Tape-disc-loop-mediated isothermal amplification (TD-LAMP) method as noninvasive approach for diagnosis of cutaneous leishmaniasis caused by L. tropica.

Cutaneous leishmaniasis (CL) is a parasitic disease caused by the bite of infectious female sand flies with high socioeconomic burdens. There is currently no non-invasive, point-of-care, diagnostic method with high sensitivity and specificity available for CL. We herein report the development of a non-invasive tape disc (TD) sampling method combined with a loop-mediated isothermal amplification (LAMP) assay using primer sets targeting kinetoplast DNA (kDNA) of Leishmania tropica (L. tropica) with a colorimetric readout for species-specific diagnosis of CL. We tested our Tape-Disc (TD)-LAMP method on a panel of skin samples collected by TD from 35 confirmed L. tropica patients, 35 healthy individuals and 35 patients with non-L. tropica infections. The detection limit of the TD-LAMP assay was determined as 1 fg (fg), and the assay sensitivity and specificity of 97 % and 100 % for L. tropica infection, respectively. This non-invasive, sensitive and rapid diagnostic method warrants further exploration of its use for differential diagnosis of CL in disease endemic settings.

Blood transcriptional profiles distinguish different clinical stages of cutaneous leishmaniasis in humans.

Cutaneous leishmaniasis (CL) is a neglected tropical disease with severe morbidity and socioeconomic sequelae. A better understanding of underlying immune mechanisms that lead to different clinical outcomes of CL could inform the rational design of intervention measures. While transcriptomic analyses of CL lesions were recently reported by us and others, there is a dearth of information on the expression of immune-related genes in the blood of CL patients. Herein, we investigated immune-related gene expression in whole blood samples collected from individuals with different clinical stages of CL along with healthy volunteers in an endemic CL region where Leishmania (L.) tropica is prevalent. Study participants were categorized into asymptomatic (LST+) and healthy uninfected (LST-) groups based on their leishmanin skin test (LST). Whole blood PAXgene samples were collected from volunteers, who had healed CL lesions, and patients with active L. tropica cutaneous lesions. Quality RNA extracted from 57 blood samples were subjected to Dual-color reverse-transcription multiplex ligation-dependent probe amplification (dcRT-MLPA) assay for profiling 144 immune-related genes. Results show significant changes in the expression of genes involved in interferon signaling pathway in the blood of active CL patients, asymptomatics and healed individuals. Nonetheless, distinct profiles for several immune-related genes were identified in the healed, the asymptomatic, and the CL patients compared to the healthy controls. Among others, IFI16 and CCL11 were found as immune transcript signatures for the healed and the asymptomatic individuals, respectively. These results warrant further exploration to pinpoint novel blood biomarkers for different clinical stages of CL.

Combination of topical liposomal amphotericin B and Glucantime in comparison with glucantime alone for the treatment of anthroponotic cutaneous leishmaniasis (ACL) caused by Leishmania tropica: study protocol for a randomized, controlled trial.

BACKGROUND AND OBJECTIVES: Cutaneous leishmaniasis (CL) treatment is a challenging issue, although numerous modalities have been introduced as candidate treatment for CL yet only antimonial agents are commonly used to treat CL, a different form of amphotericin B is used to treat visceral form of leishmaniasis but the efficacy against CL is not high. There are a few reliable clinical trials on CL, the main reason is the nature of the disease which required a well design protocol to evaluate the efficacy of any candidate treatment against CL. In this study, a protocol was developed and used to evaluate a topical formulation of a nano-liposomal form of amphotericin B in addition to glucantime to treat CL caused by L. tropica. MATERIALS AND METHODS: This study is a phase 3, randomized, double-blind, placebo-controlled trial to evaluate the efficacy and safety of topical nano-liposomal amphotericin B (SinaAmpholeish 0.4%) in combination with intralesional injections of meglumine antimoniate in the treatment of ACL caused by L. tropica. Overall, 130 patients, aged 12-60 years, with a diagnosis of ACL caused by L. tropica are recruited and treated according to the protocol. RESULTS: A total of 130 patients with CL lesion will be recruited and doubleblind randomly treated with received intralesional injections of Glucantime weekly or Glucantime plus SinaAmpholeish for 4 weeks. CONCLUSION: The results of this study showed that the protocol works well and the treatment was tolerated by both groups of patients.

