ABSTRACT

         Cardiovascular disease (CVD) is one of the most important causes of mortality worldwide. Several risk factors have been proposed to be associated with CVD. The main risk factors include family history, smoking, hypertension, history of diabetes, hyperlipidemia and obesity. However, new factors have been recently proposed to be associated with CVD. These include blood homocysteine, fibrinogen, plasma factor VII activity, lipoprotein (a) and several other factors. Nowadays, the role of homocysteine in the development and exacerbation of several diseases has been demonstrated. Extensive studies have been performed on the role of homocysteine in diabetes, preeclampsia, depression, Alzheimer's disease and multiple sclerosis. Homocysteine level increases with age. Moreover, absorption of micronutrients, especially vitamin B12, decreases in old age that could cause a further increase in homocysteine level. On the other hand, elasticity of blood vessels decline with aging, and increased levels of homocysteine accelerate atherosclerosis. The sum of these processes highlights the role of homocysteine in the development and exacerbation of several diseases.

         Keywords: Homocysteine, Cardiovascular Disease, Diabetes, Preeclampsia, Depression, Alzheimer's Disease.

ABSTRACT

          Background and Objective: Several studies have

shown the antimicrobial activity of lactobacilli against Salmonella enterica (serotype typhimurium). The aim of this study was to evaluate the inhibitory potential of metabolites produced by probiotic culture of Lactobacillus casei against S. typhimurium and its impact on S. typhimurium motility and biofilm formation.

         Methods: In order to evaluate the impact of the metabolites, L. casei cell-free culture supernatant (CFCS) was collected by centrifugation of L. casei secondary cultures. Effectiveness of the CFCS against Salmonella was evaluated by the well-diffusion method. In addition, in vitro effect of this treatment on motility in Swarm agar and biofilm formation by the bacteria was investigated.

         Results: Inhibition zone diameters of S. typhimurium were 0.83 and 12.1 mm at concentrations of 50 and 100 μl of Lactobacillus CFCS against the log4 of S. typhimurium, respectively. Moreover, CFCS treatment inhibited the motility and biofilm formation by Salmonella. Concentrations of 5% and 10% were determined as the minimum inhibitory concentrations for motility and biofilm formation by S. typhimurium. Furthermore, effectiveness of the CFCS against Salmonella was dose-dependent (P<0.05).

         Conclusion: L. casei CFCS is able to inhibit the growth, motility and biofilm formation in S. typhimurium.

         Keywords: Anti-Bacterial, Lactobacillus Casei Metabolites, Phenotypic Characteristics of S. typhimurium

ABSTRACT
        Background and Objectives: The unfavorable lipid profile in obese individuals is associated with high incidence of various diseases including cardiovascular disease, hypertension, etc. Dieting for weight loss and physical activity are among the most important factors affecting the serum lipid profile. The aim of this study was to compare the effect of diet with and without physical activity on body mass index and serum lipid profile of obese women.
      Methods: This clinical trial was performed in 2011 on 39 obese women referred to a weight loss and nutrition counseling center. Subjects were randomly divided into an active group (diet and exercise) and inactive group (diet without exercise). Blood sampling was done before the intervention and two months after the intervention. Data was analyzed using SPSS-16 and t-test.
       Results: Mean level of triglyceride reduced significantly in both groups. Mean level of cholesterol and low-density lipoprotein decreased significantly only in the active group. The Mean level of high-density lipoprotein in the two groups had no significant difference.
Conclusion: The results indicate that dieting for weight loss along with short-term physical activity improves serum levels of cholesterol and low-density lipoprotein, but does not affect serum HDL level.
        Keywords: Diet, Exercise, Triglyceride, Cholesterol, LDL, HDL.

