Antileishmanial effect of podophyllotoxin and podophyllin on Leishmania major in vitro and in vivo.

Therapeutic research is very important in the prevention and treatment of leishmaniasis due to problems such as drug resistance, scarring and disease recurrence. The aim of this study was to determine how Leishmania major responds to the anti-leishmaniasis properties of podophyllotoxin and podophyllin. Cultured Leishmania promastigotes were exposed to different concentrations of podophyllotoxin and podophyllin for 24 and 48 h. Then, during the animal phase, Balb/c mice were experimentally injected with Leishmania promastigotes. After wounding, the effects of 0.5% podophyllotoxin and 25% podophyllin on reducing wound diameter and the number of amastigotes in the wound were evaluated. Podophyllotoxin and podophyllin were 83% and 59% lethal to Leishmania major promastigotes at the highest concentrations (200 µg/ml) and time (48 h). In the in vivo study, the mean lesion diameter at the end of treatment in the negative control group was 15.10 mm compared to 14.21 mm and 11.55 mm in the 25% podophyllin and 0.5% podophyllotoxin groups, respectively. Although both agents reduced the size of mice wounds and the number of amastigotes in the wounds, podophyllotoxin was more effective in this regard. Based on the results, podophyllotoxin and podophyllin can be used as leishmaniasis drugs after further research.

Molecular detection and genotype identification of Acanthamoeba species from bronchoalveolar lavage of patients with pulmonary symptoms suspected of cancer.

Acanthamoeba spp. are the most common free-living amoeba worldwide, inducing life-threatening diseases such as Granulomatous Amoebic Encephalitis, pulmonary infection, and amoebic keratitis. This study aimed to identify the FLA and Acanthamoeba genotypes in patients with pulmonary symptoms suspected of cancer in Kashan's hospitals, Kashan, Iran. This cross-sectional study was conducted on 97 bronchoalveolar lavage samples of patients with respiratory symptoms suspected of lung cancer, who were admitted to the Shahid Beheshti Hospital of Kashan from 2019 to 2020. The samples were cultured onto 1.5% non-nutrient agar enriched with killed Escherichia coli and examined for the presence of FLA. Following amoeba isolation and DNA extraction, Acanthamoeba spp. were determined by Polymerase Chain Reaction using JDP1 and JDP2 primers, which amplified a 490 bp fragment from the 18 S rDNA gene. Eighteen Acanthamoeba isolates were sequenced, and the genotypes were identified. The prevalence of FLA and Acanthamoeba and the relationship between symptoms and demographic variables were analyzed with SPSS Software version 16. The prevalence rates of FLA and Acanthamoeba in the BAL samples was 86.6% and 73.2%, respectively. All Acanthamoeba isolates belonged to the T4 genotype. The most symptoms among Acanthamoeba-positive patients were dyspnea and cough; however, their difference was not statistically significant. The findings indicated the high prevalence of FLA and Acanthamoeba in BAL in the population suspected of cancer in Kashan. Since the T4 genotype is a pathogenic genotype of Acanthamoeba, training health and improving sanitation levels would help to prevent infection.

Scolicidal Effects of Chitosan-Curcumin Nanoparticles on the Hydatid Cyst Protoscolices.

PURPOSE: In the current era, cystic echinococcosis (CE), as larval stage of Echinococcus granulosus, is considered as a threat to human health. Scolicidal agents used in the surgery of cysts have different side effects. Therefore, the present study aimed to assess the effects of chitosan nanoparticles containing curcumin (Ch-Cu NPs) on the protoscolices of the hydatid cyst in vitro. METHODS: Ch-Cu NPs were synthesized using a simple co-precipitation method and their structural and morphological properties were characterized by X-ray diffraction (XRD), scanning electron microscopy (SEM), zeta analyzer, and Fourier transform infrared (FT-IR) spectroscopy. Then, the effects of different concentrations of Ch-Cu NPs (0.25, 0.05, 1, 2, and 4 mg/mL) on the fatality rate, and the length and width of protoscolices in different times (5, 10, 20, 30, and 60 min) were investigated. In addition, the SEM technique was used to evaluate the structure of the protoscolices after treatment. RESULTS: Based on the results, the presence of curcumin on the chitosan nanoparticles was confirmed by FT-IR analysis. Further, XRD analysis approved the crystal structure of chitosan NPs. Furthermore, the highest fatality rate was 68% in 4 mg/mL concentration of Ch-Cu NPs. The length and width of protoscolices decreased based on the high concentrations of Ch-Cu NPs, compared to the control group. CONCLUSION: Finally, Ch-Cu NPs expressed good scolicidal activities, which made them suitable to be considered as an anti-protoscolex agent.

