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Background and Aim: Alzheimerchr('39')s Disease (AD) is a neurodegenerative brain disease that gradually destroys memory and cognitive skills. The disease is caused by the formation of beta-amyloid plaques, oxidative stress, dysfunctions in the cholinergic system, neuronal killing inflammation, and ultimately brain atrophy. Donepezil and hyoscyamoside have inhibitory effects on these pathogens; therefore, their impact on the learning process of Alzheimer’s rats in the Morris Water Maze was investigated. Methods & Materials: In the present experimental study, 60 male rats of Wistar breed with approximately 7 weeks age within the control group (rats that received normal water and food), the PBS group (underwent surgery), PBS group (received solvent Aβ), the first Alzheimer›s group (animals that received beta-amyloid by Alzheimer’s surgery, second Alzheimer’s group (after Alzheimer’s surgery, they received 1 cc of normal saline daily, and treatment groups that treated the rats with beta-amyloid after Alzheimer. In the hyoscyamoside group, they received 10 mg/kg daily of hyoscyamoside for 28 days. The donepezil group received it 4 mg/kg daily for 28 days by gavage. The Morris Water Maze test was used to evaluate learning and memory. Data were analyzed by ANOVA statistical analysis and Post Hoc test. Ethical Considerations: The Ethics Committee in Biomedical Research, Islamic Azad University, Science and Research Branch approved the research (Code: IR.IAU.SRB.REC 1397.057) Results: Beta-amyloid injection caused extensive damage to memory. The treatment groups with hyoscyamoside and donepezil spent less time and distance with a significant level (P

Background: Multiple sclerosis (MS) is a common disease of the central nervous system, and the incidence of this disease in women is twice that of men. Considering the importance of miRNA polymorphisms in the expression and function of mRNAs as well as the risk of disease, this study investigated the association of rs6977848 polymorphism (a genetic variant in 19-nucleotide downstream of miR-148a coding gene) with the risk of MS disease in Isfahan City population in Iran, for the first time. Methods: In this case-control study, a population of 95 healthy individuals and 99 patients were assayed. The genotype of the individuals for the polymorphism was determined using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). To confirm the results, a number of samples were sequenced. Then, the frequency of genotypes and alleles were evaluated. Findings: Considering the total dominance of T allele, the total TT and TG genotypes significantly increased the susceptibility to MS disease compared with the GG reference genotype [odds ratio (OR) = 2.27, P = 0.043); while considering the genotype GG as reference genotype, there was a significant difference between the TG and the reference genotypes in this population with a risk of disease (P < 0.050). However, no significant difference was observed between genotype TT and GG genotype as reference genotype. Conclusion: Findings indicate that rs6977848 polymorphism in miR-148a coding gene downstream may be effective in influencing the performance of this miRNA as well as its expression level. Moreover, the incidence of the ability of individuals to affect this disease is not significantly related to the incidence of disease in the studied populations.

Background: Tumor-targeting peptides are attracting subjects in cancer therapy. These peptides, which are widely studied, deliver therapeutic agents to the specific sites of tumors. In this study, we produced a new form of recombinant listeriolysin O (LLO) with genetically fused Anti-HER2/neu peptide (AHNP) sequence adding to its C-terminal end. The aim of the study was to engineer this pore-forming toxin to make it much more specific to tumor cells. Materials and Method and Results: Two forms of the toxin (with and without peptide) were subcloned into a bacterial expression plasmid. Subcloning was performed using a polymerase chain reaction (PCR) product as a megaprimer in a quick-change PCR to introduce the whole insert gene into the expression plasmid. After expression of two recombinant forms of LLO in BL21 DE3 cells, purification was performed using Ni-NTA affinity column. MDA-MB-231 and MCF-7 cell lines (as negative and positive controls, respectively) were treated with both LLO toxins to evaluate their cytotoxicity and specificity. The IC50 of LLO on MDA-MB-231 and MCF-7 cells was 21 and 5 ng/ml, respectively. In addition, IC50 for the fusion AHNP-LLO toxin was 140 and 60 ng/ml, respectively. It was found that the cytotoxicity of the new engineered AHNP-LLO toxin has decreased by about 9x compared to the wild-type toxin and the specificity of the AHNP-LLO toxin has been also reduced. Conclusions: Results show that the C-terminal of the LLO should not be modified and it seems that N-terminal of the toxin should be preferred for engineering and adding peptide modules.

