Association between Endothelial Selectin (E-selectin) gene polymorphisms and E-selectin level with visceral leishmaniais, in an ARMS-PCR based study

Background: In the visceral leishmaniasis (VL), parasites reside in reticuluendothelial system, mainly in macrophages. Endothelial Selectin (E-selectin) might play an important role in leukocyte-endothelium interactions and inflammatory cell recruitment. The aim of this study was determining E-selectin level and its polymorphism in three groups, patients, seropositive and healthy individuals. Methods: Serum soluble E-selectin levels as well as 2 polymorphisms of E-selectin (Ser128Arg and Leu554Phe) were measured in a cohort of patients with documented VL (n=64), a healthy control group (n=74) and a seropositive for VL but without any symptoms (n=81). Circulation concentration of E-selectin levels was measured by ELIS. The amplification refractory mutation system (ARMS)-PCR procedure was used for detecting polymorphisms. Results: The mean of E-selectin levels significantly differed between three groups (P<0.026), and were increased in patients in comparison with other groups. Difference was more considerable between two groups of patients and healthy ones (patients 92.8 ng/ml; healthy individuals 71.9 ng/ml). Polymorphisms were associated with soluble E-selectin levels and altogether explained 14.4%, 7.2%, and 8.7% in patients, seropositive and seronegative healthy individuals, respectively. Distribution of polymorphisms of 128Ser/Arg and 554Leu/Phe among three groups was not different significantly; however, there was a considerable arrangement in distribution of Ser128Arg polymorphism and 128Arg allele in healthy group was more than two fold of patients (55% against 20%). Conclusion: The association between soluble E-selectin levels and visceral leishmaniasis suggests that this molecule might have significant role in the inflammatory process in VL. Moreover, frequency of 128Arg allele in healthy group was higher than patients

Identification and Genetic Variation of Fasciola Species from Tabriz, North- Western Iran

Background: Fascioliasis is considered as the most important helminthic infection of cattle and sheep. Traditional approaches using morphological and biologic characters cannot cause a cer­tainty in the accurate and precise identification and intra-specific differences of Fasciola spp. In this study, we identified Fasciola species using ITS-1 marker and described genetic variation of each species of the parasite in isolates from Tabriz slaughterhouse in West Azerbaijan Province, north- western Iran.Methods: Overall, 100 samples (50 from sheep and 50 from cattle) morphologically detected as Fasciola worms were studied for identification of Fasciola species by PCR-RFLP method and intra-species variation of the parasite using RAPD-PCR technique.Results: A region of approximately 460bp in all samples was successfully amplified. There were no identifiable variations among the size of PCR products. Two and three fragments in samples correspond to F. hepatica and F. gigantica was seen, respectively, through PCR-RFLP method. No difference was seen in digestion pattern according to host (sheep or cattle). Different types of each species of the parasite was observed using RAPD-PCR technique.Conclusion: We could have an estimate of frequency of F. hepatica and F. gigantic and different genotypes of the parasite in isolates from one locality in north- western of Iran. By extension of such studies in future to other animal hosts (buffalo and goat) and including more regions to sam­pling, the reliability of the results and their application for control programs in zoonotic diseases will be increased

Association between Endothelial Selectin (E-Selectin) Gene Polymorphisms and E-Selectin Level with Visceral Leishmaniais, in an ARMS-PCR Based Study

Background: In the visceral leishmaniasis (VL), parasites reside in reticuluendothelial system, mainly in macrophages. Endothelial Selectin (E-selectin) might play an important role in leukocyte-endothelium interactions and inflammatory cell recruitment. The aim of this study was determining E-selectin level and its polymorphism in three groups, patients, seroposi­tive and healthy individuals. Methods: Serum soluble E-selectin levels as well as 2 polymorphisms of E-selectin (Ser128Arg and Leu554Phe) were measured in a cohort of patients with documented VL (n=64), a healthy control group (n=74) and a seroposi­tive for VL but without any symptoms (n=81). Circulation concentration of E-selectin levels was measured by ELIS. The amplification refractory mutation system (ARMS)-PCR procedure was used for detecting polymorphisms. Results: The mean of E-selectin levels significantly differed between three groups (P<0.026), and were increased in pa­tients in comparison with other groups. Difference was more considerable between two groups of patients and healthy ones (patients 92.8 ng/ml; healthy individuals 71.9 ng/ml). Polymorphisms were associated with soluble E-selectin levels and altogether explained 14.4%, 7.2%, and 8.7% in patients, seropositive and seronegative healthy individuals, respectively. Distribution of polymorphisms of 128Ser/Arg and 554Leu/Phe among three groups was not different significantly; however, there was a considerable arrangement in distribution of Ser128Arg polymorphism and 128Arg allele in healthy group was more than two fold of patients (55% against 20%).  Conclusion: The association between soluble E-selectin levels and visceral leishmaniasis suggests that this molecule might have significant role in the inflammatory process in VL. Moreover, frequency of 128Arg allele in healthy group was higher than patients

