Anti-parasitic effects of resveratrol on protoscolices and hydatid cyst layers.

The main goal of the current study was to evaluate the effectiveness of resveratrol (RESV) on protoscolices and hydatid cysts of Echinococcus granolosus. Echinococcus granolosus protoscolices and hydatid cyst were exposed to RPMI, DMSO, formalin, mebendazole, and different concentrations of RESV in vitro. Then, viability, GGT, and caspase-3 activity of protoscolices were evaluated using light microscopy, colorimetric, and enzymatic assay, respectively. Tissue changes and expression of caspase-3 apoptosis were analyzed on the hydatid cyst wall by histologic and immunohistochemistry methods. The cell toxicity effect of RESV was evaluated on mouse PBMCs by Annexin V-FITC assay. The RESV-treated protoscolices showed loss of viability, increased gamma-glutamyl transpeptidase, and caspase-3 activity with significant differences compared to all control groups (P < 0.05). Dose and time dependence of mortality, GGT, and caspase-3 enzymatic activity was confirmed in the protoscolices of Echinococcus granulosus treated by RESV. Also, the tissue changes and apoptosis were prominent in RESV-treated hydatid cyst layers; however, tissue changes were only time-dependent, and RESV concentration had no apparent effect on tissue. In cell toxicity evaluation, RESV is safe without any significant apoptosis induction from 31.5 to 250 µg/ml; however, it was significant at 350 and 500 µg/ml in PBMCs.

Isolation and molecular identification of Acanthamoeba spp. from hospital dust and soil of Khomein, Iran, as reservoir for nosocomial infection.

BACKGROUND: Acanthamoeba spp. are commonest opportunistic amoebae, which ubiquitous in various environmental resources. Acanthamoeba species are the causative agents of amoebic keratitis, granulomatous amoebic encephalitis and i.e. in immunocompromised and immunocompetent patients. Moreover Acanthamoeba spp. can act as reservoir and transmission agent of bacterial pathogens. Due to this issue the aim of this study was to characterized Acanthamoeba spp. genotypes in dust and soil of hospital samples from Khomein of Iran. METHODS: In a cross sectional study, a total of 100 soil and dust samples were collected from hospital environment of Khomein Iran, and analyzed for the presence of Acanthamoeba spp. based on phenotypic and molecular methods including PCR amplification and sequence analysis of 18SrRNA. A total of 5 Acanthamoeba isolates were sequenced, and different genotypes of isolates were detected via direct sequence analysis. RESULTS: The results showed that 20% of samples (20/100) were positive for Acanthamoeba, while only 5 cases were successfully cultured in NNM medium and were subjected to molecular assay. A. lenticulata, A. castellanii and A. quina were the prevalent identified species that were belonged to T4 and T5 genotypes. CONCLUSIONS: Acanthamoeba spp. are the most prevalent free living amoeba in the dust and soil of hospital environment. Moreover, due to the presence of potentially pathogenic T4 genotypes in our hospital, it is recommended that in health and hygienic programs elimination of FLA should be considered.

Mobile Phone Radiation: Its Efficacy as Protoscolicidals.

OBJECTIVE: There are different types of radiations, such as microwaves and mobile waves. Certain types of radiofrequency were evaluated in hydatid cyst ablation or as protoscolicidals. This study aimed to assess the influence of mobile waves on hydatid cyst protoscolices. METHODS: Hydatid cysts were collected from the slaughterhouse and transferred to the laboratory. The contents of the cysts were drained in sterile conditions and the protoscolices were rinsed three times with phosphate buffered saline. Equal volumes of protoscolex suspensions were aliquoted into similar tubes. Based on the distance of the samples from the mobile generation waves, the tubes containing the parasitic suspensions were classified into three groups, each of which was further categorised into nine subgroups according to the time of the radiation exposure. The subgroup with zero exposure time was considered the control. RESULTS: It was found that the mortality rate of the protoscolices increases as the distance of the sample from the wave-generation source decreases (p<0.0001). Increasing the time of exposure also improves the mortality rate of protoscolices. CONCLUSION: The mortality rate of protoscolices was directly proportional to the time of exposure and inversely proportional to the distance from the mobile generation waves.

Fungal and parasitic contamination of indoor public swimming pools in Arak, Iran.

