The Prevalence of Blastocystis sp. and Its Relationship with Gastrointestinal Disorders and Risk factors.

Background: Blastocystis sp., located in the large intestine, is one of the most common zoonotic parasites. Risk factors affect its prevalence and pathogenicity, and it causes gastrointestinal disorders. Thus, this study aimed to investigate the Blastocystis sp. prevalence and its relationship with gastrointestinal disorders, in patients referred to laboratories, and provide some prevention strategies. Methods: In this descriptive-analytical study, 1,000 stool specimens were collected from patients referred to Ilam, Iran laboratories from 2018-2019. Wet mount method was conducted on samples, and suspected specimens were confirmed using trichrome staining. The demographic and clinical information was recorded in a questionnaire. Finally, the results were analyzed using the SPSS. Results: Blastocystis infection was detected in 81 out of 1,000 patients (8.1%) including 61 (75.3%) males and 20 (24.7%) females. and illiterate people were more at risk. The prevalence in rural was more than urban areas, and it was more in the age group of 31-50 year. Conclusion: There was a significant relationship between Blastocystis sp. and risk factors (age, sex, level of education, and residence) and clinical symptoms (stomach ache and nausea) (P<0.05), but interestingly there was no significant relationship between bloating and diarrhea.

Molecular characterization of Trichomonas infections in women of Ilam City, southwestern Iran.

Trichomoniasis is a sexually transmitted infection (STI) caused by the flagellated protozoan Trichomonas vaginalis. Little information is available on the epidemiology and genetic diversity of T. vaginalis in Ilam City, southwestern Iran. A descriptive cross-sectional investigation was carried out between July 2017 and December 2018 on the suspected women patients referred to eight gynecology clinics of Ilam City for probable Trichomonas infection. They were undergone a set of clinical, parasitological, and molecular examinations. During clinical consultation, posterior vaginal fornix secretions and urine samples were gathered from the participants. For the reasons such as physical conditions and cultural and religious constraints, most of participating women, especially young girls due to their virginity, preferred to give urine samples instead of vaginal discharge. The presence of Trichomonas was diagnosed by microscopic examination and molecular detection using conventional PCR targeting ITS1-rDNA. A total of 1765 suspected individuals were examined clinically via vaginal secretions (495 specimens) and urine samples (1270 specimens). Of them, 21 (1.18%) cases, including 13 vaginal secretions and 8 urine samples, were positive for Trichomonas infection by microscopy. Slightly more than half of the patients (11/21, 52.4%) complained of vulvar itching, burning, and frequent urination. Cervical lesions, patchy erythema, and vaginal discharge were recorded in 28.6%, 23.8%, and 19% of the patients respectively. All patients with positive microscopic identification were confirmed by amplification of 450-bp fragment of ITS1-rDNA. Phylogenetic analysis revealed a high rate of genetic homogeneity in which all our isolates together with homologous sequences from China, Philippines, Austria, and USA were clustered within the same clade. A statistically significant relationship was recorded between the patients positive for trichomoniasis and the presence of chronic disease (e.g., diabetes, immune system deficiency).

Identification of Trichomonas Vaginalis Genotypes Using by Actin Gene and Molecular Based Methods in Southwest of Iran.

BACKGROUND: Trichomonas vaginalis (T. vaginalis) is a sexually transmitted protozoan parasite and the causative agent of trichomoniasis. The genetic characterization of T. vaginalis isolates shows notable genetic variation in this parasite. In the present study, we aimed to identify the T. vaginalis genotypes based on analyzing of actin gene in women specimens referred to health centers of Ilam city, southwest Iran. METHODS: A total of 1765 female samples were collected from gynecology clinics in the city of Ilam. DNA was extracted from positive samples and nested polymerase chain reaction (Nested PCR) was used to amplify the actin gene. Then, partial sequencing and genotyping of the actin gene was performed. A phylogenetic tree was drawn using the detected genotypes of T. vaginalis and reference sequences. RESULTS: Twenty-one of the 1765 urine and vaginal samples were positive for T. vaginalis. All infected individuals were married and their age in years was between 25 to 34. Further, the majority of infected women had cervical lesions, patchy erythema, and white color discharge. According to sequencing analysis, the isolates were identified as genotype G (n= 8) and genotype E (n= 2). CONCLUSION: From the collected samples, we were able to distinguish at least two genotypes (G and E) of T. vaginalis. However, lesser is known about these genotypes in the city of Ilam. Further studies with a higher number of isolates should be performed in order to understand the implications of these results in this region.