Neural retina is the part of the diencephalon and because of the relatively simple structure in known as a suitable model for studying the molecular mechanisms of the central nervous system. Visual perception is the result of the function of six types of neurons organized in the structure of the neural retina. Neural retina develops via the proliferation of a common precursor cell in the inner layer of the optic cup. Retinal progenitor cell acquires the competent to differentiate into different cell fates by different factors in a time-dependent and protected manner in the mammals. Destruction and loss of these cells in the retina occurs in various retinal diseases and impairs the process of human vision. Lack of reconstruction of damaged nerve cells in the retina of mammals, including humans is a noted problem; and in recent decades, a wide range of research in the eye field allocated the possibility of replacing the retinal cells and many efforts is made to treat these diseases. Study and identifying the transcription factors involved in neuronal differentiation can provide a useful tool for gene therapy aiming to regenerate retinal neurons in the near future.

تلومرها، انتهای فیزیکی کروموزوم‌های خطی را تشکیل می‌دهند و عملکرد آن‌ها به منظور نگهداری پایداری کروموزومی ضروری می‌باشد. در صورت از دست رفتن عملکرد تلومرها، ظرفیت همانندسازی سلولی کاهش می‌یابد و در نهایت، منجر به پیری سلولی و بیماری‌های مرتبط با نقص در احیای اندام و بازسازی بافت (سندرم‌های تلومر) می‌گردد. از سوی دیگر، سرطان‌ها نیز می‌توانند در نتیجه‌ی ناپایداری‌های ژنومی ناشی از اختلال در عملکرد تلومر ایجاد شوند. تاکنون دو نوع مکانیسم‌های نگهداری طول تلومر (TMM یا Telomere maintenance mechanism) شناسایی شده‌اند که شامل تلومراز و ALT (Alternative lengthening of telomeres) می‌باشد. در نئوپلازی‌ها، مکانیسم‌های نگهداری طول تلومر (TMM) می‌توانند به پیش‌بینی بیماری کمک نمایند و احتمال می‌رود در آینده باعث درمان مستقیم شوند. در طی مطالعات اخیر، مشخص شد که تلومرهای یوکاریوتی رونویسی می‌شوند و به رونوشت آن‌ها TERRA (Telomeric repeat-containing RNA) می‌گویند. به طور کلی، افزایش میزان رونویسی از TERRA با کوتاه شدن طول تلومر مرتبط می‌باشد و در عملکرد تلومر تأثیرگذار است. در این مقاله، به طور خلاصه اصول کلی بیماری‌های مرتبط با اختلال در عملکرد تلومر بررسی شده ‌است. به علاوه، در مورد ارتباطات شناخته شده بین رونوشت تلومر (TERRA) و بیماری‌ها بحث شده است. در نهایت، پتانسیل TERRA در رویکرد‌های درمانی بیماری‌های مرتبط با نقص در عملکرد تلومر مورد بررسی قرار گرفته است.

مقدمه: انسولین هورمون پروتئینی است که توسط سلول‌های بتای پانکراس ترشح می‌شود. به علت معایب تولید پروتئین‌های نوترکیب در میکروارگانیسم‌ها، هزینه به نسبت بالا، امکان آلودگی با پروتئین‌های سمی و مراحل هزینه‌بر خالص‌سازی، تولید پروتئین‌های نوترکیب در گیاهان امری قابل بررسی است. توسعه‌ی سیستم بیان گذرا بر پایه‌ی نوع حذف شده‌ی RNA-2 ویروس موزائیک لوبیا چشم بلبلی (Cowpea Mosaic Virus-Hyper Translatable یا CPMV-HT)، امکان تولید سریع و سطوح بالای پروتئین‌ها را بدون استفاده از همانندسازی ویروسی فراهم کرده است. روش‌ها: در این مطالعه، سازه‌های pBI121-Proinsulin-Zera (pBI-ProZ) در بر دارنده‌ی توالی ژن پروانسولین انسانی و Zera (دومین انتهای N غنی از پرولین گاما- زئین ذرت) و pCAMBIA1304-Proinsulin-Extensin (pCAMBIA-ProE) در بر دارنده‌ی سیستم بیانی CPMV-HT، به منظور بهبود ترجمه‌ی ژن پروانسولین انسانی و توالی سیگنال پپتید اکستنسین هویج ساخته شد. این دو سازه، به واسطه‌ی باکتری Agrobacterium tumefaciens pv. C58، به صورت بیان گذرا به گیاهان کاهو و یونجه انتقال داده شدند. تحلیل آماری این پژوهش بر پایه‌ی آزمایش فاکتوریل، در قالب طرح به‌کلی تصادفی بر روی غلظت پروانسولین تولیدی در هر گرم برگ تراریخت صورت گرفت. بیان ژن پروانسولین در بافت گیاهی تراریخت در سطح رونویسی، با استفاده از واکنش Reverse transcription polymerase chain reaction (RT-PCR) و در سطح ترجمه، با استفاده از آزمون لکه‌گذاری نقطه‌ای و Enzyme-linked immunosorbent assay (ELISA) مورد تأیید قرار گرفت. یافته‌ها: میزان پروانسولین فعال تولیدی با سازه‌های pCAMBIA-ProE و pBI-ProZ، در برگ‌های یونجه تراریخت، به ترتیب 82/6 و 32/4 نانوگرم در هر گرم برگ تر و در برگ‌های کاهوی تراریخت، به ترتیب 6/6 و 8/3 نانوگرم در هر گرم برگ تر بود. نتیجه‌گیری: طبق نتایج به دست آمده از این تحقیق، میزان بیان پروتئین پروانسولین در سازه‌ی pCAMBIA-ProE در بر دارنده‌ی خصوصیات سیستم بیانی CPMV-HT، بیشتر از سازه‌ی pBI-ProZ بود.

