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Objective(s): It is worthwhile to note that, some probiotics such as Lactobacilli and Bifidobacteria isolated from dairy products have significant therapeutic effects against cancer cells. Here, we evaluated anti-proliferation and the apoptotic effects of isolated Lactobacillus fermentum Ab.RS22 from traditional dairy products on the HeLa cervical cancer cells in vitro. Materials and Methods: The viability of treated HeLa cells with supernatant of Lactobacillus in 0.5, 0.75, 1, 1.5, and 2 ng/ml concentrations, and IC50 values were detected by tetrazolium bromide. The L. fermentum Ab.RS22-induced cell death by flow cytometry was confirmed through evaluation of the expression of caspase-3, P53, PTEN, and AKT genes by quantitative reverse transcriptionpolymerase chain reactions (qRT-PCR). Results: Most cytotoxicity effects of Lactobacillus on HeLa cells were detected in 2 ng/ml at 24 hr (P<0.01); also, the IC50 value was measured as 1.5 ng/ml. The findings of the flow cytometry assay showed that L. fermentum Ab.RS22 in 1.5 ng/ml concentration at 24 hr increased the percentage of both apoptosis and necrosis cells. Lactobacillus-induced cell death was verified through results of Real-time PCR; where expression of caspase-3, P53, and PTEN genes was increased (P<0.01), and also expression of AKT gene (anti-apoptotic) was decreased (P<0.05). Conclusion: Our findings showed that L. fermentum Ab.RS22 could dose-dependently inhibit the proliferation of the HeLa cells. Its apoptotic effect was confirmed via modulating PTEN/p53/Akt gene expression and activation of the caspase-3 mediated apoptosis pathway. Therefore, L. fermentum Ab.RS22 can be considered a valuable anticancer candidate against cervical cancer progression in subsequent studies. [ABSTRACT FROM AUTHOR]

Background: Several markers have been described to characterise the population structure and genetic diversity of Fasciola species (Fasciola hepatica(F. hepatica) and Fasciola gigantica (F. gigantica). However, sequence analysis of a single genomic locus cannot provide sufficient resolution for the genetic diversity of the Fasciola parasite whose genomes are ∼1.3 GB in size. Objectives: To gain a better understanding of the gene diversity of Fasciola isolates from western Iran and to identify the most informative markers as candidates for epidemiological studies, five housekeeping genes were evaluated using a multilocus sequence typing (MLST) approach. Methods: MLST analysis was developed based on five genes (ND1, Pepck, Pold, Cyt b and HSP70) after genomic DNA extraction, amplification and sequencing. Nucleotide diversity and phylogeny analysis were conducted on both concatenated MLST loci and each individual locus. A median joining haplotype network was created to examine the haplotypes relationship among Fasciola isolates. Results: Thirty‐three Fasciola isolates (19 F. hepatica and 14 F. gigantica) were included in the study. A total of 2971 bp was analysed for each isolate and 31 sequence types (STs) were identified among the 33 isolates (19 for F. hepatica and 14 for F. gigantica isolates). The STs produced 44 and 42 polymorphic sites and 17 and 14 haplotypes for F. hepatica and F. gigantica, respectively. Haplotype diversity was 0.982 ± 0.026 and 1.000 ± 0.027 and nucleotide diversity was 0.00200 and 0.00353 ± 0.00088 for F. hepatica and F. gigantica, respectively. There was a high degree of genetic diversity with a Simpson's index of diversity of 0.98 and 1 for F. hepatica and F. gigantica, respectively. While HSP70 and Pold haplotypes from Fasciola species were separated by one to three mutational steps, the haplotype networks of ND1 and Cyt b were more complex and numerous mutational steps were found, likely due to recombination. Conclusions: Although HSP70 and Pold genes from F. gigantica were invariant over the entire region of sequence coverage, MLST was useful for investigating the phylogenetic relationship of Fasciola species. The present study also provided insight into markers more suitable for phylogenetic studies and the genetic structure of Fasciola parasites. [ABSTRACT FROM AUTHOR]

