Global Distribution of Human Protoparvoviruses.

Development of next-generation sequencing and metagenomics has revolutionized detection of novel viruses. Among these viruses are 3 human protoparvoviruses: bufavirus, tusavirus, and cutavirus. These viruses have been detected in feces of children with diarrhea. In addition, cutavirus has been detected in skin biopsy specimens of cutaneous T-cell lymphoma patients in France and in 1 melanoma patient in Denmark. We studied seroprevalences of IgG against bufavirus, tusavirus, and cutavirus in various populations (n = 840), and found a striking geographic difference in prevalence of bufavirus IgG. Although prevalence was low in adult populations in Finland (1.9%) and the United States (3.6%), bufavirus IgG was highly prevalent in populations in Iraq (84.8%), Iran (56.1%), and Kenya (72.3%). Conversely, cutavirus IgG showed evenly low prevalences (0%-5.6%) in all cohorts, and tusavirus IgG was not detected. These results provide new insights on the global distribution and endemic areas of protoparvoviruses.

The Effects of Chemotherapy on the Levels of Serum Anti-Müllerian Hormone in Patients with Gestational Trophoblastic Neoplasia.

Background: Gestational trophoblastic neoplasia (GTN) is a group of tumors highly responsive to chemotherapy. It has been suggested that cancer therapies have detrimental effects on female fertility. Anti-Müllerian hormone (AMH) is considered fertility potential and ovarian reserves in women. The aim of this study was to compare serum AMH levels between the patients with GTN treated with chemotherapy and the patients with hydatidiform mole who underwent suction curettage without receiving any chemotherapy. Methods: In 35 patients with GTN, serum AMH levels were measured before suction curettage and after the administration of chemotherapy and compared with serum AMH levels measured in 35 patients with hydatidiform mole, who did not receive any chemotherapy as a control. In controls, serum levels of AMH were measured before suction curettage and at the time when beta human chorionic gonadotrophin (ß-hCG) levels approached zero concentration. Results: The mean serum AMH levels in the GTN group were significantly lower than those measured in the control group after chemotherapy. In addition, serum AMH levels measured after intervention in each group significantly decreased compared to the basal levels (p=0.034). Serum AMH levels showed significant differences between the patients who received chemotherapy regimens with methotrexate (MTX) alone, actinomycin-D (Act-D) alone, or the combination of MTX and Act-D (p=0.001). Conclusion: Our study showed that fertility preservation is of great importance in patients with GTN treated with chemotherapy. Furthermore, both MTX and Act-D could have potential adverse effects on ovarian reserve.

Determination of virulence determinants of Escherichia coli strains isolated from patients with colorectal cancer compared to the healthy subjects.

AIM: This study determined the genes encoding the binding and receiving factors of iron and microbial biofilm in E. coli strains isolated from mucosal samples of patients with colorectal cancer and inflammation of the colorectal compared to healthy people. BACKGROUND: Colorectal cancer is one of the most important malignancies in recent years. Escherichia coli is the most important infectious agents associated with colorectal cancer that has numerous virulence factors such as iron uptake and adhesion factors included in the process of inflammation and colorectal cancer. METHODS: Of the three healthy, inflammatory bowel diseases and colorectal cancer groups, 40 Escherichia coli strains isolated after confirmation by biochemical and molecular methods. After determining the isolates phylogroups, the frequency of genes was measured by PCR method. The biofilm formation of isolates was performed using Crystal Violet method. RESULTS: In the determination of the bacteria phylogroups, the colorectal cancer isolates had a maximum incidence of phylogroups B2 and A. In the analysis of fimH, papA, papC, iutA, ireA and fyuA genes, the highest frequency was observed in these two phylogroups. The presence of ireA gene in bacterial isolates from three groups showed a significant difference (P value: 0.004). There was also no significant difference in biofilm productions in Escherichia coli strains isolated from the three groups. CONCLUSION: Unlike previous studies focusing solely on Escherichia coli toxins, we found that iron absorption and adhesion factors could be effective in developing inflammatory bowel diseases and colorectal cancer. It was also determined that biofilm formation is a specific characteristic of Escherichia coli isolated from the healthy colon.

Detection of plasmid-mediated quinolone resistance in clinical isolates of Enterobacteriaceae strains in Hamadan, West of Iran.

