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 OBJECTIVE: Stachys inflata Benth. is used as an anti-inflammatory and antiseptic agent in traditional medicine in most mountain villages of Golestan province. Therefore, this study aimed to investigate the antioxidant, ethnopharmacological and phytochemical properties of extract from different parts of S. inflata, collected from Chahar Bagh Mountain.

         METHODS: Flowering branches and root of the plant were collected from Chahar Bagh Mountain (2100 m) in July 2013. At the same time, the most important information about traditional uses of the plant (ethnopharmacology) was recorded by questioning local people. Phytochemical evaluation (total phenolic, flavonoid and anthocyanins content) of ethanolic extract of plant organs was done using spectrophotometry and folin-ciocalteu. The antioxidant activity of the extract was evaluated by DPPH test. P ≤0.05 was considered as statistically significant.

       RESULTS: The amount of chemical compounds in the extract of flowering branches and root extract was significantly different. The total phenolic (129.96 ± 5.6 mgGAE/g), flavonoid (29.62 ± 1.4 mgQUE/g) and anthocyanin (0.021 ± 0.001 µg/g) content in the extract of aerial parts of the plant was approximately 1.5 to 3 times higher than those in the root. Due to higher production of active compounds, the antioxidant activity of the aerial parts’ extract showed  a greater potential in free radical scavenging (IC₅₀= 76.33 ± 4.2 µg/ml) compared to the root extract.

        CONCLUSION: Phytochemical findings and antioxidant activity of the extract of aerial parts of the plant in free radical scavenging, confirm the traditional applications of this plant as analgesic, anti-inflammatory and antiseptic agent in treatment of rheumatism, wounds, burns and diarrhea. It is recommended that further evaluation of the plant’s traditional applications be conducted in vivo and in vitro.

       

ABSTRACT

        Background and Objective: Nutritional factors and activation of inflammatory pathways are thought to be involved in pathogenesis of preeclampsia in pregnant women. The present study aimed to compare the serum levels of vitamin D and interleukin-6 in healthy pregnant women with those of preeclampsia ones.

        Methods: This case-control study was performed on 120 healthy pregnant women and 120 women with preeclampsia referred to Imam Reza Hospital in Kermanshah. The serum levels of vitamin D and IL-6 were measured by ELISA method. The data was analyzed by SPSS software (version 20) using independent t-test, the P-value of <0.05 was considered as statistically significant.

        Results: There was no statistically significant difference between the mean level of vitamin D in the patients (37.64 ± 29.50 ng/ml) and the controls (40.06 ± 33.20 ng/ml). the serum level of IL-6 in patients with preeclampsia (21.71 ± 32.24 pg/ml) was significantly higher compared to that of  control group (15.04 ± 28.6 pg/ml) (P <0.001).

       Conclusion: Based on the findings of this study, inflammatory factors and cytokines such as IL-6 can be considered as risk factor for preeclampsia. However, more studies with larger sample sizes are required to further evaluate the association of vitamin D levels and risk of preeclampsia.

           Keywords: Pregnancy, Pre-Eclampsia, Vitamin D, Interleukin-6.

Background and Objective: Nowadays, incidence of antibiotic-resistance among pathogenic bacteria has increased due to indiscriminate use of antimicrobial drugs for treatment of diseases, especially urinary tract infections. Medicinal plants are also of great importance as antibacterial agents. Therefore, the aim of this study was to determine the antibacterial effect of ethanolic extract of nettle (Urtica dioica L.) leaves using two methods of disk diffusion and well diffusion.

         Methods: Ethanolic extract of nettle leaves was prepared by the percolation method. Effect of different concentrations of the extract on Escherichia coli (PTCC1399), Staphylococcus aureus (PTCC 1431), Staphylococcus epidermidis (PTCC 1435) and Staphylococcus saprophyticus (PTCC1440) was evaluated using the disk diffusion and well diffusion methods by measuring diameter of growth inhibition zone. Gentamicin and propylene glycol were used as positive and negative control, respectively.

        Results: In both methods, especially in the well diffusion, the ethanolic extract of nettle leaves had favorable inhibitory effect on the growth of S. aureus, S. epidermidis and S. saprophyticus. In the well diffusion method, the highest rate of susceptibility to the extract (89%) was related to S. saprophyticus and S. epidermidis.

         Conclusion: The ethanolic extract of nettle leaf has good inhibitory effect on the growth of S. aureus (especially in the well diffusion method), which confirms the traditional use of this plant for the treatment of urinary tract infections.

          Keywords: Antibacterial Effect, Staphylococcus, E. Coli, Ethanolic Extract, Nettle (Urtica dioica L.).

