ABSTRACT

          Background and Objective: Genetic variations in the gene encoding endothelial nitric oxide synthase (eNOS) enzyme affect the susceptibility to cardiovascular disease. Identification of the way these changes affect eNOS structure and function in laboratory conditions is difficult and time-consuming. Thus, it seems essential to perform bioinformatics studies prior to laboratory studies to find  the variants that are more important. This study aimed to predict the damaging effect of changes in the coding region of eNOS using homology- and structure-based algorithms (SIFT and PolyPhen).

           Methods: First, the single nucleotide polymorphisms in the coding region (cSNPs) of the human eNOS gene were extracted from dbSNP. Resulting amino acid changes were reported as primary data required for the study. Then, position and type of amino acid changes along with the complete amino acid sequence were separately entered into the SIFT and PolyPhen tools for analysis.

         Results: Of 144 single nucleotide changes, 38 changes by the SIFT, 47 changes by the PolyPhen and 18 amino acid substitutions by both tools were predicted as damaging.

          Conclusion: It is predicted that 18 amino acid changes may have damaging phenotypic effects on the structure of the eNOS enzyme that may affect its performance by potentially affecting the enzyme’s various functional regions. Therefore, computational prediction of potentially damaging nsSNPs and prioritizing amino acid changes may be useful for investigating protein performance using targeted re-sequencing and gene mutagenesis experiments.

        

ABSTRACT

       Background and Objectives: Pseudomonas aeruginosa is the most frequent opportunistic pathogen isolated from the sputum of patients with cystic fibrosis (CF). Resistance to β -lactam antibiotics may arise from over expression of the naturally occurring AmpC cephalosporinases or acquired extended-spectrum β-lactamases (ESBL). The aim of this study was to determine the antibiotic resistance profiles as well as the prevalence of ESBL and AmpC production in clinical isolates of P. aeruginosa from CF patients in Tehran.

         Methods: Antibiotic resistance of 50 non-duplicate P. aeruginosa isolates was determined by the disc diffusion method. AmpC β-lactamase production was detected by the antagonism disc test and ESBL production was detected by the phenotypic confirmatory test. The presence of ESBL and AmpC genes was assessed by PCR, followed by sequencing the PCR products.

         Results: The antibiotic resistance rates were as follows: 22% to ceftriaxone, 20% to cefotaxime, 10% to imipenem, 8% to carbenicillin and 6% to ticarcillin, 4% each to cefepime, tobramycin, amikacin and aztreonam and 2% to each piperacilin, meropenem and ceftazidime. AmpC production was observed in 47 isolates (94%) and ESBL production was observed in one isolate (2%). PCR results showed that all isolates carried the bla_AmpC β-lactamase gene. One multidrug-resistant isolate carried both bla_TEM and bla_PER-1 genes.

        Conclusion: The results showed that despite the low rate of antibiotic resistance in P. aeruginosa CF isolates,the  presence of multiple β-lactamases even in one isolate is alarming and can complicate the already difficult treatment of chronic infections in the lungs of CF patients.

         

ABSTRACT

         Background and objectives: Globally, about one third of the population has been infected with Hepatitis B virus (HBV) and more than 400 million people have become chronically infected. Nearly, 20-25% of all carriers develop serious liver diseases such as cirrhosis, chronic hepatitis and hepatocellular carcinoma (HCC). According to the World Health Organization, HBV infection causes more than one million deaths every year. Co-infection with Human Immunodeficiency virus (HIV) and HBV is common, since both viruses have the same routes of transmission. Approximately 10 -15% of HIV-infected individuals develop chronic hepatitis B. The risk of liver diseases-related deaths is also higher in the co-infected patients. According to previous studies, mutation of the pre-core (PC) and basal-core promoter (BCP) regions may play an important role in development of HBV-related HCC and severe liver disease. The aim of this study was to investigate mutations in the BCP, PC and core regions of HBV in HIV-positive patients.