Background: Patients affected by gliomas have a poor prognosis. Astrocytoma is a subtype of glioma. Identification of biomarkers could be an effective way to an early diagnosis of tumor or to distinguish more aggressive tumors that need more intensive therapy. In this study, we investigated whether the expression of miR-362 was increased or decreased in patients with different grades of astrocytoma. Materials and Methods: miR-362 expression was compared in 25 patients with astrocytoma with that of 4 normal nonneoplastic brain tissues. Results: In all tumor tissues, the expression of miR-362 was significantly decreased relative to its expression in normal brain tissues. However, there was no significant difference between miR-362 expressions in high and low grades of astrocytoma. Conclusions: In conclusion, miR-362 showed a down-regulation pattern in astrocytoma tissues that was different from the pattern obtained from previously published microarray studies.

Background: Glioblastoma (GBM) is the most common and aggressive brain tumor, which has a poor prognosis despite the advent of different therapeutic strategies. There are numerous molecular biomarkers to contribute diagnosis, prognosis, and prediction of response to the current therapy in GBM. One of the most important markers that are potentially valuable is immortalization-specific or immortalization-associated marker named “hTERT messenger ribonucleic acid (mRNA)” the key subunit of telomerase enzyme, which is expressed in more than 85% of cancer cells, in spite of the majority of normal somatic cells. In this study, we investigated the effects of resveratrol (RSV) on this mRNA marker level, leading to cancer progression. Materials and Methods: U-87MG cell line was obtained from Pasteur Institute of Iran and treated with various concentrations of 0– 160 μg/mL of RSV and at different time points (24, 48, and 72 h). To evaluate viability of U-87MG cells, standard 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay was performed. Real-time polymerase chain reaction (RT-PCR) was used for comparative and quantitative assessment of human telomerase reverse transcriptase (hTERT) mRNA copy number versus control– untreated group. Results: The results of our investigation suggested that RSV effectively inhibited cell growth and caused cell death in dose-dependent (P < 0.05) and not in time-dependent manner (P > 0.05), in vitro. Interestingly, quantitative RT-PCR analysis demonstrated that at half inhibition concentration, RSV dramatically decreased mRNA expression of hTERT, the catalytic subunit of telomerase enzyme, which leads to prevention of cell division and tumor progression. Conclusion: With regard to downregulation of this immortalization-associated marker, RSV may potentially be used as a therapeutic agent against GBM.

miRNAs(microRNAs) were discovered in 1993 in Caenorhabditis elegans as one of the most important gene expression regulatory factors. This regulatory role includes many of the important intracellular processes like genesis, differentiation, proliferation and apoptosis. miRNAs are among mRNAs and like them are transcribed by RNA polymerase II and then cap and polyA tail are added to them. The resulted pri-miRNA converts to mature miRNA by two sequential trimming reactions. miRNAs exerts their regulatory effects by translation repression, translation activation and mRNA degradation. miRNAshave an important role in regulation of immune and nervous systems and disorder in their expression can lead to separate disease in both systems.