Serum Levels of Soluble Fas and Fas Ligand in Iranian Women with Pre-eclampsia.

BACKGROUND: The precise responsible mechanism of pre-eclampsia remains controversial however, recent data suggest a main role of the abnormal activation of the adaptive immune system and Apoptosis. In this study, we have measured serum levels of Fas/Fasl as two important members of extrinsic apoptotic pathway in patient with pre-eclampsia. METHODS: 207 participants including 99 pre-eclampsia patients and 108 age and sex-matched normal pregnant women were involved in the case-control study. Plasma sample from each participant was collected and stored at -20 °C until batch processing.Serum levels of Fas and Fas ligand were measured by ELISA for each participant including 99 pre-eclampsia patients and 108 normal pregnant women. Following a test of statistical normality, nonparametric data were analyzed by Mann-Whitney. RESULTS: sFas levels in case group was significantly higher than controls; 584 (397-892) pg/ml in cases opposed to 341 (213-602) pg/ml in controls (p value< 0.01). sFasL in pre-eclampsia women was a little lower than controls; 255 (173-318) pg/ml and in case group compared to 265.5 (184-381.5) pg/ml in controls. CONCLUSION: We have found the increased levels of sFas in patients with pre-eclampsia in compare with the healthy pregnant women. It seems that abnormality in sFAS is related with pre-eclampsia.

A systematic review and meta-analysis of recent studies reporting hormone levels related to thyroid gland function in migraineurs, until April 2020.

PURPOSE: The purpose of the current study was to evaluate thyroid function in terms of serum thyroid-stimulating hormone (TSH, also known as thyrotropin), 3,5,3'-triiodo-L-thyronine (T3), and 3,5,3',5'-tetraiodo-L-thyronine (T4, also known as thyroxine) levels in migraineurs in comparison with non-migraineurs using a systematic review of literature and a meta-analysis. METHODS: This is a systematic review of case-control studies on serum TSH, T3, and T4 concentrations of migraineurs in comparison with non-migraineurs. After extracting the data from the finally included studies, the weighted overall standardized mean difference (SMD) was calculated. RESULTS: The weighted overall SMD for the impact of TSH, T3, and T4 blood levels for migraineurs in comparison with non-migraineurs was as follows: 0.804 (95% CI, 0.045-1.564), - 0.267 (95% CI, - 0.660-0.125), 0.093 (95% CI, - 0.077-0.263), respectively. It is noteworthy that only the p value for the significance of the overall SMD for serum TSH level was statistically significant (p = 0.038), as examined by the z-test. CONCLUSIONS: The results of the current study point to an association between migraine pathogenesis and changing TSH levels in comparison with those of controls.

Seroprevalence of Hepatitis E Virus Infection Among Pregnant Women in Ilam, West of Iran.

INTRODUCTION: Hepatitis E infection is commonly known as acute and self-limiting hepatitis, and therefore, less attention is paid to it, whereas hepatitis E virus is a major cause of fulminant hepatitis in pregnant women and its infection during pregnancy is associated with maternal and fetal mortality. The prevalence of anti-HEV antibodies in pregnant women in Ilam city is unknown. The aim of the present study was to investigate the prevalence of anti-HEV total and anti- HEV IgM antibodies among pregnant women in this area. MATERIALS AND METHODS: A total of 420 serum samples were collected between March 2018 and September 2019 from pregnant women, with a mean age of 29.61 years, ranging from 19 to 47 years, referred to Ilam health centers, West of Iran. Demographic data, including age and place of residence, were collected from patient records. The titers of anti-HEV total and anti-HEV IgM antibodies were measured by the ELISA method. The association between the prevalence of hepatitis E antibody and age and place of residence variables was evaluated by the chi-square test. Statistical analysis was performed using SPSS version 20. RESULTS: In total, 18 of the 420 participants (4.3%) were positive for anti-HEV total, while 2 (0/47%) tested positive for anti-HEV immunoglobulins M (IgM). Anti-HEV status had no statistically significant association with age and place of residence. CONCLUSION: The seroprevalence rate of HEV infection among pregnant women in Ilam city is relatively low. Considering that seronegative pregnant women are at risk of acquiring HEV, it is recommended that pregnant women be educated to avoid sources of HEV infection.

