N-acetylcysteine compared to metformin, improves the expression profile of growth differentiation factor-9 and receptor tyrosine kinase c-kit in the oocytes of patients with polycystic ovarian syndrome

Background: Paracrine disruption of growth factors in women with polycystic ovarian syndrome (PCOS) results in production of low quality oocyte, especially following ovulation induction. The aim of this study was to investigate the effects of metformin (MET), N-acetylcysteine (NAC) and their combination on the hormonal levels and expression profile of GDF-9, BMP-15 and c-kit, as hallmarks of oocyte quality, in PCOS patients. Materials and Methods: This prospective randomized, double-blind, placebo controlled trial aims to study the effects of MET, NAC and their combination (MET+NAC) on expression of GDF-9, BMP-15 and c-kit mRNA in oocytes [10 at the germinal vesicle (GV) stage, 10 at the MI stage, and 10 at the MII stage from per group] derived following ovulation induction in PCOS. Treatment was carried out for six weeks, starting on the third day of previous cycle until oocyte aspiration. The expression of GDF9, BMP15 and c-kit were determined by quantitative real time polymerase chain reaction (RT-qPCR) and western blot analysis. Data were analyzed with one-way ANOVA. Results: The follicular fluid (FF) level of c-kit protein significantly decreased in the NAC group compared to the other groups. Significant correlations were observed between the FF soluble c-kit protein with FF volume, androstenedione and estradiol. The GDF-9 expression in unfertilized mature oocytes were significantly higher in the NAC group compared to the other groups (P<0.001). Similar difference was not observed between the MET, NAC+MET and control groups. The c-kit expression in unfertilized mature oocytes were significantly lower in the NAC group compared to the other groups (P<0.001). Similar difference was not observed between the MET, NAC+MET and control groups (Registration number: IRCT201204159476N1). Conclusion: We concluded that NAC can improve the quality of oocytes in PCOS. © 2017, Royan Institute (ACECR). All rights reserved

Reduction of truncated Kit Expression in Men with Abnormal Semen Parameters, Globozoospermia and History of Low or Fertilization Failure

Objective: Phospholipase C zeta 1 (PLCζ) is one of the main sperm factor involved in oocyte activation and other factors may assist this factor to induce successful fertilization. Microinjection of recombinant tr-kit, a truncated form of c-kit receptor, into metaphase II-arrested mouse oocytes initiate egg activation. Considering the potential roles of tr-KIT during spermiogenesis and fertilization, we aimed to assess expression of tr-KIT in sperm of men with normal and abnormal parameters and also in infertile men with previous failed fertilization and globozoospermia. Materials and Methods: This experimental study was conducted from September 2015 to July 2016 on 30 normozoospermic and 20 abnormozoospermic samples for experiment one, and also was carried out on 10 globozoospermic men, 10 men with a history low or failed fertilization and 13 fertile men for experiment two. Semen parameters and sperm DNA fragmentation were assessed according to WHO protocol, and TUNEL assay. Sperm tr-KIT was evaluated by flow cytometry, immunostaining and western blot. Results: The results show that tr-KIT mainly was detected in post-acrosomal, equatorial and tail regions. Percentage of tr-KIT-positive spermatozoa in abnormozoospermic men was significantly lower than normozoospermic men. Also significant correlations were observed between sperm tr-KIT with sperm count (r=0.8, P<0.001), motility (r=0.31, P=0.03) and abnormal morphology (r=-0.6, P<0.001). Expression of tr-KIT protein was significantly lower in infertile men with low/ failed fertilization and globozoospermia compared to fertile men. The significant correlation was also observed between tr-KIT protein with fertilization rate (r=-0.46, P=0.04). In addition, significant correlations were observed between sperm DNA fragmentation with fertilization rate (r=-0.56, P=0.019) and tr-KIT protein (r=-0.38, P=0.04). Conclusion: tr-KIT may play a direct or indirect role in fertilization. Therefore, to increase our insight regarding the role of tr-KIT in fertilization further research is warranted. © 2019 Royan Institute (ACECR). All rights reserved

One-carbon cycle support rescues sperm damage in experimentally induced varicocoele in rats