The effect of oral miltefosine in treatment of antimoniate resistant anthroponotic cutaneous leishmaniasis: An uncontrolled clinical trial.

BACKGROUND: Recent circumstantial evidence suggests increasing number of Iranian patients with cutaneous leishmaniasis (CL) who are unresponsive to meglumine antimoniate (MA), the first line of treatment in Iran. Oral meltifosine was previously reported to be effective in visceral leishmaniasis as well CL. The current study is designed to determine efficacy and safety of oral miltefosine for the treatment of anthroponotic cutaneous leishmaniasis (ACL) cases who were refractory to MA in Iran. METHODOLOGY/PRINCIPAL FINDINGS: Miltefosine was orally administered for 27 patients with MA resistant ACL with approved L.tropica infection, at a dosage of ∼2.5 mg/kg daily for 28 days. Patients were evaluated on day 14 and 28, as well as 3, 6 and 12 month post treatment follow up sessions. Laboratory data were performed and repeated at each visit. Data were analyzed using SPSS version 17. Twenty-seven patients including 16 men (59.25%) and 11 women (40.74%) with mean age of 28.56 ± 4.8 (range 3-54 years old) were enrolled. Total number of lesions were 42 (1-4 in each patient). Most of lesions were on face (76.19%). Mean lesions' induration size was 2.38 ± 0.73 cm at the base-line which significantly decreased to1.31 ± 0.58 cm and 0.61 ±0.49 cm after 14 and 28 days of therapy, respectively (p value <0.05). At 12-months follow-up post treatment, 22 patients had definite/partial cure (81.48%) including 17 definitely cured patients, corresponding to a cure rate of 68% on per protocol analysis, and 62.96% according to intention to treat analysis. Recurrence of lesion was only occurred in one patient (3.70%). Nausea was the most subjective complication during the therapy (33.33%). CONCLUSION: Oral miltefosine could be an effective alternative for the treatment of MA-resistant ACL.

Salvage therapy with Sodium chlorosum (formerly DAC N-055) for cases of refractory lupoid cutaneous leishmaniasis: results from a compassionate use study with 0.09% Sodium chlorosum in amphiphilic basic cream.

BACKGROUND: Lupoid cutaneous leishmaniasis (LCL) is known as a rare but serious complication of anthroponotic cutaneous leishmaniasis (ACL) resistant to conventional treatments. Sodium chlorosum, a pro-oxidative preparation of pharmaceutical sodium chlorite (NaClO2), has been successfully used for the treatment of Old World cutaneous leishmaniasis lesions (OWCL) and of some LCL cases in Afghanistan. This clinical trial study aimed to evaluate the effect of a last resort therapy with topical 0.09% sodium chlorosum on LCL in Iran. METHODS: Twenty Iranian patients (12 women and 8 men) with LCL refractory to treatment were included in this salvage study. A magistral preparation of sodium chlorosum (10 mM NaClO2 in amphiphilic basic cream) was applied twice daily to the lesions for 6 weeks and continued up to 12 weeks in patients who showed a clinical response within the first 6 weeks. Responders were followed up for a maximum of 1 year. Lesions were photographed during weekly visits. Disappearance of erythema and indurated lesions were rated as complete clinical response. RESULTS: Patients with a mean age of 28.6 (±24.3) and with an ACL proven lesion history of 3.8 (±1.4) years were treated for an average of 7.9 (±1.8) weeks. At the end of the treatment period (12th week), a complete response was observed in 9 of 20 patients (45%). During the one-year follow-up period, LCL lesions recurred in 4 of these 9 patients (with one patient showing only a tiny lesion) and one case lost to follow up whereas the other four remained completely lesion-free. Mild temporary side-effects such as erythema and itching were seen in 4 of 20 patients (20%). CONCLUSIONS: Topical sodium chlorosum showed promising therapeutic results and can be considered as safe, painless, and relatively effective treatment for LCL, an ethical prerequisite for a two-armed controlled trial. TRIAL REGISTRATION: This study was registered in Iranian registry of clinical trials on 2019-02-02 with registration number IRCT20190114042356N1.

