Cell Journal (Yakhteh)

555-568

Authors: Fatemeh Daneshimehr, Zahra Barabadi, Shahrokh Abdolahi, Masoud Soleimani, Javad Verdi, Somayeh Ebrahimi- Barough, Jafar Ai

Angiogenesis is a characteristic of glioblastoma (GBM), the most fatal and therapeutic-resistant brain tumor. Highly expressed angiogenic cytokines and proliferated microvascular system made anti-angiogenesis treatments a thoroughly plausible approach for GBM treatment. Many trials have proved to be not only as a safe but also as an effective approach in GBM retardation in a certain time window as seen in radiographic response rates; however, they have failed to implement significant improvements in clinical manifestation whether alone or in combination with radio/chemotherapy. Bevasizumab, an anti-vascular endothelial growth factor-A (VEGF-A) antibody, is the only agent that exerts meaningful clinical influence by improving progression-free survival (PFS) and partially alleviate clinical symptoms, nevertheless, it could not prolong the overall survival (OS) in patients with GBM. The data generated from phase II trials clearly revealed a correlation between elevated reperfusion, subsequent to vascular normalization induction, and improved clinical outcomes which explicitly indicates anti-angiogenesis treatments are beneficial. In order to prolong these initial benefits observed in a certain period of time after anti-angiogenesis targeting, some aspects of the therapy should be tackled: recognition of other bypass angiogenesis pathways activated following antiangiogenesis therapy, identification of probable pathways that induce insensitivity to shortage of blood supply, and classifying the patients by mapping their GBM-related gene profile as biomarkers to predict their responsiveness to therapy. Herein, the molecular basis of brain vasculature development in normal and tumoral conditions is briefly discussed and it is explained how "vascular normalization" concept opened a window to a better comprehension of some adverse effects observed in anti-angiogenesis therapy in clinical condition. Then, the most targeted angiogenesis pathways focused on ligand/receptor interactions in GBM clinical trials are reviewed. Lastly, different targeting strategies applied in anti-angiogenesis treatment are discussed.

577-585

Authors: Yaser Masoumi-Ardakani, Hamid Najafipour, Hamid Reza Nasri, Soheil Aminizadeh, Shirin Jafari, Zohreh Safi

Objective: Hypertension (HTN) is among the leading causes of myocardial infarction, stroke, and kidney disease. The MitoQ supplement is a mitochondrial-targeted antioxidant that attenuates the generation of reactive oxygen species (ROS). miRNAs play an essential role in the pathophysiology of HTN. Regular aerobic exercise is recommended to decrease the risk of cardiovascular disease. We aimed to evaluate the effects of MitoQ supplementation and moderate endurance training (ET), alone and in combination, on cardiac function, blood pressure, the circulatory levels of miRNA-21 and miRNA-222, and oxidative status in individuals with HTN. Materials and Methods: In a double-blind, randomized clinical trial (except for ET group), 52 male hypertensive subjects (40-55 years old) were randomly divided into four groups (n=13): Placebo, MitoQ (20 mg/day, oral), ET (Cycle ergometer, moderate intensity, 40-60% VO2 peak, three sessions/week for six weeks), and MitoQ+ET. Cardiac echocardiography indices, serum oxidative and inflammation status, and miRNAs 21 and 222 were assessed before and after interventions. Results: Left ventricular mass [effect size (ES): -6.3, 95% confidence interval (CI): -11.2 to -1.4] and end-systolic/ diastolic diameters significantly improved in the intervention groups (ES: -0.05, 95% CI: -0.11 to 0.00 and -0.09, 95% CI: -0.16 to -0.02). Total serum antioxidant capacity (TAC) increased (ES: 36.0, 95% CI: 26.1 to 45.8), and malondialdehyde (MDA) (ES: -0.43, 95% CI: -0.53 to -0.32), IL-6 (ES: -1.6, 95% CI: -1.98 to -1.25), miR-21 (ES: -0.48, 95% CI: -0.61 to -0.35), and miR-222 (ES: -0.31, 95% CI: -0.44 to -0.18) significantly decreased in response to ET, MitoQ, and their combination. Conclusion: MitoQ and ET, individually and more pronouncedly in combination, can improve cardiovascular health in people with high blood pressure (BP) by reducing inflammation and increasing antioxidant defense, in association with reduction in circulatory miR-21 and miR-222 levels (registration number: IRCT20190228042870N1).