Background: The present study was designed to determine the prevalence of Toxoplasma gondii seronegative pregnant women, estimate the frequency of fetuses at risk of congenital toxoplasmosis in the study area, and also to investigate the awareness of the studied mothers about T. gondii disease. Methods: A total of 740 pregnant women referred to two health centers in Qazvin province were surveyed voluntarily for anti-T. gondii IgG and IgM antibodies by enzyme-linked immunosorbent assay (ELISA). IgG avidity test was used as a confirmatory test for samples showing positive or borderline results for IgM antibodies. Data on socioeconomic, demographic, and knowledge of participants about toxoplasmosis were collected using questionnaires and through face-to-face interviews. Multivariable logistic regression modeling was used to identify the potential predictor variables for T. gondii infection. Results: Overall, 21.2% (157/740) and 0.27% (2/740) of pregnant women were positive for T. gondii IgG and IgM antibodies, respectively. Only 1.7% (13/740) of expectant mothers had prior information on toxoplasmosis. Among socioeconomic and demographic variables, contact with soil was the merely cause of a significant association with T. gondii infection. Other variables including age, occupation, education level, residential area, source of drinking water, abortion history, number of delivery, consumption of raw/undercooked meat, having home cat and and consumption of raw vegetables failed to establish significant associations. Conclusion: Our results clarified that the prevalence of T. gondii has remarkably reduced in the study area. This finding indicates a low risk of congenital toxoplasmosis in this region. [ABSTRACT FROM AUTHOR]

Background: Free-living amoebae (FLA) such as Acanthamoeba spp., are considered as opportunistic and pathogenic protozoans. Acanthamoeba granulomatous encephalitis (AGE) is a serious threat for immunodeficient patients and Acanthamoeba keratitis (AK) for contact lens users. We aimed to identify the presence of free living amoebae in nasal swabs of patients and contact lens users in Qazvin, Iran. Methods: During 2019, 251 nasal and oral swabs (including the pharynx and mouth) were collected from patients with diabetes, AIDS and those under periodic dialysis in Qazvin, Iran. In addition, 27 soft contact lenses were collected from the participants. Following DNA extraction, PCR and sequencing were conducted to identify the genotypes of the amoeba. Phylogenetic analysis of the identified sequences was performed using MEGA 7 software. Results: A strain of Acanthamoeba belonging to the T3 genotype was isolated from hemodialysis patients. Two specimens of Acanthamoeba with T3 genotype were isolated from keratitis patients. Conclusion: The clinicians should pay attention to the possible complication of this organism because this amoeba is potentially pathogenic for immunocompromised patients. Since the amoeba is present in environmental resources, the use of contact lenses should be accompanied by considering proper hygiene. [ABSTRACT FROM AUTHOR]

Background: Our knowledge of the epidemiology of rodents’ parasitic agents in Iran is scarce, although some of these pathogens play an important role in human and veterinary medicine, such as Toxoplasma gondii and Neospora caninum. The purpose of this study was to determine the seroprevalence of Toxoplasma gondii and Neospora caninum in rodents of northwestern Iran between Mar and Dec 2015. Methods: Overall, 157 serum samples from rodents (101 Meriones persicus, 41 Mus musculus, and 15 Cricetulus migratorius) were assayed by the indirect fluorescence antibody test (IFAT) for antibodies to T. gondii and N. caninum. Results: We found a prevalence of 20.38% (32/157) for N. caninum, 35% (55/157) for T. gondii. Co-presence of antibodies to N. caninum and T. gondii was found in 10 (6.36%) rodents. A significant association was found between the rodents species and seropositivity to N. caninum (P<0.05) but there was no association with rodents species for T. gondii. The overall prevalence of the aforementioned parasites was higher in male versus female rodents. Conclusion: The high seroprevalence of toxoplasmosis and neosporosis in rodents in the study area has implications for translocation of these infections across wider geographical regions since these rodents are mostly preyed on by cats or dogs; hence, which can transfer the parasite to other hosts. [ABSTRACT FROM AUTHOR]

Strongyloides stercoralis is an endemic parasite in some regions including the tropical and subtropical areas with high humidity. Most infections are asymptomatic with nonspecific signs and symptoms, making the final diagnosis complicated. Here, we report a patient referred to our hospital with signs consistent with sepsis, intestine obstruction, which finally died with the diagnosis of strongyloidiasis. The patient was from northern parts of Iran which are considered as endemic areas for S. stercoralis. In conclusion, there is an important message in this history, i.e. physicians should be aware of specific and non-specific signs of strongyloidiasis especially in people living in endemic areas to make an accurate final diagnosis by proper clinical and paraclinical examinations. [ABSTRACT FROM AUTHOR]