Successful treatment by adding thalidomide to meglumine antimoniate in a case of refractory anthroponotic mucocutaneous leishmaniasis.

Mucosal leishmaniasis (ML) is mostly associated with Leishmania braziliensis; however, a few cases of Leishmania tropica induced mucocutaneous leishmaniasis have been reported. The standard treatment for leishmaniasis is pentavalent antimonials, but several other drugs for resistant cases have been proposed including amphotericin and miltefosine. Here we present a case of multiple treatment resistant mucocutaneous leishmaniasis with nasal involvement caused by L. tropica; cure was not achieved by multiple treatments and was eventually improved by adding thalidomide to Meglumine Antimoniate (Glucantime). To the best of our knowledge use of thalidomide in humans for leishmaniasis treatment is reported here for the first time.

Ocular Leishmaniasis Treated by Intralesional Amphotericin B.

Leishmaniasis is a group of diseases with varied clinical manifestations. Ocular involvement is an unusual presentation of leishmaniasis, and the eyelid is not a common site of cutaneous lesions, likely due to the mobility of the lids. Some case reports of conjunctival involvement are either a contiguous dissemination from lid margin or in the setting of disseminated leishmaniasis in an immunocompromised host. To our knowledge, isolated involvement of the bulbar conjunctiva has not been reported. We present the first case in the literature of a patient with an erythematous fibrovascular lesion in the interpalpebral zone that was clinically diagnosed as pterygium, but recurred at the site of surgical excision. After histopathologic diagnosis, the lesion was treated with intralesional injection of amphotericin B and improved completely within a few weeks. An accurate diagnosis of leishmaniasis in the eye may be challenging in many clinical settings. To our knowledge, an isolated pterygium.like lesion has not been reported in literature. In addition, intralesional injection of amphotericin B is a novel treatment method in this setting.

Efficacy of intralesional amphotericin B for the treatment of cutaneous leishmaniasis.

BACKGROUND: Antimoniate compounds have been used as gold standard treatment for cutaneous leishmaniasis since many years ago, but with increase in incidence of drug as well as individual contraindications, more attention has been given to alternative treatments. AIM: The aim of this study was to evaluate the efficacy of intralesional amphotericin B as an alternative treatment for cutaneous leishmaniasis in Mashhad, Iran, during 2007-2009. MATERIALS AND METHODS: Non-random sampling from both sexes and without any age limitation of cases eligible for this alternative treatment was done. Size and induration of lesions were measured before beginning and weakly during the treatment. Amphotericin B (2 mg/ml) was injected into lesions weekly for up to 12 weeks and the cases were followed up for the treatment responses, possible side effects and recurrence of the disease. RESULTS: A total of 93 patients with a mean age of 20.81 ± 15.26 years were included in this study. At the end of 12(th) week, 61.4% of the patients were recovered completely (more than 90% reduction in size and induration), 21.6% had partial remission (60-90% reduction in size and induration), and 17% had less than 60% reduction in size and induration of skin lesions. Injection side effects were insignificant and did not lead to premature discontinuation of treatment in any patients. CONCLUSION: Weekly intralesional injection of amphotericin B looks promising, considering the fact that most of the patients in this study were resistant to antimoniates.

Dermoscopic features of cutaneous leishmaniasis.

BACKGROUND: Previous studies have demonstrated that dermoscopy improves accuracy in diagnosing pigmented and non-pigmented tumors. Recently, there has been increasing evidence that dermoscopy can also be useful in the diagnosis of some skin infections. OBJECTIVE: We sought to describe the dermoscopic features of cutaneous leishmaniasis. METHOD: Dermoscopic examination (using the Derm Lite FOTO, 3Gen) of 144 CL lesions in 82 patients was performed. This study was conducted in the Imam Reza and Ghaem hospitals (Mashhad, Iran) from October 2010 to September 2011. RESULTS: We observed the following dermoscopic features: generalized erythema (81.9%), white starbursts (60.4%), yellow hue (43.8%), yellow teardrop-like structures (41.7%), central ulcers (59%), hyperkeratosis (33.3%), and milia-like cysts (4.9%). We also observed vascular structures, including dotted vessels (61.1%), hairpin vessels (37.5%), linear irregular vessels (30.6%), comma-shaped vessels (29.9%), glomerular vessels (22.9%), arborizing telangiectasia (10.4%), and corkscrew vessels (4.2%). LIMITATIONS: Biopsy and pathologic evaluation were not performed due to ethical considerations. CONCLUSION: Important vascular patterns seen in melanocytic and non-melanocytic tumors were frequently observed in this infection. Dermoscopy may be a promising tool to predict the clinical course in cases of cutaneous leishmaniasis.

