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Breast cancer is one of the most common causes of death due to cancer in women. More than half of hereditary breast/ovarian cancer families could be attributed to mutation in breast cancer susceptibility gene BRCA1. This study was performed on blood samples of 30 women who affected with familial breast cancer. Non-radioactive PCR-SSCP technique was utilized mutation screening in exons 3, 10, 12 of BRCA1 gene. Two shifts in exon 3 and also two in exon 12 was detected, but no shift in exon 10 was found. Due to low number of recognized mutations, the statistical analysis didn’t show a meaningful correlation between mutations and pathological characteristics. Results from this study showed that there was a low possibility of germline mutation in these three exons. Low rate of mutation in this report was concordance with the others.

BACKGROUND & OBJECTIVE: Von willebrand disease (VWD) is the most common bleeding disorder caused by von willbrand factor (VWF) deficiency and autosomal dominance inheritance pattern. It is divided into three types, one and three (quantitative) and type 2 (qualitative). Type one is known with mild bleeding symptoms in comparison with type 3. The aim of this study was to determine the mutations responsible for the type one VWD.METHODS: DNA was extracted from affected members of the family with type 1 VWD and the von willebrand gene was amplified using 63 different PCR. Then PCR fragments were analyzed using CSGE gel electrophoresis, and the fragments with the extra bands were analyzed using DNA sequencing.FINDINGS: The average of von willebrand antigen and VWF activity were found 35.18 and 31.4 unit/deciliter in 23 patients from 6 families, respectively. Mutations were found in 4 families from 6. Argenine 1205 to histidine in 2 families and two new mutations including glysine 19 to arginine in exon 2 and a nucleotide change 2821 guanine to adenine (G>A) in intron 21 splice site in two other families. No mutation was found in two other families.CONCLUSION: This study showed that different kind of mutations causing von willebrand disease in different families.

Genetics, cellular and molecular medicines are cutting-edge sciences and technologies that play an important role in improving human health and quality of life. In addition, medical and biological sciences have clearly shown that the onset of diseases differs from person to person due to their different genetic profiles and variations in molecular basis. Therefore, it is feasible that patients respond differently to a single treatment. Personalized medicine is a new field of medicine aims to tailor medical treatments to the individual characteristics of patients. In this review, in addition to genomics and personalized medicine, we touch upon common techniques in this field and the future of personalized medicine. With this development, people can get more information about the possibility of contracting diseases like cancer and early-stage treatments as well as personalized therapies based on their genetic characteristics. Technologies, such as mutation detection in the genome, microarrays or microchips, and next-generation sequencing of the human genome, have not only made molecular detection better and easier but also facilitated the integration of molecular detection with targeted drug delivery for the development of novel personalized treatment. [ABSTRACT FROM AUTHOR]

Aims: Glucantime has been considered as a drug of choice for treating cutaneous leishmaniasis for many years. In the recent years, resistance to Glucantime has been increasingly reported in some regions of Iran. In the Leishmania, Arsenate/Antimony reductase acts on the basis of thiol metabolism; it can donate the electron from reduced glutaredoxin to pentavalent (sbV) antimony and arsenate and reduce them to trivalent (sbIII) antimony and arsenite, based on its enzymatic property. It has been assumed that a functional mutation in the enzyme can result in drug resistance. In the present study, the role of Sb (V)-As (V) reductase of Leishmania tropica in drug resistant to glucantime was investigated. Materials & Methods: In the present experimental research, 15 glucantime sensitive samples and 15 glucantime resistant specimens were collected from different regions of Iran through patients with cutaneous leishmaniasis. For mutation detection, first degenerate primers were designed; then, sequencing and simulation techniques were used based on molecular dynamics method. Findings: In Leishmania tropica-resistant isolates, only one mutation was seen as replacing alanine (Ala) at position 80 instead of valine (Val). The analysis of the radius of gyration did not reveal any increase in the radius of gyration while simulation. Conclusion: Mutations in glucantime-resistant isolates did not significantly change simulated active site of antimony ion. [ABSTRACT FROM AUTHOR]

Background: Thyroid carcinoma is the most common endocrine malignancy. Medullary thyroid carcinoma (MTC) approximately accounts for 5-10% of all thyroid carcinoma. Nowadays, it is obviously, the mutations in REarranged during transfection (RET) proto-oncogene, especially, mutations in exons 10, 11 and 16 are associated with MTC pathogenesis and occurrence. Thus, early diagnosis of MTC by mutation detection in RET proto-oncogene allows to identify patients who do not have any developed symptoms. The aim of this study was to screening of germline mutations in RET proto-oncogene exons 17 and 18 in MTC patients and their first degree relatives in Iranian population. Methods: In this cross-sectional study, three hundred eleven participates (190 patients, 121 their relatives) were referred to endocrine research center, Shahid Beheshti University of Medical Science during September 2013 until September 2015. The inclusion criteria were pathological and clinical diagnosis. After whole blood sampling, genomic DNA was extracted from peripheral blood leucocytes using the standard Salting Out/Proteinase K method. Nucleotide change detection in exons 17 and 18 was performed using PCR and direct DNA sequencing methods. Results: In this study, twenty missense mutations [CGC>TGC, c.2944C>T, p.Arg982Cys (rs17158558)] which included 16 heterozygote and 4 homozygote mutations were found in codon 982 (exon 18). In the present study, 154 G>A (rs2742236) and 4 C>T (rs370072408) nucleotide changes were detected in exons 18 and intron 17 respectively. There was no mutation in exon 17. Conclusion: It seems that because of arginine to cysteine substitutions in RET tyrosine kinase protein structure and its polyphen score (0.955) and SIFT score (0.01) the mutation in codon 982 (exon 18) could be have pathogenic effects. On the other hands, the mentioned mutation frequency was 6.4% among MTC patients, so this mutation of exon 18 could be checked in genetic screening tests of RET proto-oncogene. Although this needs more study. [ABSTRACT FROM AUTHOR]

