Comparison of SPAG11A gene expression in infertile men with grade 1 and 2 varicocele before and after treatment.

OBJECTIVE: Sperm Associated Antigen 11A (SPAG11A) protein is a family of the epididymis-specific secretory proteins implicated in sperm maturation and function. Varicocele might cause pathophysiological difficulties in the testis and epididymis, with a harmful effect on the environment for spermatogenesis and sperm maturation. The aim of this study was to evaluate the expression level of the SPAG11A gene and sperm parameters in infertile men with grade 1 and 2 varicocele before and after treatment. METHODS: Semen specimens were collected from 20 infertile men with varicocele pre-and post-treatment and 10 healthy volunteers. Semen analysis was conducted according to world health organization guidelines. Real time PCR (qRT-PCR) reaction was applied for determination of SPAG11A mRNA expression. RESULTS: The results showed that there was a significant difference between the concentration and normal morphology between pre- and post-treatment groups and the controls. There were significant differences between pre-treatment and control groups in terms of progressive and non-progressive mobility. SPAG11A mRNA levels were significantly lower in the pre-treatment group than in healthy control subjects (p=0.007). There was no statistically significant difference in the expression of SPAG11A as well as semen parameters in the post-treatment group compared to the pre-treatment group. CONCLUSIONS: SPAG11A gene expression and semen parameters may be affected by varicocele. Whether varicocele treatment is an effective approach to reduce the adverse effect of this disease on SPAG11A expression and semen parameters needs further investigation.

Possibility of intrauterine transmission from mother to fetus/newborn: Systematic review and meta-analysis of diagnostic methods to detect SARS-CoV-2 infection.

Several studies have reported vertical transmission of SARS-CoV-2; however, information regarding intrauterine transmission based on diagnostic methods to detect SARS-CoV-2 infection is scarce. A systematic review and meta-analysis was conducted to identify and explore the studies that attempt to ascertain the possibility of intrauterine transmission of SARS-CoV-2 infection according to the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA 2020) statement. The results demonstrate that SARS-CoV-2 can be transmitted intrauterine, as detected by clinical manifestations (1.00, 95 % CI: 1.00 - 1.00, 0.51, 95 % CI: 0.22 - 0.80), imaging (0.50, 95 % CI: 0.24 - 0.76, 0.03, 95 % CI: 0.00 - 0.17), molecular (1. 00, 95 % CI: 1.00 - 1.00, 0.92, 95 % CI: 0.77 - 1.00), immunological (0.32, 95 % CI: 0.10 - 0.57, 0.34, 95 % CI: 0.11 - 0.61), and histological approaches (0.79, 95 % CI: 0.52 - 0.98) in maternal and fetal/neonatal specimens, respectively. The possibility of intrauterine transmission of SARS-CoV-2 from mother to fetus/newborn was 41 % (95 % CI 0.37 - 0.45). We might confirm/verify the intrauterine transmission of SARS-CoCV-2 from mother to fetus/newborn.

Quality of testicular spermatozoa improves with changes in composition of culture medium.

BACKGROUND: Spermatozoa retrieved from the testis and epididymis are deprived of the beneficial effects of seminal fluid. Thus applying an artificial medium with normal seminal fluid characteristics, known as artificial seminal fluid (ASF), may provide an appropriate condition for improving some sperm parameters in azoospermia. The objective was to investigate the impact of in vitro exposure of testicular and epididymal spermatozoa to ASF on sperm quality. The study was conducted on testicular (n = 20) and epididymal (n = 20) sperm specimens obtained from azoospermic men. Each sample was divided into two equal parts: Part I) for processing and incubation with Ham's F10 medium; Part II) for processing and incubation with ASF. RESULTS: After 2 h incubation, testicular sperm motility was significantly higher in ASF than in Ham's F10 medium. In comparison to 0 h, mitochondrial membrane potential levels of testicular spermatozoa were significantly higher after 2 h and 24 h in ASF and after 24 h in Ham's F10 medium. Furthermore, the data indicated significantly lower rates of epididymal spermatozoa with high MMP in both media after 24 h. There were no significant differences in the DNA fragmentation index of testicular and epididymal spermatozoa between ASF and Ham's F10 medium at different time points. CONCLUSION: The results demonstrated that in vitro incubation of testicular spermatozoa improved their motility more effectively than Ham's F10 medium in the short term (2 h), but had no effect on epididymal spermatozoa. Since the physiology of testicular spermatozoa is different from that of ejaculated spermatozoa, it seems that a special environment should be designed and used for each of them.