Background: Cystinuria, one of the first diagnosed inborn errors of metabolism, recognized by hyperexcretion of cystine, lysine, ornithine and arginine into the urine. So far, two genes associated with cystinuria have been identified: SLC3A1 (2p16.3) that encodes the heavy subunit rBAT of the renal b0,+ transporter and SLC7A9 (19q13.1), which encodes the light subunit b0,+AT. Patients with type A cystinuria have two SLC3A1 mutations, whereas patients with type B have two SLC7A9 mutations and finally patients with type AB have one mutation in each gene. Considering the population-specific distribution of mutations in disease, limited studies on the genetic bases of the cystinuria have been done in Middle East. This research presents the results of mutation analysis on patients with cystinuria in Iran. Methods: Thirty unrelated patients with cystinuria operated to remove kidney stones were screened by urologist .The patients were analyzed for mutation using amplification refractory mutation system (ARMS) and polymerase chain reaction-Restriction fragment length polymorphism-polymerase chain reaction (RFLP-PCR) methods. Findings: We found some variations including missense mutations, polymorphism and intron variant, but the most frequent mutations, M467T and also T216M and R333W, were not detected in our patients. Conclusion: Our research may confirm the ethnic distribution of mutations in cystinuria and this study can expand our concept of the genetic basis of cystinuria in Iranian patients.

Telomere is a special structure protects the ends of human chromosomes to degrade. Telomere becomes shorter after each cycle of cell division. However, telomerase is an enzyme activated in cancer cells and germ lines rebuild telomere. However, apparently there is no correlation between telomere length and telomerase activity. The enzymatic activity of telomerase correlates with tumor malignancy. In general, telomerase activity and histological grade are too closely correlated and the results of studies indicate that telomerase activity may be an important malignancy marker in brain tumors. As a conclusion, we can say that activity of telomerase should be considered as a prognostic biological marker of response to treatment.

چندین دهه است که موضوع آنالوگ‌های اسید‌های نوکلئیک توجه برخی دانشمندان علم بیولوژی را به خود معطوف نموده است. برخی از این مولکول‌های صناعی به عنوان ابزارهای قدرتمندی در بیولوژی مولکولی و بیوتکنولوژی، مورد توجه ویژه قرار گرفته‌اند. چهار دسته‌ی عمده‌ی این آنالوگ‌های صناعی عبارت از زنونوکلئیک اسید (XNA یا Xeno nucleic acid)، مورفولینو (Morpholino) و اسید نوکلئیک قفل شده (LNA یا Locked nucleic acid) و نوکلئیک اسید پپتیدی (PNA یاPeptide nucleic acid) که دسته‌ی چهارم این آنالوگ‌ها بود و در این میان، جایگاه برجسته‌ای دارد. PNA، پلیمری است از واحدهای ان-آمینو اتیل گلایسین که به هر یک از این واحدها بازهای پورین/ پیریمیدین اتصال یافته است. این پلیمر، خصوصیات پپتیدها و اسیدهای نوکلئیک را توأمان دارد و قادر است به طور اختصاصی به نوکلئیک اسیدهای طبیعی یا PNA‌های دیگر، هیبرید شود که این ویژگی و دیگر خصوصیات ویژه‌ی این مولکول باعث به کارگیری آن در بسیاری از روش‌های بیولوژی مولکولی در زمینه‌ی تشخیص و درمان بیماری‌ها شده است. در این مقاله، ساختار، خصوصیات بیوشیمیایی و کاربردهای زیست پزشکی PNA شرح داده می‌شود.