          Methods: DNA was extracted using commercial kits to determine the BCP, PC/core mutations in 124 HIV/HBV co-infected patients (32.4% female and 67.6% male). Polymerase chain reaction (PCR) was performed using specific primers. The positive PCR products were subjected to automated sequencing. Then, nucleotide sequences were aligned with the standard hepatitis B sequence [Gene bank, accession number: AB033559] for mutation detection and analysis.

          Results: In this study, three patients (8.1%) were HBeAg-positive and all of them were HBsAg-positive. The mean of CD4 cell count was 120 cells/mL. The mean age of the patients was 36.16 years. The important pathological mutations in HBV patients including 1752A (73%), 1773C (70.3%), 1753C (10.8%), 1896A (8.1%) and 1762T/1764A (2.7%) were detected in this study.

         Conclusion: Identification of mutations in co-infected patients is of greater importance compared to mono-infected patients, because it can be useful for prediction of HCC-related mutations. Co-infection with HIV has important effects on the natural history of HBV infection, and creates different mutational patterns compared to mono-infected patients.

           Keywords: HBV, HIV, Mutation.

ABSTRACT
            Background and Objectives: Diagnosis of hepatitis E virus (HEV) infection could be missed in some cases if serological tests are used solely. Molecular characterization of HEV is essential for diagnosis of acute and chronic HEV infections, and evaluating the chronic HEV infection status in immunocompromised patients. The aim of this study was to prepare a suitable HEV positive control, determine the limit of detection (LOD) of HEV RNA for a specific molecular test, and evaluate the efficiency and precision of the test.
           Methods: Genomic region of HEV NCBI reference sequence was constructed. LOD, intra-assay precision, and inter-assay precision were calculated to evaluate the efficiency and precision of the test. Then, tenfold serial dilutions of the HEV positive control were prepared. Real time PCR was performed three times for each dilution. Mean, standard deviation, and coefficient of variation of cycle thresholds obtained in three independent and simultaneous tests were calculated, and the results were analyzed.
          Results: The LOD of this test was determined as 1.4×104 copy/ml or 42 copy/reaction or 14 copy/µl. Intra-assay precision and inter-assay precision for all assays were lower than 2.5% and 10%, respectively.
          Conclusion: We propose that the real time PCR assay targeting the ORF2/3 overlapping conserved region is suitable for detection of a wide range of different HEV genotypes found in acute and chronic HEV infections. However, the precision of the test should be improved for detecting HEV RNA lower than 103 copy/ml.
          Keywords: Hepatitis E virus, Limit of Detection, Real Time PCR.

ABSTRACT
       Background and Objectives: Blood transfusion may induce some adverse effects on receivers. Some methods such as antibody screening and cross matching have been suggested to reduce the risk of transfusion complications. However, these methods require commercial antibody screening kits that may also need special equipment. The aim of this study was to introduce a new method for antibody screening that does not require a commercial kit, and could be used in any transfusion laboratory. 
       Methods: We examined 350 samples that contained alloantibody and 350 control samples without the antibody. A solution containing two O+ and one O- samples were used instead of screening cells.
      Results: Sensitivity and specificity of the method were 73.32% and 45.15%, respectively. Positive predictive value and negative predictive value were 58.33% and 63.88%, respectively.
       Conclusion: Our new method can be used in basic hematology laboratories with some modifications.
      Keywords: Antibodies, Antigens, Coombs test.