Giardia lamblia infection: review of current diagnostic strategies.

Giardiasis has a global distribution and it is a common cause of diarrhea in both children and adults and is transmitted via the fecal-oral route through direct or indirect ingestion of cysts. The laboratory diagnosis of Giardia spp. is mainly based on demonstration of microscopic cyst or trophozoite in stool samples but several immunological-based assays and molecular methods are also available for giardiasis diagnosis. The aim of this study was to conduct a review of the applied methods in medical laboratory and to highlight pitfalls and challenges of them for diagnosis of giardiasis. In this article we have evaluated the Giardia diagnostic methods with a broad review of literature, electronic databases and books. The search has covered the articles and some textbooks that have published up to 2018. It has been concluded that traditional microscopy combination with stool concentration method should still be held in the routine medical laboratory due to economical and high sensitivity and immunological-based assay and molecular methods which are recommended to use as a complementary test to the traditional technique.

Review of Trichomonas vaginalis in Iran, Based on Epidemiological Situation.

Trichomoniasis, which is caused by Trichomonas vaginalis, is the most common non-viral sexually transmitted infection (STI) in the world including Iran. There were roughly 250 million new cases all over the world in a year. T. vaginalis as an important disease has been associated with HIV (in terms of exposure to sexually transmitted infection, STI) which increases the number of high-risk members, and thus it is an important public health problem. Additionally, this pathogen has been associated with serious health consequences. For instance, it may cause a woman to deliver a low-birth-weight or premature infant, and increase chances of cervical cancer. Because little information is available about the prevalence of T. vaginalis infection in Iranian population, this review was carried out to determine the prevalence of T. vaginalis among Iranian population. For this systematic review, data about epidemiology of T. vaginalis in different parts of Iran with different populations were systematically collected from 1992 to 2017 through the international databases such as PubMed, Scirus, ISI Web of Science, Scopus, EMBASE, Science Direct and Google Scholar and Islamic World Science Citation Center (ISC). National database searching included Iran Medex, Iran Doc, Magiran and Scientific Information Database (SID). A total of 39 clinical and laboratory investigations about the prevalence of Trichomoniasis from different regions of Iran were analyzed. The overall prevalence rate of T. vaginalis infection in Iranian population was estimated to be minimally 0.4% and maximally 42%. The present review showed that T. vaginalis infection rate is relatively high among the Iranian population. The control strategies, including personal hygienic education, simultaneous couple treatment, the sensitivity of diagnostic methods, appropriate preventive tool (condom) in sexual contacts could lead to the disruption of transmission.

Induction of Apoptosis by Alcoholic Extract of Combination Verbascum thapsus and Ginger officinale on Iranian Isolate of Trichomonas vaginalis.

BACKGROUND: The protozoan Trichomonas vaginalis is a sexually transmitted disease (STD). Metronidazole is a chosen drug for the treatment. This study evaluated the anti trichomonal activity of alcoholic extracts of combination Verbascum thapsus and Ginger officinale. METHODS: This experimental study was conducted in the Parasitology Laboratory, Kashan University of Medical Sciences, Kashan, Iran in 2015, on 23 women with suspected trichomoniasis referring to Kashan clinical centers. Medium TYI-S-33 was used for culture of three T. vaginalis isolates. Different concentrations (25, 50, 100, 200, 400, 800 µg/ml) of V. thapsus and G. officinale ethanol extract added to Trichomonas trophozoites in 48-well plates and metronidazole considered as positive control and the negative control was TYI-S33 containing Trichomonas trophozoites without any drug. In all of mentioned groups, trophozoites number counted 12, 24, 48 h after culture. Results were analyzed using ANOVA statistical test, to evaluate the toxicity of extract, measured by MTT assay. Induced apoptosis of T. vaginalis after treatment with different concentrations of extract was determined by Flow Cytometry. RESULTS: IC50 of alcoholic extract of combination V. thapsus and G. officinale and metronidazole after 24h was 73.80 µg/ml and 0.0326 µg/ml, respectively. The toxicity percentage of 25-800 µg/ml concentrations of this combination were between 0.2-1.98. In different concentrations of extract (25,50,100,200 and 400 µg/ml) apoptosis percent after 48h was 18.97 to 77.19 and necrosis percent was calculated 1.35, 3.18, 3.10, 1.16 and 4.09, respectively. CONCLUSION: Alcoholic extract of combination V. thapsus and G. officinale induces programmed death in T. vaginalis. Due to no toxicity on macrophages, it can be examined in vivo studies.

