Prevalence of Gardnerella vaginalis infection and antibiotic resistance pattern of isolates of gynecology clinic patients at Shahriar Noor Hospital from January to June 2020 by PCR and culture methods.

Background and Objectives: Gardnerella vaginalis is one of the most important causes of prevalent genital infections that pose serious risks. This study aimed to determine the prevalence of Gardnerella vaginalis and antibiotic resistance pattern of isolates of patients referred to the gynecology clinic of Shahriar Noor Hospital by PCR and culture methods. Materials and Methods: The study was conducted on 500 patients who had suffered from a vaginal infection. The demographic data of patients were studied. For diagnosis of Gardnerella vaginalis isolates, cultivation in anaerobic conditions, biochemical tests, PCR and Gardnerella vaginalis antibiotic susceptibility test to metronidazole and clindamycin were performed. Data analysis was performed utilizing SPSS statistical software version 19 and the Chi-square test. Results: Among the 500 patients, 173 were diagnosed with Gardnerella vaginitis. There was a significant relationship between age group, level of education, and contraceptive method with Gardnerella vaginosis incidence. Performing antibiotic susceptibility tests showed that the resistance of Gardnerella vaginalis isolated strains to metronidazole and clindamycin was 86.12% and 17.34%, respectively. Conclusion: The high prevalence of Gardnerella vaginalis infections confirms the critical role of the bacterium in the occurrence of bacterial vaginosis. Therefore, it is necessary to check the prevalence of bacterial infections to recommend the correct medical treatment in different societies.

Comprehensive study of antimicrobial susceptibility pattern and extended spectrum beta-lactamase (ESBL) prevalence in bacteria isolated from urine samples.

Nowadays, increasing extended-spectrum ß-lactamase (ESBL)-producing bacteria have become a global concern because of inducing resistance toward most of the antimicrobial classes and making the treatment difficult. In order to achieve an appropriate treatment option, identification of the prevalent species which generate ESBL as well as their antibiotic susceptibility pattern is essential worldwide. Hence, this study aimed to investigate the prevalence of ESBL-producing bacteria and assess their drug susceptibility in Fardis Town, Iran. A total of 21,604 urine samples collected from patients suspected to have urinary tract infection (UTI) were processed in the current study. The antimicrobial susceptibility of the isolates was tested by the disk diffusion method. The ESBL producing bacteria were determined by Double Disc Synergy Test (DDST) procedure. Bacterial growth was detected in 1408 (6.52%) cases. The most common bacterial strains causing UTI were found E. coli (72.16%), followed by K. pneumoniae (10.3%) and S. agalactiae (5.7%). Overall, 398 (28.26%) were ESBL producer. The highest ESBL production was observed in E. coli, followed by Klebsiella species. ESBL producers revealed a higher level of antibiotic resistance compared with non-ESBLs. In conclusion, ESBL production in uropathogens was relatively high. Carbapenems and Aminoglycosides were confirmed as the most effective treatment options for these bacteria.

High risk genotype distribution of human papillomavirus (HPV) according to age groups in Iranian asymptomatic men.

BACKGROUND: The Human Papillomavirus (HPV) is one of the most common sexually transmitted viruses worldwide. HPV infection in men is a serious clinical issue as they could be considered as a reservoir for inadvertently transmitting infection to women. Moreover, genital HPV infection could be a source for anogenital cancers in men. METHODS: This cross sectional study was conducted from January 2017 to December 2018. Four hundred fifteen asymptomatic men who were visited by specialists, referred to Nilou laboratory in terms of high risk (HR) HPV test testing. HR-HPV genotypes were detected using an approved assay which could discover HPV 16, HPV 18 and a pool of other high risk HPV genotypes as well as 16+ other HR and 18 + other HR (as multiple genotypes). SPSS software was used for statistical analysis. RESULTS: The mean age was 33 ± 8.14 years. Specimens were referred to the laboratory by urologists, (n = 132, 32%, 95%CI: 25.0-39.4), dermatologists, (n = 104, 25, 95% CI: 19.1-30.9), gynecologists, (n = 75, 18, 95%CI: 13.3-29.3) and other specialists (n = 104, 25, 95% CI:19.1-30.9). The overall prevalence of other HR HPV, HPV16, HPV18 and multiple genotypes were 54.2% (45/83), 25.3% (21/83), 3.6% (3/83) and 16.8% (14/83), respectively. The frequency of HR-HPV, HPV16 and HPV18 genotypes was the highest among 30-40 years old. CONCLUSION: The prevalence of HR-HPV infection among Iranian asymptomatic males was relatively high. Investigation on HPV infection in men as reservoir and transmission vehicle of HPV in addition to screening in women will improve the national public health provisions and will contribute to the application of infection control measurements at a national level.