A historical review of the role of cytokines involved in leishmaniasis.

Leishmaniasis is an infectious disease caused by the Leishmania genus, affecting millions of persons in the world. Despite increased studies, no vaccine has been developed against leishmaniasis, and drug resistance is evolving in some Leishmania species (spp). Innate and acquired immune cells and their associated cytokines interplay together to determine the immune responses related outcomes in leishmaniasis. Interferon (IFN)-γ or macrophage activating factor (MAF) is the first effective lymphokine (LK), with a related function to leishmaniasis, discovered in 1979. This review article discussed the history of cytokines involved in Leishmania infection, and it is the first report demonstrating the involvement in the disease by focusing on cutaneous leishmaniasis. Up to now, the role of many cytokines has been determined and the literature review showed that IL-35 is the latest known cytokine involved in leishmaniasis. This review revealed that the cytokines have pleiotropic effects, depending upon the cytokine environment, generated during the infection and the host genetic background or infecting Leishmania spp. Overall, advances in our knowledge of immune cells and their secreted cytokines, contributing to the protection or pathological process of leishmaniasis may help to reach new approaches for immunotherapy.

Serum vascular endothelial growth factor (VEGF) levels in ischemic stroke patients: a systematic review and meta-analysis of case-control studies.

Serum VEGF level is regarded to be a biomarker for the diagnosis of stroke. Even though there have been published plethora of original articles describing higher blood VEGF concentrations since the 1970s, however, there is no any meta-analysis report for serum VEGF levels in the field of evidence-based medicine yet. A systematic review was performed by searching the online biomedical databases including retrieving 14 case-control studies including within-article subgroups after fulfilling the inclusion and exclusion criteria without the beginning date restriction, until 2020 for ischemic stroke patients. The Q quantity and I2% statistic index showed a high heterogeneity (84.895 and 84.687, respectively) and the random-effects model of meta-analysis was applied for further analyses. The meta-analysis on a total number of 769 stroke subjects and 621 controls found that the weighted pooled SMD for overall serum VEGF levels on different days of testing was 1.92 (95% CI, - 4.059-0.219, p value = 0.079) and the pooled SMD for overall serum VEGF levels on day 1 of testing was - 1.083 (95% CI, - 4.229-2.063, p value = 0.500). The meta-regression results demonstrated that different days of testing do not significantly affect serum VEGF concentrations in ischemic patients and actually their serum levels are time-independent. Based on the recently published studies, this meta-analysis showed that serum VEGF levels were not significantly associated with an ischemic stroke diagnosis. Thus, researchers may concern another ideal serum or cerebrospinal fluid-derived biomarker for stroke diagnosis.

Worldwide prevalence of emerging parasite Blastocystis in immunocompromised patients: A systematic review and meta-analysis.

BACKGROUND: Blastocystis is one of the most common pathogens of the human intestine, caused by an emerging parasite, which can lead to severe symptoms and even death in immunocompromised patients. We aimed to determine the global prevalence of Blastocystosis infection in people with immunodeficiency. A systematic literature search was conducted on Web of Science, Scopus, Google scholar, Science Direct and MEDLINE databases to select all observational studies reporting the prevalence of Blastocystosis infection in Worldwide, based on different diagnostic methods in immunocompromised patients of any age and published from inception to February 2019. Pooled estimates and 95% confidence intervals (95% CIs) were calculated using random effects models and in addition, the I2 statistic was calculated. The geographic distribution of studies was evaluated and the diagnosis of Blastocystis was compared by various techniques. Electronic databases were reviewed for Blastocystosis infection in HIV/AIDS, cancer and other immunocompromised patients, and meta-analyses were conducted to calculate the overall estimated prevalence. Total68 eligible studies were included. The estimated pooled prevalence rate of Blastocystosis infection in immunocompromised patients was overall 10% (95% CI, 7-13%; I2 96.04%) (P < 0.001), of whom 21% [18-25] were in Australia, 12% [4-24] in America, 11% [6-17] in Europe and 10% [5-15], 7% [3-13] in Asia and Africa, respectively. It was calculated that the estimated pooled prevalence rate of Blastocystosis infection in immunocompromised patients was overall 10% and the prevalence estimates ranged from 0.44 to 72.39. Also, overall the prevalence of parasites co-infection in immunocompromised patients was detected as 0.024%. Our finding showed that immunocompromised people show a high prevalence of Blastocystosis infection compared to the control population. Adequate information on the prevalence rate is still missing from many countries. However, current information underscore that Blastocystis should not be neglected.