Background and Objectives: Milk production and reproductive traits are important economic traits in goat. The different approaches including candidate gene mapping and genome-wide association studies (GWAS) are now implemented in goats in an attempt to identify the molecular mechanisms affecting these economically important traits. The gene ontology (GO) analysis gives a controlled vocabulary for describing attributes of genes and gene products. Kyoto Encyclopedia of Genes and Genomes (KEGG) provides the mythology for the careful examination of gene functions in respect of networks of genes and molecules. Gene network analysis can also identify paths and processes shared by candidate genes. The purpose of the present study was to bioinformatics analysis (GO, path enrichment, and network analysis of the effects of protein-protein interaction) of genes effective in litter size and milk production in goat. Materials and Methods: Candidate genes associated with studied traits were retrieved from literature review, review of candidate gene studies, GWAS and NCBI database. GO analysis and enrichment analysis of the signaling pathways of KEGG were performed using online database of G: Profiler. The String database was used to infer the network of protein-protein interactions (PPI) and the selection of performance module. Cytoscape software was used to draw the resultant networks of protein-protein interactions. Finally, cytoHubba was used to identify Hub genes. Results: GO analysis for litter size showed that, for molecular function the candidates genes enriched in signaling receptor binding, signaling receptor active activity, hormone activity, protein binding and transforming growth factor beta receptor activity. The effect of the transforming growth factor β (TGF-β) family on fertility and reproduction, embryonic development, organogenesis, and uterine growth and function is remarkable in a wide range of organisms. In mammals, even early stages of reproductive development, including male and female germline specification, are controlled by TGF-βrelated proteins. Hormone binding play an important role in fertility and reproduction. Sex hormone-binding globulin (SHBG), specifically bound to the biologically active androgens and estrogens that are key regulators of the reproductive organs as well as other sex-differentiated tissues such as muscle, adipose tissue, and bone. KEGG analysis also identified some signaling pathways including ،PI3K-Akt signaling pathway, TGF-beta signaling pathway and ovarian steroidogenesis which are significantly associated with litter size. GO analysis for 33 candidate genes related to milk production traits identified only one GO category for biological process namely linoleic acid metabolic process. Conclusion: In this study, we identified several significant biological and signaling pathways that can be used to better understand the biological processes associated with litter size and milk production in goats. [ABSTRACT FROM AUTHOR]

Introduction: In wide and developing worlds of cellular and particularly molecular sciences, the most important issue that leads to paying more attention to and use a technique, is its critical features; high sensivity, specificity, velocity and also being available and cheap. High Resolution Melting (HRM), may be one of these techniques. Its usage is growing among others significantly in investigations, for its special advantages. HRM is based on the pattern and characteristic of DNA when melting. DNA melting temperature (Tm), is the temperature that 50% of DNA molecules have become single stranded and 50% are double stranded. The pattern of getting single stranded is specific and unique for every DNA strand; and differences are shown as a graph called “melting curve". These are effective on DNA Tm: the value of bases G and C in both DNA strands (CG%), the length of fragment, the static effects of bases on each strand and the hetrozygocity. In fact, the differences among melting curves results in recognition the special fragment. The way of being single stranded can be followed by fleurcence colors which attached to double stranded DNAs. Conclusion: High-resolution melting is powerful, fast, comprehensive and useful technique for using in molecular laboratory that can be considered as simple and fast approach for genotyping, mutation detection, matching sequence and methylation study. [ABSTRACT FROM AUTHOR]

Background: Early detection and suitable drug admission are the first attempts to control of TB and MDR-TB. In this study, PCR-RFLP method was used for rapid detection of Mycobacterium tuberculosis and resistance to isoniazid. Materials and Methods: In the present study, 87clinical isolates of MTB were used in a PCR-RFLP method. The RFLP-PCR assay by specific primer was used for two purposes In the first, PCR of katG gene for diagnosis of bacteria and in the second step, PCR product by endonuclease enzyme in a RFLP reaction produced a pattern in electrophoresis for mutation detection in katG315. Sequencing method was used for confirmation of RFLP results. Results: The study showed that all studied strains produced band 620bp that confirmed MTB. From the 46 resistant strains to isoniazid, the RFLP-PCR method showed that 44 strains had mutations in the katG gene Ser315Thr. The 41 susceptible strains had not any mutation at the codon. Results of sequencing were confirmed results of the molecular method. Sensitivity of RFLP-PCR test was 95.6% (95% CI :0.85-0 .98) and specificity was 100% (95% CI :0.91-1 .0). Conclusion: RFLP-PCR test is a good tool for simultaneous diagnosis and detection of katG315 mutation in clinical isolates of MTB. [ABSTRACT FROM AUTHOR]

Aims: Medullary thyroid carcinoma accounts for 5-10% of total types of thyroid cancers. An important role of mutations in the RET proto-oncogene in MTC has been well identified. The purpose of this study was to determine the prevalence of seven common germ-line mutations of the RET proto-oncogene in exon 10 in Iranian MTC patients. Materials & Methods: This descriptive study was performed in year 2009-2010. The study population was 217 MTC patients and their first-degree relatives who were selected by simple sampling method. Genomic DNA content of peripheral blood was extracted according to standard salting out/Proteinase K method. Mutation detection in RET proto-oncogene in exon 10 was assessed through PCR-RFLP method. Results: In 217 individuals (Female: Male 1.4:1, mean age 33.4±15.8 years), 9 mutations were identified at codons 618 (7 mutations) and 620 (2 mutations) in RET proto-oncogene exon 10. The most common amino-acid change was Cys618Phe. In addition, no mutation was found in codon 611. Conclusion: The mutations frequency of the RET proto-oncogene in exon 10 is 6% in the studied population. Accordingly, existence of the common mutations in the RET proto-oncogene in all MTC patients and their families is recommended particularly in other exons (11, 12, 13, 14, 15, 16) with a direct DNA sequencing method. [ABSTRACT FROM AUTHOR]