Background and Objective: The cell division cycle 25 (CDC25)is a familyof highly conserved dual-specificity phosphatases that activate cyclin-dependent kinase complexes. These complexes are the main cell cycle regulators. Mammalian cells ,exposure to DNA damaging radiations such as ionizing radiation and ultraviolet light, prevent cell cycle progression by activation of checkpoint pathways and lead to cell death.

      Methods: In this study, mice were exposed to different doses of ionizing radiation. Their total cellular protein was extracted from the bone marrow. After determining and matching the protein concentrations, CDC25A phosphatase levels were measured by western blotting.

        Results: The results showed that exposure to different doses of ionizing radiation in vivo significantly increased the expression of CDC25A compared to control group (P <0.05).

        Conclusion: Exposure to ionizing radiation increases the expression of CDC25A phosphatase, which increases the possibility of tumorigenesis in that area by increasing bone marrow cell proliferation.

        Keywords: Cell Cycle, CDC25A, Ionizing Radiation, Cyclin-Dependent Kinase.

ABSTRACT

       Background and Objective: Leishmania major is a flagellate protozoan parasite causing cutaneous leishmaniasis. Although pentavalent antimony compounds are the first-line drugs for leishmaniasis, their application is often accompanied by numerous limitations and side effects. Therefore, it is necessary to seek drugs of herbal origin that have fast-acting benefits and few side effects without resistance. This study aimed to evaluate the in vitro effects of methanolic extract of Arctiul lappa root on promastigotes and amastigotes of L. major.

       Methods: This experimental study evaluated the effects of 10, 100, 500, and 1000 µg/ml of A. lappa root methanol extract on L. major promastigotes using direct cell counting and MTT assay. The mean number of amastigotes in infected macrophages was calculated after 24 and 48 hours.

       Results: The half maximal inhibitory concentration (IC₅₀) of A. lappa root methanolic extract was 131.25 µg/ml after 24 hours. The mean number of amastigotes in macrophages after 24 hours in the control group and in the A. lappa group with concentrations of 500 and 1000 µg/ml were 3.52, 2.02, and 1.27, respectively.

        Conclusion: The results show that the methanolic extract of A. lappa root has anti-leishmanial effects on the promastigotes and amastigotes of L. major in vitro.

        Keywords: Leishmania Major, Amastigotes, Promastigotes, Arctium.

ABSTRACT
       Background and Objectives: Cellulose is a major component of plant biomass, which can be converted into biofuels and valuable chemicals. The key step in utilization of this organic material is its hydrolysis into soluble sugars. This study evaluated cellulase production by Trichoderma harzianum under different pH values, temperatures and incubation periods with the aim to increase enzyme production and decrease its costs.
Methods: The amount of protein production and the hydrolytic activity of cellulase enzymes including exoglucanase, endoglucanase and β-glucosidase produced by T. harzianum were evaluated using various substrates such as avicel, carboxymethyl cellulose, cellobiose, Whatman grade 1 filter paper under different pH values (4, 4.8, 5.5 and 6.5), temperatures (25, 28 and 34 °C) and incubation times (48, 72, 96 and 120 h).
Results: The optimum condition for cellulase production by T. harzianum is 120 hours of incubation at 25 °C and pH of 6.5.
Conclusion: T. harzianum can be used for the production of all three classes of cellulase. This fungus is suitable for the efficient production of cellulolytic enzymes and reducing the cost of consumables.
Keywords: Cellulose, Trichoderma harzianum, Hydrolytic enzymes, Optimization.

ABSTRACT
         Background and Objectives: Exposure to ionizing radiation in modern societies is inevitable and can cause a variety of adverse health effects such as cancer and birth defects. Therefore, a reliable, repeatable and sensitive method is required for evaluation of radiation exposure. The aim of this study was to determine the amount of histone H2AX phosphorylation as an indicator of radiation exposure to evaluate the rate of double-strand DNA breakage in irradiated mice.
         Methods: In this study, 15 mice were exposed to different doses of ionizing radiation. After extraction of total protein from bone marrow cells, γH2AX protein was measured by western blotting. Data analysis was performed using ANOVA, Tukey's post hoc test, and the Pearson's correlation test.
         Results: The amount of γH2AX protein in the exposed groups increased significantly compared to the control group (P<0.05).
        Conclusion: The results of this study indicate that exposure to ionizing radiation increases the amount of γH2AX protein in bone marrow cells during the early hours. The protein can be used as a biomarker for monitoring of acute radiation or suspected local radiation exposure.
        Keywords: γH2AX Protein, Ionizing Radiation, Mouse.