سرطان یک اختلال چند عاملی می‌باشد که بررسی آن‌ها نیازمند انجام مطالعات همه جانبه بر روی سلول‌ها، DNA، RNA و پروتئین‌های تومور می‌باشد. این نوع از مطالعات به نام Omics می‌باشند که شامل ژنومیکس و پروتئومیکس هستند. بانک تومور یک واژه است که به طور معمول برای جمع‌آوری، ذخیره‌سازی و استفاده از بافت تازه در مقاصد تحقیقات زیست پزشکی سرطان استفاده می‌شود. اکثر بانک‌های تومور به جمع‌آوری نمونه‌هایی از بافت تومور می‌پردازند که نیازی به آن برای تشخیص‌های پاتولوژیک بعد از جراحی و برداشتن تومور نیست. این بافت اغلب به منظور یخ بستن به سرعت در نیتروژن مایع قرار داده می‌شود یا این که ممکن است در محلول‌های ثابت‌کننده نگهداری شود. اکثر مراکز سرطان دارای بانک توموری هستند که ذخیره‌سازی نمونه‌های توموری از بیمار و بافت سالم مجاور آن را انجام می‌دهند. تغییر در روش‌های تشخیص و شناسایی بیومارکرها نیازمند دسترسی به تعداد کافی از نمونه‌ها دارد و فراهم نمودن یک بانک از تومور‌ها یک قدم ضروری در این مورد است. سالیانه، ده‌ها و شاید صدها عمل جراحی در کشور ما انجام می‌شود که گنج بزرگی از سلول‌های سرطانی است و می‌توان با جمع‌آوری آن‌ها یک بانک تومور ایجاد نمود.

Telomeres are special structures at the ends of chromosomes, especially in eukaryotes, that include repetitive sequences of deoxyribonucleic acid (DNA) and non-nucleosomal proteins. These structures are essential to protect chromosomes from recombination and degradation. Telomeres are gene-poor repetitive sequences, but they are close to the more gene-rich subtelomeric regions. Proteins associated with telomeres are able to interact directly with the TTAGGG repeats and also can bind to other factors to form large protein complexes. These proteins are involved in DNA repair and telomere stability. Therefore, defects in the synthesis of these proteins could induce imperfections in telomere maintenance and DNA repair; and accordingly, should cause a number of genetic syndromes. Keywords: Telomere, DNA damage, Genetic diseases

Background: Among patients with diabetic polyneuropathy, the status of folic acid, homocysteine, and nerve conduction studies (NCS) variations has been associated with methylenetetrahydrofolate reductase (MTHFR) gene polymorphisms. The objective of the present study is to assess B9 vitamin supplementation associated with MTHRF C677T polymorphism can be effective on NCS variations in patients. Materials and Methods: This study is a randomized, double-blind, placebo-controlled study. Patients were randomly allocated to either intervention (1 mg of folic acid, n = 40) or placebo (n = 40) groups based on parallel group design. Blood samples were taken to determine the serum levels of folic acid and homocysteine. The NCS data were collected for the assessment of diabetic neuropathy. Genotyping was performed for C677T polymorphism of the MTHFR gene. Results: Four months after intervention, patients significantly observed change of serum folic acid and homocysteine levels based on C677T genotypes in the MTHFR gene. The amplitude of sensory peroneal nerve between intervention and placebo groups with CC genotype was significantly different (2.8 ± 1.6 vs. 1.9 ± 1.1). However, peak latency and amplitude of sensory sural nerve between CC (3.8 ± 1.8 vs. 4.0 ± 1.5 for peak latency and 3.5 ± 1.0 vs. 2.5 ± 1.0 for amplitude; and CT + TT genotypes (3.7 ± 1.7 vs. 3.9 ± 1.3 for peak latency and 3.2 ± 1.0 vs. 2.3 ± 1.1 for amplitude) were significant. Furthermore, significant difference for variables of motor tibial nerve and motor peroneal nerve amplitude was observed in different groups of MTHFR C677T genotypes (5.4 ± 2.9 vs. 4.6 ± 3.2 for onset-latency of tibial nerve between CC genotype; 4.8 ± 2.8 vs. 4.6 ± 3.2 for onset-latency of tibial nerve between CT + TT genotype; 0.6 ± 0.2 vs. 0.3 ± 0.1 for amplitude of tibial nerve between CC genotype; 0.5 ± 0.3 vs. 0.3 ± 0.2 for amplitude of tibial nerve between CT + TT genotype; 26.0 ± 13.3 vs. 23.2 ± 13.4 for velocity of tibial nerve between CC genotype; 26.0 ± 13.7 vs. 23.1 ± 9.6 for velocity of tibial nerve between CT + TT genotype; 1.6 ± 1.0 vs. 0.9 ± 0.7 for amplitude of peroneal nerve between CC genotype; 1.4 ± 0.7 vs. 0.9 ± 0.5 for amplitude of peroneal nerve between CT + TT genotype). Conclusion: The study determined that MTHFR C677T polymorphism effects the efficacy of folic acid supplementation on serum folic acid, homocysteine levels and some NCS parameters in diabetic polyneuropathy patients.

