In Vitro and in Vivo Therapeutic Evaluation of Ethanolic Extract of Sumac and Compounds of Linalool and Eugenol Against Localized Cutaneous Leishmaniasis Induced by Leishmania major.

Cutaneous leishmaniasis (CL) is a tropical disease that can cause chronic lesions and leave life-long scars, leading to social stigmatization and psychological disorders. Using growth factors and immunomodulatory agents that could accelerate wound healing and reduce the scar is highly demanded. Epidermal growth factor (EGF) plays an essential role in wound healing. It stimulates the proliferation of keratocytes and fibroblasts, and promotes re-epithelialization. Here, the effect of EGF in combination with Glucantime and nano-liposomal Amphotericin B (SinaAmpholeish) on the healing process of CL in BALB/c mice was investigated. Seventy-two mice were infected with Leishmania major parasites and randomly divided into eight treatment groups after the appearance of the lesion. The treatment was continued for five weeks, and lesion sizes were measured weekly. Parasite load was determined in the skin biopsies using qPCR. We found that subcutaneous injection of EGF at 4.5 μg/kg, combined with each of the two antileishmanial drugs, significantly reduced the wound size and parasite load; however, EGF at 1.5 μg/kg failed to be effective. Besides, the wound size and parasite loads were significantly lower in the SinaAmfoleish groups compared to the Glucantime groups. Among the treatment groups, EGF 4.5 μg/kg combined with SinaAmpholeish exhibited the most significant reduction in wound size and parasitic load. Our results suggest that EGF can potentiate the wound healing effect of antileishmanial drugs. Further studies are warranted to explore the beneficial effects of combining EGF with antileishmanial drugs in patients with cutaneous leishmaniasis in order to accelerate wound healing and reduce the scar.

Cutaneous leishmaniasis (CL) is a parasitic disease that presents a broad spectrum of clinical features. Treatment of CL is problematic. We aimed to compare the field therapeutic efficacy of topical nanoliposomes containing 0.4% amphotericin B (Nano Lip-AmB) alone and in combination with cryotherapy and/or Glucantime® on human CL in the endemic areas of Iran. This retrospective study was performed based on the results of using Nano Lip-AmB alone or with Glucantime® and/or cryotherapy in the treatment of zoonotic cutaneous leishmaniasis (ZCL) in patients referred to health centers of Isfahan, Golestan and Ilam Provinces of Iran as endemic foci of ZCL caused by Leishmania major besides Mashhad and Bam cities as endemic foci of anthroponotic cutaneous leishmaniasis (ACL) caused by with L. tropica. Two hundred and seventy-eight patients with CL were included in the current study. All of the patients (100%) who received Nano Lip-AmB alone or in combination with Glucantime® and/or cryotherapy based on guideline of Iranian national committee for the treatment of CL. Two patients with 7 skin lesions, who was resident in ACL endemic area and received Nano Lip-AmB plus Glucantime® and another patient was a resident of ZCL endemic area and received Nano Lip-AmB plus cryotherapy showed clinical relapses after treatment. Sina Ampholeish® in combination with other standard protocols of treatment of CL is well tolerated and with acceptable clinical efficacy rate.

Earlier histopathology studies suggest that parasite loads may differ between cutaneous leishmaniasis (CL) and mucosal leishmaniasis (ML) lesions and between acute and chronic CL. Formal demonstration requires highly sensitive detection and accurate quantification of Leishmania in human lesional tissue. In this study, we developed a quantitative real-time PCR (qPCR) assay targeting minicircle kinetoplast DNA (kDNA) to detect and quantify Leishmania (Viannia) parasites. We evaluated a total of 156 lesion biopsy specimens from CL or ML suspected cases and compared the quantitative performance of our kDNA qPCR assay with that of a previously validated qPCR assay based on the glucose-6-phosphate dehydrogenase (G6PD) gene. We also examined the relationship between parasite load and clinical parameters. The kDNA qPCR sensitivity for Leishmania detection was 97.9%, and its specificity was 87.5%. The parasite loads quantified by kDNA qPCR and G6PD qPCR assays were highly correlated (r = 0.87; P < 0.0001), but the former showed higher sensitivity (P = 0.000). CL lesions had 10-fold-higher parasite loads than ML lesions (P = 0.009). Among CL patients, the parasite load was inversely correlated with disease duration (P = 0.004), but there was no difference in parasite load according to the parasite species, the patient's age, and number or area of lesions. Our findings confirm that CL and recent onset of disease (<3 months) are associated with a high parasite load. Our kDNA qPCR assay proved highly sensitive and accurate for the detection and quantification of Leishmania (Viannia) spp. in lesion biopsy specimens. It has potential application as a diagnostic and follow-up tool in American tegumentary leishmaniasis.