Plasmid mediated quinolone resistance (PMQR) determinants have arisen as a significant concern in recent years. The aim of this study was screening of resistant-clinical isolates to fluoroquinolone antibiotics and detection of qnr and aac(6')-Ib-cr genes. For this purpose we collected 100 fluoroquinolone-resistant Enterobacteriaceae which were from 3 hospitals in Hamadan, west provinces of Iran, between October 2012 and June 2013. The all samples were identified by biochemical tests and confirmed by PCR method. Antimicrobial susceptibility to 14 antimicrobial agents including levofloxacin and ciprofloxacin were determined by disk diffusion methods and ciprofloxacin MIC was obtained by broth microdilution method as Clinical Laboratory Standards Institute (CLSI) recommendations. The isolates were screened for the presence of qnrA, qnrB, qnrS and aac(6')-Ib-cr genes using PCR assay. Among the screened isolates, 64 strains (64%) of Escherichia coli, 23 strains (23%) of Klebsiella pneumoniae, 13 strains (13%) of Proteus mirabilis were collected as quinolone-resistant isolates. out of 100 isolates, two (2%) were positive for qnrS, seventeen (17%) isolates were positive for qnrB and we did not find qnrA gene in any of the isolates. There were also 32 positive isolates for aac(6')-Ib-cr determinant. We described the prevalence of qnr and aac(6')-Ib-cr genes in fluoroquinolone-resistant Enterobacteriaceae in Hamadan city. The carriage rate of multidrug-resistant Enterobacteriaceae in healthy people in Hamadan City is extremely high. Moreover, genes encoding transferable quinolones, in particular aac(6')-Ib-cr, are highly prevalent in these strains.

Molecular epidemiology of hepatitis C virus and its relation with persistence or clearance of infection in Hamadan, West-Iran.

BACKGROUND AND OBJECTIVES: Hepatitis C Virus genotyping appears to be vital for predicting the response to antiviral therapy. The present study aimed to analyze the HCV genotypes in relation to persistence or clearance of the virus in residents of Hamadan Province, West-Iran. MATERIAL AND METHODS: A total of 1159 recorded questionnaires of HCV infected people were evaluated in this prospective study. Several parameters including HCV genotypes, anti-HCV antibodies, viral load, drug treatment, response to therapy and amount of ALT and AST were analyzed. RESULTS: HCV genotyping in 637 samples revealed a predominance of type 1a (52.1%) followed by 3a (42.4%), type 1b (2.7%) and type 2 (0.2%) respectively. Mixed genotypes (3a-1a) were detected in 0.9%, and 1.7% had untypable genotype. High frequency of genotypes 1a and 3a were observed in drug-resistant (group-a) and drug-sensitive (group-b) patients respectively (P<0.0001). Additionally, duration of drug treatment was significantly higher in group-a than group-b (P<0.0001). During follow-up period, 92 cases showed spontaneous clearance of HCV infection and more importantly 86 of 92 cases were positive for anti-HCV antibodies compared with 59 of 455 antibody positive cases with treatment-induced clearance of HCV infection (P<0.0001). CONCLUSION: HCV genotyping and also antibody screening could be useful for proper therapeutic intervention in HCV infected subjects.

Expression Analysis of mir-21 and mir-221 in Cancerous Tissues from Iranian Patients with Gastric Cancer.

BACKGROUND: Early detection is a key to survival for gastric cancer. Molecular markers such as miRNA (microRNA) can have great importance in the early diagnosis of gastric cancer. Expression of miR-21 and miR-221 are deregulated in many types of human cancers. This study aimed to investigate the differences in miRNA expression patterns within the Iranian population. METHODS: Total RNA was extracted from gastric cancer tissues and adjacent non-cancerous tissues from 32 patients. Expression levels of miR-21 and miR-221 were detected by Real time RT-PCR using a specific primer, with 5s rRNA as the internal reference gene. RESULTS: Our data showed that the expression levels of miR-21 and miR-221 in gastric cancer samples were significantly higher than in paired non-cancerous samples (P value less than 0.05). The receiver operating characteristic (ROC) analyses yielded the area under the curve (AUC) values of 80.30 for miR-21 and 93.30 for miR-221, and combined ROC analysis revealed the highest AUC value of 96.90 in discriminating GC patients from healthy controls. CONCLUSION: It seems that miR-21 and miR-221 expression pattern in Iranian patients with gastric cancer are similar to any other population. Considering the increased expression level of two miRNAs in cancerous tissue compared to normal tissue as well as the area under ROC curve, miR-21 and miR-221 can be used for early detection of gastric cancer.