Human papilloma virus (HPV) genotypes concordance between Iranian couples referrals.

BACKGROUND: Human Papilloma Virus (HPV) genotypes concordance among sexual couples has been evaluated in many investigations with considerable variations in the concordance. However, no such study has carried out between Iranian couples yet. METHODS: Urogenital specimen from both males and females of couples were taken and transferred to Nilou laboratory for molecular analysis. HPV DNA extraction and typing were carried out using cobas 4800 platform. Demographic and virological data were analyzed afterwards. RESULTS: One hundred fourteen couples were enrolled in the study. The mean age of participants were 36 ± 8 and 32 ± 7 for males and females, respectively. 64 (28%) of specimens were positive for at least one HPV genotype. The positive rates within genders were 30.7 and 25.4% for females and males, respectively with a considerable association (P value 0.021). Within the positive samples, 13(5.7%), 8 (7%) and 31(13.5%) were belonged to 16, 18 and other HR genotypes. 59 (51.8%) couples who were negative for HPV showed negative concordance. Of the total positive HPV patients (55 couples, 48.2%), 9 (16.3%) couples had positive concordance and the rest of 46 (83.7%) couples (either of spouse being negative and the other being positive for HPV) showed neither kinds of concordance. CONCLUSION: Recognition of the dynamics of HPV infection not only in women, but in their sexual partners could impact the implementation of preventive measures like HPV vaccination for cervical cancer and other HPV-related diseases for both sexual partners.

Evaluation of Three Different Laboratory Methods for Identification of Pneumocystis jirovecii Pneumonia (PCP) among HIV Positive Asymptomatic Prisoners.

BACKGROUND: Pneumocystis jirovecii pneumonia (PCP) remains a leading cause of mortality among HIV-infected patients. The aim of study was to find out P. jirovecii in versatile group of HIV-positive patients prisoners. METHODS: Overall, 102 HIV positive patients from Ghezel Hesar Prison, Karaj, Iran from October 2016 to March 2017 without any respiratory symptoms were selected with different medication histories against HIV and PCP. Microscopic and molecular (qualitative real-time PCR) examination were applied on sputum specimens and serological investigation (ß-D-glucan assay for fungal diseases) carried out on patient's sera. RESULTS: Only 3 and 1 patients were positive for PCP by microscopic and molecular testing, respectively. Twenty-four (23.5%) and 78 (76.5%) out of 102 patients were seropositive and seronegative for fungi disease, respectively. Seropositive patients were older than seronegative subjects (P<0.001). Most of seropositive individuals showed less mean value of CD4 counts compared to seronegative group (P<0.001). Of 54 patients who were under HIV therapy, 13 were seropositive compared to 11 out of 24 seropositives who were no adhere to treatment (P<0.001). In terms of prophylactic antibiotic therapy against PCP, of 24 patients who received prophylaxis, 3 (12.5%) and 21 (87.5%) were seropositive and seronegative, respectively (P<0.001). On the contrary, among 78 patients who did not receive prophylaxis, 21 (27%) and 57 (73%) belonged to seropositive and seronegative patients, respectively (P<0.001). CONCLUSION: There was no strong evidence for PCP infection/disease among symptomless, HIV positive patients. According to their mean CD4 counts, the hypothesis for being negative in a majority of applied tests would be the absence of severe immunosuppression in the patients.

Characterization of a multi-epitope peptide with selective MHC-binding capabilities encapsulated in PLGA nanoparticles as a novel vaccine candidate against Toxoplasma gondii infection.