Treatment of cutaneous leishmaniasis (CL) and mucosal leishmaniasis (ML) in travelers is still controversial. Over the last decade, national and international consortia have published recommendations for treating CL in travelers. These guidelines harmonize many issues, but there are some discrepancies. Leishmania parasites causing CL can now be genotyped by polymerase chain reaction techniques for detecting Leishmania DNA. Therefore, treatment recommendations can now be species based rather than based on geographical exposure. To review the evidence on which the recommendations were based, "LeishMan" (Leishmaniasis Management), a group of experts from 13 institutions in eight European countries, performed a PubMed MEDLINE) literature search and considered unpublished evidence and the experts' own personal experiences. The Oxford evidence grading system was used to evaluate the information. In this article, the authors provide practical treatment recommendations for imported CL and ML in Europe, drawn up from the review by the European experts.

Clofazimine, an antimycobacterial with potent in vitro and in vivo leishmanicidal activity, is ineffective against cutaneous Leishmania major infection in humans

Cutaneous leishmaniasis, caused by protozoa of the genus Leishmania, presents diverse clinical manifestations, and current therapeutic options have limitations, including long treatment periods, potential hospitalization, and excessive pain during treatment. Methyl gallate, a phenolic compound found in plants such as Libidibia ferrea, presents promising antileishmanial activity. Combining this compound with existing leishmaniasis medications could lead to reduced dosages and the minimization of side effects. This study aimed to assess the efficacy of a microemulsion containing methyl gallate, either on its own or in combination with Glucantime®, for the experimental treatment of cutaneous leishmaniasis in a 30-day in vivo assay using golden hamsters infected with Leishmania (Leishmania) amazonensis. The control groups included an untreated positive control and an uninfected, untreated negative control. After treatment, we evaluated clinical, parasitological, and biochemical parameters. While none of the treatments achieved clinical or parasitological cure, notable improvements were observed in the combined group, with significant reductions in snout skin lesions and parasite load when compared to the control. Biochemical parameters such as creatinine, CK-MB, GOT, and GPT remained unchanged, but urea and CPK levels significantly increased in all the experimental groups relative to the control. In conclusion, the integration of a topical methyl gallate microemulsion with intralesional Glucantime® showed potential as an effective treatment for cutaneous leishmaniasis. Further investigations into optimal dosages and therapeutic schemes are warranted in order to enhance treatment outcomes.

Background: The optimal treatment for cutaneous leishmaniasis (CL) is not known. Topical paromomycin is one of the many drugs that have been suggested for the treatment of CL caused by Leishmania major. Recently, topical photodynamic therapy (PDT) has been reported to be effective in the treatment of CL. This study aimed to compare the parasitological and clinical efficacy of PDT versus topical paromomycin in patients with old world CL caused by L. major in Iran. Materials and Methods: In this trial, sixty patients with the clinical and parasitological diagnosis of CL were recruited and were randomly divided into three treatment groups with twenty subjects in each group. Group 1 was treated with weekly topical PDT and groups 2 and 3 received twice daily topical paromomycin and placebo, respectively. The duration of treatment was four weeks for all groups. These groups were followed up for 2 months after the end of treatment. Results: 57 patients with 95 lesions completed the study. At the end of the study, complete improvement was seen in 29 of 31 (93.5%), 14 of 34 (41.2%) and 4 of 30 (13.3%) of the lesions in group 1, 2 and 3 respectively ( P <0.001). At the same time point, 100%, 64.7 and 20% of the lesions had parasitological cure in group 1, 2 and 3, respectively ( P <0.001). Conclusion: Topical PDT can be used safely as a rapid and highly effective alternative choice for treatment of old world CL in the selected patients.