BACKGROUND: Swimming is a popular exercise for different types of people at different ages. Public swimming pools are places where fungal infections can be easily transferred. The purpose of this study is to evaluate the quality of mycological, parasitological, and physicochemical parameters of swimming pools of Arak city. METHODS: This cross-sectional study was done for 12 months from April 2013 to March 2014 in six indoor active swimming pools of Arak city (A, B, C, D, E, and F). Samples were collected in four seasons, two times/season; each time, two samples were obtained from six specified locations (shallow level pool, deep level pool, dressing rooms, showers, margin of pool walls, and foot-washing sink) from each pool with a total of 576 samples. Physicochemical parameters including water temperature, pH, turbidity, and the residual chlorine were measured on-site. In order to isolate and detect the fungal agents, special filters and culture Sabouraud's dextrose agar, chloramphenicol, and mycosel agar media were applied. Furthermore, non-nutrient agar medium enriched with Escherichia coli was used to detect and to separate the eggs of the worms, cysts, and parasitic protozoa from centrifuges of samples. In order to investigate their sediment, optical microscope and culture media were used. RESULTS: We found that 456 (79.1%) samples were positive regarding the fungal elements, and 516 fungal species were isolated. The most common isolates were saprophytic species (8.74%), yeast species (25%), and dermatophyte species (2.5%). The most contaminated surfaces were foot-washing sinks and showers. In this study, Acanthamoeba parasites were isolated from one pool only. All the investigated physicochemical parameters of pool water except for the temperature were found to be in the standard range. CONCLUSIONS: Existence of saprophytic fungi and yeast in pools' water is plausible to be considered as an indicator of water resistance to the detergent agents. This high degree of contamination is due to the huge number of visitors, the complexity of construction, the choice of materials, and the long opening hours. Isolation of dermatophytes and Acanthamoeba parasite from the pools' area and foot-washing sink reveals the important role of the public swimming pools in disease transmission.

A Simplified Protocol for the Purification of Schwann Cells and Exosome Isolation from C57BL/6 Mice.

BACKGROUND: The purification of Schwann cells has proven to be a difficult process, with most methods requiring the use of special equipment. However, obtaining a sufficient number and high purity of Schwann cells is an integral aspect in their use for clinical application. Therefore, the aim of this study was to establish a simple and effective protocol for the isolation and purification of Schwann cells from the sciatic nerve of C57BL/6 mice. Furthermore, we aimed to provide a protocol for the isolation of exosomes from these cells. METHODS: To purify Schwann cells, we used a combination of in situ nerve pre-degeneration and fetal bovine serum. To determine the most effective method of cell purification, we treated the culture with varying concentrations of fetal bovine serum and examined which concentration provided the highest Schwann cell purity. Exosomes were then isolated from Schwann cells through a process of repeated centrifugation and filtration steps. RESULTS: We were able to increase the purified population of Schwann cells from C57BL/6 mice by reducing the concentration of FBS. The purity of Schwann cells at FBS concentrations of 10%, 5%, and 2% were 93.42%, 91.25%, and 97.83%, respectively. CONCLUSION: When using a concentration of 2% FBS, we obtained the highest purification yield of Schwann cells. Our protocol does not require special equipment or materials. We have created a protocol that is simple, fast, and safe while providing a high yield of purified Schwann cells. The exosome isolation method described in this paper is an appropriate approach with a high quality and yield.

Isolation and Genotyping of Acanthamoeba from Soil Samples in Markazi Province, Iran.

AIM: A previous study confirmed the contamination of water sources with this parasite in Arak, Markazi Province, Iran. The current study investigated soil sources and determined the predominant genotype of Acanthamoeba in this region of Iran. MATERIAL AND METHODS: Forty-eight soil samples, collected from different regions of Arak, Markazi province, Iran, were evaluated in this study. The samples were processed and identified by culturing on a specific medium, performing PCR assay, and sequencing the PCR products. Finally, using the NCBI database, the genotypes were determined. RESULTS: Of 48 soil samples, 33.3% and 31.25% were contaminated with Acanthamoeba according to the culture and molecular assays, respectively. The majority of these isolates belonged to the T4, T5 and T6 genotypes of Acanthamoeba. CONCLUSION: The genotypes of most isolates from soil samples in Arak similar to other regions of Iran belong to T4 genotype of this parasite. New sequence accession numbers include MG066681 and MG298785-MG298794.

Toxoplasmosis: Seroprevalence in pregnant women, and serological and molecular screening in neonatal umbilical cord blood.

Toxoplasmosis is a common zoonotic disease that can also be transmitted from the mother to the embryo, with the risk of congenital infection varying around the world. The aim of this study was to screen pregnant women and their neonates for toxoplasmosis by serologic and molecular methods and assess the impact of risk factors associated with toxoplasmosis on the rate of congenital infection. This study was conducted at a regional maternity hospital in Arak, the capital of the Markazi Province in Iran, during a period of six months. All selected pregnant women (n=261) and the corresponding cord blood samples were serologically screened for toxoplasmosis, with seropositive samples also undergoing molecular testing. Demographic data, as well as information related to the risk factors associated with the transmission of the disease, were collected from mothers and their neonates. The detection of anti-Toxoplasma antibodies and the extraction of DNA from blood samples were conducted using commercial kits. Results showed that the sera of 87 maternal blood samples (33.3%) and 40 cord blood samples (15.3%) were positive for anti-Toxoplasma antibodies (IgG and/or IgM). Molecular screening of the seropositive samples only identified one positive cord blood sample. In other words, the diagnosis of congenital toxoplasmosis was definitive in only one neonate. There was no significant association between the risk of parasite transmission and neonatal seropositivity (p >0.05). Therefore, the results showed that the prevalence of congenital toxoplasmosis in the studied area was consistent with the global rate and suggest that the implementation of newborn screening and follow-up testing could help reduce the disease risk.