596-602

Authors: Hanieh Rezaee, Mohammad Salehi, Mojgan Bandepour, Sima Kalantari, Sara Hosseini, Khosrow Agin, Bahram Kezemi

Objective: The most common mutation in cystic fibrosis (CF), (ΔF508-CFTR), results in impaired protein maturation, folding and transportation to the surface of the cell. As a consequence of impaired protein maturation and/or transport from the extracellular matrix to the cell, different systems are influenced, including gastrointestinal system and glandular system, reproductive system and respiratory systems. CF models are essential tools to provide further knowledge of CF pathophysiology. With this aim, we designed a transgenic CF model based on the homologous recombination (HR) system. Materials and Methods: In this experimental study, a specifically designed construct containing the CFTR gene with F508del was cloned into a PTZ57R cloning vector and then the construct was transformed into the male pronucleus by microinjection after in vitro fertilization (IVF). Then the rates of blastocyst formation and embryonic development at 72 hours after IVF, were evaluated using the inverted microscope and the insertion of the construct was approved by polymerase chain reaction (PCR) method. Results: The CFTR gene was successfully cloned into the PTZ57R cloning vector and overall, from 22 injected cells, 5 blastocysts were observed after pronuclear injection of the CFTR gene construct. PCR verification of the blastocyst with CFTR-specific primers represented complete recombination of CFTR into the mouse genome. Conclusion: For the first time we designed a unique genome construction that can be detected using a simple PCR method. The pronuclear injection was performed for the transformation of the genome construct into the male pronuclei using microinjection and the development of zygote to the blastocyst stage has been observed following transgenesis

612-619

Authors: Maryam Shahverdi, Vahid Akbarinejad, Azam Dalman, Mostafa Hajinasrollah, Mehdi Vodjgani, Nima Tanhaei Vash Mohammad Hossein Nasr-Esfahani, Poopak Eftekhari-Yazdi

Objective: Scarcity of oocytes for assisted reproduction in endangered species can be bypassed by interspecies somatic cell nuclear transfer (iSCNT). In Felids, domestic cat (Felis catus) oocytes can serve as recipients for the nucleus of the endangered Persian leopard (Panthera pardus saxicolor). However, in vitro oocyte maturation is still suboptimal in cats, whereas it has been reported to benefit from micro-vibration in non-felid species. Therefore, the present study is aimed to determine whether micro-vibration, applied during in vitro maturation (IVM), improves the embryogenic potential of cat oocytes transplanted with fibroblast nuclei of the Persian leopard. Materials and Methods: In the experimental study, cat cumulus-oocyte complexes (COCs) were randomly assigned to the treatment group (micro-vibration) or control group (static culture). Resultant metaphase II (MII) oocytes were enucleated and reconstructed with nucleus transplants from leopard fibroblasts, followed by artificial oocyte activation and embryo culture under the same condition (static) for 7 days. Results: While cumulus cell expansion and oocyte maturation profited from micro-vibration (P<0.05), the quantity and quality of blastocysts were significantly lower in micro-vibration than in the control group (P<0.05). The total number of blastocyst cells tended to be lower in the micro-vibration than in the control group (P=0.075). Nevertheless, the proportion of ICM and TE cells did not differ between the micro-vibration and control groups (P>0.05). Conclusion: The present study indicated that micro-vibration at a frequency of 44 Hz for 5 secs per hour enhanced nuclear maturation and cumulus cell expansion of cat oocytes. However, exposure to micro-vibration during IVM impaired the survival rate of reconstructed oocytes during the iSCNT process and their developmental competence toward the blastocyst stage.

628-636

Authors: Shahrzad Aghajani, Ali Salehzadeh, Fatemeh Ghasemian, Marzieh Mehrafza, Ahmad Hosseini

Objective: Preimplantation genetic testing for aneuploidies (PGT-A) is used to determine chromosomal normality and achieve a successful live birth in infertile couples. There is a possible correlation between chromosomal aneuploidy, embryo development and pregnancy rate. This study evaluated the influence of single blastomere biopsy (SBB) on embryo development and pregnancy rates during frozen embryo transfer (FET) and fresh cycles. Materials and Methods: This quasi-experimental study evaluated 115 intracytoplasmic sperm injection (ICSI) cycles, including 443 embryos (6-8 cells) with a grade A on day three, following PGT-A in the fresh or FET cycles from February 2018 to June 2020. In addition, the fresh cycles without PGT were included as a control group (n=166 embryos). SBB was done on day three and was grouped as FET-PGT (n=149) and the fresh-PGT (n=128). Results: There is a more aneuploidy rate in the FET-PGT group compared to the fresh-PGT cycle (36.60% vs. 20.38%, P<0.001). There is a rate of higher development and blastocyst in the control group. While the embryos of PGT groups showed higher degrees of expansion (expansion 5) on day five. 8.6, 8.59, and 9.37% of expansion 3, 4, and 5 in the fresh-PGT embryos, 12.58, 2.78, and 14.84% of expansion 3, 4, and 5 in the FET-PGD embryos compared to 10.84and 33.73% of expansion 3 and 4 in the control group (without expansion 5; P<0.001). There was no significant relationship between 13, 18, and 21 chromosome aneuploidies with blastocyst development competence among the groups (P<0.1). Following embryo transfer (n=97), the spontaneous abortion rate was higher in the FET-PGT cycles compared to the fresh-PGT and control groups (50 vs. 22 and 11%, respectively; P<0.04). Conclusion: The process of SBB following vitrification significantly decreased embryo development and pregnancy outcomes. Therefore, a morphological analysis could not be reliable in selecting chromosomally normal embryos