Bed Bugs (Hemiptera, Cimicidae): Overview of Classification, Evolution and Dispersion.

The bed bugs (Cimex lectularius and C. hemipterus) have undergone a significant resurgence worldwide since the 1990s. A compilation of findings from a database, including 2650 scientific publications from seven major medical databases, allowed us to document main evolutionary events, from fossil evidence, dating from 11,000 years ago, until the present that has led to the current worldwide expansion of Cimicid species. We present the hypotheses on the possible dispersion pathways of bed bugs in light of the major historical and evolutionary events. A detailed classification of the Cimicidae family and finally, an illustrative map displaying the current distribution of known Cimex species in each geographical ecozone of Asia, Europe, Africa, the Americas, and Australia are presented.

Noninvasive Biological Samples to Detect and Diagnose Infections due to Trypanosomatidae Parasites: A Systematic Review and Meta-Analysis.

Unicellular eukaryotes of the Trypanosomatidae family include human and animal pathogens that belong to the Trypanosoma and Leishmania genera. Diagnosis of the diseases they cause requires the sampling of body fluids (e.g., blood, lymph, peritoneal fluid, cerebrospinal fluid) or organ biopsies (e.g., bone marrow, spleen), which are mostly obtained through invasive methods. Body fluids or appendages can be alternatives to these invasive biopsies but appropriateness remains poorly studied. To further address this question, we perform a systematic review on clues evidencing the presence of parasites, genetic material, antibodies, and antigens in body secretions, appendages, or the organs or proximal tissues that produce these materials. Paper selection was based on searches in PubMed, Web of Science, WorldWideScience, SciELO, Embase, and Google. The information of each selected article (n = 333) was classified into different sections and data were extracted from 77 papers. The presence of Trypanosomatidae parasites has been tracked in most of organs or proximal tissues that produce body secretions or appendages, in naturally or experimentally infected hosts. The meta-analysis highlights the paucity of studies on human African trypanosomiasis and an absence on animal trypanosomiasis. Among the collected data high heterogeneity in terms of the I2 statistic (100%) is recorded. A high positivity is recorded for antibody and genetic material detection in urine of patients and dogs suffering leishmaniasis, and of antigens for leishmaniasis and Chagas disease. Data on conjunctival swabs can be analyzed with molecular methods solely for dogs suffering canine visceral leishmaniasis. Saliva and hair/bristles showed a pretty good positivity that support their potential to be used for leishmaniasis diagnosis. In conclusion, our study pinpoints significant gaps that need to be filled in order to properly address the interest of body secretion and hair or bristles for the diagnosis of infections caused by Leishmania and by other Trypanosomatidae parasites.

Molecular identification of Entamoeba histolytica from stool samples of Ilam, Iran.

INTRODUCTION: Amoebiasis is a multifactorial, life-threatening public health issue and the third parasitic disease cause of mortality in worldwide, particularly in low- and mid-income countries. The aim of this study was to attempt to explore genetic encoding differences of CP8 (conserved gene) of Entamoeba histolytica/Entamoeba dispar in its various infectious properties isolated from Ilam located at a southwest part of Iran. MATERIALS AND METHODS: A total of 2023 stool samples were collected between 2016 and 2018 from the hospital in Ilam, of which only 30 isolates were identified as E. histolytica/E. dispar. These isolates were collected from the intensive care unit, infectious disease, and surgery settings. The isolates were identified and the polymerase chain reaction (PCR) was performed to detect the CP8 gene. In all stages, Entamoeba histolytica HM1: IMSS was used as a positive control. RESULTS: In genotype confirmation, only two isolates had the CP8 gene found in the PCR technique. The sequencing results confirmed the mentioned gene with 99%-100% specificity. CONCLUSION: It is concluded that PCR is highly sensitive to detect E. histolytica and indicating this important role as screening tools in direct DNA extraction from stool samples and valuable technique in early detection of symptomatic and asymptomatic E. histolytica patients.