ABSTRACT
       Background and Objective: Klebsiella pneumoniae is one of the most common causes of bacterial infections. Presence of plasmid-mediated quinolone resistance genes causes low level of resistance in K. pneumoniae. This study investigated the prevalence of resistance to quinolones and fluoroquinolones, and the frequency of qnrA, qnrB and qnrS genes among K. pneumoniae strains.
        Methods: The study was performed on 100 K. pneumoniae strains isolated from hospitals in city of Borujerd (Iran) during April to September 2014. Susceptibility of the isolates to nalidixic acid, ciprofloxacin, norfloxacin and ofloxacin was evaluated. Minimum inhibitory concentration (MIC) of ciprofloxacin was determined using ciprofloxacin Etest strips. Polymerase chain reaction was performed to detect qnrA, qnrB and qnrS genes in quinolone-resistant isolates using specific primers.
      Results: The results showed that 38% of the isolates were resistance to both nalidixic acid and ciprofloxacin. The prevalence of ofloxacin- and norfloxacin-resistant isolates was determined to be 18% and 15%, respectively. The MIC values for ciprofloxacin were ranging from 0.064 to ≥256 μg/ml. In addition, four ciprofloxacin-resistant isolates (10%) had MIC of ≥256 μg/ml. The qnrA gene was not detected in any of the quinolone-resistant isolates. Moreover, 23.6% (n=9) and 5.2% (n=2) of the quinolones-resistant isolates contained the qnrB and qnrS genes, respectively.
      Conclusion: Although 38 isolates were ciprofloxacin-resistant, the qnrB, qnrS genes were detected in a small number of isolates. This indicates the involvement of factors other than the qnr genes in resistance of these isolates to quinolones.
       Keywords: Klebsiella Pneumoniae, Qnr protein, Borujerd.

ABSTRACT
         Background and Objectives: The present study aimed to investigate effects of eight weeks of aquatic exercise and resistance training on plasma neurotrophin-4 (NT-4) levels and NT-4 expression in peripheral blood mononuclear cells (PBMCs) of women with multiple sclerosis (MS).
         Methods: Thirty women with MS were randomly assigned to a resistance-training group, an aquatic exercise group and a control group. After separating plasma and PBMCs from blood samples, level of plasma NT-4 and NT-4 expression was measured after eight weeks of exercise via ELISA and Real-time PCR, respectively.
        Results: The level of plasma NT-4 and NT-4 expression increased significantly following eight weeks of aquatic training and resistance training. 
        Conclusion: Based on the results of the present study, both resistance training and aquatic exercise can increase the level of plasma NT-4 and NT-4 expression in female MS patients. It can be concluded that such trainings could have protective and positive effects on the nervous system of MS patients.
        Keywords: Exercise, Multiple Sclerosis, Neurotrophic Factors, Women.

ABSTRACT
         Background and Objectives: We studied effects of eight weeks of resistance training and IGF-1 injection on serum level of IGF-1, IGFBP-3 and IGF-1/IGFBP-3 ratio in Wistar rats.
          Methods: We randomly divided 28 male Wistar rats into four groups of saline-injected control (C), resistance training+saline injection (RS), resistance training+IGF-1 injection (RI) and IGF-1 injection (II). Resistance training protocol consisted of climbing a ladder (three days/week with 5 reps/3 sets) while carrying a weight suspended from the tail for eight weeks. IGF-1 and saline (1.5 µg/kg/day) were injected before and after exercise sessions. Serum IGF-1, IGFBP-3 and IGF-1/IGFBP-3 ratio and morphology of colorectal tissue were evaluated.
          Results: Serum IGF-1 level and IGF-1/IGFBP-3 ratio decreased in the RS group compared to the other groups (P< 0.05). Rats in the RS group had higher IGFBP-3 level than those in the II and RI groups (P< 0.05). IGF-1 injection had no effect on morphology of colorectal tissue.
          Conclusion: The resistance training reduces IGF-1 and increases IGFBP-3 levels, which might represent a link between resistance training and lower risk of colorectal cancer.
         Keywords: Exercise, Colorectal Neoplasms, Insulin-Like Growth Factor-1, Insulin-Like Growth Factor Binding Protein-3.