AbstractBackground and Purpose: β-thalassaemia is the most frequent inherited disorder in the world, especially in Iran and Mazandaran Province. It is caused by mulation in β-globin gene on chromosome 11 with more than 150 different mulations causing β-thalassaemia, has been identified in the β-globin gene to date. Hydroxyurea, is one of the drugs used in Thalassemia patient’s treatment, however, it is not effective in all patients. The mechanisms of the hydroxyuea effect in not clear yet. This study compared different β-globin gene mutations in β-thalassaemia patients who were referred to the Thalassemia Research Center in Sari in two groups, good responder and non-responder, to the hydroxyurea.Materials and Methods: This was a case-control study, comparing two groups of 30 thalassaemic patients who received hydroxyurea. Two groups were included, 30 good responders to hydroxyurea treatment (control) and 30 who did not respond to the treatment (case). First, DNA was extracted from peripheral blood. Then, two different methods for mutation detection were used. In the Thalassaemia Research Center in Sari, mutations in 60 patients were identified using ARMS-PCR. Also the results were confirmed in Genetic laboratory of Amirkola, using two mutation detection methods, reverse-dot blot hybridization and ARMS-PCR.Results: In the group of good responder (control), the average patient’s age were 28/1 ± 7/78 years, and the average age at the onset of blood transfusion was reported to be 8/5 ± 8/56 year. In this group, the mean comparison of the hemoglobin level and red blood size (MCV) prior and after drug consumption were statistically significant. In the group of non-responder (case), the mean age was 21.3 ± 6.43, the mean age starting blood transfusions was 3.3 ± 3.75, and the mean of drug consumption was 2.3 ± 0.8 months. From the mutations identified, IVSII-1G>A was the most common type in both case and control group, while of 30 of control group, 22 individuals were homozygous, and 7 individuals were heterozygous for this mutation (frequency % 42.5). For the 30 case patients, 11 individuals were homozygous, while 11 were heterozygous (frequency % 27.5). Comparison between two groups, case and control group, were statistically significance (PA mutation in good responder patients to hydroxyurea as compared to the non responder group, is significant and similar to the previous findings. Key words: β-thalassaemia, Hydroxyurea, MutationJ Mazand Univ Med Sci 2008; 18(64): 1-10 (Persian)

Medullary thyroid cancer accounts for 5-10% of thyroid carcinomas. RET protooncogene mutations occur in all of the hereditary MTCs and about 66% of the sporadic MTCs. So, the detection of the RET mutations is necessary for rapid and proper diagnosis and treatment. This systematic review seeks to find a comprehensive list of RET gene mutations in the diagnosis of medullary thyroid cancer. The previous studies on RET proto-oncogene mutations in the diagnosis of medullary thyroid cancer were searched in the major databases including PubMed, Scopus, Medline, Embase and NCBI between 2010 and 2021. Missense mutations in exons 10, 11, 13, 14, 15, and 16 of the RET proto-oncogene have the highest frequency in MTCs. The most common mutations in FMTC, are in codons 609, 611, 618, and 620 in exon 10, codon 768 in exon 13, codon 804 in exon 14, and codon 634 in exon 11. In the case of MEN2A, RET gene mutations have been observed in exons 5, 8, 10, 11, with the highest mutations in exons 10 (codons 609, 611, 618, and 620) and exon 11 (codons 630 and 634). Moreover, M918T mutation in exon 16 and A883F mutation in exon 15 have been detected in 95% and 5% of the patients with MEN2B respectively. In the case of MTC, the M918T mutation in exon 16 is the most common mutation, which is associated with a poor prognosis. RET genetic screening is crucial for an exact approach to the diagnosis and treatment of MTC. Anyone with MTC, even without a family history of MEN2, should be genetically tested for the RET mutations to confirm or rule out the inherited disease and, if necessary, preventive thyroidectomy. This systematic review provided a comprehensive list of the reported mutations in the RET gene for the diagnosis of medullary thyroid cancer. [ABSTRACT FROM AUTHOR]

Background: Campylobacter species are among foodborne pathogens that are known as the main cause of inflammatory diarrhea in humans. In this cross-sectional study, we investigated the prevalence of Campylobacter spp. and their drug resistance status in fecal samples of patients with community-acquired gastroenteritis in Tehran. Materials and Methods: In this survey, infection with Campylobacter spp. was evaluated in 400 stool samples of patients with diarrhea. Accordingly, microscopic examination to show the presence of exudative diarrhea, rejection of fungi and parasitic infections, enrichment and culture in a specific medium under microaerophilic conditions, determination of biochemical identity, and molecular confirmation at genus and species levels for C. jejuni, C. coli, C. lari, and C. upsaliensis were performed. Antibiotic resistance patterns were determined by E-test and disk diffusion methods, and the presence of the dominant gyrA gene mutations in the quinolone-resistance domain of C. jejuni isolates was determined by using Mismatch Amplification Mutation Assay-polymerase chain reaction (MAMA-PCR)i 1. Results: A total of 28 strains of Campylobacter were isolated from the samples obtained from the patients with diarrhea. The most common species were C. jejuni (6%), C. coli (0.5%), C. lari (0.25%), and other unknown species (0.25%). The highest antibiotic resistance rate was observed against tetracycline and ampicillin (53.5% and 50%, respectively), and the lowest rate was detected for nalidixic acid and ciprofloxacin (28.5% and 28.5%). Multiple drug resistance (MDR) phenotype was detected among 51.8% of the strains. Conclusion: Results of this study indicated C. jejuni as the main Campylobacter species responsible for community-acquired diarrhea among the studied patients. The high rate of resistance to antibiotics and MDR phenotype in these strains compared with other countries is considered a risk, especially in the treatment of invasive infections. [ABSTRACT FROM AUTHOR]