Leishmania tropica infection of the ear treated with photodynamic therapy

Background: Cutaneous leishmaniasis (CL) is an endemic disease in Iran. Pentavalent antimonials including meglumine antimoniate (MA) are still the gold standard treatment. Trichloroacetic acid (TCA) is used in treatment of acne scar, photodamaged skin, and some other dermatologic diseases. Considering the TCA potential to induce collagen synthesis and its efficacy in the treatment of papular CL lesions, this study was conducted to assess the efficacy of combined intralesional MA and TCA in comparison to MA alone in treatment of CL. Objectives: This study was designed to compare the efficacy of the combination therapy of intralesional MA and TCA with intralesional MA alone in the treatment of CL. Patients and Methods: A total of 200 patients with confirmed CL were selected and randomized into two treatment groups. Both groups were treated with intralesional injection of MA twice a week until complete resolution of the lesions or end of the eighth week. In combination therapy group, TCA was applied to the lesions fortnightly for eight weeks. Data were analyzed statistically using chi-square, student t, and Kaplan-Meier tests. Results: Complete resolution of the lesions was achieved in 79.2% of patients treated with MA and 85.7% of patients receiving combination therapy (P > 0.05). There was a significant difference in time to achieve the complete resolution between the groups with an accelerated resolution rate in the combination therapy group (P < 0.05). Conclusions: According to the results of this study, combination therapy with intralesional MA and TCA 50% could accelerated the resolution of CL lesions with no significant difference in complete resolution rate in comparison to the patients treated with intralesional MA alone. Topical application of TCA 50% as an adjuvant therapy might have some advantages in decreasinge the resolution time of the lesions in patients with CL.

Leishmania aethiopica is the etiological agent of cutaneous leishmaniasis (CL) in Ethiopia and can cause severe and complicated cases such as diffuse CL (DCL), mucocutaneous leishmaniasis or extensive CL, requiring systemic treatment. Despite the substantial burden, evidence-based treatment guidelines are lacking. We conducted a systematic review of clinical studies reporting on treatment outcomes of CL due to L aethiopica in order to help identify potentially efficacious medications on CL that can be taken forward for clinical trials. We identified a total of 24 records reporting on 506 treatment episodes of CL presumably due to L aethiopica. The most commonly used drugs were antimonials (n = 201), pentamidine (n = 150) and cryotherapy (n = 103). There were 20 case reports/series, with an overall poor study quality. We only identified two small and/or poor quality randomized controlled trials conducted a long time ago. There were two prospective non-randomized studies reporting on cryotherapy, antimonials and pentamidine. With cryotherapy, cure rates were 60–80%, and 69–85% with antimonials. Pentamidine appeared effective against complicated CL, also in cases non-responsive to antimonials. However, all studies suffered from methodological limitations. Data on miltefosine, paromomycin and liposomal amphotericin B are extremely scarce. Only a few studies are available on DCL. The only potentially effective treatment options for DCL seem to be antimonials with paromomycin in combination or pentamidine, but none have been properly evaluated. In conclusion, the evidence-base for treatment of complicated CL due to L aethiopica is extremely limited. While antimonials remain the most available CL treatment in Ethiopia, their efficacy and safety in CL should be better defined. Most importantly, alternative first line treatments (such as miltefosine or paromomycin) should be explored. High quality trials on CL due to L aethiopica are urgently needed, exploring group sequential methods to evaluate several options in parallel.

