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.
Nowadays, increasing extended-spectrum β-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.
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.
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.
Background:Intestinal parasites of humans are one of the most important health problems worldwide, especially those located in tropical and subtropical areas. Objectives: The aim of this study was to determine the frequency of intestinal parasites in patients with gastrointestinal disorders, in different parts of Iran. Patients and Methods: A total of 1520 stool samples were collected from patients with gastrointestinal disorders. The stool specimens were examined by direct wet mount, formalin-ether concentration and a modified version of the Ziehl-Neelsen staining technique. Amoeba-positive samples were cultured for further differentiation of Entamoeba histolytica, E. dispar, E. moshkovskii. DNA-based methods were used to differentiate these amoebas and to detect Cryptosporidium-positive samples. Statistical analysis was carried out by SPSS ver.
16.Results: Out of the 1520 individuals studied, 153 (10.06%) were infected at least with one intestinal parasite. 781 (51.4%) of patients were male and 738 (48.6%) were female. The prevalence of protozoan parasites 148 (9.7%) was significantly higher than helminth parasites 5 (0.3%) (P < 0.001). The frequency of intestinal parasites was as follows: Blastocystis sp., 72 (4.73%); Giardia intestinalis, 35 (2.30%); Entamoeba coli 21 (1.38%); Endolimax nana 10 (0.92%); Cryptosporidium spp., 1 (0.06%); Entamoeba dispar, 1 (0.06%); Dientamoeba fragilis, 1 (0.06%); Hymenolepis nana, 3 (0.19%); Dicrocoelium dendriticum, 2 (0.13%). In five (0.32%) of the positive samples, co-infections with two parasites were found. G. intestinalis was more prevalent in male 22/35 (62.86%) than female 13/35 (37.14%) as well as in 0-9 years old group. In one sample Heterodera ova contained larva were seen. Conclusions: Blastocystis and G. intestinalis were the predominant intestinal parasites detected in patient with gastrointestinal disorders. The results indicated that the intestinal parasites, particularly helminth infections have been significantly declined in recent years.
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.
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