Objectives: To investigate whether micronutrients in support of the one-carbon cycle and glutathione synthesis are effective in improving sperm damage after surgical varicocoele induction in rats and whether any effect is achieved without a rebound reductive stress as seen with oral antioxidants. Materials and Methods: Surgical varicocoele was induced in adult male Wistar rats and resulted in significant damage to the testis and sperm cells measured at 2 and 4 months after surgery. At 2 months after surgery, rats received a 2-month oral supplementation in support of the one-carbon cycle containing B vitamins (B2, B3, B6, folic acid and B12), N-acetyl-cysteine, zinc, small amounts of vitamin E, and a natural source of betalains and quercetine (Condensyl ® ; Parthenogen SAGL, Lugano, Switzerland and Nurilia SARL, Lyon, France). Results: One-carbon cycle supplementation, compared to untreated controls, significantly improved the morphometric characteristics of testis (P &lt; 0.05), sperm concentration, motility and abnormal morphology (P &lt; 0.001), sperm chromatin condensation (aniline blue staining, P &lt; 0.05), sperm DNA damage (acridine orange staining, P &lt; 0.05) and sperm lipid peroxidation (BODIPY C11, P &lt; 0.001). The improvement in both nuclear condensation and DNA damage and the lack of excessive inhibition of lipid peroxidation confirmed that no reductive stress had occurred. Conclusions: Micronutrients in support of the one-carbon cycle are effective in the treatment of surgically induced varicocoele in rats, probably by activating natural antioxidant defences and epigenetics. These results support the idea that essential micronutrients including B vitamins may also have a positive influence in clinical varicocoele, which should be tested in prospective clinical trials. © 2018 The Authors BJU International © 2018 BJU International Published by John Wiley & Sons Lt

The role of reactive oxygen species in the vitro development of mouse embryos

SIGLEAvailable from British Library Document Supply Centre- DSC:D062067 / BLDSC - British Library Document Supply CentreGBUnited Kingdo

Expression of sperm PLC? and clinical outcomes of ICSI-AOA in men affected by globozoospermia due to DPY19L2 deletion

Globozoospermia is characterized by the presence of 100% acrosomeless round-headed spermatozoa in an ejaculate. Failed fertilization after intracytoplasmic sperm injection (ICSI) is commonly reported for globozoospermic couples and can be overcome by artificial oocyte activation (AOA). Phospholipase C zeta (PLC?) is one of the main sperm factors involved in oocyte activation and its low expression levels mainly account for fertilization failure. Deletion of the DPY19L2 gene is reported as a main genetic cause in over 70% of infertile men with globozoospermia. The current study assesses the expression profile of sperm PLC? at RNA and protein levels in 32 DPY19L2 deletion-mediated globozoospermic men and reports corresponding clinical outcomes following ICSI with AOA. The expression of PLC? relative to GAPDH at RNA (0.78 � 0.16 versus 1.65 � 0.24; P = 0.02) and protein (0.39 � 0.12 versus 0.83 � 0.13; P = 0.01) levels in globozoospermic men with DPY19L2 deletion was significantly lower compared with fertile men (n = 32). Fertilization rate in globozoospermic couples following ICSI-AOA was significantly lower compared with fertile men (53.14 � 5.13% versus 87.64 � 2.38%, P < 0.001). However, implantation (26.2%) and pregnancy (53.8%) rates were not jeopardized by DPY19L2 deletion in these couples. - 2017 Reproductive Healthcare Ltd.This study was supported by the Royan Institute ( 94000127 ) and we would like to express our gratitude to the staff of Isfahan Fertility and Infertility for their full support. Mohammad Hossein Nasr-Esfahani received his PhD from the University of Cambridge and is Professor at the Royan Institute, and Laboratory Director of the Isfahan Fertility and Infertility Centre. His special interests are male infertility and animal cloning. In addition, he has over 350 publications in international and national journals. Key message Unlabelled box ICSI-AOA resulted in a high fertilization rate in globozoospermic men with low expression of PLC? and DPY19L2 deletion.Scopu

DETECTION AND ISOLATION OF CD59 FROM HUMAN SEMINAL PLASMA

Introduction. CD59 is one of the complement regulatory proteins (CRPs) which exert inhibitory function by blocking the formation of C5b-9 complex or Membrane Attacks complex (MAC). Regarding the therapeutic role of CD59 in treatment of pathological effects in uncontrolled activation of complements system and its efficiency to overcome the hyper-acute rejection, CD59 was suggested for maintenance of transplanted organ. In this study We determined and isolated CD59 from seminal plasma.&#13; Methods. Six normospermic sample according to WHO standards were chosen. Plasma of samples was separated and to remove the postasomes, the seminal plama was ultra centrifuged. CD59 was detected by Dot-Blot using CD59 mAb (MEM43). The molecular weight and purity of protein was detected by SOS-PAGE method follwed by Westerm Blot.&#13; Results. Protein was present in the 6.5 ml and 15ml of gel fitration fractions. Molecular weight based on marker size in these two fractions was 65 and 21KD respectively. Discussion. CD59 had previously beem purified by lysis of erythrocyte cell membrane. Because of use of detergent and preservative agents, this method decreased physiologic effects of the protein. In this study the isolation was performed from prostasome granules" without using of any detergent and preservative agents

Bio-functionalized PCL nanofibrous scaffolds for nerve tissue engineering

10.1016/j.msec.2010.06.004Materials Science and Engineering C3081129-113

Electrical stimulation of nerve cells using conductive nanofibrous scaffolds for nerve tissue engineering

10.1089/ten.tea.2008.0689Tissue Engineering - Part A15113605-361