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Background: Olive oil as a mjor source of fat in diet, besides having a high level of unsaturated fatty acid contains biological components, such as the antioxidant phenolic compounds that can prevent the destructive effect of free radicals and their resulting mutation on cellular structures. The main goal of this study was to investigate the antioxidant and anticancer effects of olive oil through Salmonella typhimurium and microsome. Materials and Methods: In this study, 16 Iranian olive oil samples and one Spanish sample were used. The determination test of anti-mutagenic potential was based on the method proposed by Ames et al. using mutant strain of Salmonella typhimurium TA100 and a well-known carcinogenic material (sodium azide) which by adding microsome of rat liver (S9), its anticancer effect was examined. Positive and negative controls which respectively contained sodium azide and distilled water were studied. Each test was simultaneously done three times and the percentage of inhibition was determined according to (1-T/M) × 100. Results: The inhibitory percentage in its highest level in darkness was equal to 63.64% based on the variety of olive oil and in light was equal to 60.70%. Conclusion: The findings of this study show that the antioxidant and anticancer properties of olive oil decrease with exposure to light

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Background: Nucleos(t)ide analogues, such as lamivudine and adefovir, are effective drugs for treatment of hepatitis B patients. However, long-term treatment with these drugs leads to the emergence of the nucleos(t)ide analogue resistant strains. The impact of nucleos(t)ide analogues on the emergence of HBsAg escape mutations is not clarified. Hence, the aim of this study was to determine HBsAg escape mutations in chronic hepatitis B patients treated with nucleos(t)ide analogues. Materials and Methods:A cross-sectional study was performed on 50 patients with chronic hepatitis B under treatment with nucleos(t)ide analogues (lamivudine and/or adefovir) and 50 naive chronic hepatitis B patients. HBV DNA was extracted from plasma and S gene of virus was amplified by Nested-PCR followed by direct sequencing. HBsAg gene sequence of the samples was evaluated for detection of HBsAg escape mutations. Results: Among the 100 patients, the following HBsAg escape mutations were identified: sQ101H, sG119R, sP120S, sP127S, sA128V, sG130N, sG130R, sT131I, sM133I, and sY134N. The frequency of HBsAg escape mutations in patients under treatment of nucleos(t)ide analogues was 16% and in naïve patients was 6% (p=0.2, OR=2.98). Conclusion:According to the obtained results, there seems to be no association between using nucleos(t)ide analogues and emergence of HBsAg escape mutations.

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Background: Human &beta-defensin 126 (12kDa) is a small cationic glycoprotein that is highly rich of cysteine. DEFB126 gene is located on the subtelomeric end of 20p1.3 in human. High expression of this protein is reported in epididymis. This polypeptide coats the plasma membrane of sperm during epididy‌mal transit. It is likely that &beta -defensin 126 might have role in unexplained male infertility since it involves in sperm maturation and capacitation. The current research designed to investigate if there is relation between &beta-defensin 126 gene mutation and unexplained male infertility.

Materials and Methods: In this case-control study we followed a two cytosine nucleotides deletion of &beta-defensin 126 gene in 35 Iranian men with unexplained infertility and 40 fertile men with normal spermogeram as control group. Standard PCR, SSCP(Single strand conformational polymorphism), and sequencing were used to detect genetic alteration of &beta-defensin 126. ELISA was performed for the assessment of the protein expression on sperm cells.

Results: Analysis of genetic data revealed 28.6% homozygote deletion in unexplained infertile men while this deletion was detected in 7.5% of controls. The deletion frequency was statistically higher in infertile patients than normal control group (p<0.05). The protein expression was less in men with del/del genotype compare to the other genotypes (p<0.005).

Conclusion: Our study shows that this common sequence variation of &beta-defensin 126 takes part in impairment of male reproductive function. Consequently, men with the del/del genotype are significantly less fertile than men who carry the wild type allele.

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Background: Hypertrophic cardiomyopathy (HCM) is a various collection of heart diseases with autosomal dominant inheritance affecting 0.2% of the global population. HCM is also the most common cause of sudden cardiac death in individuals younger than 35 years old. Approximately, 40% of affected cases are associated with MYBPC3 gene. The aim of this study is to investigate the possible presence of mutation in 15 and 18 exons of MYBPC3 gene in patients with HCM in Chaharmahal Va Bakhtiyari province.