 

ABSTRACT
            Background and objectives: Pterygium is a non-cancerous growth of conjunctival tissue that can extend onto the corneal surface. The presence of some oncogenic viruses in pterygium and the neoplastic nature of these lesions led us to the postulated involvement of the viruses in the etiology of pterygium. Given the association of human herpesvirus 6 (HHV-6) with ocular diseases, we aimed to investigate presence of this virus in pterygium.
            Methods: Fifty tissue specimens were collected from patients with pterygium who underwent pterygium surgery between February 2013 and May 2015. The specimens were tested by real-time PCR using Maxima SYBR Green/ROX qPCR Master Mix (2X) kit. Demographic and clinical data were collected and analyzed using SPSS software (version 18).
            Results: Six (12 %) specimens were positive for HHV-6 DNA. There was no statistically significant correlation between pterygium and presence of HHV-6.
            Conclusion: Based on the results, a direct association between HHV-6 and development of pterygium seems less probable, which suggests that other etiologic agents must be involved in the multistep process of the disease.
            Keywords: Human Herpesvirus 6; pterygium; Real-time PCR.

ABSTRACT
           Background and Objective: Strongyloides infection is usually asymptomatic in healthy individuals; however, immunocompromised patients are subject to potential hyperinfection involvement. Hereby, we report disseminated and hyperinfection strongyloidiasis in two immunosuppressed patients from north of Iran, an endemic region for Strongyloides stercoralis.
           Case description: After assessment of clinical symptoms, disseminated strongyloidiasis            was identified in the first case with pemphigus vulgaris by the presence of abundant larvae in direct stool smears and gastric biopsy. The second patient had metastatic carcinoma and was hospitalized with complicated symptoms. The infection was diagnosed by detection of numerous first-stage larvae in wet mount stool smears and agar plate analysis. Despite antiparasitic treatment, both patients died during the hospitalization.
           Conclusion: Delayed diagnosis of active strongyloidiasis in immunosuppressed patients can lead to hyperinfection syndrome. Thus, screening for S. stercoralis infection in these patients is recommended in endemic regions to reduce mortalities.
           Keywords: Strongyloides stercoralis, Disseminated strongyloidiasis, Hyperinfection.

ABSTRACT
Background and objectives: Nonalcoholic fatty liver disease (NAFLD) is a chronic hepatic disease characterized by fat accumulation inside hepatocytes. The aim of this study was to evaluate effects of exercise training and resveratrol supplementation on activin A and follistatin levels in rats with NAFLD under a high-fat diet.
Methods: Fifty-six old (40-50 weeks) male Wistar rats were assigned to a healthy control group and seven experimental NAFLD groups: 1. high-fat diet, 2. saline, 3. resveratrol supplementation, 4. continuous exercise, 5. interval exercise, 6. continuous exercise+ resveratrol supplementation and 7. interval exercise+ resveratrol supplementation. Rats in the resveratrol supplementation groups were given 25 mg/kg of body weight intraperitoneal injection of resveratrol daily. Exercises were performed five days a week for eight weeks. Data were analyzed with SPSS (version 21) using one-way analysis of variance (ANOVA) and Tukey's post hoc test at significance of 0.05.
Results: Exercise training and resveratrol supplementation significantly decreased plasma activin A level and increased activin A expression (P<0.05). Plasma level of follistatin was significantly higher in rats under a high-fat diet compared to healthy control animals. All exercise and supplementation groups alone and combined lowered follistatin levels. However, follistatin mRNA expression increased significantly after resveratrol supplementation alone, continuous exercise+ resveratrol supplementation, and interval exercise+ resveratrol supplementation.
Conclusion: Resveratrol has a beneficial effect on activin A and follistatin levels in rats with NAFLD. In addition, resveratrol supplementation combined with exercise training may have greater health benefits for NAFLD patients compared to resveratrol supplementation or exercise training alone.
Keywords: Exercise, Resveratrol, Activin A, Follistatin, Nonalcoholic Fatty Liver Disease.

Background and Objectives: Human parvovirus B19 (B19V) is a widespread virus with various manifestations depending on the immunologic and hematologic status of the host. Infection with the virus can cause a wide range of complications in fetus of infected pregnant women, especially those with hemoglobinopathies. This study aimed to determine association of hemoglobin electrophoretic patterns and risk of B19V infection in pregnant women.
 