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: Synthesis and characterization of several metal complexes (metal ions: Co2+, Ti4+ and Ce3+) of macroacyclic Schiff base ligand have been reported. The Schiff base ligand is prepared from the condensation reaction of 1, 4-di-(4-fluoro-2-aminophenoxy) butane with salicylaldehyde in ethanol. In addition, antibacterial activity of metal ions, Schiff base ligand and their complexes have been investigated. Recent studies show that many Schiff base complexes have antibacterial activity against Gram-positive and Gram-negative bacteria. Therefore, we aimed to synthesize new Schiff base complexes and evaluate their antibacterial activity against a number of Gram-positive and Gram-negative bacteria.
         Methods: Schiff base ligand and their complexes were characterized by mass spectrometry and IR, H-NMR and C-NMR spectroscopy. The in vitro antibacterial activity of the Schiff base ligand, metal ions and their complexes were evaluated against some Gram-positive and Gram-negative bacteria by disk diffusion method and determining minimum inhibitory concentration.
          Results: In this study, the Schiff base complexes had good antibacterial activity, but the Schiff base ligand and metal ions did not show any antibacterial effect. In some cases, the antibacterial effect of the complexes was higher than that of the standard antibiotics tetracycline and gentamycin. The titanium complex showed the highest antibacterial activity in both methods. This complex created the largest growth inhibition zone (diameter: 100mm) against Staphylococcus aureus, and had the lowest minimum inhibitory concentration against Bacillus subtilis (6.75 mg/ml).
          Conclusion: The compounds synthesized in our study have strong antibacterial activity.
          Keywords: Schiff base complex, Schiff base ligand, antibacterial activity, Staphylococcus aureus, Bacillus subtilis.

ABSTRACT
       Background and Objective: MiRNAs are small RNAs that are expressed in most eukaryotes, and can regulate gene expression by attaching to the 3’ end of target mRNA. MicroRNA-101 (miR-101) post-transcriptional regulation is important for host-virus interactions. In addition, miR-101 has a tumor suppressive role in liver cancer and metastasis, and induces apoptosis in tumor cells. We examined miR-101 expression in patients with chronic hepatitis B, hepatitis B virus (HBV)-associated cirrhosis and healthy individuals.
       Methods: The study was performed on 108 whole blood samples (36 samples from each group) collected in EDTA tubes. RNA was extraction by RNX-plus kit according to the manufacturer’s protocol. Finally, miRNA expression was evaluated using relative real time PCR.
         Results: A 2.4-fold increase was observed in miR-101 expression in patients with chronic hepatitis B, while there was a 3.5-fold increase in miR-101 expression in patients with HBV-associated cirrhosis compared with healthy controls (P=0.003). MiR-101 overexpression in patients with HBV-associated cirrhosis was more notable that in patients with chronic hepatitis B.
         Conclusion: According to the results, evaluating miR-101 expression may predict disease progression from chronic hepatitis B to HBV-associated cirrhosis.
         Keywords: MicroRNAs, Chronic Hepatitis B, Liver Cirrhosis, MiR-101.

ABSTRACT
         Background and Objectives: Capparis spinosa has been used in traditional medicine for various applications including treatment of liver disorders and cancer. We studied the effects of this plant on cell proliferation and morphological characteristics of PLC/PRF/5 liver cancer cell line.
        Methods: After preparing ethanolic extract of the plant, the inhibitory effect of the extract was assessed using MTT assay, and morphological changes were assessed by an inverted microscope.
       Results: C. spinosa ethanolic extract exhibited anti-cancer effects in a concentration-dependent manner. Half-maximal inhibitory concentration of the extract was 1051±4.21 mg/mL. Morphological changes including cell shrinkage, reduction of cell volume and nuclear condensation confirmed the inhibitory effect of C. spinosa on PLC/PRF/5 cells.
        Conclusion: According to the results of this study, extract of C. spinosa seems to be suitable for prevention and treatment of liver cancer. Further studies on animal models could verify the efficiency of the extract against cancer cells.

KEYWORDS: Plants, Medicinal, Capparis spinosa, Liver Neoplasms.       

ABSTRACT
          Background and Objectives: C-C chemokine receptor type 5 (CCR5) is a chemokine receptor expressed at high levels on the surface of T-cells. A 32-bp deletion in the coding region of the CCR5 (CCR5Δ32) leads to production of an incomplete protein that is not expressed on the cell surface. CCR5Δ32 may be involved in development of autoimmune disease, such as systemic lupus erythematosus. We investigated frequency of the CCR5Δ32 polymorphism in SLE patients and healthy controls, and evaluated the relationship between the CCR5Δ32 polymorphism and susceptibility to SLE in Golestan Province, Iran.
          Methods: Whole blood samples were taken from 80 SLE patients admitted to Shahid Sayyad Shirazi hospital and 80 healthy controls (from a blood bank) in the Golestan Province, in 2016. Baseline clinical and laboratorial characteristics were evaluated regarding the CCR5Δ32 genotypes. The CCR5Δ32 polymorphism was determined from genomic DNA by polymerase chain reaction.
          Result: Genotype frequencies of both groups were in the Hardy-Weinberg equilibrium. The frequencies of the CCR5 and the CCR5Δ32 alleles were 98.13% and 1.88% among the patients, and 98.75% and 1.25% among the controls, respectively. Homozygote CCR5Δ32 was not observed in the subjects. The frequency of heterozygous Δ32 was 3.8% and 2.5% among the SLE patients and controls, respectively (P-value>0.05). There was no significant association between the CCR5 status and clinical signs of SLE (P>0.05).
          Conclusion: Our data suggest that the CCR5Δ32 polymorphism has no correlation with SLE in our study population. In addition, the frequency of the Δ32 polymorphism in SLE patients and controls does not follow the Hardy-Weinberg equilibrium
          Keywords: CCR5, Homozygote CCR5Δ32, Heterozygote CCR5Δ32, CCR5Δ32 allele, SLE.