Materials and Methods: In this experimental study, 30 HCM patients were selected. DNA was extracted using standard phenol-chloroform method. Certain exons were amplified by PCR method. And then, SSCP and HA methods were run.

Results: Significant differences were observed between the positive control samples and other samples. However, there were no difference in studied exons or shift in the bands.

Conclusion: Mutations in the exons of MYBPC3 gene may cause the HCM disease, and change in other exons may be the causative agent in this geographical region and change in this studied exons may not have contributed to the HCM disease. However, it is necessary to study more patients for getting a better conclusion.

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Background: Streptokinase is one of the most common and cost effective fibrinolytic drugs for treatment of heart attacks and vein thrombosis. Unlike many advantages over other thrombolytic drugs, administration of streptokinase can produce some complications such as immunologic reactions, hemorrhage and incomplete treatment due to relative short half life. Pegylation is one of the most common methods for improving of these shortcomings.

Materials and Methods: In this study, designing a proper candidate for specific pegylation with cysteine was done by means of SPDBviewer software. After a meaning ful mutation by SOEing PCR method, mutated (sk45cys) and intact SK (ski) genes were cloned in pET26-b vector and the structures were transformed in E.coli. Clones, Afrer growing, were expressed by IpTG and exptression of proteins was confirmed by SDS-PAGE and western blotting. The proteins were purified by affinity chromatography with NiNTA columns and amidolytic activity of purified proteins was assayed using chromogenic method and different concentrations of S2251 substrate.

Results: Results of activity assays showed that amidolytic activity of SK45cys had about 10% increase in comparison to Ski, after 30 minutes of complex formation with plasminogen.

Conclusion: Generally, it was concluded that, considering cys45 as a superficial aminoacid and also relative increase of activity, SK45cys can be considered a suitable protein for specific pegylation.

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Background: Azoospermia is defined as the absence of sperm in the semen and is divided in two types; obstructive and non-obstructive azoospermia. Non-obstructive azoospermia include approximately 60% of azoospermia patients. Several genetic and environmental factors can be involved in the development of non-obstructive azoospermia. Until now, several genes have been introduced as the causing factor of the azoospermia that are involved in spermatogenesis and testicular development. These genes are located on Y and/or autosome chromosomes .The aim of the present study was to investigate Y chromosome microdeletions and STAG3 gene mutations in Iranian males with non-obstructive azoospermia.

Materials and Methods: In this study, peripheral blood samples were obtained from 122 men with idiopathic non-obstructive azoospermia and 100 Normo-sperm men who had at least one child and DNA was extracted. Samples were investigated for the presence of Y chromosome microdeletions by Multiplex PCR. Then, existence of probable mutations in exon 7 of STAG3 gene was investigated using MSSCP (multi-temperature single-strand conformational polymorphism) method.

Results: 13 patients (10.66%) had Y chromosome microdeletions, but none of the subjects showed mutation in exon 7 of STAG3 gene. The Y chromosome microdeletions were found in none of the control individuals.

Conclusion: The results showed that Y chromosome microdeletions are the most important cause of non-obstructive azoospermia and should be considered as the main candidate for male infertility diagnostic tests. Mutations in the STAG3 gene are not common among non-obstructive azoospermia patients.

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Background: Pseudomonas aeruginosa is a major nosocomial pathogen that due to its intrinsic and acquired resistance to a wide spectrum of antibiotics poses a threat in clinical settings. One of the drug resistance mechanisms in P. aeruginosa is mutation in negative regulators of efflux pump systems such as nalC. The aim of this study was investigation of nalC mutations in P. aeruginosa isolates from some Rasht hospitals and Lahijan laboratories.  

Materials and Methods: In this cross-sectional study, forty-five P. aeruginosa strains was isolated from several Rasht hospitals and Lahijan laboratories between 2013 to 2014 and identified by biochemical tests. The antibiotic resistance and susceptibility of isolates was determined by Kirby Bauer method and microdilution method. Then PCR-sequencing was carried out to assess nalC mutations in ciprofloxacin resistant isolates.

Results: In this study, the most P. aeruginosa strains was isolated from urine sample (53%), followed by burned strains (31%). The most resistance was seen to erythromycin (100%) and the lowest resistance was seen to ciprofloxacin (~31 %). The highest MIC of ciprofloxacin was determined in some strains >512 μg/ml. Sequencing results showed that 12 ciprofloxacin resistant isolates had one or several missense mutations G71E, S209R and  E153Q in nalC gene.