ABSTRACT
           Background and Objectives: Development of ecofriendly processes for the synthesis of metal nanoparticles is of great importance in the field of nanotechnology. Microorganisms such as bacteria could be suitable candidates for bioproduction of nanoparticles due to their simplicity and high compatibility with the environment. The aim of this study was to use bacteria isolates from the effluent of wastewater treatment plants to produce silver nanoparticles.
         Methods: For identifying silver-resistant microorganisms, we used the agar diffusion method using PHG II medium containing 0.5 mM silver to determine minimum inhibitory concentration. Bacterial identification was done with biochemical testing and polymerase chain reaction (colony PCR). Finally, silver nanoparticles were produced in the desired bacteria, and the properties of these nanoparticles were studied.
         Results: We found five silver-resistant bacteria among which Stenotrophomonas maltophilia strain MS8 showed the highest resistance (MIC= 6 mM). The bacterium was able to synthesize silver nanoparticles in spherical shapes. The results obtained from visual observations using UV-VIS, TEM and XRD showed that the bacterium was able to reduce silver ions into silver nanoparticles with maximum size of 20 nm.
Conclusion: Based on our findings, this bacterium could be useful for biosynthesis of silver nanoparticles.
          KEYWORDS: Bacteria, Biosynthesis, Minimum Inhibitory Concentration. 

ABSTRACT
          Background and Objectives: The aim of this study was to evaluate the effect of 12 weeks of intense endurance training and bee pollen consumption on ABCA1 gene expression in small intestine, liver and gastrocnemius muscle tissues of male rats.
           Methods: In this study, 24 male Wistar rats (aged 6-8 weeks and weighing 90-110 g) were randomly divided into four groups of saline-control (n=6), saline-training (n=6), bee pollen-control (n=6) and bee pollen-training (n=6). The training groups exercised on a treadmill for 12 weeks (30 m/min, 90 min/day, five days/week). The bee pollen groups were given bee pollen orally (500 mg/Kg) for 12 weeks. Data were analyzed using two-way ANOVA at significance level of 0.05.
          Results: ABCA1 gene expression was highest in the liver, gastrocnemius muscle and small intestine, respectively. The findings also revealed that the intense endurance training caused a non-significant increase in ABCA1 gene expression in the small intestine and liver. However, the training caused a non-significant decrease in ABCA1 gene expression in the gastrocnemius muscle. In addition, consumption of bee pollen significantly increased ABCA1 gene expression in the small intestine and gastrocnemius muscle of male rats. However, the effect of bee pollen on the gene’s expression in the liver was not statistically significant.
           Conclusion: Based on our findings, it can be concluded that consumption of bee pollen has more beneficial effects on the ABCA1 gene expression and reverse cholesterol transport compared with the intense endurance training.
           Keywords: ABCA1 protein, Pollen, exercise.

ABSTRACT
            Background and objectives: Cardiovascular disease is the leading cause of death worldwide. This study examined the effects of cardiac rehabilitation exercise on lipid profile and expression of peroxisome proliferator-activated receptor alpha (PPAR-α) gene in patients who underwent coronary artery bypass grafting.
           Methods: In this quasi-experimental study, after screening, patients who underwent coronary artery bypass grafting (CABG) were randomly divided into an experimental group (n=12) and a control group (n=12). After the surgery and discharge from hospital, the experimental group performed rehabilitation exercise for two months, while the control group did not perform any exercise after discharge from the hospital and the initial phase of rehabilitation. Fasting blood samples were collected before and after the last training session to evaluate biochemical variables and PPAR-α gene expression of lymphocytes. PPAR-α expression level was assessed by qRT-PCR. Statistical analysis was done in the SPSS software (version 20) using repeated measures.
            Results: In the follow-up after the two-month cardiac rehabilitation exercise, the PPAR-α gene was significantly overexpressed and plasma HDL levels increased significantly in the training group compared with the control group (P<0.05). Although the concentrations of LDL and triglycerides decreased in the experimental group, this reduction was not statistically significant (P>0.05).
            Conclusion: The results indicate that the protocols carried out in the study could be utilized for improving HDL levels and cardiovascular function in CABG patients.
            keywords: Cardiac Rehabilitation, Gene Expression, PPAR-α.

ABSTRACT
         Background and objectives: 3' untranslated  region  (3'UTR) single  nucleotide  polymorphisms (SNPs) represent genetic variations that may potentially affect binding of miRNA to coding genes, potentially leading to complex disorders. We aimed to perform in silico analysis of the potential phenotypic effect of 3'UTR SNPs on human astrocyte elevated gene-1 (AEG-1), a newly identified candidate cancer gene.
         Methods: We gathered a list of all 3'UTR SNPs located in the human AEG-1 gene from the SNP database. Analysis of the potential effects was done using MirSNP and MicroSNiper.
         Results: Analysis by the MirSNP estimated that rs187728237 might increase the affinity of two miRNAs and decrease the affinity of 10 other miRNAs to the AEG-1 transcript. Moreover, MicroSNiPer showed that rs80320514 might affect 24 putative miRNA binding sites in the 3'UTR of AEG-1.
         Conclusion:  Based on our findings, it can be concluded that the 3'UTR SNPs located in the human AEG-1 gene may be within the miRNA targets of the transcript, therefore affecting the stability of putative miRNA-target interactions.
         Keywords: AEG-1, miRNA, SNPs, 3' Untranslated Region.

