Antileishmanial Activity of Niosomal Combination Forms of Tioxolone along with Benzoxonium Chloride against Leishmania tropica

In this study, we carried out extensive in vitro studies on various concentrations of tioxolone along with benzoxonium chloride and their niosomal forms against Leishmania tropica. Niosomes were prepared by the hydration method and were evaluated for morphology, size, release study, and encapsulation efficiency. This study measured leishmanicidal activity against promastigote and amastigote, apoptosis and gene expression levels of free solution and niosomal-encapsulated tioxolone along with benzoxonium chloride. Span/Tween 60 niosome had good physical stability and high encapsulation efficiency (more than 97%). The release profile of the entrapped compound showed that a gradual release rate. The combination of niosomal forms on promastigote and amastigote were more effective than glucantime. Also, the niosomal form of this compound was significantly less toxic than glucantime (P≤0.05). The flowcytometric analysis on niosomal form of drugs showed that higher number of early apoptotic event as the principal mode of action (89.13% in 200 μg/ml). Also, the niosomal compound increased the expression level of IL-12 and metacaspase genes and decreased the expression level of the IL-10 gene, which further confirming the immunomodulatory role as the mechanism of action. We observed the synergistic effects of these 2 drugs that induced the apoptotic pathways and also up regulation of an immunomodulatory role against as the main mode of action. Also, niosomal form of this combination was safe and demonstrated strong anti-leishmaniasis effects highlights further therapeutic approaches against anthroponotic cutaneous leishmaniasis in future planning.

A Novel Niosomal Combination of Selenium Coupled with Glucantime against Leishmania tropica

There is no effective treatment modality available against different forms of leishmaniasis. Therefore, the aim of this study was to improve the penetration and efficacy of selenium and glucantime coupled with niosomes and compared them with their simple forms alone on in vitro susceptibility assays. In this study, the niosomal formulations of selenium and in combination with glucantime were prepared. The size and morphology of the niosomal formulations were characterized and the effectivity of the new formulation was also evaluated using in vitro MTT assay, intra-macrophage model, and gene expression profile. From the results obtained, no cytotoxicity effect was observed for niosomal and simple forms of drugs, as alone or in combination. Niosomal formulations of the drugs significantly showed more inhibitory effects (P≤0.001) than the simple drugs when the selectivity index was considered. The gene expression levels of Interleukin (IL-10) significantly decreased, while the level of IL-12 and metacaspase significantly increased (P≤0.001). The results of the present study showed that selenium plus glucantime niosome possess a potent anti-leishmanial effect and enhanced their lethal activity as evidenced by the in vitro experiments.

Tioxolone niosomes exert antileishmanial effects on Leishmania tropica by promoting promastigote apoptosis and immunomodulation

Objective: To explore the antileishmanial effect of tioxolone and its niosomal form against Leishmania tropica. Methods: Tioxolone niosomes were prepared by the hydration method and were evaluated for morphology, size, release study, and encapsulation efficiency. The cytotoxicity of tioxolone and its niosomal form was measured by MTT assay, leishmanicidal activity against promastigote and amastigote by MTT assay, apoptosis by flow cytometry, IL-12, IL-10 and metacaspase gene expression levels by q-PCR. Results: Span/Tween 40 and Span/Tween 60 niosomes had good physical stability as depicted in their size distribution curves and high encapsulation efficiency (>99%). The release profile of the entrapped compounds showed Fickian's model of tioxolone delivery based on diffusion through lipid bilayers. With the IC

Antileishmanial and Cytotoxic Effects of Essential Oil and Methanolic Extract of Myrtus communis L

Plants used for traditional medicine contain a wide range of substances that can be used to treat various diseases such as infectious diseases. The present study was designed to evaluate the antileishmanial effects of the essential oil and methanolic extract of Myrtus communis against Leishmania tropica on an in vitro model. Antileishmanial effects of essential oil and methanolic extract of M. communis on promastigote forms and their cytotoxic activities against J774 cells were evaluated using MTT assay for 72 hr. In addition, their leishmanicidal activity against amastigote forms was determined in a macrophage model, for 72 hr. Findings showed that the main components of essential oil were alpha-pinene (24.7%), 1,8-cineole (19.6%), and linalool (12.6%). Findings demonstrated that M. communis, particularly its essential oil, significantly (P<0.05) inhibited the growth rate of promastigote and amastigote forms of L. tropica based on a dose-dependent response. The IC50 values for essential oil and methanolic extract was 8.4 and 28.9 mug/ml against promastigotes, respectively. These values were 11.6 and 40.8 mug/ml against amastigote forms, respectively. Glucantime as control drug also revealed IC50 values of 88.3 and 44.6 mug/ml for promastigotes and amastigotes of L. tropica, respectively. The in vitro assay demonstrated no significant cytotoxicity in J774 cells. However, essential oil indicated a more cytotoxic effect as compared with the methanolic extract of M. communis. The findings of the present study demonstrated that M. communis might be a natural source for production of a new leishmanicidal agent.