Conclusion: Given that mutation was defined in most isolates in this study, it seems that mutation in nalC gene plays an important role in ciprofloxacin resistance of nosocomial P. aeruginosa isolates in Guilan province.

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Abstract

Background: Uterine myomas are benign tumors of the uterus which are derived from smooth muscle cells of the myometrium. Genetic factors play a major role in the progression of the disease. One of the most important genes which have impression in the mechanism of formation of the myoma is epidermal growth factor receptor (EGFR) that plays a basic role in the process of cell growth, differentiation, proliferation and mitogenesis. The aim of this study is survey of EGFR gene common mutations in Iranian women with uterine myomas. In this test, the common mutations of the exon 21 and 19 in the EGFR gene were surveyed.

Materials and Methods: In this case-control study, 80 women with myoma and 80 healthy women were studied as control. For checking deletion mutation of the Exon 19 (rs121913438), the Tetra ARMS/PCR method has been used and also for checking point mutation of the exon 21(rs121434568), the ARMS/PCR method has been used and results of the experiments were analyzed via χ 2 test.

Results: The comparison of the genotypes frequency of exon 21 (TT, TG, GG) and exon 19 (WW, WD, DD) related to EGFR gene in two groups of patients and control with using statistical test respectively represents the significant difference (p=1.320e-16) and (p=3.053e-13) in the different genotypes frequency among the patients and control groups.

Conclusion: The results of research indicate a significant relationship between EGFR gene mutations in exon 19 and exon 21 and potential for myoma in the studied population.

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Introduction: Hereditary spastic paraplegia is a rare disease with different inheritance patterns. The prevalence of this disease is about 1.8 per 100 thousand people. Most of the affected patients are the result of consanguineous marriages. Weakness and muscle spasm is the main manifestation of this disease. The purpose of this study was to investigate the mutation analysis of a person with hereditary spastic paraplegia by the whole exome sequencing method.
Methods: Peripheral blood samples were obtained from a person suffering from gait disorder and lower limb spasm with autosomal recessive inheritance pattern. DNA extraction and whole exome sequencing was performed. After analyzing the data related to the whole exome sequence, the disease-causing mutation in the affected person was confirm using Sanger sequencing method. Also, parents were investigated to separate mutation and carrier status. This study was approved by Arak University of Medical Sciences (code IR.ARAKMU.REC.1401.039). Ethical principles have been followed in accordance with the guidelines of the National Ethics Committee and COPE regulations.
Results: In the present study, a homozygous pathogenic mutation (NM_030954.4): c.304T>C (p.Cys102Arg) in the RNF170 gene was identified in the patient, therefor, hereditary spastic paraplegia type 85 was confirmed in him. Also, the parents of the affected person were heterozygous for the mutation.
Conclusions: Homozygous mutation in RNF170 gene was detected using whole exome sequencing method. A mutation in this gene causes hereditary spastic paraplegia. Considering the consanguineous marriage of carrier parents, this finding can be used for preventive measures in future children.

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Introduction: Oculocutaneous Albinism is a hereditary disease with an autosomal recessive pattern. The incidence of this disease is about 1 in every 17 thousand births. Most of the affected people in Iran are the result of consanguineous marriages. White hair, fair skin, and reduction of iris pigments are the main manifestations of this disease. Also, exposure to sunlight increases the susceptibility of these patients to skin cancer. This study aimed to investigate the genetic cause of a person with Oculocutaneous Albinism by whole exome sequencing.
Methods: A 6cc peripheral blood sample was obtained from a child with oculocutaneous Albinism with an autosomal recessive inheritance pattern. DNA extraction and whole exome sequencing were performed. After analyzing the exome sequencing data, the pathogenic mutation was identified. Then, the Sanger sequencing method was used to confirm and segregate.
Results: The affected case showed homozygous pathogenic mutation (NM_000372.5): c.286dupA p.(Met96AsnfsTer73) in exon 1 of the TYR gene. Oculocutaneous albinism IA was determined according to the mutated gene. Also, the parents of the affected person were heterozygous for the mutation.
Conclusions: The mutation causing oculocutaneous albinism was identified in the affected person using the high-efficiency whole exome sequencing method and then confirming the mutation through Sanger sequencing. Considering the parents' consanguineous marriage of the parents, this finding can be used for preventive measures in the future.