ABSTRACT
          Background and Objectives: Mycobacterium tuberculosis is the causative agent of pulmonary tuberculosis, a main public health problem that results in 1.5 million deaths annually. A number of epidemiological studies suggested that host genetic factors could play a main role in susceptibility to tuberculosis infection.
SP110 is an interferon-induced nuclear body protein with vital roles in apoptosis, cell cycling and immunity. SP110 gene has been suggested to be a suitable candidate for limiting TB infections. Thus, we investigated the possible association between SP110 gene polymorphisms and susceptibility to tuberculosis in the Golestan Province, Iran.
          Methods: We investigated the frequency of rs1135791 polymorphism of the SP110 gene among 100 pulmonary tuberculosis patients and 100 healthy individuals who were referred to the health centers in the Golestan Province (Iran) between 2014 and 2015. Frequency of genotypes was evaluated using amplification refractory mutation system-polymerase chain reaction.
          Results: The frequency distribution of TT, TC and CC genotypes among the patients was 65%, 31% and 4%, respectively. In the control group, the frequency distribution of TT, TC and CC genotypes was 56%, 46% and 7%, respectively. There was no significant difference in the frequency of rs1135791 between the patients with pulmonary tuberculosis and the healthy controls (P=0.42).
          Conclusion:  Based on the results, the SP110 rs1135791 variant is not a genetic risk factor for development of pulmonary tuberculosis in Golestan Province, Iran.
          Keywords: rs1135791T, Pulmonary tuberculosis, Golestan Province.

ABSTRACT
            Background and Objectives: Non-synonymous single nucleotide polymorphisms are typical genetic variations that may potentially affect the structure or function of expressed proteins, and therefore could be involved in complex disorders. A computational-based analysis has been done to evaluate the phenotypic effect of non-synonymous single nucleotide polymorphisms in the gene encoding the human hypoxanthine-guanine phosphoribosyltransferase (HGPRT-1). HGPRT-1 is an enzyme involved in purine recycling pathway and its deficiency is associated with several human genetic disorders.
            Methods: We provide a list of all amino acid replacements in the human HGPRT-1 from the dbSNP, Uniprot and dbEST databases. Sorting intolerant from tolerant (SIFT) and PolyPhen softwares were also used in our study.
            Results: Of 94 amino acid substitutions, rs 267606863 was predicted to be the most deleterious. Substitutions of S110L and S104A in flexible loop and D194N, D201Y, H204R, Y195C, F199V and H204D in hood domain were predicted as functionally damaging.
            Conclusion: It could be concluded that these intolerant changes may lie within a functional region of the protein and may affect the stability and folding of HGPRT-1. These variants could be used for future functional and molecular epidemiology studies of HGPRT-1-related disorders.
            Keywords: Polymorphism, Single Nucleotide, Amino acid substitution, Hypoxanthine Phosphoribosyltransferase.

ABSTRACT
           Background and objectives:  Sirtuin-1 (SIRT1) is a conserved a NAD⁺-dependent protein deacetylase essential for cellular metabolism and defence against oxidative stress and endothelial dysfunction. This study aimed to compare effects of 8 weeks of aerobic and combined training on serum level of SIRT1 and PGC1-α in coronary artery bypass graft patients.
           Methods: In this semi-experimental study, 45 non-active men (mean age 47.4 ± 3.3 years) who had undergone coronary artery bypass graft surgery were randomly divided into three groups: aerobic training, combined resistance and aerobic training and control. Trainings were performed at 80% of maximum heart rate, three sessions a week for 8 weeks. Blood samples were collected 24 hours before the first exercise session and 48 hours after the last training session. T-test and one-way analysis of variance were used for assessment of within-group and between-group differences, respectively. P-values less than 0.05 were considered as statistically significant.
           Results: The 8-week aerobic and combined training increased serum levels of SIRT1 and PGC-1α compared with the control group, and significantly decreased the weight of subjects (p=0.001). There was no significant difference in serum levels of SIRT1 and PGC-1α between the two training groups (P≥0.05).
           Conclusion: Both combined and aerobic training significantly improve the SIRT1 and PGC-1α levels in coronary artery bypass graft patients.
           KEYWORDS: Combined training, Aerobic training, PGC-1α, SIRT1, CABG patient.