Background: The aim of this study was the seroepidemiological survey for detecting the status of human fasciolosis in Lorestan Province, western Iran. Methods: This cross-sectional study was conducted in 2015-16. Based on statistical estimations, 1256 serum samples were collected from different parts of Lorestan Province, western Iran, and stored at -20 °C until use. The collected serum samples were analyzed at Tehran University of Medical Sciences, Tehran, Iran using indirect ELISA method. Results: Anti-Fasciola antibodies were detected in 16 (1.3%) individuals. Regarding the seropositivity to fasciolosis, no significant differences were found between age groups, sex, level of education and occupation; however significant differences were observed regarding location, consuming local freshwater plants and water resources (P<0.02.). Conclusion: Local freshwater plants and unfiltered water resources were probably the main sources of the infection. Health education by local health centers to elevate awareness of people, and providing facilities for safer drinking water, especially in rural areas may help decrease the risk of fasciolosis infection in this region. [ABSTRACT FROM AUTHOR]

Background: Leishmaniasis is a sand fly-borne disease caused by the protozoan parasites belonging to the genus Leishmania. Because of the preventing and controlling methods, clinical course, prognosis and choice of treatment are differing from species; differentiation of species is critical. The present study was aimed to detect the parasite species using the PCR-RFLP method. Methods: A total of 130 Giemsa-Stained slides from suspected Cutaneous leishmaniasis (CL) patients were examined under a light microscope at×1000. DNA from each slide was extracted PCR method was undertaken with HSP70 genes and the PCR products were digested with a restriction enzyme HaeIII (BsuR1). The study was conducted in the laboratory of Zahedan University of Medical Sciences in the Sistan and Baluchestan Province, southeastern Iran in 2015. Results: From 130 suspected samples, 59 (45.3%) were positive by the microscopic examination, meanwhile 64 (49.2%) were positive by PCR-RFLP, Leishmania species were recognized, and L. tropica was introduced as predominant species in current study. Conclusion: PCR-RFLP is a valuable technique for distinguish of Leishmania species. Furthermore, anthroponotic CL is the dominant cause of CL in Sistan and Baluchestan Province. [ABSTRACT FROM AUTHOR]

Due to irregular and non-normative extension of cities, a lack of appropriate model for consumption and increasing growth of waste production, sanitary landfill is the most logical and economical method for municipal waste disposal. Due to the high sensitivity of the issue, detection of potential waste landfill sites requires comprehensive techniques. Therefore, this study aimed to select a suitable location for waste sanitary landfill of Shahriar by using multi-criteria evaluation techniques of fuzzy analytic hierarchy process and FTOPSIS (Fuzzy Technique for Order Preference by Similarity to Ideal Solution). First, the effective criteria for the selection of waste landfill were collected from the relevant organizations and they were analyzed and standardized by geographic information systems. Then, criteria weights were calculated by fuzzy analytic hierarchy process and after weights were applied in corresponding layers, layers were overlaid by fuzzy functions. Next, options were prioritized by FTOPSIS method. Thirty-one appropriate options were obtained by implementing above methods. Then prioritizing led to one option as the best site, which is located in the southwest of Shahriar. The results showed that selected options are located in a better site than the current landfill and this combined approach has a better performance than other methods because of considering the imprecise nature of phenomena. [ABSTRACT FROM AUTHOR]

Aim: Cutaneous leishmaniasis (CL) is one of the most important health problems that are capable of involving both tropical and subtropical areas, especially in Iran. This cross-sectional study aimed to differentiate the species that are able to cause CL in Zahedan city by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method. Materials and Methods: It was conducted on 145 suspected CL patients in Zahedan city between 2014 and 2016. The smears were initially prepared, air-dried, fixed with absolute methanol, and stained with 10% Giemsa. Then, we examined the stained samples by a light microscope under 1000× magnifications. PCR assay targeted cytochrome b (cyt b) gene using LCBF1 and LCBR2 primers and the products digested by Ssp1 enzymes. Results: From 145 suspected CL patients, 76 (52.4%) were positive in microscopic examination. In addition, we detected gene of interest (cyt b) in 98 (67.5%). The results of PCR-RFLP indicated that 53/98 (54%) cases were Leishmania major and 45/98 (46%) were Leishmania tropica, and the main species in these areas was L. major. Conclusion: We concluded that the microscopic examination is not sensitive enough and is not able to distinguish between different Leishmania species. Instead, molecular methods like PCR-RFLP can be appropriately used with promising results. [ABSTRACT FROM AUTHOR]