Background: Cardiac arrhythmias are responsible for several cases of syncope and sudden cardiac death annually worldwide. Due to overlapping clinical symptoms in some cardiac arrhythmias genetic studies would help to confirm the primary clinical diagnosis made on the basis of solely clinical findings. In addition clinical management of the patient, family screening and provide appropriate counseling and risk assessment for the family members are other advantages of genetic study. Materials and Methods: Totally nine patients from a family included in this study. The primary diagnosis on the basis of clinical findings was second-degree atrioventricular (AV) block for this family. Mutation in SCN5A gene is frequently reported for second-degree AV block and hence the gene was analyzed using whole gene sequencing but no mutation was detected. Subsequently, the samples were subjected to customized Ampliseq 77 gene panel using next generation sequencing to detect the underlying molecular defects. Results: We found c. 5570T>A missense mutation in ANK2 gene for this family. Based on the Online Mendelian Inheritance in Man, ANK2 gene and the mutation detected correspond to long QT syndrome type 4. Conclusion: This mutation, although already known in other populations, but is reported for the first time in Iranian patients with cardiac arrhythmias. As the case with this family, genetic analysis of patients with cardiac arrhythmias would be helpful in reassessment of clinical diagnosis and therefore would help for patients' management and in some cases re-evaluation of ongoing treatment may be needed.

Background: Migraine is the most common chronic neurological disorders that may be associated with vasodilatation. According to the role of prostaglandin I2 (prostacyclin) receptor (PTGIR) in migraine as a receptor, which acts in vasodilatation, we decided to study the changes of PTGIR expression in migraine patients in relation to a suitable control group. Materials and Methods: Extracted mRNA from lymphocytes of 50 cases and 50 controls was used to synthesize cDNA. Real-time polymerase chain reaction was performed, and the data were analyzed. Our results show that PTGIR mRNA expression in cases was significantly higher than the control group (P = 0.010). Results: In conclusion, mRNA expression of PTGIR in the blood of people with migraines could be considered as a biomarker. Conclusion: In addition, repression of PTGIR gene expression by methods such as using siRNA is probably suitable for therapy of migraine patients.

A brain tumor is an intracranial neoplasm within the brain or in the central spinal canal. Primary malignant brain tumors affect about 200,000 people worldwide every year. Brain cells have special characters. Due to the specific properties of brain tumors, including epidemiology, growth, and division, investigation of brain tumors and the interpretation of results is not simple. Research to identify the genetic alterations of human tumors improves our knowledge of tumor biology, genetic interactions, progression, and preclinical therapeutic assessment. Obtaining data for prevention, diagnosis, and therapy requires sufficient samples, and brain tumors have a wide range. As a result, establishing the bank of brain tumors is very important and essential.

Background: Thalassemias are the most common monogenic disorders in many countries throughout the world. The best practice to control the prevalence of the disease is prenatal diagnosis (PND) services. Extensive practicing of PND proved effective in reducing new cases but on the other side of this success high abortion rate is hided, which ethically unfair and for many couples, especially with a previous experience of a therapeutic abortion, or moral concerns, is not a suitable choice. Preimplantation genetic diagnosis (PGD) is a strong alternative to conventional PND. At present PGD is the only abortion free fetal diagnostic process. Considering the fact that there are more than 6000 single gene disorders affecting approximately 1 in 300 live-births, the medical need for PGD services is significant. Materials and Methods: In the present study development of a PGD protocol for a thalassemia trait couple using nested multiplex fluorescent polymerase chain reaction (PCR) for the combination of polymorphic linked short tandem repeat (STR) markers and thalassemia mutations is described. Restriction fragment length polymorphism used to discriminate between wild and mutated alleles. Results: In PGD clinical cycle, paternal and maternal alleles for D11S988 and D11S1338 STR markers were segregated as it was expected. PCR product for IVSII-1 mutation was subsequently digested with BtscI restriction enzyme to differentiate normal allele from the mutant allele. The mother's mutation, being a comparatively large deletion, was detectable through size differences on agarose gel. Conclusion: The optimized single cell protocol developed and evaluated in this study is a feasible approach for preimplantation diagnosis of β-thalassemia in our patients.

