IVF Technology

en 26140180 IVF Technology The Parliament of Great Britain approved three-parent IVF project with 382 votes for and 128 votes against it on 3rd February, 2015. In case of the project approval in House of Lords, Great Britain would be the first country which legalizes the action. The purpose of this procedure is prevention of genetic diseases with mitochondrial defects in which the defective mitochondria in the cytoplasm of the mother’s egg is transferred to the embryo. The consequence is the death of the child, muscle weakness, blindness and heart diseases. If the healthy mitochondria of another person are used in this procedure, the defects would disappear. This is done in two ways: 1. Nucleus transfer from the mother's egg with mitochondrial defects or egg cytoplasm to the donated egg in which the nucleus has been removed, 2. Transfer of parents’ nuclei from the early embryo (zygote) containing the cytoplasm to a donated early embryo (zygote) in which the parents’ nuclei have been removed. In each of these cases, the mitochondria in the egg or the donated zygote replace defective mitochondria and protect the child from related diseases. The advancement of IVF technology brought up the possibility in replacing the egg of patients with mitochondrial diseases with healthy donated eggs. In this case, the child produced has no genetic relationship with the patient, while in the current practice, 23 pairs of parental chromosomes construct the genetic essence of the child. In this practice, 37 mitochondrial genes, which comprise 1.10% of the total genetic essence of the child and play key roles in production of energy in cytoplasm of the egg, can be replaced with donated mitochondria in case of any defects, and thereby culminate in preventing mitochondrial defects in 1200 cases of children in Great Britain. The support of majority of reproduction scientists from the action and the concern and opposition of the Catholic Church together with some law and ethics specialists for authorizing the birth of engineered children denote the contrary position of the parties in applying this preventive and therapeutic action. The advent and development of reproductive technologies have always been intermingled with theological, ethical, legal and social issues and consequences. While great emphasis has been placed on ethical, legal and social dimensions and implications of cutting edge advances in reproduction, it seems that under supervision and surveillance of law makers and regulations, the possibility for amending inappropriate decisions is always provided. This is the focal point that the Parliament of Great Britain adhered to. However, there is no doubt that if necessary practical measures are not taken for discussing problems and compiling rules and regulations in treatment of reproductive diseases with higher incidence (such as application of IVF in indications for surrogacy in Iran), more theoretical and practical vexing issues will emerge which are complicated enough to be handled optimally. Although further studies are required to evaluate the action approved by the Parliament of Great Britain, the legal aspect of the action is worthy of attention since timely evaluation of it has been done. Also, as a prominent advancement in the field of biotechnology, the potential for its modification and amendment is permitted by the law. 49 49 https://www.ajmb.org/En/Article.aspx?id=201 https://www.ajmb.org/PDF/En/FullText/201.pdf Mohammad MehdiAkhondi13