ABSTRACT
          Background and Objectives: Nonviral carriers including those based on synthetic cationic lipids, offer several advantages over the viral counterparts. These carriers are able to form complexes with nucleic acids and deliver genes into the cells via the cellular endocytosis pathway, without significant toxicity. The level of transgenes expression depends on some experimental variables including cell type and density, Lipofectamine and DNA concentrations and Lipofectamine-DNA complexing time. The main objective of this study was to optimize transfection of SW480 colon cancer cells with Lipofectamine 2000.
          Methods: In this study, SW480 cells were transfected with plasmid containing green fluorescent protein reporter gene using Lipofectamine 2000. Green fluorescent protein expression was studied under a reverse fluorescence microscope and the results were analyzed with the ImageJ software. Effect of different quantities of plasmid DNA and different Lipofectamine 2000 volumes on cell transfection efficiency was evaluated.
          Results: The optimal volume of Lipofectamine and quantity of plasmid was 2 µl and 1µg, respectively, which showed 59% efficiency for the transfection of SW480 cells at 24 hours post-transfection.
          Conclusion: This study shows that Lipofectamine 2000 is an efficient reagent for the delivery of genes into SW480 cells. According to the results, the quantity of DNA per transfection and reagent concentrations are essential factors for a successful transfection.
          Keywords: Optimization; pEGFP-NI; Lipofectamine; SW480.

ABSTRACT
             Background and Objectives: Cancer and its conventional treatments may bring about some psychological and health-related symptoms leading to unstable cortisol level. Since evidence has shown that mindfulness based approaches can be helpful to reduce cortisol level, we aimed at investigating the efficacy of   mindfulness-based cognitive therapy (MBCT) on decreasing salivary cortisol level in the women with breast cancer. 
             Methods: in this one- group pretest posttest study, the women with early stage breast cancer were assessed to select eligible ones (N = 15). They were first trained for collecting salivary cortisol sample (before, after and follow-up)   and then were asked to participate in MBCT, an eight-week session approach.  Participants provided salivary samples in the morning (7: 30- 8:30), afternoon (1-3) and night (9-11).  To measure cortisol, Cortisol Enzyme Immunoassay Kit was used. Analysis was performed on the data of 12 participants remained, using repeated measurement.
              Results: the results of repeated measure ANOVA revealed that cortisol level of posttest (after 8-weeks of MBCT and follow-up period) in comparison with pretest was significantly reduced for morning and afternoon cortisol while it was not true for night cortisol.  The post-treatment cortisol was not significantly correlated with age groups, education levels and socio-economic status using repeated measure ANCOVA.   
             Conclusion: Given that the women with breast cancer are at the high risk of psychological distresses and unstable cortisol level, we recommend mindfulness-based approaches esp. MBCT to health providers specifically those working with breast cancer patients.
             Keywords: Mindfulness, MBCT, Cortisol , Salivary Cortisol, Breast cancer.

ABSTRACT

            Background and Objectives: Psychiatric disorders such as schizophrenia and bipolar disorder pose a high burden among the general population. Etiological factor(s) of such disorders remain unknown. Borna disease virus (BDV) is a neurotropic virus that has been suggested as an etiological agent for psychiatric disorders. Therefore, we aimed to investigate the prevalence of BDV among patients with schizophrenia and bipolar disorder.
            Methods: Peripheral blood mononuclear cells (PBMCs) of schizophrenic (n=75) and bipolar (n=55) patients and healthy blood donors (n=125) were extracted form whole blood samples. RNA was extracted from PBMCs and the presence of BDV P40 RNA was assessed by reverse transcription-polymerase chain reaction.
            Results: The BDV genome was not detected in any of the subjects. Positive family history of disease was significantly more frequent among patients (P=0.0001). There was a significant association between contact with animals and psychiatric illnesses (P<0.05). Moreover, education level differed significantly between the two groups (P<0.05).
            Conclusion: The results indicate no evidence of BDV genome among patients with psychiatric disorders. Serological examination for BDV antigens or antibodies could provide further information in this regard. In addition, contact with cats is significantly more prevalent among patients with mental illnesses, which might be due to infection with Toxoplasma gondii.
            Keywords: Borna disease virus, Psychiatric disorder, Schizophrenia, Bipolar disorder, Risk factors.