Background: Y chromosome is one of the two sex chromosomes and is male specific. Due to limited genetic exchange, the main part of that is passed virtually unchanged from one generation to next generation. The short tandem repeats (STRs) are almost constant on chromosomes that make them as an appropriate factor for use in population genetic studies. In this study, we used the STRs of Y chromosome markers in Sadat families and comparison with other families was investigated. Materials and Methods: In this study, sampling was done from fifty unrelated males of Sadat families and fifty unrelated males of non-Sadat families. After the extraction of DNA from blood samples and primer design, polymerase chain reaction (PCR) was performed for each primer pairs separately. The PCR products were run on agarose gel that followed by running on polyacrylamide gel for better resolution. In addition, some sequenced samples were used as identified markers to determine the length of other alleles in polyacrylamide gel. Results: The survey of six STR in two case and control groups was carried out, and analysis revealed that the frequency of some alleles is different in case group compared to control group. Allele frequency of the markers DYS392, DYS393, DYS19, DYS390, DYS388, and DYS437 on the Y chromosome in Sadat families was quite different in comparison with other families. Conclusions: The reason for these differences in allele frequencies of the Sadat family in comparison with other families is having a common ancestor.

Background: Considering a few studies on the genetic basis of the cystinuria in the Middle East and the population-specific distribution of mutations in the SLC3A1 , we tried to find genetic variants in three exons (1, 3, and 8) of SLC3A1 . Materials and Methods: In this study, exons 1, 3, and 8 of SLC3A1 gene of 25 unrelated cystinuria patients searched for genetic variations by polymerase chain reaction and sequencing. Results: There were five different variations in our studied population. We found one mutation in the SLC3A1 gene including missense variant M467K and identified three polymorphisms: nonsynonymous variant G38G, c. 610 + 169C>T and c. 610 + 147C>G within the SLC3A1 gene, and one new variant. Conclusion: Our results confirm that cystinuria is a heterogeneous disorder at the molecular level and more studies are needed to identify the distribution and frequency of mutations causing cystinuria in the Iranian population. Key word: Aminoaciduria, cystinuria, rBAT, SLC3A1 , transport

Background: Teicoplanin is a member of vancomycin-ristocetin family of glycopeptide antibiotics. It mediated wound healing by increasing neovascularization possibly through activation of MAP kinase signaling pathway. The aim of this study is an evaluation of c-myc and c-fos genes expression after treatment of cells by teicoplanin and determines whether this glycopeptide antibiotic exerts its proliferation effects by influencing the expression of these genes. Hence, this study was designed to elucidate one possible mechanism underlying teicoplanin effects on cell proliferation using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay. Materials and Methods: Breast cancer cell line, MCF-7, was cultured, and three different concentrations of teicoplanin were added to the plates. We measured the cell proliferation rate by MTT assay. After cell harvesting, total RNA was extracted to synthesize single-stranded cDNA. Real-time polymerase chain reaction was performed, and the data were analyzed. Results: It was observed that the level of c-fos and c-myc genes' expressions was decreased at all three different concentrations of teicoplanin. Conclusion: it could be concluded that although teicoplanin is considered as an enhancing cell growth and proliferation, but probably its effect is not through MAP kinase signaling pathway or perhaps even has inhibitory effect on the expression of some genes such as c-myc and c-fos in this pathway. Hence, the mechanism of action of teicoplanin for increasing cell propagation, through cell signaling pathways or chromosomal abnormalities, remains unclear, and further studies should be conducted.

Background: Tetralogy of Fallot (TOF), the most common cyanotic heart defect and one of the most common congenital heart diseases, occurs mostly sporadically and nonsyndromically. The underlying molecular genetic mechanism is not known. Therefore, the existence of mutations in the homeodomain-encoding region of NKX2.5 gene in Iranian patients with tetralogy of Fallot is evaluated. Materials and Methods: In the present study, we analyzed the peripheral blood samples of27 patients in order to find any mutation in the 180 bp homeodomain-encoding region of NKX2.5 gene, which is known to be involved in heart development and diseases. DNA was extracted and all the samples were amplified by polymerase chain reaction (PCR) and sequenced. Results: Twenty-seven patients were included in the study. Twenty-five of them were infants and children (6 days to 11 years of age), one was a teenager (14-years of age), and another was a 33-year-old man [mean ± standard deviation (SD): 5.80 ± 3.90 years]. Thirteen patents were males (mean ± SD: 6.587077 ± 5.02 years) and 14 were females (mean ± SD: 5.0726 ± 2.81 years). One synonymous variant, i.e., c.543G>A was identified in one patient. Conclusion: Mutations in the homeodomain-encoding region of NKX2.5 gene may not have an outstanding role in etiology of tetralogy of Fallot patients in Iran. Key words: Cardiac defect, Iranian patient, NKX2.5 gene, tetralogy of Fallot (TO

