visual examination of the associated skin lesions by medical experts. Here, a mobile application was developed to aid this pre-diagnosis using an automatic image recognition software based on a convolutional neural network model.Material and Methods: A total of 2022 images of cutaneous diseases taken from 2012 to 2018 were used for training. Then, in 2019, machine learning techniques were tested to develop an automatic classification model. Also, a mobile application was developed and tested against specialized human experts to compare its performance.Results: Transfer learning using the VGG19 model resulted in a 93% accuracy of the classification model. Moreover, on average, the automatic model performance on a randomly selected skin image sample revealed a 99% accuracy while, the ensemble prediction of seven human medical expert’s accuracy was 83%.Conclusion: Mobile skin monitoring applications are crucial developments for democratizing health access, especially for neglected tropical diseases. Our results revealed that the image recognition software outperforms human medical experts and can alert possible patients. Future developments of the mobile application will focus on health monitoring of Cutaneous Leishmaniasis patients via community leaders and aiming at the promotion of treatment adherence.

Genomic DNA extraction and amplification of Leishmania donovani using polymerase chain reaction (PCR) from archived, Giemsa- stained slides

In Sri Lanka, diagnosis of cutaneous leishmaniasis (CL) is usually based on clinical features and direct microscopy examinations. Polymerase chain reaction (PCR) for Leishmania is usually performed on tissue samples. In this study, we extracted DNA from archived Giemsa-stained slides which were prepared from cutaneous lesions. A total number of 85 Giemsa-stained slides fixed between 2008-2017 were selected. All the slides were examined using light microscopy and the number of amastigotes in positive smears was recorded. A nested PCR was carried out to amplify the 385 bp fragment of L. donovani kinetoplast mini-circle sequence. All 40 positive slides had only 1-2 amastigotes per slide. Of these, only 20% were PCR positive. Of the 45 negative slides, only one gave positive PCR result. Further studies are required to confirm the efficacy of PCR on Giemsa-stained smears in our setting.

Improved Performance of ELISA and Immunochromatographic Tests Using a New Chimeric A2-Based Protein for Human Visceral Leishmaniasis Diagnosis.

Methods A total of 1028 sera samples were used for the development and validation of ELISA (321 samples from L. infantum-infected patients, 62 samples from VL/AIDS coinfected patients, 236 samples from patients infected with other diseases, and 409 samples from healthy donors). A total of 520 sera samples were used to develop and validate ICT (249 samples from L. infantum-infected patients, 46 samples from VL/AIDS coinfected patients, 40 samples from patients infected with other diseases, and 185 samples from healthy donors). Findings. Using the validation sera panels, DTL-4-based ELISA displayed an overall sensitivity of 94.61% (95% CI: 89.94-97.28), a specificity of 99.41% (95% CI: 96.39-99.99), and an accuracy of 97.02% (95% CI: 94.61-98.38), while for ICT, sensitivity, specificity, and accuracy values corresponded to 91.98% (95% CI: 86.65-95.39), 100.00% (95% CI: 96.30-100.00), and 95.14% (95% CI: 91.62-97.15), respectively. When testing sera samples from VL/AIDS coinfected patients, DTL-4-ELISA displayed a sensitivity of 77.42% (95% CI: 65.48-86.16), a specificity of 99.41% (95% CI: 96.39-99.99), and an accuracy of 93.51% (95% CI: 89.49%-96.10%), while for DTL-4-ICT, sensitivity was 73.91% (95% CI: 59.74-84.40), specificity was 90.63% (95% CI: 81.02-95.63), and accuracy was 82.00% (95% CI: 73.63-90.91). Conclusion DTL-4 is a promising candidate antigen for serodiagnosis of VL patients, including those with VL/AIDS coinfection, when incorporated into ELISA or ICT test formats.

Trends in the Epidemiology of Leishmaniasis in the City of Barcelona (1996-2019).