Detection of Specific Antibody Reactivity to Toxocara Larval Excretory-secretory Antigens in Asthmatic Patients (5-15 Years).

BACKGROUND & PURPOSE: Humans act as an intermediate host for Toxocara canis and Toxocara cati. Toxocara may be an important risk factor for asthma in humans. The aim of the present study was to evaluate immunoglobulin G (IgG) anti-Toxocara canis antibody, using enzyme-linked immunosorbent assay (ELISA) in asthmatic patients (aged 5-15 years), referring to a clinic of pulmonary diseases in Arak, Iran. MATERIALS & METHODS: In this bi-group cross sectional study, serum samples were collected from 110 children with confirmed asthma and 70 children without asthma within one year. IgG anti-Toxocara antibody was detected viaELISA method. The collected data were analyzed, using SPSS. RESULTS: The seroprevalence of antibodies against Toxocara species was estimated at 1.8% (two males) in asmathic children viaELISA method; however, no antibodies against Toxocara canis were detected in the control group. There was no significant correlation between the frequency of antibodies against Toxocara and variables such as age, gender, or place of residence (P>0.05). Moreover, the frequency of antibodies against Toxocara was not significantly correlated with contact with dogs, consumption of unwashed fruits and vegetables, or use of raw/undercooked sheep liver (P>0.05). CONCLUSION: The present study showed anti-Toxocara antibody in 1.8% of asthmatic children and determined the seroprevalence of toxocariasis in asthmatic children and adolescents in Arak, Iran. Based on the findings, the low rate of infection with Toxocara among asthmatic children may be attributed to acceptable personal hygiene and religious considerations.

Identification of Malassezia Species Isolated from Patients with Pityriasis Versicolor Using PCR-RFLP Method in Markazi Province, Central Iran.

BACKGROUND: The lipophilic yeasts of Malassezia species are members of the normal skin microbial that are cause of pityriasis versicolor. Pityriasis versicolor is a common superficial fungal infection with world-wide distribution. The phenotypic methods for identification of Malassezia species usually are time consuming and unreliable to differentiate newly identified species. But DNA-based techniques rapidly and accurately identified Malassezia species. The purpose of this study was isolation and identification of Malassezia Species from patients with pityriasis versicolor by molecular methods in Markazi Province, Central Iran in 2012. METHODS: Mycologic examinations including direct microscopy and culture were performed on clinical samples. DNA extraction was performed from colonies. The ITS1 region of rDNA from isolates of Malassezia species were amplified by PCR reaction. The PCR were digested by Cfo I enzyme. RESULTS: From 70 skin samples, were microscopically positive for Malassezia elements, 60 samples were grown on culture medium (85.7%). Using PCR-RFLP method, that was performed on 60 isolates, 37(61.6%) M. globosa, 14(23.3%) M. furfur, 5(8.4%) M. sympodialis and 4(6.7%) M. restrictawere identified. In one case was isolated M. globosa along with M. restricta. CONCLUSION: The PCR-RFLP method is a useful and reliable technique for identification of differentiation of Malas-sezia species.

Effects of Toxoplasma gondii Infection in Level of Serum Testosterone in Males with Chronic Toxoplasmosis.

BACKGROUND: Toxoplasma gondii is an intracellular protozoan parasite that infects human and animals. Toxoplasma parasites are isolated from different parts of animals even from semen but there are little information about the effect of toxoplasmosis on fertility in animals and humans. In present study, the effect of chronic toxoplasmosis on serum levels of testosterone in men was studied. METHODS: In this case-control study, 1026 men referred to Arak Post Marriage Center were selected. Three ml of blood samples were collected and sera separated by centrifugation at room temperature. These sera were analyzed for detection of anti-T. gondii IgG antibody. Next 365 positive sera were selected as cases and also the same number of negative sera (365) as controls. Finally the level of testosterone was analyzed for the cases and controls samples. RESULT: Serological tests on the sera of 1,026 men in Arak City showed that 365 of them had anti-Toxoplasma antibody. Comparison of testosterone concentration in case and control groups showed that testosterone concentration in case group was less than control group and this difference was statistically significant (P<0.05). CONCLUSION: The chronic toxoplasmosis could affect reproductive parameters in men.