Epidemiology and economic loss of fasciolosis and dicrocoeliosis in Arak, Iran.

AIM: Fasciolosis and dicrocoeliosis are important parasitic diseases worldwide, causing significant financial losses due to decrease in production and viscera condemnation in animals. We performed the current research to assess the epidemiology of these infections and determine their significance from an economic perspective in Arak, Iran. MATERIALS AND METHODS: In total, we evaluated 118,463 sheep, 207,652 goats, and 43,675 cattle through necropsic analysis at the slaughterhouses. The average weight of sheep, goat, and cattle liver was 1000, 900, and 5000 g, respectively. The average price of liver in the market was 8 USD/kg. Moreover, the elimination of fundamental nutrients and vitamins was evaluated in infected livers. The prevalence of fasciolosis and dicrocoeliosis was determined. Analysis of variance test was applied for the statistical analysis, and the significance level was <0.05. RESULTS: In total, Fasciola hepatica and Dicrocoelium dendriticum were detected in 0.56% (confidence interval CI, 0.54-0.59) and 0.77% (CI, 0.75-0.81) of the animals, respectively (p=0.1). The annual economic loss attributed to fasciolosis and dicrocoeliosis was 26698.4 and 30479.2 USD, respectively. The total economic loss was 10,880, 9079.2, and 10,520 dollars in sheep, cattle, and goats, respectively. On the other hand, financial loss resulting from fasciolosis was 7160, 6098.4, and 13,440 dollars in sheep, goats, and cattle, respectively. In addition, economic loss due to dicroceliasis was 10,880, 9079.2, and 10,520 dollars, respectively. CONCLUSION: Overall, fasciolosis and dicrocoeliosis in Iran always remain common in sheep, goats, and cattle that afford major economic loss of all the country also exist in Arak province. The present study could provide basic information for further examination of liver fluke infections in Iran.

Chitosan-titanium dioxide-glucantime nanoassemblies effects on promastigote and amastigote of Leishmania major.

The purpose of the present study was to design nanoassemblies of chitosan-titanium dioxide (TiO2) nanoparticles (NPs) loaded with glucantime for using their synergistic effects and enhancing the toxic effects of glucantime on Leishmania parasites. The nanoassemblies were prepared by electrostatic interactions and optimized by a response surface central composite design. The effects of glucantime, chitosan and TiO2 NPs amounts were studied on the particle size, zeta potential, loading efficiency, and release efficiency of drug from nanoassemblies. The conjugation of TiO2/chitosan-glucantime was verified by UV spectroscopy and changes in surface charge of NPs. The anti-promastigots effect of glucantime loaded in TiO2/chitosan nanoassemblies was studied by tripan blue dye test and their anti-amastigotes effect by counting the average number of parasites per infected J774 macrophages in 100 cells. The optimized formulation obtained by using 12.5mg glucantime, 25mg chitosan and 6mg TiO2 NPs. Although TiO2 NPs alone were effective more than negative control in reduction of promastigots and amastigotes but they didn't show significant difference compared with free glucantime (p>0.05). However, at the concentration of 50µg/mL and after 72h exposure nanoassemblies decreased the proliferation of L. major promastigotes and amastigotes 13 and 4-fold, respectively compared with glucantime alone.

Genetic Variation of Giardia lamblia Isolates from Food-handlers in Kashan, Central Iran.