Background: Leishmaniasis is a neglected zoonosis produced by 20 different flagellated parasites of the Leishmania genus, a protozoan transmitted to humans and other vertebrates by the bite of dipteran insects of the Phlebotominae subfamily. It is endemic in Mediterranean countries and the number of cases is expected to increase due to climate change and migration. Prioritizing public health interventions for prevention and control is essential. The objective was to characterize the epidemiology and temporal trends in the incidence of human leishmaniasis in the city of Barcelona, between the years 1996 and 2019.Methods: A population-based, analytical observational study among residents in the city of Barcelona was conducted of all the cases of leishmaniasis reported between 1996 and 2019 to the Public Health Agency. The epidemiological survey contains clinical, diagnostic, and epidemiological data, including contact with suspicious mammals or insects. Annual incidence-rates were calculated by sex, age, and country of origin. Chi-square tests were used to assess association between studied risk factors, periods of time and type of leishmaniasis.Results: During the study period a total of 177 cases of leishmaniasis were reported in Barcelona, being 74.6% (n = 132) of the total cases in Spanish born, although within the foreign-born population the incidence was higher. Median age was 34 years (IQR = 10–48) and 121 (66.8%) were male. The main type was cutaneous (46%) followed by visceral (35.1%). The cumulative incidence was 0.47 per 100,000 inhabitants, with the highest incidence found in 2017 (1.60 per 100,000 inhabitants). A higher incidence was observed in the 0–4-year-old group (1.73 per 100,000 inhabitants), but increased during the study period for all age groups. There was an increase of foreign origin cases, and a decrease in the number of cases associated to any immunosuppression.Conclusion: In Barcelona, leishmaniasis incidence continues to be higher in people under 5 years of age, and 25–64 years old males, but it has also increased in population from foreign country of birth. There is an increase of the cases since 2016, probably due to the changes in the notification system, increasing the diagnosis of cutaneous leishmaniasis. Improvements in the current surveillance system are needed. Notification of the disease, vector, and reservoir control activities are also essential for the control of the disease.

Real-Time Fluorimetry Loop-Mediated Isothermal Amplification for Diagnosis of Leishmaniasis and as a Tool for Assessment of Cure for Post-Kala-Azar Dermal Leishmaniasis.

Despite the dwindling number of visceral leishmaniasis (VL) cases in India, there is an urgent need for early and unequivocal diagnostics for controlling and preventing the reemergence of VL. Post-kala-azar dermal leishmaniasis (PKDL), a dermal sequela of VL, serves as a reservoir of the parasite. Diagnosis of PKDL, especially the macular variant, is challenging and poses impediment toward attainment of VL elimination. In this study, a real-time fluorimetry loop-mediated isothermal amplification (RealAmp) assay has been established for the detection of different clinical manifestations of leishmaniasis. The study included 150 leishmaniasis patients (25 VL, 25 cutaneous leishmaniasis [CL], and 100-PKDL) along with 120 controls. The assay demonstrated sensitivity of 100% (95% CI: 86.68-100) for diagnosis of VL and PKDL (95% CI: 79.61-100) and 96% (95% CI: 86.68-100) for CL with 100% specificity. Moreover, considering the cardinal role of PKDL, diagnosis using minimally invasive slit aspirate was explored, which demonstrated remarkable sensitivity of 96% (95% CI: 87.64-98.47). As a test of cure for PKDL, RealAmp successfully detected parasite in two of posttreatment cases who later reported relapse on follow-up. Also, direct sample lysis using slit aspirate was attempted in a small group that yielded sensitivity of 89% (95% CI: 67.20-96.90). RealAmp depicted excellent diagnostic accuracy in the diagnosis of leishmaniasis in concordance with the established SYBR Green I-based (Molecular Probes, Eugene, OR) visual loop-mediated isothermal amplification (LAMP) and the reference comparator real-time PCR. The study endorsed the employment of LAMP either as visual-LAMP or RealAmp for an accurate and expeditious diagnosis of PKDL and as a tool for assessment of cure.