Identification of Visceral Leishmaniasis-Susceptible Areas using Spatial Modelling in Southern Caucasus

This study explores the application of spatial modelling techniques to generate susceptibility maps for a neglected zoonotic disease, visceral leishmaniasis (VL), in an endemic area in southern Caucasus that includes Iran, Armenia and Azerbaijan. The social and physical environment of southern Caucasus has been mainly characterized by the presence of several factors that are strongly associated with VL, which has caused a significant number of infections during the past decade. Three popular spatial modelling techniques, consisting of the weights of evidence, logistic regression and fuzzy logic methods, were evaluated and trained using a study area in north-western Iran where an inventory of highly infected areas and high-quality evidential factors was available. Model performance was assessed using the receiver-operating characteristic (ROC) approach. According to the results of these assessments, the fuzzy logic method with γ = 0.5 was chosen for the prediction of VL incidence in southern Caucasus. The susceptibility map generated using the fuzzy logic method indicated that VL followed a spatial pattern at the conjunction of the three countries, which suggests that the prevalence of VL in southern Caucasus is socio-ecologically dependent

Comparison of Resistant and Susceptible Strains of Trichomons vaginalis to Metronidazole Using PCR Method

Background: Metronidazole is drug of choice recommended by WHO for treatment of trichomoniasis, however, some reports claims drug resistance in Trichomonas vaginalis isolates recently. The objective of this study was to determine the minimum lethal concentration (MLC) of metronidazole in resistant and sensitive strains, as well as genetic patterns of these stains by PCR method. Methods: From February 2006 to March 2007, in a cross sectional study, clinical and wet mount examination of vaginal smear along with culture were performed on 683 women attending to public and private outpatient clinics in Hamadan. Trichomoniasis marked based on major clinical symptoms. Diagnosis confirmed using wet mount microscopically and culture in Diamond medium. A serial concentration of metronidazole was provided and all isolated Trichomonas strains (resistant and sensitive) tested by standard method. Finally, all sensitive and resistant strains examined by PCR technique. Results: Only 15/683, (2.2%) of patients clinically diagnosed trichomonal vaginitis were positive for T. vaginalis by wet smear and culture. The minimum lethal concentration (MLC) for clinically sensitive isolates was 25 µg/ml; however, this concentration for resistant isolates was 200 µg/ml after 24 h and 100 µg/ml after 50 h. The results of PCR examination of DNA from sensitive and resistant isolates had same pattern. The lanes appeared by two primers were 98 bp and 261 bp for both clinically sensitive and resistant strains. Conclusion: Resistance to metronidazole in T. vaginalis has not relation to genetic variations and might be related to some physiologic pathways of organism

Diversity of Merozoite Surface Protein-3α Gene of plasmodium vivax in isolates from Sistan and Bluchestan provinces in Iran

Approximately 90 million people are suffered from vivax malaria annually. This disease imposes a heavy economic cost in endemic areas of Asia and the Americas. This study was design and implemented to reveal the extent of polymorphism of the Plasmodium vivax Merozoite Surface Protein -3αgene. 37 blood samples (20 Iranian and 17 non-Iranian) collected from patients attending to malaria clinics in the Sistan and Baluchistan province. The DNA was amplified by nested PCR and the products were digested with HhaI enzyme through PCR/RFLP technique. Three biotypes of the gene based on the size of PCR products, including A (about 1900bp), B (about 1400bp) and C (about 1100bp) and 10 allelic groups after digesting of PCR products with HhaI enzyme was observed. A significant difference between the two groups of patients was not detected.We observed that P. vivax isolates of Sistan and Bluchestan were extremely diverse, and the results are almost compatible with the results of other studies performed previously. Based on the results of our study RFLP method using HhaI (small fragments from 50- 500bp) is suitable technique for describing diversity of the gene

Survey of feline visceral leishmaniasis in Azarshahr area, north west of Iran, 2013

Leishmania infantum is a causative agent of visceral leishmaniasis or kala-azar, which is endemic in some part of Iran. Azarshahr city located in East Azerbaijan province, North West of Iran, which is endemic for visceral leishmaniasis. This study aimed to investigate the possible reservoir role of cats for visceral leishmaniasis in the Azarshahr area. Totally 65 cats have been trapped alive from villages of Azarshahr county and their serum samples subjected to direct agglutination test (DAT) for L. infantum antibodies. Giemsa stained impression smears have been prepared for parasitological examination of spleen and liver tissue. Also liver and spleen samples of the cats have been cultured in Novy-MacNeal-Nicolle (NNN) medium and also used for PCR. None from 65 samples was positive in NNN culture, PCR and microscopic examination. Fifteen (23.07 ) out of 65 serum samples showed Leishmania specific antibody agglutination at 1:320 dilution or above, but all considered as negative because none of them confirmed by Giemsa stained smears, PCR and NNN culture. According to the findings of the present study, cats are not a reservoir for visceral leishmaniasis in the Azarshahr area. © 2014, Indian Society for Parasitology