Background: The liver flukes of the Fasciola species and Dicrocoelium spp. are recognised as parasites of domestic and wild herbivores. Both species of F. hepatica and F. gigantica as well as D. dendriticum are distributed in Iran. The present study aimed to identify Fasciola spp. and Dicrocoelium spp. using mitochondrial Cox1 (cytochrome c oxidase I) gene by HRM method., Methods: Totally, thirty infected liver specimens were collected from the sheep (n:23) and cattle (n:7) at the abattoirs of Qazvin Province, northwest Iran in 2022. DNA extraction and PCR amplification of Cox1 gene were conducted by HRM technique. DnaSP v.5.0 was used for compression of diversity indices of ribosomal 28S rDNA and mitochondrial Cox1 markers of Dicrocoelium spp. The taxonomic status of Dicrocoelium spp. was performed by sequencing and p hylogenetic analysis., Results: Overall, 26 and 4 isolates were identified as F. hepatica and F. gigantica , respectively. D. dendriticum was the sole infecting species of Dicrocoelium revealed by HRM analysis. Genomic analysis showed a moderate ( 28S rDNA genes: 0.600±0.215) to high ( Cox1 : 0.733±0.155) haplotype diversity for D. dendriticum ., Conclusion: The parasite-dependent mitochondrial gene ( Cox1 ) could identify a higher genetic diversity of D. dendriticum compared to nuclear 28S rDNA gene. HRM technique in the present study found to be a reliable technique for identification and genetic diversity of liver flukes but more comprehensive and in-depth studies in different parts of the country are needed., Competing Interests: Conflicts of interest The authors declare no conflicts of interest., (Copyright © 2023 Hajialilo et al. Published by Tehran University of Medical Sciences.)

Background: We aimed to detect the genetic diversity of samples identified morphologically as Fasciola spp. from sheep, cattle and goat from Lorestan Province, western Iran using PCR-RFLP method. Besides, we evaluated the genetic diversity indices, sequencing and phylogenetic analysis using mitochondrial gene ( ND1 and CO1 )., Methods: PCR-RFLP analysis of ribosomal ITS1 fragment by RsaI restriction enzyme to investigate the genetic characteristics of Fasciola species obtained from different hosts (18 sheep, 21 cattle, and 17goats) was conducted. The samples were sequenced. Sequences were evaluated using BLAST software and the parasite species were identified with similarity percentage and overlap with the species registered in the gene bank. Then similarity and diversity of intra-species and intra-species diversity of Fasciola species were calculated., Results: In Lorestan, based on RFLP pattern, 93% (52) of the Fasciola spp. isolates had a RFLP pattern related to F. hepatica and 7% (4) were F. gigantica . No hybrid forms were detected. The CO1 gene could clarify 19 haplotypes against ND1 gene that found 22 haplotypes among livestock. Sequencing results of the mtDNA showed intra-species identity 98. 5%-100% and Intra-species-diversity: 0-1.5% compared to the GenBank sequences., Conclusion: Using PCR-RFLP method, two species of F. hepatica and F. gigantica , were present in Lorestan Province, but F. hepatica was more prevalent. Mitochondrial genes could better test variability indices in different hosts than ribosomal genes, consequently among mitochondrial genes, the ND1 gene could better examine differences and similarities than CO1., Competing Interests: Conflict of Interest The authors declare there is no conflict of interest., (Copyright © 2022 Heydarian et al. Published by Tehran University of Medical Sciences.)

Introduction: Fascioliasis and dicrocoeliasis are the most frequent zoonotic diseases with increasing human health problems in different parts of Iran. Two species, Fasciola hepatica (F. hepatica) and Fasciola gigantica (F. gigantica), are spread in the country. Molecular approaches have a decisive role in identifying both the species. The aim of this study was to detect Fasciola spp. and Dicrocoelium spp. by amplifying the ITS-2 and 28S rDNA gene sequence., Methods: Overall, 30 infected liver samples were collected from the livestock of Qazvin, Iran. The adult flukes were collected from different livestock. DNA extraction and PCR amplification of ribosomal RNA gene region (ITS2) and 28S rDNA gene fragment were conducted and a phylogenetic tree was constructed., Result: All the isolates obtained from the cattle (No: 7) and 82.6% (No: 19) of sheep isolates were infected with F. hepatica species, whereas 17.4% (No: 4) of sheep isolates were infected with F. gigantica. It was also shown that F. hepatica was the predominant species of Fasciola present in the region. All the specimens were infected with Dicrocoelium dendriticum (D. dendriticum)., Conclusion: Both the species of Fasciola were found in Qazvin. D. dendriticum was the sole infecting species of the Dicrocoelium genus in the livestock of the city of Qazvin. Further research studies are needed to determine the intermediate host of the parasites in the region., (Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.net.)