Are Nanobiosensors an Improved Solution for Diagnosis of Leishmania?

Leishmaniasis is one of the deadliest neglected tropical diseases affecting 12–15 million people worldwide, especially in middle- and low-income countries. Rapid and accurate diagnosis of the disease is important for its adequate management and treatment. Several techniques are available for the diagnosis of leishmaniasis. Among these, parasitological and immunological tests are most widely used. However, in most cases, the utilized diagnostic techniques are not good enough, showing cross-reactivity and reduced accuracy. In recent years, many new methods have been reported with potential for improved diagnosis. This review focuses on the diagnosis of Leishmania exploring the biosensors and nanotechnology-based options for their detection. New developments including the use of nanomaterials as fluorophores, fluorescence quenchers as reducing agents and as dendrimers for signal improvement and amplification, together with the use of aptamers to replace antibodies are described. Future research opportunities to overcome the current limitations on the available diagnostic approaches are also discussed.

POS-170 LEISHMANIASIS COMPLICATE SYSTEMIC LUPUS ERYTHEMATOSUS: ABOUT 7 CASES

The association between leishmania (Lsc) and systemic lupus erthematosus (SLE) is rare. It is particularly serious in cases of visceral leishmania (VL). It is a parasitic infection whose hallmarks may mimic SLE symptoms especially when the two pathologies appear concomitantly. The prognosis of SLE depends on the one hand on the rapidity of diagnosis and on the other hand on the quality of treatment. The aim of this study was to analyze the characteristics of infection in leishmania in patients affected by SLE. It was a retrospective study including 07 patients hospitalized for leishmania and systemic lupus erthematosus (SLE) in the Internal Medicine Department of the Charles Nicolle hospital during the period between January 1976 and November 2020. All patients fulfilled four or more criteria defined by the American College of Rheumatology1982, revised in 1997. The diagnosis of cutaneous leishmaniasis (CL) was confirmed by a biopsy specimen. The diagnosis of VL was confirmed by the association of clinical and biology sings associated with positive bone marrow smear and/or positive serologic tests for Leishmania antibodies. Seven patients were enrolled in the study. Their mean age was 27, 85 years [14-41].There were six women and one man. The diagnosis of and SLE was concomitant in three patients, while in four cases the diagnosis of leishmania was after the diagnosis of LES. The time interval between the date of discovery of leishmaniasis and the date of confirmation of the diagnosis of SLE was 36 months[2-24]. For the diagnosis of LES, malar rash, photosensitivity, nephritis, arthralgia and positive antinuclear antibody were observed in six cases. Anti-double stranded DNA antibody was positive in two cases. For the diagnosis of CL, it was based on clinical manifestation in one case and was based in cutaneous biopsy in two cases. Five patients had nephrotic syndrom and one patient had proteinuria. Patients who had renal biopsy had active lupus nephritis class III in one case and lupus nephritis class IV in three cases. Two patients had not renal biopsy because one of them had severe hypertension and the other had severe sepsis. The diagnosis of CL was clinical in one case and histological in two cases. Clinical manifestations of VL were dominated by fever, pale mucous membranes and splenomegaly. There was pancytopenia in all patients with VL. The diagnosis of VL was confirmed by serology(n=3) andbone marrow smear (n=4). Meglumine antimonite was prescribed in the three cases of CL, in the other cases pentosan polysulfate was prescribed in a single patient while in the other two cases, they required the addition of liposomal amphotericin B. One patient was died by tamponade before treatment. Two patients were died by septic shock and only one was cured. Patients with CL had scar in two cases. The last patient died of it. The distinction between the diagnosis of SLE or its exacerbation and VL may be a clinical dilemmaresponsible for diagnostic delay. All the cases diagnosed so far have been by conventional techniques, which may underestimate the incidence of this association. The results of this study highlight a high mortality rate in cases of VL with SLE accordingly with several studies in literature.