Emerging issues associated with HIV patients seeking advice from health food stores.

OBJECTIVES: To ascertain the recommendations, training and education of health food store employees and determine how they communicate the costs, benefits and risks associated with natural health products for the HIV/AIDS community. METHODS: Four male research assistants, posing as asymptomatic HIV-positive individuals, inquired of employees of all retail health food stores in a major Canadian city as to what is recommended for their condition. The research assistants asked about product costs, side effects, potential drug interactions and efficacy. They also inquired as to employee education related to Complementary and Alternative Medicine (CAM) and noted whether employees asked about which conventional medications they were taking and whether they recommended that the subjects seek physician or CAM provider advice. RESULTS: A total of 32 stores were included. Eight store employees (25%) offered no advice; eight (25%) inquired whether the subjects were currently taking medications; six (19%) suggested visiting a physician; and eight (25%) suggested visiting a CAM provider. A total of 36 different products (mean 2.3 per employee) were recommended with considerable variability in product evidence and cost. The education of the employees varied from postgraduate education (n=3), to undergraduate degree (n=3), college level (n=5) in CAM, or no formal education in CAM (n=21). CONCLUSION: There was considerable heterogeneity in advice on natural food products provided by employees of natural food stores and, in general, these individuals had limited formal training in CAM. The products they recommended had limited evidence supporting their efficacy and in some instances were potentially harmful and had considerable costs. The findings of this study support the need to further examine how best to regulate this growing component of the health care system.

Evaluation of Helicobacter pylori vacA and cagA Genotypes and Correlation With Clinical Outcome in Patients With Dyspepsia in Hamadan Province, Iran.

BACKGROUND: Helicobacter pylori is known to be a causative agent of chronic active gastritis, peptic ulcer and gastric cancer in human. Diverse genotypes of H. pylori strains have different virulence potency and geographic distribution. OBJECTIVES: The purpose of this study was to investigate the association between the cytotoxin-associated gene (cagA), and the various vacuolating cytotoxin (vacA) genotypes of H. pylori strains and clinical outcomes in patients referred to Shahid-Beheshti Hospital in Hamadan, Iran. PATIENTS AND METHODS: In this cross-sectional study, biopsy samples were collected consecutively from 153 patients with gastric cancer (GC), peptic ulcer dyspepsia (PUD) and non-ulcer dyspepsia (NUD) in the gastroenterology department of Shahid-Beheshti Hospital in Hamadan province, the west of Iran. H. pylori infection was confirmed in 83 patients (3 with GC, 27 with PUD, and 53 with NUD) by histology, rapid urease test (RUT) and culture. Genomic DNA was extracted from the bacterial isolate and was further confirmed with 16S rRNA gene sequencing as H. pylori, and characterized based on cagA and vacA genotyping using the polymerase chain reaction (PCR) method. RESULTS: In this study, vacA genotypes s1/m2, s1/m1, s2/m2 and s2/m1 were determined in 43.4%, 19.3%, 13.2% and 6% of the isolated H. pylori, respectively. The vacAs1 genotype was detected in 52 (62.6%) isolates, of which the vacAs1a genotype was detected in 45.2, 40.7, and 66.6% of the isolates from patients with NUD, PUD, and GC, respectively. The cagA-positive genotype was determined in 73 (87.9%) isolates and 10 (12.1%) were negative. The frequency rates of cagA gene were 84.9, 92.6 and 100% in isolates of patients with NUD, PUD, and GC, respectively. The cagA-positive genotype is strongly associated with s1a/m2 and s1a/m1 vacA genotypes. CONCLUSIONS: The most predominant VacA genotypes in our areas were s1/m2 and s1/m1, which regard as the genotypes with more virulence intensity. The H. pylori vacAs1a, cagA genotypes have a significant relationship with the presence of PUD and GC in Iranian patients with dyspepsia.