BACKGROUND: Based on genotyping study of human isolates of Giardia lamblia; humans are mainly infected by two assemblages A and B. The present study was carried out to determine the sub-assemblages of G. lamblia isolated from food handlers referred to Kashan health centers, central Iran, 2015. METHODS: In this cross-sectional study, 3653 stool samples collected from food-handlers that annually refer to health center for getting a health certification and examined microscopically for G. lamblia cyst. Totally, 44 isolates were selected from 47 Giardia positive samples. Cysts were partially purified by the sucrose density gradient method. After freeze-thaw cycles, genomic DNA was extracted using QIAamp Stool Mini kit. A single step PCR-RFLP method was used to amplify a 458bp fragment at the glutamate dehydrogenase (gdh) locus, restriction enzymes BspLI and RsaI were used for distinguish between genotypes A and B and their subgroups. RESULTS: Of 44 isolates, 24(54.5%) were sub-assemblage AII, 9(20.5%) group B including 7(15.9%) BIII and 2(4.6%) BIV sub-assemblage and 11(25%) isolates showed a mixed pattern of AII and B. Sub-assemblage AI was not detected in this study. CONCLUSION: The higher rate of sub-assemblage AII demonstrated an anthroponotic origin of the infection so infected food-handlers could directly transmit this protozoan to consumers via contaminated food and water. For finding of pattern of transmission and distribution of Giardia assemblages and sub-assemblage, more studies in human and animal population in different regions are necessary.

Molecular and Morphological Characterizations of Echinococcus granulosus from Human and Animal Isolates in Kashan, Markazi Province, Iran.

BACKGROUND: One of the most important zoonotic helminths in the world is known as Echinococcus granulosus. Different strains of the E. granulosus have been described based on morphological and molecular characterizations, however, there is limited information regarding the characteristics of the phenotypes and genotypes of E. granulosus in Iran. METHODS: The present study was prepared to evaluate the phenotypic and genotypic diversity of E. granulosus isolates collected from human, goat, sheep, and cattle based on 19 standard morphometric parameters and mitochondrial and nuclear genes (CO1, ND1, and ITS1) in Kashan, Markazi Province, Iran during 2013-2014. RESULTS: The biometric analysis for the 19 characters revealed that the 19 morphometric values of cattle isolates were exceptionally higher than human, goat, and sheep isolates (P<0.05). Molecular analysis confirms the morphological findings. Phylogenic analysis of the CO1, NAD1 and ITS1 genes for all isolates, independent of the host, revealed that the common sheep strain (G1) is traveling among livestock in Kashan and the strains are highly adapted to goats, cattle, sheep, and humans. CONCLUSION: Both morphological and molecular results of this study indicated that the only genotype G1 of E. granulosus travels between humans and other intermediate hosts of this parasite in the area study.

Therapeutic effects of Iranian herbal extracts against Trichomonas vaginalis

Trichomonas vaginalis is a flagellated parasite affecting about 276 million people annually worldwide. Tricomoniasis is associated with different complications in pregnant women and infants. 5'-nitroimidazole derivatives (metronidazole, ornidazole, and tinidazole) are FDA approved drugs recommended for trichomoniasis treatment. Treatment with metronidazole 5'-nitroimidazole derivatives is associated with many side effects, and drug resistance to metronidazole has been reported in some cases. Recently, many attempts have been made to evaluate the effects of plants on causative agents of vaginal infections. In our research, the national and international databases were searched and the effects of various herbal extracts on T. vaginalis in Iran were reviewed from 2006 to 2016. In articles investigated, some plants had favorable antitrichomonal effects and needed to be further investigated. All the plant extracts have only been evaluated in vitro. Surveys of different articles in this review show that the active ingredients present in different parts of plants, including aerial parts, leaves, flowers, stems, and root can be suitable sources for introducing and developing new antitrichomonal compounds.

Ginger (Zingiber officinale) induces apoptosis in Trichomonas vaginalis in vitro.