Leishmaniose visceral canina em Rondônia: relato de um caso autóctone

Visceral Leishmaniasis (VL), also known as kala-azar, is a disseminated protozoan infection caused by the Leishmania donovani complex. Traditionally, the definitive diagnosis is made by detecting amastigotes in biological tissue samples. In August 2015, an apparently healthy, young adult, female, mongrel dog from the Zoonosis Control Center of the Rolim de Moura,Rondônia, was subjected to the canine visceral leishmaniasis diagnosis. The diagnosis was conducted using bone marrow aspirate smears and popliteal lymph node smears by Fine-needle aspiration biopsy (FNAB); stained slides with Diff-Quick screened for Leishmania amastigotes by means of direct optical microscopic examination (100×). Lymph node and bone marrow aspirates were used to investigate the presence of Leishmania infantum chagasi DNA by real-time PCR. The popliteal lymph node positive result was observed in typical amastigotes, presenting nucleus and kinetoplasts associated with lymphoblastic proliferation, reactive macrophages, plasmocytes (commonly Mott cells), eosinophils, and lymphoglandular hyperplasia. A positive bone marrow sample resulted from the observation of amastigotes in the monocyte cytoplasm or free on the smear background. The presence of the L. infantum kDNA was detected in lymph node and bone marrow FNAB samples. This research note describes the autochthonous case of CVL recorded in the state of Rondônia, Brazil, a non-endemic area for VL. According to the data, future studies must include a larger number of animals to elucidate the parasite’s epidemiological resource in Rondônia.

Pediatric visceral leishmaniasis: diagnosis and therapy

Visceral leishmaniasis is a generalized infection of the reticuloendothelial system, it is fatal if left untreated in most cases. Early diagnosis and effective therapy can reduce the risk of disability and mortality. However, in non-endemic areas diagnosis may be delayed or missed. In the article we presented a case report of a child with imported visceral leishmaniasis. The gradual onset of the disease with irregular fever, growing weakness, fatigue, loss of appetite, anemia, cytopenic syndrome required differential diagnosis with oncological diseases. The diagnosis of leishmaniasis was made by microscopy of a bone marrow smear and the detection of leishmania. Recovery occurred after the prolonged course of amphotericin B and supportive treatment.

A Review of Leishmaniasis: Current Knowledge and Future Directions.

Purpose of ReviewThe goal of this review is to summarize the current knowledge of the epidemiology, clinical manifestations, diagnosis, and treatment of cutaneous, mucosal, and visceral leishmaniasis. We will describe the most recent findings and suggest areas of further research in the leishmaniasis field.Recent FindingsThis article reviews newer leishmaniasis tests (including rapid diagnostic tests using rK39 antibodies), vaccine candidates, and updated treatment recommendations.SummaryWhile leishmaniasis is a complex disease, learning the prominent clinical manifestations and major parasite species can guide the recommendations for diagnosis and treatment.

Loop-Mediated Isothermal Amplification Allows Rapid, Simple and Accurate Molecular Diagnosis of Human Cutaneous and Visceral Leishmaniasis Caused by Leishmania infantum When Compared to PCR.

Loop-mediated isothermal amplification allows the rapid, sensitive and specific amplification of DNA without complex and expensive equipment. We compared the diagnostic performance of Loopamp™ Leishmania Detection Kit (Eiken Chemical Co., Ltd., Tokyo, Japan) with conventional and real-time polymerase chain reaction (PCR) for human cutaneous and visceral leishmaniasis caused by L. infantum. A total of 230 DNA samples from cutaneous (CL) and visceral (VL) leishmaniasis cases and controls from Spain, characterized by Leishmania nested PCR (LnPCR) were tested by: (i) the Loopamp™ Leishmania Detection Kit (Loopamp), run on Genie III real-time fluorimeter (OptiGene, UK); and (ii) real-time quantitative PCR (qPCR). The Loopamp test returned 98.8% (95% confidence interval—CI: 96.0–100.00) sensitivity and specificity of 97.7% (95% CI: 92.2–100) on VL samples, and 100% (95% CI: 99.1–100) sensitivity and 100.0% (95% CI: 98.8–100.0) specificity on CL samples. The Loopamp time-to-positivity (Tp) obtained by real-time fluorimetry showed excellent concordance (C = 97.91%) and strong correlation (r = 0.799) with qPCR’s cycle threshold (Ct). The performance of Loopamp is comparable to that of LnPCR and qPCR in the diagnosis of cutaneous and visceral leishmaniasis due to L. infantum. The excellent correlation between the Tp and Ct should be further investigated to determine the accuracy of Loopamp to quantify parasite load in tissues.