Background: Following human radiation exposure in hospital or accidents, dose assessments are of prime importance in radiation accidents. These issues are of continuing importance with respect to socioeconomic policy relating to the industrial and medical uses of ionizing radiation, and also for risk assessment among people who are occupationally exposed to low and/or high linear energy transfer (LET) radiation, such as astronauts, pilots, stewardesses, nuclear power plant workers, and victims of radiation accidents. Materials and Methods: In this study, an assay for assessing radiation dose based on the induction of apoptosis in human T-lymphocytes was done to examine T-lymphocyte cells isolated from the fresh blood of 16volunteers, cultured and exposed to gamma rays. Radiation-induced apoptosis (RIA) was assessed by flow cytometric identification of cells displaying apoptosis-associated DNA condensation. Results: Dose-response experiments showed that at 2Gy dose level of radiotherapy programs, the RIA frequency was significantly above control. Apoptotic levels significantly depend on the dose of radiation rather than the donor. Conclusion: The results demonstrate the potential use of this assay as a biological indicator of radiation toxicity, optimizing patient dose in radiotherapy and biological dosimetry process.

Background: A major bottleneck in the production of recombinant proteins in conventional linear method is the heterogeneity in number of transgene copies in the genome of the host cell that lead to variable levels of transgene expression. Aside from the low efficiency of the random integration, other phenomena such as positional effect contribute to low efficiency of transgene expression. PiggyBac is a class of DNA transposons which can transpose through “cut and paste” mechanism in host genome. Some specific characteristic of this transposon makes it promise vector in gene transfer studies. High capacity of transgene transposition and flexibility in molecular engineering of transposase are characteristics of PiggyBac transposons that are important in recombinant protein and gene therapy approaches. The aim of this study was estimation of transgene copy number based on helper-dependent and independent PiggyBac transposition system in human embryonic kidney (HEK) cells. Methods: Plasmid containing interferon gamma inducible protein 10 (IP-10) coding sequence flanked by PiggyBac terminal repeat element and plasmid containing transposase system and a unit plasmid containing both transposae and IP-10 coding sequence were used for generating recombinant cells in helper-dependent and independent system, respectively. Human embryonic kidney cells were transfected by each group. After clonal selection, absolute quantitative real-time polymerase chain reaction was used for estimation of transgene copy number. Findings: Estimation of IP-10 copy number has revealed 5 and 2 copies per cell in helper-independent and dependent system, respectively. Conclusion: Here, we report activity of PiggyBac transposition in human embryonic kidney cell line. PiggyBac helper-independent and dependent system was used for generation of stable cell line producing mouse IP-10 protein and our data confirmed permanent expression of this transgene with the mean of about 5 transgene copies per each cell.

Infertility is one of the most common reproductive disorders occurring in approximately 15% of the couples. Male factor accounts for about half of these cases. The causes of reproductive defects in infertile men are multifactorial and many environmental and genetic factors affect male infertility. Genetics factors cause an account for 10-15% of male infertility, including chromosomal aberrations and single gene mutations. The current review will focus on genetics aspect of male infertility, including chromosomal disorder, single gene mutation and polymorphism, role of mitochondrial DNA and microRNA. We also take a look at last reported new genes causes of infertility.

Telomeres consist of repetitive DNA sequences and a variety of non-nucleosomal proteins which are essential to survive chromosome. Telomeres protect the ends of chromosomes, but can also inhibit the expression of nearby genes, called telomere position effect. The telomeric cap is a dynamic structure between a fully capped closed and a partially uncapped or open conformation. The protection of chromosome termini from being sensed by the cell as broken DNA is an important function of the telomeres as the capping function. Functional telomeres have a special role in response to DNA damage. The presence of telomere and the length of telomere are two important determinants for binding of chromosomes to the nuclear envelope in meiosis. Future studies about the characteristics of epigenetic factors in telomere length will lead to a better understanding of telomere regulation and its role in human cancers. It seems that telomere dysfunction, rather than telomere length alone, may create insights not only into the pathogenesis, but could also have significant impact on the diagnosis of cancer. Keywords: Role, Telomere, Tumorigenesis, Telomere dysfunction