en 26140181 Comparison between Platinum-Azidothymidine and Azidothymidine Effects on Bcl-2 and Telomerase Gene Expression in Rats with Hepatocellular Carcinoma Background: High expression of telomerase and Bcl-2 are reported in hepatocellular carcinoma. Some anticancer drugs show their effects through reduction of these factors. In this study, it was aimed to investigate the effects of a new synthetic compound, platinum azidothymidine, on inhibition of telomerase and Bcl-2 expression in hepatocellular carcinoma compared to azidothymidine. Methods: To study the effects of Pt-AZT on hepatocellular carcinoma and compare its effects with AZT in inhibition of telomerase and Bcl-2 gene expression, pathogen-free male Wistar rats (n=100) were used. They were randomly divided to 4 groups (n=25). Group A as the control group contained 25 healthy rats; in the rest of animals, preneoplastic lesions were induced in their livers (groups B, C, and D) using Solt-Farber resistant hepatocyte protocol. Cancer development was approved by a pathology laboratory. Group B was negative control (untreated), groups C and D were treated by intraperitoneal injection (IP) of Pt-AZT (0.9 mg/kg/day) and AZT (0.3 mg/kg/day), respectively for 14 days. At the end of the protocol, all rats were sacrificed and Bcl-2 and telomerase gene expression was determined using real -time PCR. Results: No tumor in the livers was found in group A at any point of the study, but it was present in livers of all animals in B, C and D groups. Results showed that telomerase and Bcl-2 expression was significantly lower in group C compared with group B (0.473±0.231 vs. 5.137±5.08, p<0.001, for telomerase expression, and 0.41±0.276 vs. 7.25±11.6, p<0.001, for Bcl-2 expression) and also compared with group D (0.473±0.231 vs. 3.48±4.02, p<0.001, for telomerase expression, and 0.41±0.276 vs. 4.93±18, p<0.001, for Bcl-2 expression). Conclusion: For the first time, it was demonstrated that Pt-AZT has more inhibitory effect on telomerase and Bcl-2 expression than AZT. It effectively inhibits the growth of liver tumor in rats by extending apoptosis. Gene expression, Hepatocellular carcinoma, Platinum, Telomerase 50 56 https://www.ajmb.org/En/Article.aspx?id=202 https://www.ajmb.org/PDF/En/FullText/202.pdf AbdolrezaSabokrouhDepartment of Clinical Biochemistry, Hamadan University of Medical Sciences, Hamadan, Iran793AsadVaisi-RayganiMolecular Diagnostic Research Center, Kermanshah University of Medical Sciences, Kermanshah, Iran795Mohammad TaghiGoodarziResearch Center for Molecular Medicine, Hamadan University of Medical Sciences, Hamadan, Iran794ShohrehKhatamiDepartment of Clinical Biochemistry, Pasteur Institute, Tehran, Iran796MasoudTaghizadeh-JahedReproductive Biotechnology Research Center, Avicenna Research Institute, ACECR, Tehran, Iran797NahidShahabadiDepartment of Chemistry, Razi University of Kermanshah, Kermanshah, Kermanshah, Iran881NiknamLakpourReproductive Biotechnology Research Center, Avicenna Research Institute, ACECR, Tehran, Iran66YadollahShakibaMolecular Diagnostic Research Center, Kermanshah University of Medical Sciences, Kermanshah, Iran803

en 26140182 The Effects of Ultraviolet Light and Riboflavin on Inactivation of Viruses and the Quality of Platelet Concentrates at Laboratory Scale Background: This study investigated the effects of Riboflavin (RB) combined with different doses of UV on Platelet Concentrate (PC) which was infected by three models of virus. Platelet quality after treatment was also assessed. Methods: Three models of virus used in this study were Vesicular Stomatitis Virus (VSV), Herpes Simplex Virus (HSV), and Polio virus, which were added to PC. After photochemical treatment with RB and UV light, residual viral infectivity was titrated using 50% Tissue Culture Infective Dose (TCID50)/ml. This treatment was done with concentration of 50 µM of RB and different doses of UV light (0.24, 0.48, 0.97, 1.29 J/cm2). Platelet quality was assessed by measuring pH, Lactate Dehydrogenase (LDH), MTT assay and cell count after treatments and during 4 days of storage against control groups. Results: Concentration of 50 µM RB with combination of 1.29 J/cm2 dose of UV resulted in the highest titer reduction of VSV (4 log 10) and HSV (4.26 log10) and lowest titer reduction of Polio virus (2.6 log10). No significant difference was observed between different doses in comparison with control groups. In all treatment groups, the storage stability of platelets in PC was in the acceptable range in comparison with control group. Conclusion: This study indicated that RB/UV treatment was a promising pathogen reduction technique in PC and had limited effects on platelet quality. However, further