Tuberculosis is one of the most dangerous infectious diseases in the world and causes nearly two million deaths each year, especially in developing countries. Meanwhile, multidrug resistance tuberculosis (MDR-TB) is due to the resistance of Mycobacterium tuberculosis (Mtb) strains to two effective first-line drugs, isoniazid and rifampin, which is increasing worldwide. MDR-TB strains are mainly caused by inadequate treatment of TB patients. The emergence and spread of these strains is an obstacle to the control and management of tuberculosis as well as a threat to the World Health Organization's goal of eliminating the disease by 2050. Proper management of MDR-TB relies on early recognition of the disease. Recently, phenotypic and genotypic diagnostic methods have been developed to rapidly identify MDR strains in tuberculosis patients. Some of them are also economically suitable to use in developing countries. Proper treatment of patients with drug-resistant TB requires the rapid detection of resistant strains and appropriate drug administration. Regular monitoring of patients' side effects of medications as well as enhancing the quality of bacterial tests is essential to identify resistant strains. Therefore, in this review, we will describe the available phenotypic and genotypic tests for drug-resistant tuberculosis detection and discuss their advantages and limitations. [ABSTRACT FROM AUTHOR]

Background: Thyroid carcinoma is the most common endocrine malignancy. Medullary thyroid carcinoma (MTC) approximately accounts for 5-10% of all thyroid carcinoma. Nowadays, it is obviously, the mutations in REarranged during transfection (RET) proto-oncogene, especially, mutations in exons 10, 11 and 16 are associated with MTC pathogenesis and occurrence. Thus, early diagnosis of MTC by mutation detection in RET proto-oncogene allows to identify patients who do not have any developed symptoms. The aim of this study was to screening of germline mutations in RET proto-oncogene exons 17 and 18 in MTC patients and their first degree relatives in Iranian population. Methods: In this cross-sectional study, three hundred eleven participates (190 patients, 121 their relatives) were referred to endocrine research center, Shahid Beheshti University of Medical Science during September 2013 until September 2015. The inclusion criteria were pathological and clinical diagnosis. After whole blood sampling, genomic DNA was extracted from peripheral blood leucocytes using the standard Salting Out/Proteinase K method. Nucleotide change detection in exons 17 and 18 was performed using PCR and direct DNA sequencing methods. Results: In this study, twenty missense mutations [CGC>TGC, c.2944C>T, p.Arg982Cys (rs17158558)] which included 16 heterozygote and 4 homozygote mutations were found in codon 982 (exon 18). In the present study, 154 G>A (rs2742236) and 4 C>T (rs370072408) nucleotide changes were detected in exons 18 and intron 17 respectively. There was no mutation in exon 17. Conclusion: It seems that because of arginine to cysteine substitutions in RET tyrosine kinase protein structure and its polyphen score (0.955) and SIFT score (0.01) the mutation in codon 982 (exon 18) could be have pathogenic effects. On the other hands, the mentioned mutation frequency was 6.4% among MTC patients, so this mutation of exon 18 could be checked in genetic screening tests of RET proto-oncogene. Although this needs more study.

Background and Objectives Hemophilia is one of the chronic disorders of blood coagulation which due to the lack of coagulation factors exposes the person to a variety of physical problems which affect patient. The effectiveness of cognitive-behavioral therapy and treatment based on acceptance and commitment in adherence to treatment, illness perception and quality of life in hemophilic patients was studied in Isfahan city in 1396. Materials and Methods This research was semi-experimental with pre-test and post-test design with control group and follow-up periods. To do the research, 60 hemophilic patients were selected based on eligibility criteria in three groups. The research instrument was a questionnaire for Broadbent questionnaire of illness perception, quality of life and Mariski treatment adherence questionnaire. The statistical analysis was done by SPSS 22 and at two descriptive and inferential levels. Results The results of repeated analysis of variance showed that the difference between the mean scores of the two groups of cognitive-behavioral therapy and the control group was significant in the three variables of treatment adherence, illness perception and quality of life (p < 0.05) and the difference between the mean scores of the two groups of the treatment based on acceptance and commitment and the control group were similarly significant in the three variables of treatment adherence, illness perception and quality of life (p < 0.05). Conclusions The results showed that the two cognitive-behavioral treatments and acceptance and commitment therapy could be used to increase adherence to treatment, illness perception and quality of life in patients with hemophilia. [ABSTRACT FROM AUTHOR]