No effective human vaccine against Toxoplasma gondii (T. gondii) has yet been developed; however, a protective vaccine using immunogenic peptides in a safe delivery vehicle system offers promise. Here, we employed bioinformatics to design a multimeric recombinant T. gondii vaccine using predicted T and B cell epitopes of SAG1, AMA1, ROP2, and GRA4 proteins based on their binding capabilities to common major histocompatibility complex (MHC) molecules. Furthermore, we encapsulated the expressed protein in poly lactic-co-glycolic acid (PLGA) nanoparticles as a delivery vehicle and also used alum as an adjuvant to determine the vaccine potency of this multimeric antigen. BALB/c mice were vaccinated and then challenged with T. gondii RH strain, and the survival rate and cytokine profiles were studied. Mice vaccinated with the multi-epitope-based vaccine, both with and without PLGA, had greater Th1 immune responses, survival rates, specific antibody titers, and IFN-γ and IL-2 levels than controls, while the alum-adsorbed vaccine stimulated a Th2-type humoral immune response.

Detection of anti-Toxoplasma gondii antibodies in chronic myeloid leukemia and acute myeloid leukemia patients.

IN: Background and Aim: Infection of Toxoplasma gondii is a worldwide distribution. Toxoplasmosis in patients who are immunocompromised by virtue of underlying leukemia disease has received relatively little attention. This study was aimed to evaluate IgG and IgM antibodies of T. gondii and to minimize the role of T. gondii and opportunistic infection complication at the early stage of infection in leukemia patients. MATERIALS AND METHODS: The purpose of this assay was to measure anti-T. gondii IgG and IgM antibodies by enzyme-linked immunosorbent assay (ELISA) technique in leukemia patients. RESULTS: IgG antibodies against T. gondii were detected by ELISA in 96 (56.4%) leukemia patients and 72 (42.4%) control group. IgM antibodies were found in 10 patients (5.9%) with leukemia and 3 (1.8%) in the corresponding. CONCLUSION: Our finding indicated that leukemia patients under immunosuppressive condition should not be neglected. Toxoplasmosis in leukemia patients as a main risk factor is considered, meanwhile in some patients, due to possibility of the presence of secondary infection that leads to severe toxoplasmosis.

T Helper 1 (Th1), Th2, and Th17 Responses to Leishmania major Lipophosphoglycan 3.

Leishmania major is the main causal agent of cutaneous leishmaniasis (CL) that remains a serious public health concern in many tropical and subtropical countries. A long-lasting protective vaccine against leishmaniasis remains as a medical unmet need. Lipophosphoglycan 3 (LPG3) is one of the class II LPG genes from HSP90 family involved in the host immune responses. The aim of the present study is to investigate the capability of recombinant LPG3 (rLPG3) to induce Th1, Th2, Th17 responses. The results showed that rLPG3 in moderate and high concentrations significantly induced expression of Th1 lineage-specific transcription factor (T-bet) and cytokine (IFN-γ)(P < 0.05). Moreover, the Th1-stimulating effect of rLPG3 was confirmed by significant induction of IFN-γ secretion from treated T cells (P < 0.01). However, no significant effect of rLPG3 on Th2 and Th17 lineage cells was observed even in high concentration. Our findings demonstrate that rLPG3 induces Th1, but not Th2 and Th17, lineage responses. Further studies are needed to investigate adjuvant properties of rLPG3 for leishmania therapy.

Simultaneous detection and differentiation of Entamoeba histolytica, E. dispar, E. moshkovskii, Giardia lamblia and Cryptosporidium spp. in human fecal samples using multiplex PCR and qPCR-MCA.

Entamoeba histolytica, Giardia lamblia and Cryptosporidium spp. are common causes of diarrheal and intestinal diseases all over the world. Microscopic methods are useful in the diagnosis of intestinal parasites (IPs), but their sensitivity was assessed approximately 60 percent. Recently, molecular techniques have been used increasingly for the identification and characterization of the parasites. Among those, in this study we have used multiplex PCR and Real-time PCR with melting curve analysis (qPCR-MCA) for simultaneous detection and differentiation of E. histolytica, E. dispar, E. moshkovskii, G. lamblia and Cryptosporidium spp. in human fecal samples. Twenty DNA samples from 12 E. histolytica and 8 E. dispar samples and twenty stool samples confirmed positive for G. lamblia and Cryptosporidium spp. were analyzed. After DNA extraction from the samples, multiplex PCR was done for detection and differentiation of above mentioned parasites. QPCR-MCA was also performed for the detection and differentiation of 11 isolates of above mentioned parasite in a cycle with a time and temperature. Multiplex PCR was able to simultaneous detect and differentiate of above mentioned parasite in a single reaction. QPCR-MCA was able to differentiate genus and species those five protozoa using melting temperature simultaneously at the same time and temperature programs. In total, qPCR-MCA diagnosed 7/11 isolation of E. histolytica, 6/8 isolation of E. dispar, 1/1 E. moshkovskii Laredo, 10/11 G. Lamblia and 6/11 Cryptosporidium spp. Application of multiplex PCR for detection of more than one species in a test in developing countries, at least in reference laboratories has accurate diagnosis and plays a critical role in differentiation of protozoan species. Multiplex PCR assay with a template and multi template had different results and it seems that using a set of primers with one template has higher diagnostic capability in compare with multi template. The results of this study showed that the use of the qPCR-MCA can be an effective method to simultaneous distinguish of the above mentioned parasites.