Antileishmanial and Cytotoxic Effects of Essential Oil and Methanolic Extract of Myrtus communis L

Plants used for traditional medicine contain a wide range of substances that can be used to treat various diseases such as infectious diseases. The present study was designed to evaluate the antileishmanial effects of the essential oil and methanolic extract of Myrtus communis against Leishmania tropica on an in vitro model. Antileishmanial effects of essential oil and methanolic extract of M. communis on promastigote forms and their cytotoxic activities against J774 cells were evaluated using MTT assay for 72 hr. In addition, their leishmanicidal activity against amastigote forms was determined in a macrophage model, for 72 hr. Findings showed that the main components of essential oil were alpha-pinene (24.7%), 1,8-cineole (19.6%), and linalool (12.6%). Findings demonstrated that M. communis, particularly its essential oil, significantly (P<0.05) inhibited the growth rate of promastigote and amastigote forms of L. tropica based on a dose-dependent response. The IC50 values for essential oil and methanolic extract was 8.4 and 28.9 mug/ml against promastigotes, respectively. These values were 11.6 and 40.8 mug/ml against amastigote forms, respectively. Glucantime as control drug also revealed IC50 values of 88.3 and 44.6 mug/ml for promastigotes and amastigotes of L. tropica, respectively. The in vitro assay demonstrated no significant cytotoxicity in J774 cells. However, essential oil indicated a more cytotoxic effect as compared with the methanolic extract of M. communis. The findings of the present study demonstrated that M. communis might be a natural source for production of a new leishmanicidal agent.

Canine visceral leishmaniasis in Kerman, southeast of Iran: a seroepidemiological, histopathological and molecular study

Canine visceral leishmaniasis [CVL] is a systemic disease with a high mortality rate, caused by a diphasic protozoan parasite, Leishmania infantum/chagasi in the world. The objective of the present study was to determine the presence of CVL in the city and suburbs of Kerman, using a range of serological, histopathological and molecular methods. Blood samples were taken from 80 clinically symptomatic stray dogs All the collected blood samples were tested by direct agglutination test [DAT] to detect the anti-Leishmania antibodies in dogs, using a cut-off value of >/= 1:320. Pathological specimens including spleen, liver and lymph nodes were prepared for paraffin blocks, sectioning, staining and final microscopic examination in the pathology laboratory. PCR amplification of kDNA from 9 samples of DAT positive stray dogs was studied. The anti-Leishmania antibody was detected in 9 dogs [11.25%] of the total 80 studied dogs. No significant difference was found between VL infection and gender. In contrast, there was a significant difference between seropositivity and age [P<0.05]. Pathological samples showed changes including hyperplasia of infected macrophages and inflammatory cells that occupied sinusoids and splenic cords. Among the samples which was characterized by PCR, only one specimen revealed to be mixed infection between L. infantum and L. tropica. The results revealed a high prevalence of L. infantum infection in stray dogs in Kerman. This kind of information is needed for implementation of future control programs

In vitro study of leishmanicidal activity of biogenic selenium nanoparticles against Iranian isolate of sensitive and glucantime-resistant leishmania tropica

Sensitive and glucantime [MA] resistance Leishmania tropica are referred to those isolates, which are responsive, or non-responsive to one or two full courses of treatment by MA systematically and/or intra-lesionally, respectively. In this study, we evaluated the antileishmanial activity of biogenic selenium nanoparticles [Se NPs] alone and in combination with MA against sensitive and glucantimeresistant L. tropica on in vitro model. The Se NPs were synthesized by employing the Bacillus sp. MSh-1. The antileishmanial effects of Se NPs alone and in combination with MA on promastigote and amastigote stages of sensitive and glucantime-resistant L. tropica strains have been investigated using a colorimetric MTT assay and in a macrophage model. In addition hemolytic activity in type O+ human red blood cells and infectivity rate of the promastigotes before and after treatment with the Se NPs was evaluated. In the promastigote stage, various concentrations of Se NPs significantly inhibited [P<0.05] the growth of promastigotes of both strains in a dose-dependent manner. Similarly, Se NPs especially in combination with MA significantly reduced the mean number of amastigotes of both strains in each macrophage. Se NPs showed no hemolytic effect on human RBCs at low concentrations. Moreover, infection rate of macrophages by promastigotes significantly [P<0.05] was reduced when promastigotes pre-treated with Se NPs. The findings of this study suggest a first step in the search of Se NPs as a new antileishmanial agent. Further experiments are needed to investigate antileishmanial effects of biogenic Se NPs on L. tropica using a clinical setting