ABSTRACT
            Background and objectives: Coronaviruses are the main causes of respiratory tract infections in humans. They are also the second leading cause of common cold after rhinoviruses, and can lead to otitis media and asthma. The aim of this study was to investigate the molecular detection of coronaviruses in clinical samples of patients with flu-like symptoms.
            Methods: Specimens were taken from 297 patients with flu-like symptoms who were referred to the influenza laboratory of Golestan University of Medical Sciences during 2012-2014. RNA was extracted from the specimens using an RNA extraction kit. Accordingly, RNA was used for cDNA synthesis and GAPDH was used as the internal control. Synthesized cDNA was investigated for presence of human coronaviruses genome with real-time polymerase chain reaction using specific primers. Data were analyzed by SPSS 16.0 software. 
            Results: The coronavirus genome was not detected in the specimens of patients with flu-like symptoms.
            Conclusion: Genome of human coronaviruses is absent in samples from patients with upper respiratory tract infections and influenza-like symptoms, which may indicate the low prevalence of the virus in the Golestan Province, Iran.
            KEYWORDS: Human coronaviruses, Upper respiratory tract infection, Golestan Province.

ABSTRACT
            Background and Objectives: Leptospirosis is a widespread zoonotic disease that is transmitted directly or indirectly from animals to humans. Humans mainly acquire pathogenic leptospires through mucosal or percutaneous exposure to environment contaminated with urine from an infected animal. We aimed to identify pathogenic leptospiral serovars by detection of the ompL37 gene using polymerase chain reaction (PCR).
            Methods: Sixteen pathogenic leptospiral serovars and a saprophytic serovar, L. biflexa were cultured in modified semisolid Ellinghausen-McCullough-Johnson-Harris medium containing 5% rabbit serum. Genomic DNA extraction was done using the phenol-chlorophorm method. The ompL37 gene was amplified using specific primers. PCR products were analyzed by agarose gel electrophoresis.
            Results: The ompL37 gene was amplified only in the pathogenic leptospiral serovars. We detected no amplified fragment for the saprophytic serovar.
Conclusion: Leptospirosis may be confused with other infectious diseases, and therefore, its early and accurate diagnosis is crucial. We showed that molecular detection of pathogenic leptospires based on the ompL37 gene could be used for laboratory diagnosis of leptospirosis.
            Keywords: Leptospirosis, PCR, ompl37 Gene, Pathogenic Leptospires.

ABSTRACT
           Background and Objectives: Candida albicans is one of the most common fungal pathogens that can form biofilm, particularly on surface of medical devices. In recent years, C. albicans has shown increased resistance to antifungal agents. In this experimental study, we aimed to study effects of superparamagnetic iron oxide nanoparticles (Fe₃O₄ nanoparticles or SPION) on biofilm formation by C. albicans.
           Methods: First, the SPION were synthesized by chemical co-precipitation. The formation of nanoparticles was confirmed by Fourier-transform infrared spectroscopy and X-ray diffraction. Minimum inhibitory concentration (MIC) and minimum fungicidal concentration (MFC) of SPION were determined. Then, antibiofilm effects of the nanoparticles were investigated by enzyme-linked immunosorbent assay. Finally, data were analyzed using SPSS 22.0 at significance level of 0.05.
           Results: According to the results of X-ray diffraction, the SPION had a mean diameter of about 70 nm. MIC and MFC values of SPION against C. albicans were 100 ppm and 200 ppm which reduced biofilm formation by 87.2% and 100%, respectively. SPION showed significant inhibitory effects on C. albicans growth and biofilm formation.
           Conclusion: Based on the findings, SPION may be considered as a novel family of fungicidal compounds. However, further studies are necessary to evaluate the safety of these nanoparticles for treatment of fungal infections in humans.
           Keywords: Candida albicans; Biofilms; SPION; Nanoparticles.