SYBR green-based detection of Leishmania infantum DNA using peripheral blood samples.

Parasitological methods for the diagnosis of visceral leishmaniasis (VL) require invasive sampling procedures. The aim of this study was to detect Leishmania infantum (L. infantum) DNA by real time-PCR method in peripheral blood of symptomatic VL patient and compared its performance with nested PCR, an established molecular method with very high diagnostic indices. 47 parasitologically confirmed VL patients diagnosed by direct agglutination test (DAT > 3200), bone marrow aspiration and presented characteristic clinical features (fever, hepatosplenomegaly, and anemia) and 40 controls (non-endemic healthy control-30, Malaria-2, Toxoplasma gondii-2, Mycobacterium tuberculosis-2, HBV-1, HCV-1, HSV-1 and CMV-1) were enrolled in this study. SYBR-green based real time-PCR and nested PCR was performed to amplify the Kinetoplast DNA minicircle gene using the DNA extracted from Buffy coat. From among 47 patients, 45 (95.7 %) were positive by both nested-PCR and real time-PCR. These results indicate that real time-PCR was not only as sensitive as a nested-PCR assay for detection of Leishmania kDNA in clinical sample, but also more rapid. The advantage of real time-PCR based methods over nested-PCR is simple to perform, more faster in which nested-PCR requires post-PCR processing and reducing contamination risk.

Methodology and Early Findings of the Assessment of Determinants of Weight Disorders among Iranian Children and Adolescents: The Childhood and Adolescence Surveillance and PreventIon of Adult Noncommunicable Disease-IV Study.

BACKGROUND: This paper presents the methodology and primary findings of a national project on determinants of weight disorders among Iranian children and adolescents at national and sub-national levels. METHODS: This nationwide study was conducted in 2011-2012 in Iran as part of the fourth phase of a national surveillance program entitled Childhood and Adolescence Surveillance and PreventIon of Adult Noncommunicable disease-IV study. It had two phases of qualitative and quantitative study. This multicentric study was conducted among 25,000 students aged 6-18 years, living in urban and rural areas of 30 provinces of Iran. Students were selected by multistage cluster sampling method. Data regarding weight disorders including sociodemographic variables, perinatal factors, lifestyle factors, family and student dietary habits, quality of life, and family history of chronic diseases as well as body image were gathered via validated questionnaires. RESULTS: Overall, 23043 students completed the survey (participation rate: 92.17%). The mean age of participants was 12.55 ± 3.31 years; 50.8% were boys, and 73.4% were from urban areas. Underweight was found in 10.4% of boys and 9.2% of girls, the corresponding figure for overweight and obesity was 21% and 18.3%. Abdominal obesity was found in 17.6% of students. Among parents, obesity was more frequent than other weight disorders, with higher prevalence in parents of girls than boys (24.5% vs. 21.5%, respectively, P < 0.001). Overweight and obesity were more prevalent in urban than in rural parents (66.7% vs. 59.7%, respectively, P < 0.001). CONCLUSIONS: This survey serves as confirmatory evidence on the prevalence of dual burden of weight disorders in Iran. Its findings on determinants of weight disorders would help policymakers to implement relevant programs at national and sub-national levels.

In vitro activity and cytotoxicity of Crocus sativus extract against leihmania major (MRHO/IR/75/ER).