Clinical and immunological characteristics of tegumentary leishmaniasis cases in Bolivia.

BackgroundTegumentary leishmaniasis (TL) is a parasitic disease that can present a cutaneous or mucocutaneous clinical form (CL and MCL, respectively). The disease is caused by different Leishmania species and transmitted by phlebotomine sand flies. Bolivia has one of the highest incidences of the disease in South America and the diagnosis is done by parasitological techniques. Our aim was to describe the clinical and immunological characteristics of CL and MCL patients attending the leishmaniasis reference center in Cochabamba, Bolivia, in order to gain updated clinical and epidemiological information, to evaluate the diagnostic methods used and to identify biomarkers related to clinical disease and its evolution.Methodology/Principal findingsThe study was conducted from September 2014 to November 2015 and 135 patients with lesions compatible with CL or MCL were included. Epidemiological and clinical data were collected using a semi-structured questionnaire. Two parasitological diagnostic methods were used: Giemsa-stained smears and culture of lesion aspirates. Blood samples obtained from participants were used to measure the concentrations of different cytokines. 59.2% (80/135) were leishmaniasis confirmed cases (CL: 71.3%; MCL: 28.7%). Sixty percent of the confirmed cases were positive by smears and 90.6% were positive by culture. 53.8% were primo-infections. Eotaxin and monokine induced by IFN-γ presented higher serum concentrations in the MCL clinical presentation compared to CL cases and no-cases. None of the cytokines presented different concentrations between primo-infections and secondary infections due to treatment failure.Conclusions/SignificanceIn Bolivia, parasitological diagnosis remains the reference standard in diagnosis of leishmaniasis because of its high specificity, whereas the sensitivity varies over a wide range leading to loss of cases. Until more accurate tools are implemented, all patients should be tested by both smears and culture of lesion aspirates to minimize the risk of false negatives. Our results showed higher concentrations of several cytokines in MCL compared to CL, but no differences were observed between CL and no-cases. In addition, none of the cytokines differed between primary and secondary infections. These results highlight the need of further research to identify biomarkers of susceptibility and disease progression, in addition to looking at the local cellular immune responses in the lesions.

Immunoproteomics characterization of Leishmania panamensis proteins for potential clinical diagnosis of mucosal Leishmaniasis.

Diagnosis of leishmaniasis based on antibodies detection represents a challenge due to cross-reaction of sera with other infectious agents, which co-exist in endemic areas of Leishmania sp, especially patients with Trypanosoma cruzi. This work is aimed at searching for immunogenic proteins in sera from patients with cutaneous and mucosal leishmaniasis that may be potential candidates for the development of diagnostic tests and/or vaccines that help control the infection. Total protein extracts of L.panamensis promastigotes were put in contact with sera from patients with cutaneous and mucosal leishmaniasis (immunoblots). Immunoreactive proteins were identified by mass spectrometry and bioinformatics tools. 81 proteins were identified. One of these was uniquely recognized by the sera from patients with ML but not from sera from either CL or Chagas disease patients. MS analysis of this band pointed to the putative leishmanial 3-oxoacyl-(Acylcarrierprotein) reductase.