BACKGROUND: Trichomoniasis is the most common sexually transmitted protozoan diseases in the worldwide. Metronidazole is the choice drug for trichomoniasis treatment, however, metronidazole resistant Trichomonas vaginalis (T.vaginalis) has been reported. Natural products are the source of most new drugs, and Zingiber officinale (Ginger) is widely used ingredient in the traditional medicine. OBJECTIVE: The aim of the present study was to determine the effect of different concentrations of the ginger ethanol extract on the growth of T.vaginalis trophozoites in vitro. MATERIALS AND METHODS: In this experimental study, 970 women who were attend in Kashan health centers were examined for T. vaginalis. Of them, 23 samples were infected with T.vaginalis. Three T. vaginalis isolates were cultured in a TYI-S-33 medium. The effect of ginger ethanol extracts and its toxicity in different concentrations (25, 50, 100, 200, 400, 800 µg/ml) on mouse macrophages were measured in triplicate exam by MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] assay. The effect of ginger on apoptosis induction was determined by Flow cytometry. RESULTS: The IC50 of ginger and metronidazole were 93.8 and 0.0326 µg/ml, respectively. 12, 24 and 48 hr after adding different concentrations of extract on mouse macrophages, fatality rates in maximum dose (800 µg/ml) were 0.19, 0.26 and 0.31 respectively. Flow cytometry results showed the apoptosis rate following treatment with different concentrations of the extract after 48 hr were 17, 28.5, 42.1, 58.8, 76.3 and 100% respectively, while in the control group was 2.9%. CONCLUSION: Ginger ethanol extract induces programmed death in T. vaginalis. It is recommended that due to the known teratogenic effect of metronidazole, ginger can be considered as an alternative drug for metronidazole.

Tamoxifen Induces Apoptosis of Leishmania major Promastigotes in Vitro.

Tamoxifen is an antagonist of the estrogen receptor and currently used for the treatment of breast cancer. The current treatment of cutaneous leishmaniasis with pentavalent antimony compounds is not satisfactory. Therefore, in this study, due to its antileishmanial activity, effects of tamoxifen on the growth of promastigotes and amastigotes of Leishmania major Iranian strain were evaluated in vitro. Promastigotes and amastigotes were treated with different concentrations (1, 5, 10, 20, and 50 µg/ml) and time periods (24, 48, and 72 hr) of tamoxifen. After tamoxifen treatment, MTT assay (3-[4,5-dimethylthiazol-2-yl]-2,5 biphenyl tetrazolium bromide assay) was used to determine the percentage of live parasites and Graph Pad Prism software to calculate IC50. Flow cytometry was applied to investigate the induction of tamoxifen-induced apoptosis in promastigotes. The half maximal inhibitory concentration (IC50) of tamoxifen on promastigotes was 2.6 µg/ml after 24 hr treatment. Flow cytometry analysis showed that tamoxifen induced early and late apoptosis in Leishmania promastigotes. While after 48 hr in control group the apoptosis was 2.0%, the 50 µg/L concentration of tamoxifen increased it to 59.7%. Based on the in vitro antileishmanial effect, tamoxifen might be used for leishmaniasis treatment; however, further researches on in vivo effects of tamoxifen in animal models are needed.

Temporal Dynamics of Phlebotomine Sand Flies Population in Response to Ambient Temperature Variation, Bam, Kerman Province of Iran.

BACKGROUND: Variations in climate condition may have changed the dynamic of zoonotic cutaneous leishmaniasis (ZCL) and its agents such as sand flies and reservoir in the Bam Kerman the dry region of Iran. OBJECTIVES: In this study we intend to examine the seasonal and interannual dynamics of the phlebotomine mosquito as a function of ambient temperature in Bam, Kerman one of the main leshmaniasis prevalence area in Iran. METHODS: The MODIS land surface temperature product (LST; MODIS/Terra LST/E Monthly L3 Global 0.05Deg CMG [MOD11C3]) and land-based climatic data were used as explanatory variables. Monthly caught mosquitoes in Bam, Kerman, were used as a dependent variable. The temporal associations were first investigated by inspection of scatterplots and single-variable regression analysis. A multivariate linear regression model was developed to reveal the association between ambient temperature and the monthly mosquito abundance at a 95% confidence level (P < 0.05). FINDINGS: The findings indicated that the monthly variation of 0-10 cm of soil depth temperature is the main driver of phlebotomine mosquito temporal dynamics. The developed multivariate model also indicated that the ambient temperature variation was responsible for >0.80 of temporal dynamics of phlebotomine mosquitos in Bam.