RéSUMé: CONTEXTE: Les spermatozoïdes prélevés dans les testicules et les épididymes sont privés des effets bénéfiques du liquide séminal. Ainsi, l'utilisation d'un milieu artificiel avec des caractéristiques normales du liquide séminal, connu sous le nom de liquide séminal artificiel (ASF), peut constituer une condition appropriée pour améliorer certains paramètres des spermatozoïdes obtenus dans l'azoospermie. L'objectif était d'étudier l'impact de l'exposition in vitro de spermatozoïdes testiculaires et épididymaires à l'ASF sur la qualité du sperme. L'étude a été menée sur des échantillons de spermatozoïdes testiculaires (n = 20) et épididymaires (n = 20) obtenus chez des hommes azoospermiques. Chaque échantillon a été divisé en deux parties égales: Partie I) pour le traitement et l'incubation avec le milieu F10 de Ham; Partie II) pour la transformation et l'incubation avec l'ASF. RéSULTATS: Après 2 h d'incubation, la mobilité des spermatozoïdes testiculaires était significativement plus élevée dans l'ASF que dans le milieu F10 de Ham. Par rapport à 0 h, les niveaux du potentiel de membrane mitochondriale (PMM) des spermatozoïdes testiculaires étaient significativement plus élevés après 2 h et 24 h dans l'ASF, et après 24 h dans le milieu F10 de Ham. En outre, les données ont indiqué des taux significativement plus faibles de spermatozoïdes épididymaires avec un PMM élevé dans les deux milieux après 24 heures. Il n'y avait pas de différences significatives dans l'indice de fragmentation de l'ADN des spermatozoïdes testiculaires et épididymaires entre l'ASF et le milieu F10 de Ham aux différents temps. CONCLUSION: Les résultats ont montré que l'incubation in vitro de spermatozoïdes testiculaires dans l'ASF améliorait leur mobilité plus efficacement que le milieu F10 de Ham à court terme (2 h), mais n'avait aucun effet sur les spermatozoïdes épididymaires. Étant donné que la physiologie des spermatozoïdes testiculaires est différente de celle des spermatozoïdes éjaculés, il semble qu'un environnement spécial devrait être conçu et utilisé pour chacun d'eux.

Improvement of human sperm properties with platelet-rich plasma as a cryoprotectant supplement.

PURPOSE: Platelet-rich plasma (PRP) is a remarkable substance, which involves the growth and proliferation of all cell types. As a source of growth factors, we evaluated whether sperm cryopreservation supplemented with PRP improves the rates of sperm motility, viability, and DNA integrity after vitrification compared with conventional cryo-medium. MATERIALS AND METHODS: 20 normal semen specimens were collected from healthy men. After swim-up preparation, each sample was divided into four aliquots. One, as control, received no treatment, and the other three experimental samples were treated with three different concentrations of PRP as cryoprotectant. Sperm parameters were examined before and after freezing procedure. RESULTS: PRP had no significant effect on sperm count. Meanwhile, the percentage of sperm progressive motility and viability in the PRP treated samples with 1×105 /µL concentration was significantly higher than control group. Besides, the rate of immotile sperms in these samples was significantly lower than the control. Sperm viability was significantly higher in the PRP samples at 1×105/µL concentration. In the case of DNA integrity, CMA3 staining showed that the lower PRP concentration was correlated with the higher rate of abnormal spermatozoa. SCD showed that the rate of abnormal sperms in the PRP samples with 1×105 /µL concentration was significantly lower than control group. CONCLUSIONS: This study showed a protective effect of PRP on human sperm quality at an optimized concentration after vitrification. Besides, the effects of PRP supplementation of sperms on successful fertility following sperm preservation will be of interest.

Single sperm vitrification with permeable cryoprotectant-free medium is more effective in patients with severe oligozoospermia and azoospermia.