Improving The Principles Of Treatment In Patients With Zoonotic Leishmaniasis With The Immunomodulator Gepon And Methylene Blue Using The Alt Vostok Apparatus

The object of the study was 102 patients with zoonotic leishmaniasis aged 2 to 65 years. The diagnosis of zoonotic leishmaniasis was established by clinical and epidemiological data and confirmed by the results of parasitological studies. We studied the duration of the clinical periods of zoonotic leishmaniasis against the background of complex therapy with methylene blue using the ALT Vostok apparatus in the control group of 51 patients and complex therapy with methylene blue using the ALT Vostok apparatus and the immunomodulator Gepon in the main group of 51 patients. The results of the study showed that in patients of the main group, on the background of treatment with the ALT Vostok apparatus using the immunomodulator Gepon, the duration of the main periods was significantly reduced than in the control group, who were treated only with methylene blue using the ALT Vostok model-03 apparatus.

Redox regulation of adventitious root formation through downstream changes in hormonal system in mung bean [Vigna radiata (L.) R. Wilczek

<em>In vitro</em> cultivation of Leishmania parasites plays an important role in diagnosis and treatment of leishmaniasis and in vaccine and drug development studies. There is no information about the effects of urine within culture on the infectivity of Leishmania parasites. In this study we used two culture media with different overlays to enhance the <em>In vitro</em> growth rates of Leishmania donovani.

Defining the Molecular Signal Pathways and Upstream Regulators in Cutaneous Leishmaniasis with Transcriptomic Data Approach

Leishmaniasis is a disease caused by the genus Leishmania spp., which are intracellular parasites. Depending on parasite species and host immune response, there are three basic clinical forms of the disease: cutaneous, mucocutaneous, and visceral leishmaniasis. Cutaneous leishmaniasis is a chronic disease and characterized by the presence of ulcerated skin lesions. The type of skin pathology seen during disease is determined in part by the infecting Leishmania spp., but also by a combination of inflammatory and antiinflammatory host immune response factors resulting in diverse clinical outcomes. In this study, it was aimed to determine the genes, molecular signaling mechanisms and biological functions of the molecules that play a role in the pathogenesis of the disease and immune response and determine host-parasite interactions in mice that are naturally resistant and susceptible to Leishmania major and Leishmania braziliensis. For this, transcriptomic series GSE56029 was downloaded from "Gene Expression Omnibus"(GEO) data base, including expression profiling of twenty-four tissue samples that were recovered from both naive mice and mice (BALB/c, C57BL/6) infected with L.major and L.braziliensis. Then, "Differentially Expressed Genes" (DEGs) were identified by limma package in R script. FDR q<0.05 and absolute log2FC> 2 as threshold values were accepted in the analysis. Subsequently, functional and pathway enrichment analyses were performed for the DEGs by "Ingenuity Pathway Analysis" (IPA). For each of DEGs, p<0.01, FDR q<0.01, and absolute log2FC> 1 were used and analyzed with the software program IPA 8.0. Ingenuity Pathway Analysis revealed the most enrichment pathways to be the inflammation, dendritic cell maturation and "Triggering Receptor Expressed on Myeloid Cells 1" (TREM-1) signal mechanisms and that the DEGs related to the regulation of immune system process were closely associated with the progress of cutaneous leishmaniasis. The upstream regulator analysis predicted that TNF-α, IFNy, IL-1 β, IL-10RA and "Signal Transducer and Activator of Transcription-1" (STAT-1) are the regulators that explained gene expression changes causing biological activities in the tissues. Chemical compounds that may have anti-leishmanial effects were also identified in the study. In this study, the mechanisms belonging to the parasite species and host that determine the resistance/susceptibility phenotype were attempted to elucidate. Assessment of gene expression patterns, cytokine/chemokines, and signaling pathways in BALB/c and C57BL/6 mice infected with L.major and L.braziliensis will provide a better understanding of the potential mechanisms underlying infection from a genetic perspective. These results may guide for the future studies in terms of developing potential biomarkers for the diagnosis and prognosis prediction of cutaneous leishmaniasis and providing information about new treatment targets.