Causes of pediatric visceral leishmaniasis in southeastern Iran

Leishmania infantum is the most frequent cause of visceral leishmaniasis and L. tropica has been rarely linked to the disease in Iran. In this study, bone marrow aspirates were collected from 10 child patients, suspected with visceral leishmaniasis referred to the Pediatric Wards of Kerman Medical Hospitals, Kerman, Iran during 2002- 2011. Leishmania species were identified by using nested PCR in all slides. The PCR samples from nine patients indicated L. infantum as principal causative agent of visceral leishmaniasis and one L. tropica as a minor species

vitro inhibitory effect of berbcris vulgaris [berberidaceae] and its main component, berberine against different leishmania species

Leishmaniasis has been identified as a major public health problem in tropical and sub-tropical countries. The present study was aimed to investigate antileishmanial effects of various extracts of Berberis vulgaris also its active com-poenent, berberine against Ieishmania tropica and L infantum species on in vitro experiments..In this study in vitro antileishmanial activity of various extracts of B. vulgaris also its active compoenent, berberine against promastigote and amastigote stages of L. tropica and L. infantum was evaluated, using MTT assay and in a macro-phage model, respectively. Furthermore, infectivity rate and cytotoxicity effects of B. vulgaris and berberine in murine macrophage cells were investigated. The findings of optical density [OD] and IC[50] indicated that B. vulgaris particulary berberine significantly [P<0.05] inhibited the growth rate of promastigote stage of L.tropica and lL.infantum in comparison to meglumine antimoniate [MA]. In addition, B. vulgaris and berberine significantly [P<0.05] decreased the mean number of amastigotes in each macrophage as compared with positive control. In the evaluation of cytotoxicity effects, it could be observed that berberine as compared with B. vulgaris exhibited more cytotoxicity against murine macrophages. Results also showed that when parasites were pre-incubated with B. vulgaris their ability to infect murine macrophages was significantly decreased. Conclusion. B.vulgaris particularly berberine exhibited potent in vitro leishmanicid-al effects against L tropica and L.infantum. Further works are required to evaluate the antileishmanial effects of B. vulgaris on Leishmania species using clinical settings

Evaluation of a possible synergistic effect of meglumine antimoniate with paromomycin, miltefosine or allopurinol on in vitro susceptibility of leishmania tropica resistant isolate

Pentavalent antimonials are still the first choice treatment for leishmaniasis, but with low efficacy and resistance is emerging. In the present study, the effect of meglumine antimoniate [MA, Glucantime] combined with paromomycin, miltefosine or allopurinol on in vitro susceptibility of Leishmania tropica resistant isolate was evaluated. The drugs were obtained from commercial sources and diluents of each drug in medium were prepared on the day of experiment. J774 A.1 murine macro-phage cell lines were attached to the cultured on slide and incubated at 37 [degree sign]C with 5% CO[2] for 24 h. Then the stationary phase promastigotes were added to the cells and after 4 hrs of incubation different concentrations of MA, paromomycin, miltefosine or allopurinol were added and incubated for an additional of 72 h. Then the slides were dried and fixed with methanol, stained by Giemsa and studied under a light microscope. Drug activity was evaluated by assessing the macrophage infection rate and the number of amastigotes per infected macrophage was done by examining 100 macrophages. The experiment was done in triplicates. Various concentrations of MA along with paromomycin, miltefosine or allopurinol significantly inhibited [P<0.01] the proliferation of L. tropica amastigote stage in the macrophage cell line as compared with MA alone or positive control. Combination of Glucantime with paromomycin, miltefosine or allopurinol showed a synergistic effect on the clinical isolate of L. tropica in vitro. Use of combination therapy is a new hope and a logical basis for therapy of the patients with cutaneous leishmaniasis. Further investigations are needed to evaluate the therapeutic effects of these drugs on the CL patients

present status of cutaneous leishmaniasis in a recently emerged focus in south-west of Kerman province, Iran

The objective of this study was to assess the magnitude of the cutaneous leishmaniasis [CL] disease and identification of the causative agent by nested-PCR for current control strategy. This study was carried out as descriptive house-to-house visits in Orzoieh district in Kerman Province, south-east Iran, during 2011-2012. A questionnaire was completed for each individual consisting of demographic and clinical data. Suspected individuals were examined by direct smear microscopy and subsequent identification by nested-PCR. Chi[2]-test was used for any significance [P<0.05]. A total of 18308 inhabitants [mean age; 22.7 yr] consisting of 9011 males [49.2%] and 9297 females [50.8%] were examined for the presence of active or chronic lesions. the overall prevalence was 4.7%, including 30 cases of active and 839 cases of scar, distributed more significantly [P<0.01] in females [5.2%] than males [4.3%]. Individuals <10 years of age showed the highest [6.3%] and >50 years the lowest rate of CL disease, respectively [P<0.001]. The proportion of infection was the highest in Soltanabad [14.7%], followed by Vakilabad [6.8%], Dolatabad [3.2%] and Shahmaran [2.8%]. The majority of cases had 2 lesions [mean; 2.1 lesions]. Hand was the most common site of involvement [35%], and then face [26%], and multiple locations [39%]. Nested-PCR displayed 29 isolates as Leishmania major and one isolate L. tropica. The CL disease first emerged in 1998 as epidemic in the area and appeared endemics, thereafter. L. major was the sole species caused ZCL. These findings are necessary for future control programs and strategic planning