ABSTRACTAmBisome (LAmB), a liposomal formulation of amphotericin B (AmB), is a second-line treatment for the parasitic skin disease cutaneous leishmaniasis (CL). Little is known about its tissue distribution and pharmacodynamics to inform clinical use in CL. Here, we compared the skin pharmacokinetics of LAmB with those of the deoxycholate form of AmB (DAmB; trade name Fungizone) in murine models of Leishmania major CL. Drug levels at the target site (the localized lesion) 48 h after single intravenous (i.v.) dosing of the individual AmB formulations (1 mg/kg of body weight) were similar but were 3-fold higher for LAmB than for DAmB on day 10 after multiple administrations (1 mg/kg on days 0, 2, 4, 6, and 8). After single and multiple dosing, intralesional concentrations were 5- and 20-fold, respectively, higher than those in the healthy control skin of the same infected mice. We then evaluated how drug levels in the lesion after LAmB treatment relate to therapeutic outcomes. After five administrations of the drug at 0, 6.25, or 12.5 mg/kg (i.v.), there was a clear correlation between dose level, intralesional AmB concentration, and relative reduction in parasite load and lesion size (R2 values of >0.9). This study confirms the improved efficacy of the liposomal over the deoxycholate AmB formulation in experimental CL, which is related to higher intralesional drug accumulation.

Background: Cutaneous leishmaniasis is an endemic disease in developing countries. The first-line drugs for its treatment are the pentavalent antimony compounds such as meglumine antimoniate (MA). High cost, side effects, multiple injections and incomplete efficacy are limitations of this therapy. Zinc sulfate (ZS) has been reported to be effective in the treatment of cutaneous leishmaniasis. Objective: To compare the efficacy of intralesional injections of ZS 2% with those of MA in the treatment of cutaneous leishmaniasis. Methods: This was a prospective, double-blind, case-control clinical study. 104 patients with typical lesions of acute cutaneous leishmaniasis (ACL) were included. The duration of treatment was 6 weeks. Improvement was evaluated by clinical examination and direct smear. Results: The study was completed in only 66 patients: 35 patients received MA and 31 received ZS. The cure rates were 60% for MA and 83.8% for ZS. After the second and fourth weeks, the efficacy of treatment with ZS was higher than that with MA (p < 0.01), but after 6 weeks no significant differences were observed between the two groups (p > 0.05). Conclusion: On the basis of this trial and despite the high number of drop-outs, we consider that the intralesional injection of ZS 2% is an alternative treatment in ACL.

Leishmaniasis is still a serious neglected tropical disease that may cause death in infected individuals. At present, the clinical diagnosis and treatment monitoring still rely on parasitological culture and microscopy that needs experienced technicians. The low sensitivity and inconvenience of microscopic examination could cause misdiagnosis and relapse of leishmaniasis. There is an urgent need for developing a sensitive and easily operated diagnostic method for the diagnosis and disease management of leishmaniasis. Thus, a quantitative real-time PCR (qPCR) based on the conversed regions of kinetoplast minicircle DNA (mkDNA) of Leishmania spp. was developed to detect different species of Leishmania. The designed mkDNA-based qPCR was able to detect as low as one copy of Leishmania mkDNA or DNA from single parasite. It also detected Pan-Leishmania protozoa including Leishmania donovani, Leishmania infantum and Leishmania major without cross-reaction with other pathogen DNAs available in our lab. This method was clinically applied to quantitatively detect skin lesion samples from 20 cutaneous leishmaniasis (CL) and bone marrow and/or PBMC samples from 30 current and cured visceral leishmaniasis (VL) patients, and blood samples from 11 patients with other infections and 5 normal donors as well. Total 20 skin lesion samples from current CL patients and 20 bone marrow and/or PBMC samples from current VL patients were all detected as positive with qPCR without cross-reaction with samples from patients with malaria, brucellosis and dengue or normal donors. Two VL patients with parasite converted to microscopically negative after treatment were detected positive with qPCR. The patients with bigger skin lesion in CL and higher level of immunoglobulin or splenomegaly in VL, had the higher parasite load detected by qPCR. The parasite load was significantly reduced after treatment. In conclusion, the mkDNA-based qPCR assay that we developed in this study can be used not only for diagnosis of both cutaneous and visceral leishmaniasis with high sensitivity and specificity, but also for evaluating the severity and treatment efficacy of this disease, presenting a rapid and accurate tool for clinical surveillance, treatment monitoring and the end point determination of leishmaniasis.

Cutaneous leishmaniasis due to Leishmania aethiopica: A therapeutic challenge

Immunodetection and molecular determination of visceral and cutaneous Leishmania infection using patients' urine