Introduction: Soil contamination to heavy metals such as lead and zinc in and around mines causes a change in structure, complexity, diversity and activity of soil microbial communities like bacteria. Materials and methods: In the present research, PCR-DGGE approach was used to investigate the effects of Pb and Zn-contamination in Bama mine near Isfahan city on bacterial diversity, structure and complexity. Basal Respiration (BR) and Substrate Induced Respiration (SIR) was also used to assess microbial activities. Nine samples from three locations (3 for each) with different levels of heavy metal contamination were taken (from low to high), then their DNA were directly extracted. Also a 468 base pair of their 16S rRNA genes were amplified using specific primers, and their fingerprints were obtained by denaturing gradient gel electrophoresis (DGGE). BR and SIR were measured, and metabolic quotient was calculated. Finally, soil microbial activity in polluted conditions was achieved. Results: Our findings illustrate that heavy metal contamination has negative effects on bacterial diversity. By increasing the bioavailability of Pb and Zn, the complexity and diversity of bacterial communities decreased and the frequency of resistant bacteria increased. By increasing Pb and Cd contamination, SIR reduced and this shows the reduction in microbial biomass. In these conditions, SIR and metabolic quotient was more sensitive than BR, so they are better ecological indicators in polluted soils. Discussion and conclusion: Although bacterial diversity showed reduction in polluted soils, diversity is still relatively high. Bacterial ability to adapt in heavy metal contamination, bacterial resistance and their important functional roles in such conditions are valuable in soil ecosystem suggesting further researches on them. [ABSTRACT FROM AUTHOR]

Introduction: Medullary thyroid carcinoma (MTC) accounts for 5- 10% of thyroid malignancies. The orig in of medullary thyroid cancer is para-follicular cells of the thyroid. Several studies showed that relationship between adipokines and cancers. Omentin is an adipokine secreted from adipose tissue. The aim of this study was to investigate the relationship between plasma levels of omentin in patients with medullary thyroid cancer. Materials and Methods: In this case-control study, 90 individuals were studied (45 cases and 45 controls). Omentin plasma levels of these individuals were measured by ELISA method. Results: In general, plasma levels of omnentin in all patients with MTC was significantly higher than the control group (P = 0.002). Omentin plasma levels in both women and men with MTC were ignificantly higher than the corresponding control group (P = 0.0371, and P = 0.0240, respectively). Conclusion: The results of this study showed that plasma levels omentin in patients with MTV were higher than the healthy group. Thus omentin may be considered as a novel and promising biomarker for diagnosis or confirmation of MTC. [ABSTRACT FROM AUTHOR]

Introduction One of the most important sources of the red meat in Iran is the meat produced from sheep. Increasing lamb per ewe considered as a strategy for improving the efficiency for sheep production, although reproduction traits have low heritability. Several genes associated with reproduction were investigated in the recent years. The BMP 15 gene and its paralog GDF 9 and receptor, BMPR-IB, are related to fecundity in sheep and attracted the interest of breeders recently. All these genes that are members of TGFβ super family are functionally closely related together and they affect expression and secretion of hormones affecting follicle growth and ovulation rate in mammals. BMP15 plays a key role in regulating many processes in granulosa cells and ovulation rate. Mutations in some candidate genes such as BMP 15 proved to affect the lambing rate. Since 2008, introgression of the BMP Receptor IB mutant (FecB) from Booroola Merino (from New Zealand) into Afshari sheep was initiated. Thereafter, several genes that proved to have an effect on reproductive traits were studied in this breed. This study was conducted to identify possible polymorphism(s) in BMP 15 and to compare its expression in ovaries of pregnant and non-pregnant ewes. Materials and Methods To study these, blood samples were collected from 35 and 45 Afshari and Afshari × Booroola Merino ewes, respectively. DNA was extracted from all samples using phenol-chloroform procedure and Total RNA was extracted using the RNA extraction kit, CinnaPure RNA Kit (Cinnagen Inc®, Iran), extraction was performed according to the manufacturer’s instruction. To remove any possible residual DNA contamination, RNA samples were treated with 1 unit of DNase (Vivantis Inc®, Malaysia). The specific primers were designed for three areas of BMP 15, namely promotor (581 bp), exon one (325 bp) and exon two (857 bp) and the targets were amplified using PCR. The PCR products were sequenced using forward and reverse primer for all of the samples. Result and discussion There was no difference among sequences of the promoter and the first exon among samples. But, a nucleotide in position 134 of the second exon, C was replaced by A, was observed in two samples with heterozygote genotypes AC instead of CC. Nonetheless, the codon of amino acid encoding proline is remained unchanged. This mutation occurred in two Afshari × Booroola Merino ewes. This mutation, to our knowledge, was not reported to date. Parents of these ewes were not available and also due to the low frequency of the mutation, detection and identification breed of the origin for the mutation was not possible. To date, such a mutation neither was reported in Afshari nor in Booroola Merino breeds. Obviously, the promoter of the gene is conserved and it shows high similarity amongst related species. Nevertheless, in our study sample size was limited to conclude this well. Considering conservation of the promoter of the gene within the species and closely related species, it appears that the regulatory regions were very protected and required for its sustained action. Given that a number of animals used in the study were twine bearing Afshari × Booroola Merino crosses, but there was no difference between them and Afshari pure breed in terms of BMP15 gene expression and gene sequences. Thereupon, the results of this study indicate that this gene plays no role in litter size of this new genetic component. In order to assess the expression of this gene in ovaries of the ewes, after slaughtering, ovary samples of 22 pregnant ewes and those of 8 none pregnant were collected. Total RNA was extracted from the samples and mRNA converted to cDNA using oligo d (T) primer and reverse transcriptase. Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) was used as the endogenous control for normalization. Results of real time PCR using designed specific primers showed no difference in BMP 15 gene expression between pregnant and non-pregnant ewes. Conclusion It is possible that this gene plays its role in relation to other genes include their receptors and its expression is needed in different steps of reproduction. This result and reports of other studies suggest that more data on BMP15 gene with a simultaneous expression of other genes in the ovary is needed to clarify the integral role of BMP 15 in reproduction.