Molecular Diagnosis of Clinical Isolates of Cutaneous Leishmaniasis Using ITS1 and KDNA Genes and Genetic Polymorphism of Leishmania in Kashan, Iran.

Cutaneous leishmaniasis is a common skin disease caused by leishmania parasite. An accurate diagnosis of parasites species is possible using molecular techniques. This study was carried out to compare internal transcribed spacer (ITS1) and kinetoplast deoxyribonucleic acid (KDNA) genes for identifying Leishmania species by Polymerase Chain Reaction (PCR), furthermore, genetic diversity of isolates was studied. This research examined 130 patients who were suspected of cutaneous leishmaniasis and referred to Kashan's health centers from 2011-2014. After DNA extraction from serosity, PCR were performed using ITS1 and KDNA primers. Cutaneous Leishmaniasis was diagnosed by the observation of 320 bp band in the ITS1-PCR. The PCR products were digested with restriction enzyme HaeIII and then leishmania species were identified by patterns of enzymatic digestion. The diagnostic criteria of Cutaneous Leishmaniasis (CL) in KDNA-PCR were based on the observation of 760 and 650 bp for Leishmaniasis tropica and Leishmaniasis major, respectively. Twelve isolates of leishmania were sequenced and the phylogenetic tree was traced using the results of sequencing by Mega 4 software. Out of 130 suspected patients to CL, 70 (53.8%) and 98 (75.4%) isolates were positive by Restriction Fragment Length Polymorphism (RFLP) of ITS1 and KDNA, respectively. Using ITS1 PCR, 60 samples (85.7%) and 10 samples (14.3%) were identified as L. tropica and L. major, respectively, ITS1-PCR had 25.3% false negative, compare to microscopy. While, microscopy had false negative in 13 cases compare to KDNA-PCR. Due to the lower sensitivity of the PCR-RFLP of ITS1, KDNA-PCR is recommended for diagnosis of CL. The L. tropica and L. major are the causative agents of CL.

Evaluation of Apoptotic and Antileishmanial Activities of Artemisinin on Promastigotes and BALB/C Mice Infected with Leishmania major.

BACKGROUND: In leishmaniasis, some drugs prescribed for treatment have toxic effects and there are reports about drug resistance in some countries. Due to this fact, using herbal drugs such as artemisinin with good efficacy and low toxic effect might be suitable. METHODS: We evaluated the apoptotic effect of artemisinin on Leishmania major in vitro and the antileishmanial activities of artemisinin on leishmaniasis in BALB/c mice and at the end INF-γ and IL-4 cytokines levels were detected by ELISA in spleen cell culture supernatants. During treatment the lesion size and survival rate were measured each four and ten days, respectively. RESULTS: Percentage of early and late apoptosis in promastigotes of control group and promastigotes treated with 10, 25, 50 and 100 µg/ml of artemisinin after 48 h were 0.13, 16.04, 41.23, 49.03 and 81.83, respectively. The IFN-γ in ointment treated group were higher than those of other groups (P<0.05). The in vivo results showed that ointment compounds healed the lesions more effectively rather than intraperitoneal injection method (P<0.05). The survival rate of mice 150 days after challenge in treated group with ointment of artemisinin was 66% while all mice in control groups were died. CONCLUSION: All of in vitro results represented that this drug had antileishmanial effects and these results were confirmed by evaluation effects in vivo condition of leishmaniasis. Interestingly, according to these results it can be concluded that this drug has antileishmanial effects in vitro and in vivo conditions. Artemisinin induces cytotoxic effect on L. major via apoptosis-related mechanism.

Effect of Verbascum thapsus ethanol extract on induction of apoptosis in Trichomonas vaginalis in vitro.