Molecular Identification and Phylogenetic Classification of Leishmania spp. Isolated from Human Cutaneous Leishmaniasis in Iran: A Cross-sectional Study.

BACKGROUND: In Iran, both forms of cutaneous (CL) and visceral leishmaniasis (VL) have been reported; so the accurate species identification of the parasite(s) and the analysis of genetic diversity are necessary. METHODS: The smears were collected from lesions samples of 654 patients with CL, who attended local health centers in 12 provinces of Iran during 2013-2015. The smears were checked for the presence of amastigotes by light microscopy. DNA of 648 Leishmania isolates, amplified by targeting a partial sequence of ITS (18S rRNA-ITS1-5.8S rRNA-ITS2) gene. Twenty-five of all the amplicons were sequenced and analyzed with restriction fragment length polymorphism (RFLP) using the Taq1 enzyme. RESULTS: All the smears were positive microscopically. The PCR-RFLP analysis revealed that 176 (27%) CL patients were infected with L. tropica and, 478 (73%) with L. major. The dominant species in all over Iran is L. major. The sequencing results of all CL patients and RFLP analysis confirmed each other. Based on our phylogenetic tree, 25 ITS DNA sequences were grouped into two clusters representing L. major and L. tropica species. Phylogenetic tree derived from the ITS sequences supports a clear divergence between L. major from the other species. CONCLUSION: Discrimination of Iranian Leishmania isolates using ITS gene gives us this opportunity to detect, identify, and construct the phylogenetic relationship of Iranian isolates.

Immunodetection and molecular determination of visceral and cutaneous Leishmania infection using patients' urine.

The diagnosis of leishmaniasis relies mainly on the use of invasive processes, to collect the biological material for detecting Leishmania parasites. Body fluids, which can be collected by non-invasive process, would greatly facilitate the leishmaniasis diagnosis. In the present study, we investigated the potency of urine immunoblotting to diagnose cutaneous and visceral leishmaniasis and we compared with routine molecular methods. A total of 80 samples, including 40 sera and their 40 corresponding urine samples were collected from 37 suspected patients with cutaneous and visceral leishmaniasis, and 3 healthy individuals (as control), in Ilam and Ardabil provinces of Iran. All sera and urine samples were analyzed, using immunoblotting. The confirmation of leishmaniasis infection was performed, using conventional and quantitative PCRs as well as by sequencing the amplicons. Among 37 suspected patients, 23 patients presented cutaneous lesions (CL) and 14 exhibited clinical symptoms reminiscent of visceral leishmaniasis (L. infantum). Among cutaneous patients, 15 were positive for zoonotic cutaneous leishmaniasis (L. major), and eight for anthroponotic cutaneous leishmaniasis (L. tropica). Molecular quantification of Leishmania parasites was performed on sera, urines and cutaneous biopsies of CL and VL patients, demonstrating that parasite load is lower in urines, compared to sera or biopsy. DNA can be detected in 20 out of 23 (86.9%) CL urine samples and in 13 out of 14 (92.8%) VL urine samples. Immunodetection analysis demonstrates that 22 out of 23 (95.6%) sera from CL patients and all patients suspected with VL are positive. For urine samples, 18 out of 23 (78.2%) urine of CL patients and 13 out of 14 (92.8%) urine of VL patients were positive, using Western blot. Therefore, immunodetection and molecular analysis using urine samples can be used as a diagnostic tool for surveying cutaneous and visceral leishmaniasis.

Determination of genetic diversity of Leishmania species using mini-circle kDNA, in Iran-Iraq countries border.