optimization of this method is necessary to deal with blood-borne viruses like non-enveloped viruses. Blood platelet, Riboflavin, Ultraviolet light, Virus inactivation 57 63 https://www.ajmb.org/En/Article.aspx?id=203 https://www.ajmb.org/PDF/En/FullText/203.pdf HamidehMirshafieeDepartment of Microbiology, Faculty of Biological Sciences, Shahid Beheshti University, Tehran, Iran638ZohrehSharifiBlood Transfusion Research Center, High Institute for Research and Education in Transfusion Medicine, Tehran, Iran640Seyed MasoudHosseiniDepartment of Microbiology, Faculty of Biological Sciences, Shahid Beheshti University, Tehran, Iran639FatemehYariBlood Transfusion Research Center, High Institute for Research and Education in Transfusion Medicine, Tehran, Iran882HamedNikbakhtLaser and Plasma Research Institute, Shahid Beheshti University, Tehran, Iran883HamidLatifiLaser and Plasma Research Institute, Shahid Beheshti University, Tehran, Iran884

en 26140183 Cobalt Zinc Ferrite Nanoparticles as a Potential Magnetic Resonance Imaging Agent: An In Vitro Study Background: Magnetic Nanoparticles (MNP) have been used for contrast enhancement in Magnetic Resonance Imaging (MRI). In recent years, research on the use of ferrite nanoparticles in T2 contrast agents has shown a great potential application in MR imaging. In this work, Co0.5Zn0.5Fe2O4 and Co0.5Zn0.5Fe2O4-DMSA magnetic nanoparticles, CZF-MNPs and CZF-MNPs-DMSA, were investigated as MR imaging contrast agents. Methods: Cobalt zinc ferrite nanoparticles and their suitable coating, DMSA, were investigated under in vitro condition. Human prostate cancer cell lines (DU145 and PC3) with bare (uncoated) and coated magnetic nanoparticles were investigated as nano-contrast MR imaging agents. Results: Using T2-weighted MR images identified that signal intensity of bare and coated MNPs was enhanced with increasing concentration of MNPs in water. The values of 1/T2 relaxivity (r2) for bare and coated MNPs were found to be 88.46 and 28.80 (mM–1 s–1), respectively. Conclusion: The results show that bare and coated MNPs are suitable as T2-weighted MR imaging contrast agents. Also, the obtained r2/r1 values (59.3 and 50) for bare and coated MNPs were in agreement with the results of other previous relevant works. Magnetite nanoparticles , MR imaging, Prostatic neoplasm 64 68 https://www.ajmb.org/En/Article.aspx?id=204 https://www.ajmb.org/PDF/En/FullText/204.pdf ZeinabGhasemianDepartment of Medical Physics, Faculty of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran885DaryoushShahbazi-GahroueiDepartment of Medical Physics, Faculty of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran886SohrabManouchehriDepartment of Physics, Nano-center, Maleke-ashtar University of Technology, Shahin-shahr, Isfahan, Iran887

en 26140184 Release Studies on Ciprofloxacin Loaded Non-ionic Surfactant Vesicles Background: Development of new drug carriers would be an interesting approach if it allowed increased efficacy of antibiotics and a reduction in doses, thus reducing the risk of developing resistance. As with most drug carriers, niosomes have been used to improve the selective delivery and the therapeutic index of antimicrobial agents. Methods: In this study, different formulation of niosomes containing ciprofloxacin (CPFX), Span (20, 60 or 80), Tween (20, 60 or 80) and cholesterol were prepared by film hydration method. The release of the drug from different formulations was studied by using Franz diffusion cell. The niosomes were further characterized by optical microscopy and particle size analysis, and their antimicrobial activity was evaluated. Results: Size of the niosomes was significantly dependent on the amount of cholesterol and surfactant type and varied from 8.56 to 61.3 μm. The entrapment efficiency of CPFX niosomes prepared by remote loading was more than 74%. Niosomes composed of Span/Tween 60 provided a higher CPFX release rate than other formulations. The obtained results indicated a diffusion-based mechanism for drug leakage through bilayers. All formulations presented more antibacterial activity as