Testicular sperm extraction (TESE) is an invasive surgery for achieving the spermatozoa in cases with azoospermia. In these patients, the number of retrieved spermatozoa is limited and the optimal cryo-storage is very critical for their fertility preservation. Therefore, single sperm vitrification has been introduced for preservation of low number of spermatozoa. The goal was to assess the efficacy of sperm freezing medium (SFM) and sucrose medium as cryoprotectants for single sperm vitrification in cases with severe oligozoospermia and azoospermia. A total of 20 ejaculates from severe oligozoospermia and 20 testicular samples from azoospermia were processed. Twenty-five sperm cells were collected using ICSI injection pipette and transferred to a cryoprotectant droplet placed on the Cryotech, then vitrified by plunging in liquid nitrogen. Sperm motility, viability, fine-morphology, mitochondrial activity and DNA fragmentation index (DFI) were assessed before and after vitrification. Sperm motility, viability and the percentage of cells with mitochondrial activity were significantly decreased after vitrification in both severe oligozoospermic and testicular samples in either cryoprotectants. However, the rates of post-warm sperm motility and the cells with mitochondrial activity increased significantly in sucrose medium in both severe oligozoospermic and testicular samples compared to SFM. In testicular samples, the DFI of spermatozoa vitrified in SFM was significantly higher than those vitrified with sucrose medium. Sperm motility, viability, mitochondrial activity, and DNA integrity were better preserved in sucrose medium than SFM after single cell vitrification. The presented method may be a useful candidate for successful freezing of individual sperm cells in clinical setting.

Impact of biological and artificial seminal fluids on sperm parameters and DNA status in asthenozoospermic ejaculates.

The chemical composition and physiological properties of seminal fluid (SF) affect sperm quality. The objective was to investigate the effects of in vitro exposure of artificial seminal fluid (ASF) and biological seminal fluid (SF) on sperm quality. Asthenozoospermic ejaculates (n = 20) were divided into two aliquots. The first aliquot was centrifuged for obtaining asthenozoospermic SF. The second aliquot was processed with density gradient centrifugation (DGC), and the pellet was diluted separately with following media: (a) ASF; (b) Ham's F10; (c) normozoospermic SF; and (d) asthenozoospermic SF. Sperm parameters and DNA status were assessed after DGC, as well as 2 and 24 hr after incubation. The data showed that sperm progressive motility, viability and DNA integrity were significantly higher in ASF than Ham's F10 medium immediately after DGC. At 2 and 24 hr, the progressive motility was significantly decreased in biological SF compared with ASF and Ham's F10. DNA fragmentation index (DFI) was significantly lower in normozoospermic SF than asthenozoospermic SF and Ham's F10 at time 2 hr. In conclusion, normal SF showed the protective role on sperm DNA structure. Moreover, ASF preserved sperm motility better than biological SF during 24 hr, despite being similar to normal SF regarding DNA integrity preservation in short time.

Impairment of sperm efficiency in mice following short-term nano-titanium dioxide exposure: An experimental study.

BACKGROUND: Titanium dioxide nanoparticles (TiO 2 NPs) are widely used in many compounds. Recent evidence has displayed some cytotoxic effects of TiO 2 NPs on male reproduction. OBJECTIVE: The effects of TiO 2 NP administration on sperm parameters and chromatin and seminiferous histopathology of male mice were investigated. MATERIALS AND METHODS: In this experimental study, 32 NMRI male mice (35 ± 3 gr, 8-12-week-old) were divided into four groups (n = 8/each): treated groups were fed orally with 2.5 (group I), 5 (group II) and 10 (group III) mg/kg/day TiO 2 NPs for 40 days and the control group received phosphate buffered saline. Sperm parameters, DNA integrity and chromatin quality were assessed using chromomycin A3, aniline blue, toluidine blue staining and TUNEL. Hematoxylin eosin staining was performed to measure spermatogenic cells and the total diameter of seminiferous tubules. Also, sex hormone and malondyaldehyde levels were measured. RESULTS: Abnormal sperm tails rose in group III (28.87 ± 4.91) in comparison with the control group (12.75 ± 3.95). However, chromomycin A3 staining and TUNEL showed higher levels in group III in comparison with the control group, whereas aniline blue and toluidine blue staining showed no differences. A significantly lower spermatogenesis index and lumen parameters were observed in group III. Leydig cell numbers, cellular diameters and the area of the seminiferous tubules were lower in the treated groups. The testosterone level was also lower in these groups and the percentage of malondyaldehyde in the seminal fluid was higher. CONCLUSION: Exact mechanisms of TiO 2 NPs are not clear; however, cytotoxic and genotoxic effects of TiO 2 NPs may relate to oxidative stress. Given their widespread use, TiO 2 NPs should be a public health focus of attention.