      Methods: Blood samples were collected from 200 pregnant women attending University of Abuja Teaching Hospital, Abuja, Nigeria. The samples were screened for anti-B19V IgM and IgG. Hemoglobin patterns were determined using commercial enzyme-linked immunosorbent assay kits and Minicap Flex Piercing Electrophoresis system. Structured questionnaires were used to collate sociodemographic variables and associated risk factors of B19V.
 
      Results: Of 200 participants, 12 (6.0%) were positive for B19V IgM, 45 (22.5%) were positive for IgG and two (1.0%) were positive for both antibody, while 145 (72.5%) had no detectable B19V antibody. Twenty-six subjects (28.3%) with HbAA hemoglobin pattern had B19V IgG of whom, nine (12.5%) had HbAS and 11 (30.6%) had HbSS electrophoretic patterns. There was a significant association between prevalence of anti-B19V IgG and hemoglobin electrophoretic pattern of participants (P=0.037). However, no association was found between prevalence of B19V IgM and hemoglobin electrophoretic pattern, age and parity of pregnant women (P>0.05).
 
      Conclusion: Our findings revealed a high prevalence of B19V infection among pregnant women in the studied area. In addition, acute B19V seems to be associated with hemoglobin electrophoretic patterns of pregnant women. It is recommended to follow up newborns of anti-B19V IgM positive pregnant women.

Background and objectives: Intestinal parasitic infections are a major public health problem worldwide, especially in developing countries. It is estimated that around 3.5 billion people are infected with intestinal parasites. Human intestinal parasites (HIP) are clinically important due to broad epidemiological distribution, reinfection and drug resistance. In the last decades, bioactive compounds from herbs were used against a wide variety of microorganisms including parasites. We aimed to perform a systematic review on studies on the effects of medicinal herbs on HIPs in Iran.
 
      Methods: Relevant scientific publications until April, 2015 were extracted from five English databases (PubMed, Google Scholar, Ebsco, Science Direct and Scopus) and four Persian databases (Magiran, Irandoc, IranMedex and the Scientific Information Database).
 
      Results: A total of 18 papers and two dissertations met the inclusion criteria. Overall, 22 different plant extracts were used against Giardia lamblia, Entamoeba histolytica, Cryptosporidium and Hymenolepis nana. Based on the results, the extracts could exert time- and dose-dependent inhibitory effects against the tested HIPs. Five plants types including Allium, Chenopodium botrys, Carum copticum, F. asafoetida and Artemisia annua were able to completely inhibit the tested parasites, while Thymus vulgaris and A. paradoxum showed the lowest inhibitory effect (7%).
 
      Conclusions: Given the findings, it is recommended to conduct in vivo studies on medicinal herbs with favorable in vitro effects against HIPs.

Background and objectives: Cardiac apoptosis is one of the most important cardiovascular complications of diabetes. We aimed to investigate the changes of Bax, Bcl2 and caspase 3 in cardiac tissue of diabetic rats after six weeks aerobic exercise.
Methods: Thirty two male Wistar rats were randomly divided into healthy control, diabetes control and diabetes + exercise groups. Diabetes was induced by intraperitoneal injection of streptozotocin solution (55 mg/kg). Two weeks after the injection, fasting blood glucose levels were measured to confirm induction of diabetes. The exercise program was performed five days a week for six weeks. Variables were evaluated by ELISA and western blot analysis. All statistical analyses were performed in SPSS (version 22) using ANOVA and at significance of 0.05.
Results: The induction of diabetes in the control groups resulted in a significant increase in Bax, Bax/Bcl2 ratio and a significant decrease in Bcl2 levels (P=0.024). The six-week training exercise in diabetic groups significantly decreased Bax and Bax/Bcl2 ratio and significantly increased Bcl2 (P=0.018).
Conclusion: Our finding showed that diabetes could increase apoptosis in cardiac tissue. In addition, the six-week aerobic exercise can be used as a non-pharmacological strategy to reduce diabetes-related apoptosis in cardiomyocytes.