Evaluation of the effect of formic acid and sodium formate on hair reduction in rat.

Hirsutism is a common problem in dermatology that imposes high socioeconomical costs on medical care. Consequently, researchers are actively searching for cheaper and safer methods for therapeutic treatment. The objective of the present study is to evaluate formic oil, enriched from formic acid, for the removal of unwanted hair. In this study, 32 female rats (150-200 g) were randomly divided into four groups and maintained with normal water and food availability. A patch of skin was shaved on each rat for application of test solutions. The control group was treated with local once-daily applications of normal saline. The formic acid, acetic acid, and sodium formate groups were treated with once-daily applications of formic acid (pH 5.5), acetic acid (pH 5.5), or sodium formate, respectively. After 2 weeks, horizontally cut sample biopsies were removed, and the numbers of hair follicles were counted under high field microscopy by a specialist blinded to the treatments. Kolmogorov-Smirnov test results indicated a nonparametric distribution for the rat groups. ANOVA analysis indicated no statistically significant differences between groups (P < 0.05). There weren't any side effects or evidence for toxicity during the study period. However, hair follicle counts showed a descending order of control, acetic acid, formic acid, and sodium formate. Although the sodium formate group had the lowest hair follicle numbers, the difference was not statistically significant (P > 0.05). Formic acid was not effective in reducing hair follicle numbers in rats.

Evaluation of the relationship between androgenetic alopecia and demodex infestation.

INTRODUCTION: Androgenetic alopecia (AGA) is one of the most common dermatologic disorders with a multifactorial etiology. Inflammatory activators such as Demodex infestation may play a role in the pathogenesis of some cases of androgenetic alopecia that do not respond to common treatments such as minoxidil and finasteride. The goal of this study is to evaluate the relationship between Demodex infestation and AGA. MATERIALS AND METHODS: In this case-control study, 41 patients with AGA referred to the Dermatology Clinic of Imam Reza Hospital and 33 healthy individuals were evaluated as control. All of them were between 20 and 40 years old men. In order to identify Demodex infestation they were referred to the Parasitology laboratory. RESULTS: Demodex was detected in 19.5% of patients and 15.2% of controls; therefore, there was no significant relationship between them statistically (P = 0.0787). Most of patients (85.4%) had greasy hair. The most common pattern of baldness was II degree in Hamilton scale. CONCLUSION: There is no relation between AGA and Demodex.

Clinicopathologic evaluation of mammary Paget's disease.

UNLABELLED: Mammary and extramammary Paget's diseases are rare neoplasms of epidermis and mucosal epithelium. Due to their nonspecific and variable clinical view, they have differential diagnosis with eczema, melanoma, Bowen's disease, etc. To the best of our knowledge, no such study has been performed in Iran regarding the prevalence, clinical aspects, underlying disease and pathological characteristics of these two diseases. In this study, we have evaluated the clinical and histopathological aspects of this disorder. MATERIALS AND METHODS: In this retrospective study, all Paget's biopsied samples referred to the Pathology Department of Imam-Reza hospital, Mashhad, since 1984 till 2004 were evaluated. Collected data were analyzed by descriptive statistical methods. RESULTS: Among 98925 specimens, there were 29 cases of Paget's disease. All cases were married women suffering from mammary Paget. The mean age was 53 +/- 11 years. Left and right breast involvement was observed in 17 and 12 cases, all unilateral. The most common clinical view was ulcerated (27%) and then erythematosus exudative plaques. More than 50% of patients were symptomatic. Most common symptoms were itching, pain and burning. The exclusive underlying pathological diagnosis was ductal carcinoma (55%). DISCUSSION: In most cases, the clinical view of mammary Paget's disease was helpful. Unilateral ulcerated plaque was the most common clinical sign. Majority of the accompanying pathology was ductal carcinoma. We had no cases of extramammary Paget's disease in our study.