BACKGROUND AND OBJECTIVE: Cutaneous leishmaniasis (CL) is one of the most important diseases worldwide, with a different range of prevalence in endemic areas. Anthroponotic and zoonotic CL are two epidemiological forms of CL, in Iran. Although Ilam Province in the west of Iran is one of the main endemic areas of the disease, there is no inclusive study to determine the genetic variations of parasite in these areas. The objective of this study was to determine the genetic diversity of Leishmania species in Ilam Province, using mini-circle kDNA gene. MATERIALS AND METHODS: Direct smears were taken from skin lesions of 200 suspected cases of CL. Smears were stained, screened under light microscope. Polymerase chain reaction (PCR) was performed, using specific kinetoplast DNA primers. Data were analyzed, using the molecular bio-software. RESULTS: All the samples were positive by direct examination. PCR results showed all cases were positive for Leishmania major. Although all isolated cases belong to a different county of Ilam province, all were positive for L. major with intra-species genetic diversity, divided into four clades in the dendrogram. INTERPRETATION AND CONCLUSION: This variation can affect drug resistance and controlling strategies of parasite. It is possible that different species of sand flies and rodents are the vector and reservoir of parasite, respectively; however, further studies are needed to validate this.

Molecular Identification and Intra-species Variations among Leishmania infantum Isolated from Human and Canine Visceral Leishmaniasis in Iran.

BACKGROUND: In Iran, both forms of cutaneous (CL) and visceral leishmaniasis (VL) have been re-ported; so the accurate species identification of the parasite(s) and the analysis of genetic diversity are necessary. METHODS: The investigation was conducted from 2014 to 2015 in the northwest and south of Iran, where VL is endemic (7 provinces). Blood samples of patients and infected dogs were collected and sera separated for serologic examinations (DAT, rK39). Spleen or bone marrow samples from infected dogs were also collected to confirm the infection. DNAs of 70 samples amplified by targeting a partial sequence of ITS (18S rRNA-ITS1-5.8S rRNA-ITS2) gene. All the amplicons were sequenced and analyzed with restriction fragment length polymorphism (RFLP) using the TaqI enzyme. RESULTS: The cause of all 70 VL cases, were L. infantum, so, the dominant specie is L. infantum. The sequencing results of all VL cases and RFLP analysis corroborate each other. Discrimination of Iranian Leishmania isolates using ITS gene gives us this opportunity to detect, identify and construct the phylogenetic relationship of Iranian isolates. In addition, detection and differentiation of Leishmania spp. DNA was confirmed by amplification of variable area of the minicircle kDNA (conserved sequence blocks (CSB)). CONCLUSION: Low divergence and high likelihood were seen among L. infantum isolates of human and dogs from Iran with a very slight divergence was seen between isolates from northwest and south of Iran, thus grouped in a unique clad. No correlation was observed between intraspecies divergence and geographic distribution of the isolates.

Contribution of Blastocystishominis subtypes and associated inflammatory factors in development of irritable bowel syndrome.

Blastocystis hominis with worldwide distribution is a human intestinal protozoa found in all countries. There have been differences in the severity of the pathogenesis of various Blastocystis spp. and a concomitant variation in the plasma concentration of the cytokines in patients with irritable bowel syndrome. In the present study, we aimed to demonstrate the contribution of B. hominis subtypes in the development of irritable bowel syndrome. Stool samples were collected from patients with gastrointestinal disorders. All samples were evaluated through native-lugol method. Total DNA was extracted. A PCR protocol was developed to amplify a specific region of the SSU ribosomal DNA (rDNA) gene. Serum levels of IL-6 and TNF-α were determined by immunoassay methods. The ClustalW algorithm was applied to align and blast the nucleotide sequences of the amplified region of the SSU rDNA gene. To evaluate the phylogenetic and molecular evolutionary of the nucleotide sequences, we used the MEGA software. In this study, we found 26 haplotypes of B. hominis in the studied samples which were collectively belong to five subtypes (ST1, ST2 in patients without irritable bowel syndrome vs. ST3 and two unknown subtypes in patients with irritable bowel syndrome). Result of ELISA showed a high level of IL-6 and TNF-α in the serum of patients with irritable bowel syndrome. The genetic heterogeneity of B. hominis and the existence of different subtypes of the protozoan in patients with IBS may shed light to the fact that some subtypes of parasites may involve in the pathogenesis of IBS.

Elevated level of microRNA-21 in the serum of patients with colorectal cancer.