compared to free CPFX solution. Conclusion: Niosomal CPFX appears to be a promising approach in the management of bacterial infections, especially ophthalmic ones, and should be further evaluated by in vivo experiments. Ciprofloxacin, Niosomes, Release 69 75 https://www.ajmb.org/En/Article.aspx?id=205 https://www.ajmb.org/PDF/En/FullText/205.pdf VajiheAkbariDepartment of Pharmaceutical Biotechnology and Isfahan Pharmaceutical Research Center, Faculty of Pharmacy, Isfahan University of Medical Sciences, Isfahan, Iran273DaryoushAbediDepartment of Pharmaceutical Biotechnology and Isfahan Pharmaceutical Research Center, Faculty of Pharmacy, Isfahan University of Medical Sciences, Isfahan, Iran299AbbasPardakhtyKerman Pharmaceutics Research Center and Department of Pharmaceutics, Kerman University of Medical Sciences, Kerman, Iran888HojjatSadeghi-AliabadiDepartment of Pharmaceutical Biotechnology and Isfahan Pharmaceutical Research Center, Faculty of Pharmacy, Isfahan University of Medical Sciences, Isfahan, Iran889

en 26140185 TP53 Binding to BRCA1 and RAD51 in MCF7 and MDA-MB-468 Breast Cancer Cell Lines In vivo and In vitro Background: Tumour suppressor genes such as TP53, BRCA1 and RAD51 are involved in DNA repair and their malfunctions result in genomic instability and cancer. Wild type (WT) TP53 binds to BRCA1and RAD51 in vivo and in vitro. However, mutated TP53 in tumours can interfere with WT TP53 function. We studied how mutation of TP53 in MDA-MB-468 cell line could affect its binding capacity and interfere with WT TP53 interaction with these DNA repair proteins. Methods: Binding capacity of mutated TP53 in MDA-MB-468 breast cancer cell line to BRCA1 and RAD51 proteins in comparison to WT TP53 in MCF7 cell line was studied by Immunoprecipitation. In vitro studies were performed by GST-WT p53 pull-down assays in these cell lines to assess the interaction of GST-WT p53 with BRCA1 and RAD51 proteins. Results: The results showed that mutated TP53 in MDA-MB-468 cells interacted with BRCA1 protein in vivo and did not effect WT TP53 binding to this protein in vitro. The Immunoprecipitation assays revealed that the mutated TP53 did not bind to RAD51 in comparison to WT TP53. However, this mutated protein could not interfere with binding of RAD51 to GST-WT p53 in MDA-MB-468 cell line by in vitro experiment. Conclusion: It was found that WT TP53 interactions with BRCA1 and RAD51 did not interfere with mutated TP53 in MDA-MB-468 cell line. In addition, RAD51 did not bind to TP53 with R273C mutation in vivo. BRCA1 protein, Immunoprecipitation, RAD51 protein, TP53 76 79 https://www.ajmb.org/En/Article.aspx?id=206 https://www.ajmb.org/PDF/En/FullText/206.pdf MozhganRastiRecombinant Lab, Department of Biochemistry, Faculty of Medicine, Shiraz University of Medical Sciences, Shiraz, Iran808TayebehAzimiRecombinant Lab, Department of Biochemistry, Faculty of Medicine, Shiraz University of Medical Sciences, Shiraz, Iran890

en 26140186 Maternal Supplementary Folate Intake, Methylenetetrahydrofolate Reductase (MTHFR) C677T and A1298C Polymorphisms and the Risk of Orofacial Cleft in Iranian Children Background: The purpose of this study was to describe the association of MTHFR gene single nucleotide polymorphisms (C677T and A1298C) and maternal supplementary folate intake with orofacial clefts in the Iranian population. Methods: In this case-control study, peripheral venous blood was taken from 65 patients with orofacial clefts and 215 unaffected controls for DNA extraction and kept in EDTA for further analysis. The genotyping was carried out using Polymerase Chain Reaction (PCR) followed by Restriction Fragment Length Polymorphism (RFLP) and gel electrophoresis. Data were analyzed using Chi square test and logistic regression tests. Results: Genotype frequencies of 677TT were reported to be 13.5 and 36.1% in controls and CL/P patients, respectively, which showed a significant difference compared to CC as reference (OR=4.118; 95% CI=1.997-8.492; p=0.001). Conversely, 1298CC with frequencies of 10.8 and 12.7% in controls and patients, respectively, showed no significant