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

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

Efficiency of Sperm Gradient Processing and HA-binding Test for the Selection of Sperm with Normal Protamine, Intact DNA and Normal Morphology

Objective: The aim of this study was to examine the efficiency of HA-binding test and sperm gradient processing method for the selection of sperm with normal protamine,intact DNA and normal morphology.Materials and Methods: The study was conducted on 29 patients undergoing ICSI. Semen analysis was carried out according to WHO criteria. Semen samples were divided into three equal portions. One portion was left untreated (control), the secondportion was used for HA-binding (HA: Hyaluronic acid) and the third portion underwent PureSperm gradient processing. Then, the three portions were evaluatedto examine protamine deficiency (Chromomycin A3 staining), sperm morphology(Papanicolaou staining) and DNA integrity (Sperm Chromatin Dispersion Test). Coefficients of correlation and student t-test were carried out using SPSS 11.5 and P-value lower than 0.05 was considered as significant.Results: The mean number of sperm abnormality, protamin deficiency, and DNA fragmentation in HA binding and sperm gradient processing groups were significantlydecreased as compared to the control group (p<0.05). In addition, sperm gradient processing was superior to HA-binding method in the selection of sperm with normal protamine, intact DNA and normal morphology.Conclusion: Both HA-binding and sperm gradient processing methods were effectivein the selection of sperm with better quality in terms of normal morphology, normalprotamine content and DNA integrity. However, sperm gradient processing was superior to HA-binding method in the selection of normal sperm

The effect of vitamin E & vitamin B on sperm function in rat varicocele model

We assessed the effect of vitamin E and vitamin B (Vit E & Vit B) and their combination on sperm functional parameters in the rat varicocele model. Male rats (n = 120) were divided into control (n = 30), sham (n = 30) and varicocele induction (n = 60) groups. After 2 months, 10 rats from each group were sacrificed to verify varicocele model. This part of results showed that sperm parameters, DNA damage, lipid peroxidation and residual histone were adversely effected in the varicocele group. From the 50 remaining rats in varicocele group, 10 rats received Vit B complex (6, 9.6, 30.4, 9.6 and 0.006 mg/kg for B6, B2, B9, B1 and B12 respectively), 10 rats received Vit E (40 mg/kg), 10 rats received Vit B & E, 10 rats only received water and 10 rats were only received sesame oil as a solvent for Vit E, for 2 months. From 40 remaining rats in control and sham groups, 20 rats only received water and other 20 rats only received sesame oil for 2 months. Then, all the aforementioned parameters were assessed. These results showed that Vit B antioxidant was more efficient in improvement of sperm parameters, chromatin integrity and lipid peroxidation in varicocelized rats compared with Vit E. © 2019 Blackwell Verlag Gmb

Could high DNA stainability (HDS) be a valuable indicator of sperm nuclear integrity?

International audienceBackground: The Sperm Chromatin Structure Assay (SCSA®), in addition to identifying the DNA Fragmentation Index (DFI) also identifies High DNA satiability (HDS), supposed to reflect the nuclear compaction of spermatozoa. However, data on what exactly this parameter reveals, its relevance and usefulness are contradictory. In order to shed light on this situation, spermatozoa of a cohort (N = 397) of infertile men were subjected to the SCSA®, TUNEL (terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate-biotin nick end labeling) and CMA3 (Chromomycin A3) tests. In a smaller subcohort (N = 100), aniline blue (AB) and toluidine blue (TB) staining were performed in addition. The objective of this study was thus to answer the question of whether HDS is a relevant and reliable parameter to be taken into account?Results: HDS does not appear to be a reliable indicator of nuclear immaturity because it shows a weak correlation with the CMA3, AB and TB stains. The low correlation of HDS with sperm DNA fragmentation (TUNEL and SCSA®) and DNA condensation (CMA3, AB and TB) tests suggests that these two parameters could be decoupled. Unlike DFI and TUNEL, HDS has not been shown to correlate with classic clinical situations of male infertility (asthenozoospermia, teratozoospermia or astheno-teratozoospermia).Conclusion: HDS correlates poorly with most tests that focus specifically on the level of maturity of the sperm nucleus. To our knowledge, this study is the first to compare SCSA®, TUNEL, AB, TB and CMA3 assays on identical samples. It shows the potency, consistency and limitations of each test and the care that must be taken in their interpretation

Maternal Health-Related Quality of Life and Its Predicting Factors in the Postpartum Period in Iran

Background/Purpose. Postpartum period is accompanied by many physical, emotional, and social changes in women's health. The aim of this study was to examine the mothers' quality of life in postpartum period. In addition, it also sought to recognize the variables that predict their quality of life. Methods. This cross-sectional, descriptive study was undertaken among 380 women in 10 urban health centers in Ilam province in west of Iran. They were selected using proportional random sampling method. The SF-36 questionnaire was used to identify women's health-related quality of life (HRQoL). Data were analyzed using SPSS version 15. Results. Women who were employed, aged less than 30 years, had college degree, have no history of disease in pregnancy, and had given birth more than 3 months ago had higher quality of life scores. Independent predictors for lower physical HRQoL were being with history of disease in pregnancy; being with high school diploma or lower education; and giving birth less than 3 months ago. Also, independent predictor for lower mental HRQoL was being housewife. Conclusion. According to study findings, greater attention must be paid to providing postpartum healthcare for housewife and less educated women as well as those with history of disease in pregnancy