Improved real-time rt-PCR assays of two colorectal cancer peripheral blood mRNA biomarkers: a pilot study.

BACKGROUND: Efficient screening for detection of colorectal cancer (CRC) at earlier stages reduces its mortality. The purpose of this study was to investigate expression of carcinoembryonic antigen (CEA) and human telomerase reverse transcriptase (hTERT) mRNA in peripheral blood of CRC patients and to present strategies for early detection screen test. METHODS: Twenty seven patients in non-metastatic stage and 27 healthy individuals were studied. Expression of CEA, hTERT mRNA and 18srRNA (18s subunit of ribosomal RNA, as reference gene) were determined based on real-time RT-PCR on 3 µg of total RNA from blood in 3 separate vials (1 µg per vial). RESULTS: Positive expression rate of CEA mRNA (78%) and hTERT mRNA (81%) were higher in patient group (P<0.001). These rates were meaningfully higher than the results of individual vials containing only 1 µg of total RNA. Difference between Ct values of markers with 18srRNA ΔCt) was higher in healthy group than patient one. Therefore, a ΔCt cut-off value was determined for distinguishing between true- and false-positive results. Concurrent expression of both markers was found in 67% of the patients, which was higher than healthy cases (11%). Combination of concurrent marker expression with cut-off point strategy increased specificity to 100%. CONCLUSION: These results showed that concurrent evaluation of marker expression and performing the test on 3 µg of samples in 3 separate vials may increase specificity and sensitivity of real-time RT-PCR for early detection of non-metastatic CRC. However, more investigations with larger numbers of samples are needed to verify these results.

Association of polymorphism of ser311cys paraoxonase-2 gene with type 2 diabetes mellitus in iran.

BACKGROUND: It is believed that paraoxonase-2 gene polymorphism is associated with type 2 diabetes. This study is aimed to investigate the association between paraoxonase-2 gene polymorphism and type 2 diabetes in an Iranian population. METHODS: This study was performed on 200 individuals including 100 diabetics and 100 healthy subjects. Genomic DNA was extracted from peripheral blood leukocytes, and PCR-RFLP was carried out. Palindromic sequence in PON2 gene was recognized by Dde1 restriction endonuclease. In order to visualize restriction products, electrophoresis was carried out using polyacrylamide gel (8%) and ethidium bromide staining. RESULTS: The expected PCR product of 331 bp was obtained. Digestion of this product with DdeI showed four Ser homozygotes, three Cys homozygotes, and five Ser311 Cys heterozygotes. The gene frequency of Cys (C) in diabetic subjects was significantly higher than in healthy subjects. CONCLUSIONS: This study suggests that an association exists between Ser311 Cys polymorphism and type 2 diabetes mellitus.

Seroprevalence of hepatitis D virus and its risk factors in the west of Iran.

Despite the decreasing trend in hepatitis D virus (HDV) infection worldwide, the importance of this disease cannot be underestimated. The aim of this study was to evaluate patients positive for HBsAg with respect to HDV infection and related factors. Patients with chronic hepatitis B who presented at Hamedan Province Hepatitis Community Center in 2002-2007 were included. A questionnaire covering demographic variables and history of hepatic disease was completed for each patient. Necessary tests were performed and antibodies to HDV were measured using an enzyme-linked immunosorbent assay. Of 81 HBsAg positive patients, 14 (17.3%) contained anti-HDV IgG. Only one of the patients with anti-HDV IgM was positive for HBsAg. Of the anti-HDV IgG positive patients, two (14.3%) were women. Among the women examined in this study, 24 (35.8%) were anti-HDV IgG negative (p = 0.21), and of these, six (42.8%) were HBeAg positive while 17 (25.4%) of the anti-HDV IgG negative women were positive for HBeAg (p = 0.16). The prevalence of chronic hepatitis B among anti-HDV IgG positive and negative patients was 28.6% and 39.2% respectively (p = 0.31). Because of the relatively high rate of hepatitis B virus (HBV) and HDV co-infection in our study subjects, it is vital that healthcare providers and policy makers to recognize the risk factors associated with this HBV and HDV co-infection as well as the reasons for this increased anti-HDV serology in HBV carriers.