Leishmaniasis is a complex protozoan disease comprising a wide range of clinical manifestations that is usually divided into visceral leishmaniasis, cutaneous leishmaniasis, and muco-cutaneous leishmaniasis depending on leishmania parasite species and host's immune system responses. Most of the drugs produced for the treatment of leishmaniasis, from the first used to the most recently accepted, are toxic, resistance issues and poorly tolerated. The purpose of this study is to evaluate the effectiveness of saffron (Crocus sativus) and its apoptotic activity against Leishmania major (MRHO/IR/ 75/ER) promastigotes. MTT assay was used to find viability of L. major promastigotes and the achieved results were explicated as IC50 (50% inhibitory concentration). ED50 (50% effective doses) for L. major amastigotes were also analyzed. Annexin-V FLUOS staining was performed to study the cell death properties of saffron by using FACS analysis. Qualitative analysis of the DNA fragmentations was accomplished by agarose gel electrophoresis and light microscopy was used to observe morphological changes of promastigotes. Our results revealed that L. major promastigotes and amastigotes are sensitive to saffron at different concentrations and time dependent manner with apoptotic features including DNA laddering, cytoplasmic shrinkage, and externalization of phosphatidylserine. IC50 and ED50 of this extract after 48 h of incubation was 0.7mg/ml and 0.5 mg/ml respectively. Finally, C. sativus has shown anti-leishmanial activity against L. major promastigote and amastigote and may induce apoptosis.

Development and Assessment of Loop-Mediated Isothermal Amplification (LAMP) Assay for the Diagnosis of Human Visceral Leishmaniasis in Iran.

BACKGROUND: Parasitological methods for the diagnosis of Visceral leishmaniasis (VL) require invasive procedures, so serological and molecular approaches have been developed but are not generally applicable in the field. We evaluated a loop mediated isothermal amplification (LAMP) assay using blood from VL patients and compared it to nested PCR. METHODS: Forty-seven subjects with clinical features (fever, hepatosplenomegaly and anemia) were confirmed positive for VL by the direct agglutination test (DAT) at titers >3200. Forty DAT negative individuals from non-endemic areas with no clinical signs or symptoms of VL served as controls. A LAMP assay was performed using a set of six primers targeting Leishmania infantum kinetoplast DNA (kDNA) minicircle gene under isothermal (64 °C) conditions. For nested PCR we used primers targeting the kDNA minicircle gene. RESULTS: The LAMP assay provided a detection limit of 1 parasite in 1 ml of peripheral blood and detected L. infantum DNA in 44 of 47 DAT-confirmed VL cases, with diagnostic sensitivity of 93.6% (95% CI). No L. infantum DNA was amplified in controls, indicating a specificity of 100%. The nested PCR yielded sensitivity of 96% (95% CI) and a specificity of 100% (95% CI). CONCLUSION: The LAMP assay gave results similar to those of nested PCR but in a shorter time. The LAMP method is simple; requires no sophisticated equipment; has a short reaction time; and results, indicated by turbidity of the reaction mixture, are observable with the naked eye.

Application of Multiplex PCR for Detection and Differentiation of Entamoeba histolytica, Entamoeba dispar and Entamoeba moshkovskii.

BACKGROUND: Entamoeba moshkovskii and E. dispar are impossible to differentiate microscopically from the pathogenic species E. histolytica. Multiplex polymerase chain reaction (Multiplex PCR) is a widespread molecular biology technique for amplification of multiple targets in a single PCR experiment. METHODS: For detection and differentiation of the three-microscopy indistinguishable Entamoeba species in human, multiplex PCR assay using different DNA extraction methods was studied. A conserved forward primer was derived from the middle of the small-subunit rRNA gene, and reverse primers were designed from signature sequences specific to each of these three Entamoeba species. RESULTS: A 166-bp PCR product with E. histolytica DNA, a 580-bp product with E. moshkovskii DNA and a 752-bp product with E. dispar DNA were generated in a single-round and multiplex PCR reaction. CONCLUSION: We recommend this PCR assay as an accurate, rapid, and effective diagnostic method for the detection and discrimination of these three Entamoeba species in both routine diagnosis of amoebiasis and epidemiological surveys.