Background and Aim: Thalassemia is considered as the most important monogenic disorders around the world. So far, about 60 mutations of this type have been reported in Iranian patients. Comparison between different provinces of the country reveals that the dispersion of the mutations is significantly various with respect to their types and frequencies. The current study aimed at assessing prevalent molecular mutations in β Thalassemia carriers in Birjand and Amirabad at the suburb of the city. Materials and Methods: In this descriptive in-vitro study, 34 carriers (15 females and 19 males), who had been identified through marriage screening tests at the health centers in the South Khorasan province were assessed. After obtaining written informed consent of the subjects and completing the respective questionnaires, 2 cc of intravenous blood sample from each subject were collected into EDTA tubes. Salting out and Arms-PCR methods were used for DNA extraction and mutation detection, respectively. Finally, the obtained data was analysed by means of SPSS software (V;19) using Fisher’s test at the significant level of P

Background: Early detection and suitable drug admission are the first attempts to control of TB and MDR-TB. In this study, PCR-RFLP method was used for rapid detection of Mycobacterium tuberculosis and resistance to isoniazid. Materials and Methods: In the present study, 87clinical isolates of MTB were used in a PCR-RFLP method. The RFLP-PCR assay by specific primer was used for two purposes In the first, PCR of katG gene for diagnosis of bacteria and in the second step, PCR product by endonuclease enzyme in a RFLP reaction produced a pattern in electrophoresis for mutation detection in katG315. Sequencing method was used for confirmation of RFLP results. Results: The study showed that all studied strains produced band 620bp that confirmed MTB. From the 46 resistant strains to isoniazid, the RFLP-PCR method showed that 44 strains had mutations in the katG gene Ser315Thr. The 41 susceptible strains had not any mutation at the codon. Results of sequencing were confirmed results of the molecular method. Sensitivity of RFLP-PCR test was 95.6% (95% CI :0.85-0 .98) and specificity was 100% (95% CI :0.91-1 .0). Conclusion: RFLP-PCR test is a good tool for simultaneous diagnosis and detection of katG315 mutation in clinical isolates of MTB.

Single-strand specific (sss) nucleases (S1/P1 family) are widely used in molecular genetics studies. The most widely used sss-nucleases are those plant endonucleases with the ability of specific cleavage of mismatch loops in heteroduplex DNA molecules. Because of the valuable activity of these endonucleases, enzymatic cleavage of heteroduplex DNA molecules is one of the most common techniques of point mutation detection in genomes. In this study, gene sequences encoding single-strand specific endonucleases were isolated from parsley (Petroselinum crispum L.) using heterologous primers in PCR reactions. The isolated gene sequences, after cloning and molecular identification, were designated as pars I and pars II. Physicochemical, structural and phylogenetic analysis using software tools, verified PARS I and PARS II as two members of S1/P1 family with a substantial homology to heteroduplex DNA specific endonucleases. However, it is essential to produce pure enzyme preparations as native or recombinant forms to determine mechanisms of activity and substrate preferences of these nucleases on heteroduplex DNA substrates.

Background: Thyroid carcinoma is the most common endocrine malignancy and approximately accounts 2% of all cancer cases. Medullary thyroid cancer (MTC) is an endocrine tumor with differentiation of Parafollicular or C-cells and is categorized into hereditary or sporadic types. Medullary thyroid carcinoma approximately accounts for 5-10% of all thyroid carcinoma. Germ-line and somatic mutations in exons 10 and 11 RET (Rearranged during Transfection) proto-oncogene are responsible for the occurrence of the familial and sporadic types, respectively. Calcitonin is a key marker in MTC diagnose and has been demonstrated to be highly sensitive for differential diagnosis prognostic assessment, follow-up and evaluation of MTC treatment. The aim of this study was to investigate the relationship between plasma levels of calcitonin in MTC patients with or without RET mutation. Methods: In this cross-sectional study, the population consist of MTC patients who have referred to the endocrine and metabolism research center of Shahid Beheshti University of medical sciences since October 2013 till October 2016. Genomic DNA was extracted from peripheral blood leucocytes using the standard salting out/proteinase K method. Nucleotide change detection in exons 10 and 11 was performed using polymerase chain reaction (PCR) and direct DNA sequencing methods. Participants were then divided into two groups with or without mutation (43 individuals in each group). Plasma calcitonin levels were determined by enzyme-linked immunosorbent assay (ELISA) method in both groups. Results: Evaluation of the level of plasma calcitonin in 43 patients with a molecular mutation in RET proto-oncogene (mean age 31 years) and 43 patients without molecular mutations in RET proto-oncogene (mean age 43 years) were 7.6 pmol/mL and 3.07 pmol/mL respectively. This difference is statistically significant (P=0.0014). Conclusion: Routine measurement of calcitonin has been investigated as a screening method for the diagnosis of medullary thyroid carcinoma patients. Nevertheless, additional data are required to definitely support routine measurement of calcitonin due to the role of RET proto-oncogene. [ABSTRACT FROM AUTHOR]

Introduction: Mucopolysaccharidosis I (MPS-I) is an autosomal recessive lysosomal storage diseases, caused by α-L-iduronidase (IDUA) enzyme deficiency. The clinical manifestations of MPS-I patients are variable ranging from severe to mild, and therefore prediction of disease severity is difficult. From when IDUA gene has been cloned more than 109 distinct mutations have been identified in it and this number is increasing. This mutation analysis has provided some molecular explanations for the range of MPS-I phenotypes. The aim of this study was identification and molecular characterization of IDUA gene mutations in our subset of MPS I patients. Methods: The present study performed on 5 Iranian families, each with a suspected child for MPS-I. Initially by using enzyme activity assay, the Hurler syndrome was verified and then presence of L123R mutation was evaluated by PCR-SSCP. Finally by PCR amplification of all 14 exons of the gene, SSCP and sequencing the mutations underlying the disease were identified and characterized. Results: The detected mutations turned to be L123R (in 2 patients), W402X, P533R and G51D mutations in other 3 patients. Discussion: L123R mutation, which was reported for the first time from our centre, was also present in 2 of the patients of this study but other 3 mutations were not novel. From our results, as well others, it can be concluded that the range of mutations in IDUA gene differ in different geographical areas. This should be considered when designing mutation detection strategies for MPS-I.