Background: Fasciolosis is a shared disease between humans and livestock caused by hepatic trematodes; Fasciola hepatica and F. gigantica . Differentiate between the two species of this genus is essential. High-Resolution Melting (HRM) Analysis represents a new approach to this issue. This method can be performed right after termination of Real-Time PCR. This technique has not been used for identification of adult F. hepatica and F. gigantica genotypes. The aim of this study was to determine Fasciola species by using HRM in isolates taken from Iran, respectively., Methods: Ninety-three Fasciola spp. samples were collected from infected slaughtered animals in different regions of Iran, including North West (Ardebil Province) and South East (Zahedan Province) during 2016. Genomic DNA from the samples was extracted using a DNA extraction kit and then after Real-Time PCR amplification, HRM was done., Results: Overall, 59 and 34 isolates were identified as F. hepatica and F. gigantica , respectively. The percentages of each species from animals were as follows: sheep ( F. hepatica , 80.39% and F. gigantica , 19.61%), cattle ( F. hepatica , 42.85% and F. gigantica , 57.15%)., Conclusion: HRM technique developed in the present study is a powerful, rapid and sensitive technique for epidemiological survey and molecular identification between F. hepatica and F. gigantica ., Competing Interests: Conflict of interests The authors declare that there is no conflict of interests.

Background: We evaluated the anti-malarial activity of Heracleum persicum individually and in combination with chloroquine., Methods: This study was conducted at the School of Public Health, Tehran University of Medical Sciences, Tehran, Iran in 2015-2016. The Peter's method was used for determining fifty percent effective dose (ED50) of the H. persicum extract and chloroquine individually against chloroquine sensitive P. berghei in small white mice. Six experimental groups for H. persicum and 6 groups for chloroquine and two control group (positive and negative) were considered for determination of ED50. Interaction between H. persicum and chloroquine also was evaluated based on fixed ratios method. Ratios of 0/100, 20/80, 40/60, 60/40, 80/20, 100/0 of ED50 of chloroquine and H. persicum respectively were tested against the parasite. Then inhibitory effects of two drugs were calculated and plotted in the relevant graphs., Results: Overall, 1500 mg/kg, and 1000 mg/kg concentrations of H. persicum against P. berghei resulted in ED50 and ED74 respectively. ED50 of chloroquine against the parasite was obtained as 1.4 mg/kg of mouse body weight. Moreover, combination of H. persicum and chloroquine showed a weak potentiation in ratios of 40/60 (chloroquine + H. persicum ) with 64% inhibition, but not in other ratios., Conclusion: Although H. persicum individually showed a reasonable antimalarial efficacy against chloroquine sensitive P. berghei , in combination with chloroquine it showed additive or antagonism result except in ratios of 40%CQ+60%HP., Competing Interests: Conflict of interests The authors declare that they have no conflict of interests.

Fascioliasis is a foodborne zoonotic disease caused by the two parasite species Fasciola hepatica and F. gigantica . In spite of the presence of both species of Fasciola in the livestock, to our knowledge, to date, no cases of human F. gigantica infection have been reported in Iran officially. Here, we report such a case in a 25 yr old woman referred to The Department of Medical Parasitology and Mycology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran in 2016. CT imaging and MRCP revealed an ill-defined lesion of segments of liver. Specific ELISA produced a positive result besides detecting egg of the parasite via stool exam. The identification of parasite species was performed by the DNA extracted from the eggs and sequencing ITS-1, in addition to comparison to GenBank retrieved sequences, using the BLAST search tool. The sample showed 100% identity with F. gigantica . She was treated for fasciolosis with a single dose of Egaten® 10 mg/kg with positive response. This is the first case of human fasciolosis due to F. gigantica reported in Iran., Competing Interests: Conflict of interest We have no conflict of interest related to this work.