Miltefosine induces metacaspase and PARP genes expression in Leishmania infantum

OBJECTIVES: Apoptosis is the process of programmed cell death (PCD) that occurs in both animal and plant cells. Protozoan parasites possess metacaspase and these caspase-related proteases could be involved in the PCD pathways in these organisms. Therefore we analyzed the activities of metacaspase and PARP genes in Leishmania infantum (MCAN/IR/96/LON49) treated with miltefosine. MATERIALS AND METHODS: Anti-leishmania activity of miltefosine was studied by treatment of cultured promastigotes with various concentration of miltefosine. MTT assay and Annexin-V FLUOS staining by using FACS flow cytometry methods were used. Cytotoxic potential of HePC on the amastigots of L.infantum was evaluated in J774 cell line. In addition, metacaspase and PARP genes expression of treated L. infantum were studied. RESULTS: Miltefosine led to dose-dependent death of L. infantumwith features compatible with apoptosis. Over expression of metacaspase and PARP was seen 6 hr after treatment. CONCLUSIONS: Our study showed that miltefosine exerts cytotoxic effect on L. infantum via an apoptotic-related mechanism.

Alteration of rat hippocampal neurogenesis and neuronal nitric oxide synthase expression upon prenatal exposure to tamoxifen.

The present study delineates the effect of tamoxifen on neuronal density and expression of neuronal nitric oxide synthase (nNOS) in hippocampal nerve cells during prenatal and postnatal periods in rats. Pregnant rats were administered with tamoxifen one day prior to labor (E21) and on the childbirth day (E22). Hippocampi of embryos at E22 and newborns at postnatal days of 1, 7, and 21 (P1, P7, and P21) were investigated. Density of the neurons in areas of the developing hippocampus including cornu ammonis (CA1, CA3), dentate gyrus, and subiculum were studied. Our findings show that the number of pyramidal neurons was significantly decreased in CA1 and subiculum of tamoxifen-treated rats in E22, P1, and P7. We found that cellular density was lower in early stages of development, however, cellular density and thickness gradually increased during the development particularly in the third week. We found that nNOS expression was decreased in E22, P1, and P7 in animals treated with tamoxifen. The present study shows that tamoxifen affects development and differentiation of postnatal rat hippocampus, CA1 neurons, and nNOS expression.

Miltefosine-induced apoptotic cell death on Leishmania major and L. tropica strains.

The aim of this study was to assess the cytotoxic effects of various concentrations of miltefosine on Leishmania major (MRHO/IR/75/ER) and L. tropica (MHOM/IR/02/Mash10) promastigotes and to observe the programmed cell death features. The colorimetric MTT assay was used to find L. major and L. tropica viability and the obtained results were expressed as 50% inhibitory concentration (IC50). Also, 50% effective doses (ED50) for L. major and L. tropica amastigotes were also determined. Annexin-V FLUOS staining was performed to study the cell death properties of miltefosine using FACS analysis. Qualitative analysis of the total genomic DNA fragmentation was performed by agarose gel electrophoresis. Furthermore, to observe changes in cell morphology, promastigotes were examined using light microscopy. In both strains of L. major and L. tropica, miltefosine induced dose-dependent death with features of apoptosis, including cell shrinkage, DNA laddering, and externalization of phosphatidylserine. The IC50 was achieved at 22 µM and 11 µM for L. major and L. tropica after 48 hr of incubation, respectively. ED50 of L. major and L. tropica amastigotes were 5.7 µM and 4.2 µM, respectively. Our results indicate that miltefosine induces apoptosis of the causative agent of cutaneous leishmaniasis in a dose-dependent manner. Interestingly, L. major did not display any apoptotic changes when it was exposed to miltefosine in concentrations sufficient to kill L. tropica.

Miltefosine induces metacaspase and PARP genes expression in Leishmania infantum.

OBJECTIVES: Apoptosis is the process of programmed cell death (PCD) that occurs in both animal and plant cells. Protozoan parasites possess metacaspase and these caspase-related proteases could be involved in the PCD pathways in these organisms. Therefore we analyzed the activities of metacaspase and PARP genes in Leishmania infantum (MCAN/IR/96/LON49) treated with miltefosine. MATERIALS AND METHODS: Anti-leishmania activity of miltefosine was studied by treatment of cultured promastigotes with various concentration of miltefosine. MTT assay and Annexin-V FLUOS staining by using FACS flow cytometry methods were used. Cytotoxic potential of HePC on the amastigots of L.infantum was evaluated in J774 cell line. In addition, metacaspase and PARP genes expression of treated L. infantum were studied. RESULTS: Miltefosine led to dose-dependent death of L. infantum with features compatible with apoptosis. Over expression of metacaspase and PARP was seen 6 hr after treatment. CONCLUSIONS: Our study showed that miltefosine exerts cytotoxic effect on L. infantum via an apoptotic-related mechanism.