Background: Thyroid cancer is the most common endocrine malignancy. Accounting for approximately 1-2% of all cancers. Thyroid cancers have been divided into four main types: papillary, follicular, medullary and anaplastic. The active form of vitamin D (1,25- (OH) 2-vitamin D3) by binding to its receptor, using genomic and non-genomic mechanisms inhibits the proliferative effect of TSH on thyroid cells. Therefore, vitamin D may have a role in regulating of thyroid gland cell proliferation. Many studies have shown anti-cancer effects of vitamin D in cancers. Polymorphisms of Vitamin D receptor can influence the prevalence to various cancers. In the present study, serum level of vitamin D and FokI, BsmI and Tru9I polymorphism of vitamin D receptor was investigated. Methods: This case-control study was performed in the summer of 2015 in Endocrinology and Metabolism Center of Shahid Beheshti University of Medical Sciences, Tehran, Iran. Forty medullary thyroid cancer patients and 40 healthy controls were investigated. Genomic DNA of subjects was extracted with saturated salt/proteinase K and polymorphisms of vitamin D receptor gene investigated by polymerase chain reaction-sequencing. Serum level of vitamin D evaluated by ELISA technique. The results were analyzed by SPSS, ver. 20 (Chicago, IL, USA) and GraphPad Prism, ver. 5 (GraphPad, Inc., CA, USA) softwares. Results: Genotypic and allelic abundance of FokI and BsmI polymorphisms between test and control groups have not shown significant different. In Tru9I polymorphism, Tt genotype abundance in test group were 45 percent and in control group were 17.5 percent and t allelic abundance in test group were 25 percent and in control group were 8.7 percent which this different were significant. Average serum level of vitamin D in test group was 23.32 ng/ml and in control group was 18.95 ng/ml which was statistically significant. Conclusion: Unexpectedly, serum levels of vitamin D in test group were higher than control group. Tru9I polymorphism is significantly correlated to medullary thyroid carcinoma prevalence. [ABSTRACT FROM AUTHOR]

Background and aims: HCM is a common form of hereditary heart disease with Mendelian inheritance that is a frequent cause of sudden cardiac death in the people younger than 35 years. In more than 50% of cases the cause of HCM identified as gene mutations. Mutations in the gene MYBPC3 (which encodes the cardiac Myosin binding protein C) are about 40% of clinical cases. This study was aimed to investigate the presence of mutations in exons 30 and 33 MYBPC3 gene in patients with hypertrophic cardiomyopathy in Chaharmahal and Bakhtiari. Methods: 30 probands of hypertrophic cardiomyopathy were selected from patients referred to the cardiac clinic of the Shahrekord Medical University. DNA extraction was done by using of standard phenol - chloroform protocol from blood samples of patients. The exons 30 and 33 were amplified by PCR and were converted to single-stranded with SSCP and they electrophoresed with double-stranded sample on polyacrylamide gels. Results: All of selected patients had hypertrophic cardiomyopathy with genetic cause. Qualities of extracted DNA were certified by electrophoresis and determination of their absorption ratio. By investigation of obtained results from SSCP/HA electrophorese in polyacrylamide, we did not find any change in exons 30 and 33. Conclusion: None of the patients had mutations in exons 30 and 33 gene MYBPC3. Based on the results of this study, there is no change in exons 30 and 33 of MYBPC3 in hypertrophic cardiomyopathy patients with autosomal dominant heritage. As the result, the mentioned exons do not consider as suspicious exons for prognosis and cause of HCM in Chaharmahal and Bakhtiari province. [ABSTRACT FROM AUTHOR]

Background and aims: Hypertrophy cardiomyopathy (HCM) is the most common type of heart disease with monogenic inheritance distinguished by thickening of left ventricle, contractile dysfunction and potentially fatal arrhythmias. Great progress in clarifying the genetic basis of HCM obtained. It was identified more than 900 unique mutations in 20 genes. Mutations in the gene MYBPC3 (which encodes the cardiac Myosin binding protein C) are about 40% of clinical cases. This study aimed to investigate the presence of mutations in exons 27 and 29 of MYBPC3 gene in patients with hypertrophy cardiomyopathy in Chahar Mahal and Bakhtiari province. Methods: 30 probands of hypertrophy cardiomyopathy were selected from patients referred to cardiac clinic of the Shahrekord Medical University. To extract DNA from blood samples of patients we used standard phenol - chloroform protocol. The exons 27 and 29 were amplified by Polymerase Chain Reaction and were converted to single-stranded with Single Strand Conformation Polymorphism and they electrophoresed with double-stranded samples on polyacrylamide gels. Results: Extracted DNA was electrophoresed and the attraction ratio was examined and the outcomes indicate that the DNA extraction and quality of samples are respectable. By investigation of obtained results from SSCP/HA electrophorese in Polyacrylamide, no change was found in exons 27 and 29 of MYBPC3. Conclusion: Any of the patients had no mutation in exons 27 and 29 gene MYBPC3. Based on the results of this study there is no mutation in exons 27 and 29 gene MYBPC3 of HCM patients with autosomal dominant inheritance. It concluded that exons 27 and 29 of MYBPC3 do not have any role to cause HCM in examined patients. [ABSTRACT FROM AUTHOR]