[Comparison of the efficacy of weekly vs. twice a week intralesional injections of meglumine antimoniate in the treatment of anthroponotic cutaneous leishmaniasis: a randomized clinical trial]

Treatment of cutaneous leishmaniasis, especially when caused by L. tropica, is challenging. Meglumine antimoniate [Glucantime[R]] is used as the standard treatment, but multiple injectiond are necessary. The objective of this study was to compare the efficacy of weekly intralesional injections with twice weekly injections of Glucantime for the treatment of anthroponotic cutaneous leishmaniasis [ACL]. This randomized open clinical trial was conducted, in Bam, Kerman province, Iran. 96 eligible patients according to inclusion and exclusion criteria who were willing to participate were included. The included patients were randomly assigned into two groups, one group treated with weekly intralesional injections of Glucantime[R] and the other group treated with intralesional Glucantime[R] twice a week. Type and size of each lesion [induration, ulcer and scar] were recorded weekly. Complete healing was defined as complete re-epithelialization and absence of induration in all lesions and was considered as the primary outcome measure. A total of 48 patients completed the study; complete cure was seen in 24 of 27 [89%] patients who received weekly intralesional MA with a mean duration of healing equals to 70 +/- 10 days. Complete cure was seen in 24 of 31 [77%] patients who received intralesional MA twice a week, the mean duration of healing in the latter group was 58 +/- 5 days. There was no significant difference between the two groups [P=0.23]. It seems that the efficacy of intralesional injections of Glucantime[R] once a week is similar to efficacy of twice a week Glucantime[R] injections

prevalence, incidence and reinfection rate of Giardia lamblia in primary school children in rural suburb of Kerman city in southern Iran

Giardiasis is one of the most common parasites in young children in both developed and developing countries. This disease is very prevalent in Iran, especially in Kerman city, southeast Iran. To determine the prevalence, incidence and reinfection rate of Giardia lamblia, this study has been conducted in primary schoolchildren in rural suburb of Kerman city in southern Iran. A total of 574 schoolchildren were selected randomly and their stools were examined by formalin-ether concentration method. Infected children were weighted and treated with suitable doses of metronidazole or furazolidone. Post treatment stool specimens were collected after two weeks for assessment of complete cure, and follow-up specimens were collected six months after the last treatment. The prevalence rate in both boys and girls was generally equal, while it was lower in 8.9 years old children than 10-11 years old ones in both sexes. The incidence rate was higher in girls [21.4%] than the corresponding boys [14.5%], and this different was significant [P<0.05]. In both sexes younger children were more likely to become infected than older children. The reinfection rate was 54.5% in boys, whereas it was 79.3% in girls. As indicated, the younger children acquired the infection in a much higher rate than the older ones, particularly in female groups [P<0.05]. The overall reinfection rate was 66.8%. On the basis of this study we would suggest that in endemic areas, children infected with giardiasis should be treated in a regular manner

prevalence of intestinal parasites in afghan refugees in Kerman City, Iran

Single stool specimens were obtained from the subjects at the referral center for refugees in Kerman city during spring and fall seasons of 1993. Specimens were processed within 1-2 hours of receipt by the formalin-ether concentration method. The overall prevalence of infection was 23.7% for one or more species of intestinal parasites. The common parasites were Giardia lamblia, 15.2%; Hymenolepis nana, 5.2%, Entamoeba histolytica, 2.5%, and Ascoris lumbricoides, 0.8 perecnt. In general the difference in the distribution of parasites between females and males was not significant

Comparative study between efficacy of metronidazole and furazolidone in 7 to 11 year old children with giardiasis

Metronidazole and furazolidone were compared for efficacy in 268 seven to 11 year old children infected with giardiasis. With metronidazole the cure rate was 82.3% and 80.0% in boys and girls respectively, while furazolidone showed an efficacy of 74.4% and 71.7% in the corresponding groups. The overall efficacy was 81.1% with metronidazole and 73.3% with furazolidone. There were no significant differences between metronidazole and furazolidone efficacies in regard to the age groups or sexes