difference compared to AA (OR=2.359; 95% CI=0.792-7.023; p=0.123). Comparing patients whose mothers did not report the folate supplement intake during pregnancy, to controls, it was observed that lack of folate intake was a predisposing factor for having a child with oral clefts (OR=5/718, p=0.000). Conclusion: Children carrying the 677TT variant of the MTHFR gene may have an increased risk of CL/P. In addition, the finding that the risk associated with this allele was obviously higher when the mothers didn't use folic acid, supports the hypothesis that folic acid may play a role in the etiology of CL/P. Cleft lip, Cleft palate, Genes, Polymorphism 80 84 https://www.ajmb.org/En/Article.aspx?id=207 https://www.ajmb.org/PDF/En/FullText/207.pdf AsgharEbadifarDentofacial Deformities Research Center, Research Institute of Dental Sciences, Shahid Beheshti University of Medical Sciences, Tehran, Iran891Hamid RezaKhorram KhorshidGenetic Research Center, University of Social Welfare and Rehabilitation Sciences, Tehran, Iran42KooroshKamaliReproductive Biotechnology Research Center, Avicenna Research Institute, ACECR, Tehran, Iran89MehdiSalehi ZeinabadiPediatric Department, Dental School, Semnan University of Medical Sciences, Semnan, Iran893TayyebehKhoshbakhtGenetic Research Centre, University of Social Welfare and Rehabilitation Sciences, Tehran, Iran894NazilaAmeliOrthodontic Department, Dental school, Semnan University of Medical Sciences, Semnan, Iran895

en 26140187 Evaluation of a Modified DNA Extraction Method for Isolation of Cell-Free Fetal DNA from Maternal Serum Background: Discovery of short cell free fetal DNA (cffDNA) fragments in maternal plasma has created major changes in the field of prenatal diagnosis. The use of cffDNA to set up noninvasive prenatal test is limited due to the low concentration of fetal DNA in maternal plasma therefore, employing a high efficiency extraction method leads to more accurate results. The aim of this study was to evaluate the efficiency of Triton/Heat/Phenol (THP) protocol in comparison with the QIAamp DNA Blood mini Kit for cffDNA purification. Methods: In order to evaluate the efficiency of THP protocol, DNA of Rhesus D (RhD) negative pregnant women's plasma was collected, then real-time PCR for RHD exon 7 was performed. The Ct value data of real time PCR obtained by two different methods were compared and after delivery serology test on cord blood was done to validate the real time PCR results. Results: The results indicated significant differences between two extraction methods (p=0.001). The mean±SD of Ct-value using THP protocol was 33.8±1.6 and 36.1±2.47 using QIAamp DNA Blood mini Kit. Conclusion: our finding demonstrated that THP protocol was more effective than the QIAamp DNA Blood mini Kits for cffDNA extraction and lead to decrease the false negative results. Fetus, Prenatal diagnosis, Real time polymerase chain reaction, THP protocol 85 88 https://www.ajmb.org/En/Article.aspx?id=208 https://www.ajmb.org/PDF/En/FullText/208.pdf ZeinabKeshavarzStudent Research Committee, Shiraz University of Medical Sciences, Shiraz, Iran896LeiliMoezziDiagnostic Laboratory Sciences and Technology Research Center, School of Paramedical Sciences, Shiraz University of Medical Sciences, Shiraz, Iran668RezaRanjbaranDiagnostic Laboratory Sciences and Technology Research Center, School of Paramedical Sciences, Shiraz University of Medical Sciences, Shiraz, Iran667FarzanehAboualizadehDiagnostic Laboratory Sciences and Technology Research Center, School of Paramedical Sciences, Shiraz University of Medical Sciences, Shiraz, Iran669AbbasBehzad-BehbahaniDiagnostic Laboratory Sciences and Technology Research Center, School of Paramedical Sciences, Shiraz University of Medical Sciences, Shiraz, Iran664MasoomaAbdullahiDiagnostic Laboratory Sciences and Technology Research Center, School of Paramedical Sciences, Shiraz University of Medical Sciences, Shiraz, Iran900SedighehSharifzadehDiagnostic Laboratory Sciences and Technology Research Center, School of Paramedical Sciences, Shiraz University of Medical Sciences, Shiraz, Iran901