Objective: Neuromuscular disordres are a gorup of heterogenous inherited diseases. More than 150 types of these group of disorders have been known.The criticals that were used for classification of these disease include: age of onset, clinical course, disease progression and type of inheritance. Neuromuscular disorders have been classified in four categories: Myopaties (Muscular dystrophies) Neuropathy (Charcot-Marie-Tooth dieseas), Motor neuron junction (Myasthenia Syndrome) Motor neuron disorder (Spinal muscular atrophy). Objective of this study was classification of neuromuscular disorders based on clinical criteria, molecular and immunohistochemical analysis that reffered to Genetic Research Center. As a result of neuromuscular disorders was the second most common disabilities, more investigation in this group of disorders is necessary in Iranian population so this study was performed for the first time in Iranian patients. Materials & Methods: Total of 143 patient with neuromuscular deficiency that clinical, EMG and muscle enzymes have been tested and also in some cases molecular and immunohistochemical analysis was performed. Results: 82 patients with myotonic dystrophy, 19 patients with DMD/BMD, 6 patients with congenital muscular dystrophy, 3 patients with FSHD, 10 patients with SMA, 2 patients with CMS and 21 patients with LGMD. For the other cases we need more molecular and definite clinical analysis. Conclusion: For some cases that DM1 was not confirmed, study of DM2 is recommended. In patients with LGMD that analysis with 5 Abs were normal we advised multiple western blot for them (including calpain, sarcoglycans, dysferlin). Study of congenital muscular dystrophy besides merosin analysis, investigation for the other involved protein and molecular testing for mutation detection is considrable.

Background and Objective: HIV-1 and HCV infections especially in co-infected forms are among the most important infections transferred during blood transfusion.The screening of the blood products is valuable for preventing the transmission of infections. The aim of this study was to evaluate multiplex RT-PCR assay for detection of Co-infection HIV-1 and HCV Viruses in plasma samples. Materials and Methods: This laboratory study was done to evaluate the use of multiplex RTPCR assay for simultaneous detection of HIV-1 and HCV genomes in plasma samples. The amplified genomes were detectable in 3% agarose gel base on difference in the numbers of nucleotides. The sensitivity and specificity of this assay was determined on healthy and infected subjects whome simultanously exhibit HIV-1 and HCV co-infection using plasma samples. Results: The specificity results showed that the primers used in this assay have no interaction with each other and other possible interfering agents. The clinical sensitivity and specificity of the assay has been considered as 90% and 100%, respectively. Conclusion: Multiplex RT-PCR can be used for screening of blood donors due to high sensivity and specificity. [ABSTRACT FROM AUTHOR]

Background: JAK2 is a nonreceptor tyrosine kinase that plays a major role in myeloid disorders. This mutation is characterized by a G to T transverse at nucleotide 1849 in exon 12 of the JAK2 gene, located on the chromosome 9p, leading to a substitution of valine to phenylalanine at amino acid position 617 in the JAK2 protein. In this study we compared the amplification refractory mutation (ARMS) assay and allele- specific (ASPCR) to evaluate JAK2V617F mutation patients with non-CML myeloproliferative neoplasms (MPNS). Methods: In this experimental study we evaluated JAK2 mutation in 58 patients with a known or suspected diagnosis of a myeloproliferative neoplasm by simple randomized sampling. The mutation was detected by ARMS-PCR and AS-PCR in patients. In order to verify the methods, amplified products from some patients were sequenced. Results: The JAK2 V617F mutation was detected in 86.6%(26/30) of patients with polycythemia vera and 61.5%(8/13) of patients with idiopathic myelofibrosis by ARMSPCR and AS-PCR. 46.6%(7.15) of essential thrombocythemia patients were positive using ARMS- PCR method while 53%(8.15) of then were positive when AS- PCR were used. The mutation was confirmed by sequencing. Conclusions: The incidence of JAK2 mutation using above PCR methods is similar to previous studies. The different results may depend on the molecular technique used. [ABSTRACT FROM AUTHOR]

Background: Cystic fibrosis is a monogenic recessive disorder founds predominantly in caucasian population causes exocrine glands function defect. This disease arises from mutations in the cystic fibrosis transmembrane conductance regulator (CFTR) gene. Because of heterogeneity of the mutations in CFTR gene, phenotypic symptoms in this disease are very variable. In this study we consider poly T polymorphism (T5, T7, T9) in the intron 8 of CFTR gene in normal individuals and cystic fibrosis patients in mazandaran province. Methods: Forty cases of cystic fibrosis patients and 40 normal individuals were screened for poly T polymorphism in intron 8 of CFTR gene using Reverse Dot Blot method. Results: T7 allele is the most prevalent in normal individuals and CF patients and it's abundance is approximately 75%. T9 and T5 represent approximately 20% and 5% of normal or mutant alleles respectively. T7/T7 genotypes in normal individuals and CF patients are the most prevalent with 72.5% and 60% prevalence rate, respectively. T5/T9 and T5/T5 genotypes were not found. 22.5% of normal individuals and 30% of CF patients had heterozygote genotypes. Conclusion: The abundance of T5, T7, T9 alleles and the presence of 22.5-30% heterozygote genotypes in normal individuals and CF patients indicates that poly T polymorphism in intron 8 of CFTR gene can be used as a marker for detection of normal and mutant alleles in prenatal diagnosis or can be used in carrier assessment in families with previous history of the disease. [ABSTRACT FROM AUTHOR]