Trichomoniasis is a sexually transmitted disease (STD) caused by a tiny parasite called Trichomonas vaginalis. Metronidazole is used as routine treatment of disease. Some reports have confirmed the potential carcinogenic and teratogenic effects of this drug on fetus and indication of drug resistance. Verbascum thapsus belongs to the family of Scorphulariaceae. Its antiinflammatory properties, disinfectant and skin healing effects are well known. This plant has been used to treat diarrhea and genitourinary infection in traditional medicine. Effects of different concentrations of the Verbascum thapsus extract were tested on the growth and motility of T. vaginalis trophozoites. To evaluate the toxicity of extract, their effects on mice macrophages were measured by MTT([3-(4,5-dimethyl thiazolyl-2)- 2,5-diphenyle tetrazolium bromide ])assay. In this experimental study the effect of Verbascum thapsus ethanol extract on induction apoptosis in T. vaginalis was determined by Flow Cytometry. Results were analyzed by Flow Jo software and the degree of apoptosis was determined. Toxicity percentage of 25-800 µg/ml concentrations of Verbascum thapsus alcoholic extract for mice macrophages was observed between 0.17-0.25 after 12 hours and they were between 0.25-0.42 and0.45-0.95 after 24 and 48h respectively. IC50 (inhibitory concentration, 50%) of Verbascum thapsus ethanol extract and metronidazole after 24h was 39.17 and 0.0326 µg/ml respectively. Flow cytometry results showed the percent of apoptosis following treatment of trophozoites with different concentrations of Verbascum thapsus ethanol extract (25, 50,100,200,400 µg/ml), were 20.7, 37.04, 47.5, 62.72 and 86.35 respectively, while in control group was 2.9. According to this study, Verbascum thapsus extract induces programmed death in T. vaginalis. It is recommended that Verbascum thapsus extract can be considered as a suitable choice for Medical Studies.

In vitro and in vivo antileishmanial effects of aloe-emodin on Leishmania major.

Cutaneous leishmaniasis is a common parasitic disease that is endemic in some parts of Iran. The drugs of choice used for leishmaniasis therapy are associated with a risk of recurrence and serious adverse effects. Therefore, finding a safe and effective treatment is of great importance. In the present study, the effect of aloe-emodin on the growth of Leishmania major amastigotes was evaluated under in vitro conditions. In addition, the efficacy of a topical of aloe-emodin ointment was investigated in BALB/c mice with cutaneous leishmanial ulcers. Different concentrations (40 µg/mL, 80 µg/mL, 120 µg/mL, and 160 µg/mL) of aloe-emodin were tested on Leishmania amastigotes twice: 24 hours and 48 hours. The induced apoptosis and necrotic effects of two concentrations (40 µg/mL and 120 µg/mL) of aloe-emodin on promastigotes were investigated by flow cytometry. Under the in vivo condition, aloe-emodin ointment efficacy was evaluated at two concentrations (i.e., 0.1% and 1%). Serum indicator factors of the test and control groups were tested to evaluate the toxic effects of this compound on the liver and kidney. Results showed that aloe-emodin inhibited the growth of Leishmania amastigotes and induced apoptosis in promastigotes. Topical application of aloe-emodin ointment likewise reduced the ulcer size. No significant differences in biochemical analysis were observed between the control and treated groups. In conclusion, aloe-emodin showed antileishmanial effects under in vitro and in vivo conditions and may be used in clinical trials.

Developing a climate-based risk map of fascioliasis outbreaks in Iran.

The strong relationship between climate and fascioliasis outbreaks enables the development of climate-based models to estimate the potential risk of fascioliasis outbreaks. This work aims to develop a climate-based risk map of fascioliasis outbreaks in Iran using Ollerenshaw's fascioliasis risk index incorporating geographical information system (GIS). Using this index, a risk map of fascioliasis outbreaks for the entire country was developed. We determined that the country can be divided into 4 fascioliasis outbreak risk categories. Class 1, in which the Mt value is less than 100, includes more than 0.91 of the country's area. The climate in this class is not conducive to fascioliasis outbreaks in any month. Dryness and low temperature in the wet season (December to April) are the key barriers against fascioliasis outbreaks in this class. The risk map developed based on climatic factors indicated that only 0.03 of the country's area, including Gilan province in the northern region of Iran, is highly suitable to fascioliasis outbreaks during September to January. The Mt value is greater than 500 in this class. Heavy rainfall in the summer and fall, especially in Rasht, Astara and Bandar Anzaly (≥ 1000 mm/year), creates more suitable breeding places for snail intermediate hosts.