In patients with colorectal cancer, circulating micro RNA-21 (miR-21) is overexpressed and may act as a potential diagnostic and prognostic biomarker. In the present study, serum miR-21 level was determined in patients with colorectal cancer and control subjects in an attempt to explore its potential clinical diagnostic and prognostic value. Serum levels of miR-21 were measured in 40 patients with colorectal adenocarcinoma and 40 control subjects using a quantitative reverse-transcriptase polymerase chain reaction (qRT-PCR) assay. Serum miR-21 levels were compared in the colorectal cancer patients and control subjects. Furthermore, the association between serum miR-21 level and the clinical stages of tumors was also examined in the patients. Serum miR-21 level was significantly elevated in colorectal adenocarcinoma patients relative to control subjects (P=0.0001), and it was revealed as a potential diagnostic biomarker for differentiating the patients from control subjects. Increased levels of serum miR-21 were associated with clinical stages of tumors in the patients (P=0.01). These results indicated that serum miR-21 levels could serve as a reliable diagnostic and prognostic biomarker for colorectal adenocarcinoma.

Diversity of Leishmania species and of strains of Leishmania major isolated from desert rodents in different foci of cutaneous leishmaniasis in Iran.

BACKGROUND: Zoonotic cutaneous leishmaniasis (ZCL) is a polymorphic disease which may show various symptoms. Genetic diversity of the parasite is suggested to be one of the factors influencing the clinical manifestation of the disease. METHODS: This study used PCR for the detection and identification of leishmanial parasites at the species level and applied a multilocus microsatellite typing approach for investigating the genetic diversity of Leishmania major isolated from captured rodents in two foci of ZCL in Iran: Turkemen Sahara and Fars province. RESULTS: ITS1-rDNA amplification and subsequent RFLP analyses were performed using DNA extracted from the rodents' ears. Approximately one third of the rodents tested positive for Leishmania; in all rodents L. major was the predominating infecting agent. Seven Rhombomys opimus were positive for L. turanica DNA and one for both L. major and L. turanica. DNA of L. infantum was identified in one Rh. opimus. Seventeen strains of L. major, 15 from Turkemen Sahara and two from Fars province, isolated from different rodents were tested for variation at nine polymorphic microsatellite loci. Ten different MLMT genotypes were observed. They were compared to 89 previously published microsatellite profiles obtained for strains of L. major of different geographical origin. Bayesian model-based and genetic distance based approaches confirmed that strains from Turkemen Sahara and from Fars are genetically different and belong to different genetic groups, largely corresponding to their geographical origins. DISCUSSION: The considerable genetic variability of L. major might be related to differences in reservoir host and/or to the existence of different populations of the vector, Phlebotomus papatasi.

Novel identification of Leishmania major in Hemiechinus auritus and molecular detection of this parasite in Meriones libycus from an important foci of zoonotic cutaneous leishmaniasis in Iran.

BACKGROUND: One of the well-known foci of zoonotic cutaneous leishmaniasis (ZCL) in Iran is Turkemen Sahara, which is located in north eastern Iran. ZCL is a disease of mammals, and humans can become infected as accidental hosts. Many researchers have argued that Rhombomys opimus is the only main reservoir host of ZCL in this region of the Golestan province. No other rodents or mammals are thought to host or have been reported to host Leishmania parasites in this region. This research was designed and developed to isolate, detect and firmly identify Leishmania parasites in mammals and rodents other than R. opimus. METHODS: Wild mammals were caught from gerbil burrows. Leishmania parasites were detected to assess the infection of reservoir hosts in 2010. Each genomic DNA sample was screened for Leishmania infection via nested PCR and sequencing using the internal transcribed spacer ribosomal DNA (ITS-rDNA) identification protocol for parasites. RESULTS: The greatest number of infections (8/19) were found in Meriones libycus. One in three infections was found in Hemiechinus auritus, and this is the first report of infection in this species. Only Leishmania major was definitively identified and unambiguously typed in M. libycus and H. auritus. The infection rates in these two wild mammals were not significantly different, and no other gerbil parasites were detected in M. libycus or H. auritus at our study site. CONCLUSIONS: Recent findings of Leishmania turanica in R. opimus and failures to detect L. turanica in M. libycus may be attributable to unidentified Leishmania infections in two M. libycus due to unreadable sequences. These cases may represent mixed infections by L. major and L. turanica. The assumptions that gerbil parasites can be co-infectors provide a starting point for the identification of the causative and potential parasites responsible for the frequent infections that are mainly mediated via sandfly vectors.