Analysis of GSTM1, GSTT1, and CYP1A1 in idiopathic male infertility.

Enzymes belonging to the glutathione-S-transferase (GST) and cytochrome P450 (CYP) families are involved in a 2-stage detoxification process of a wide range of environmental toxins and carcinogens. In order to investigate whether there is a genetic association of the biotransformation enzymes and idiopathic male fertility, we studied GSTT1, GSTM1, and CYP1A1*2A polymorphisms in 150 infertile men and 200 healthy men as controls from Northern Iran. Genotyping of the GSTT1 and GSTM1 genes were performed using the multiplex polymerase chain reaction (PCR). However, the CYP1A1 polymorphism was determined using PCR-restriction fragment length polymorphism (RFLP). The GSTM1 and GSTT1 null genotypes were present at frequencies of 0.61 and 0.34 in infertile cases, whereas in controls the frequencies were 0.33 and 0.17, respectively. Double-null genotype was found to be elevated among infertile men (odds ratio [OR] = 3.75, 95% confidence interval [CI] = 2.42-6.45; P < .0001). The frequency of TT, TC, and CC genotypes of CYP1A1 polymorphism in the controls were 42.5%, 45.5%, and 12%, respectively, while those in the infertile men were 38.7%, 48%, and 13.3%. The CYP1A1*2A did not display any association with male infertility. We observed an association between male infertility and the GSTM1 and GSTT1 null deletion, but not with the CYP1A1 polymorphism in North Iranian men with idiopathic infertility.

Administration of hepatocyte growth factor increases reelin and disabled 1 expression in the mouse cerebral cortex: an in vivo study.

Hepatocyte growth factor (HGF) and its receptor, c-Met, are widely expressed in the developing brain. HGF also known as scatter factor enhances cell proliferation and cell growth, and stimulates cell migration and motility. Neurons and glia produced in the neuroepithelium migrate along radial glial fibers into the cortical plate. Reelin, a glycoprotein which is produced by Cajal-Retzius cells in the marginal zone directs neuronal migration indirectly via the radial glial cells. It has been demonstrated that Disabled 1 functions downstream of reelin in a tyrosin kinase signal transduction pathway that controls appropriate cell positioning in the developing brain. In this study, administration of HGF on reelin and Disabled 1 expression in the cerebral cortex has been studied. Using Western blot, it was shown that the expression of reelin and Disabled 1 is increased in response to infusion of HGF when compared to control group. It is concluded that HGF is essential for reelin and Disabled 1 expression in the cerebral cortex of the newborn mouse. Moreover, this method may be applied to the other factors, allowing identification of molecules involved in neural cell migration.

Administration of anti-c-kit antibody into the cerebrospinal fluid leads to increased cell death in the developing cerebral cortex.

It is generally believed that during development, neurons are usually produced in excess. Cell death occurs in the developing nervous system. The survival of the developing neurons depends on many factors derived from the target sites, of which the neuronal trophic factors are by far the best known. Stem cell factor (SCF) and its receptor, c-kit, is expressed in cells of nervous system during development and adulthood. Although the role of SCF/c-kit in the nervous system is so far not clear, in vitro studies indicate that SCF/c-kit is trophic to certain neurons derived from neural crest and cerebral cortex. In this study the effects of anti-c-kit antibody on cell death in the newborn chick cerebral cortex have been investigated. Injection of anti-c-kit antibody into the cisterna magnum increased the number of cell death and resulted in thinning of the cerebral cortex as compared to that from the control group. It is concluded that SCF/c-kit is essential for cortical progenitor cell survival in the cerebral cortex. Moreover, this method may be applied to the other factors and different CNS regions, allowing identification of factors involved in cell death. It additionally re-emphasizes the importance of further investigations into the potential roles of SCF/c-kit signaling in neurodegenerative diseases.