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1.
Transl Androl Urol ; 10(1): 397-407, 2021 Jan.
Article in English | MEDLINE | ID: mdl-33532327

ABSTRACT

BACKGROUND: Conventional cryopreservation methods induce chemical and mechanical damage to the sperm membranes. The cryoprotectant potential of phospholipids of vegetal origin as soybean lecithin has been investigated as a substitute for egg yolk in diluents used for the cryopreservation of human spermatozoa. Therefore, the objective of this study was comparing the efficacy of a synthetic cryoprotectant supplemented with L-α-phosphatidylcholine (PC) and L-acetyl-carnitine (ANTIOX-PC) and the standard egg-based TEST-yolk buffer (TYB) in preserving sperm motility and chromatin quality in cryopreserved semen samples. METHODS: Prospective experimental study in which semen samples from 63 men with normal sperm motility and 58 men with low sperm motility were included and analyzed both before and after cryopreservation using ANTIOX-PC or TYB freezing media. Sperm quality was evaluated by routine semen analysis and DNA fragmentation index using the Terminal deoxynucleotidyl transferase dUTP nick end labeling assay. RESULTS: Differences in the post-thaw progressive motility and DNA fragmentation index were not detected between TYB and ANTIOX-PC cryoprotectants in both normal and low sperm motility groups (P>0.05). However, ANTIOX-PC medium retained higher non-progressive motility and lower percentage of immotile sperm when compared to TYB medium, resulting in a greater total motile sperm count (P<0.05), regardless baseline values of motility characteristic of the normospermic or asthenozoospermic samples. CONCLUSIONS: ANTIOX-PC medium was effective to protect human sperm during a freeze-thaw cycle compared to the TYB medium. A clinically relevant advantage in better preserving kinetic parameters as higher total motility and lower immotile post-thawed sperm from ANTIOX-PC, in normal and low motility semen samples, demonstrated the positive impact of phospholipid and antioxidant treatment on sperm cryotolerance with high potential for egg yolk lipids replacement and biosafety.

2.
Fertil Steril ; 114(4): 770-778, 2020 10.
Article in English | MEDLINE | ID: mdl-32709382

ABSTRACT

OBJECTIVE: To study the global DNA methylation pattern in spermatozoa of patients with varicocele as well as investigate their semen quality. DESIGN: Prospective observational case-control study. SETTING: University-affiliated hospital. PATIENT(S): A total of 26 men with varicocele and 26 fertile men without the disorder. INTERVENTIONS: Analysis of semen quality and sperm DNA methylation patterns. MAIN OUTCOME MEASURE(S): Semen quality evaluated by semen analysis, and sperm DNA methylation patterns investigated using the Infinium MethylationEPIC BeadChip platform. RESULT(S): Men with varicocele displayed decreased semen quality. The sperm DNA methylation analysis showed that men with varicocele exhibit global hypomethylation in comparison with the control group. A total of 59 differentially methylated CpG sites were identified, most of them hypomethylated in the varicocele group. In regional analyses, 1,695 DNA regions were differentially methylated in men with varicocele. These regions show associations with gamete generation, meiotic and meiosis cell cycle, and semen quality based on gene ontology analysis. CONCLUSION(S): Gene ontology results suggest that changes in methylation may be associated with the low semen quality phenotype observed in some varicocele patients because the observed differentially methylated regions in varicocele patients are related to male reproductive pathways. Additionally, the varicocele grade may influence the magnitude of global sperm DNA methylation change. To our knowledge, this is the first report analyzing changes at a regional or CpG-specific level in men with varicocele.


Subject(s)
DNA Methylation/physiology , Semen Analysis/methods , Spermatozoa/physiology , Varicocele/diagnosis , Varicocele/genetics , Adult , Humans , Male , Spermatozoa/cytology , Varicocele/physiopathology
3.
Syst Biol Reprod Med ; 65(2): 95-104, 2019 Apr.
Article in English | MEDLINE | ID: mdl-30601031

ABSTRACT

Varicocele pathophysiology is related to increased oxidative stress, which might result in loss sperm DNA integrity as well as in genomic instability. Sperm telomere shortening and loss of global DNA methylation are the main features of genomic instability, leading to cell senescence and death, whereas sperm DNA fragmentation (SDF) characterizes the loss of chromatin integrity. We hypothesize that sperm genomic stability and DNA integrity is reduced in infertile men with moderate and large-sized varicoceles, thus being candidate markers of sperm quality in varicocele-related infertility. Here, we assessed the sperm global DNA methylation, telomere length, and SDF in men with and without clinically palpable varicoceles. While the rates of SDF and telomere length were not statistically different between varicocele patients and controls, global sperm DNA methylation seems to be lower in men with varicocele (49.7% ± 20.7%) than controls (64.7% ± 17.1%). A negative correlation between SDF and sperm motility and a positive correlation between sperm morphology and telomere length were observed. Our results suggest that varicocele may result in genomic instability, in particular, global DNA hypomethylation. However, a large sample size may confirm these findings. The understanding of the molecular mechanisms involved in the pathophysiology of varicocele-related infertility may help to better select candidates for varicocele repair.


Subject(s)
DNA Methylation , Telomere , Varicocele/genetics , Adult , Cross-Sectional Studies , DNA Fragmentation , Humans , Infertility, Male/genetics , Male , Spermatozoa/metabolism
4.
J Assist Reprod Genet ; 34(7): 839-847, 2017 Jul.
Article in English | MEDLINE | ID: mdl-28523408

ABSTRACT

Varicocele is found in approximately 20% of adults and adolescents and in 19-41% of men seeking treatment for infertility. It is associated with a decrease in sperm count as well as sperm motility and morphology. The currently accepted description of the pathophysiology of varicocele does not explain all its clinical manifestations; therefore, other factors such as genetic and epigenetic changes, associated with the environment, might be involved in causing infertility and decrease in sperm quality. It has been reported that the varicocele-induced deterioration of testicular function is progressive and interferes with fertility; hence, early and efficient assessment of the genetic manifestations in patients would be important for developing future medical interventions. Chromosomal disorders, mutations, polymorphisms, changes in gene expression, and epigenetic changes have all been reported to be associated with varicocele. Several studies are underway to unravel the genetic basis of this disease, as it is important to understand the origin and the aggravating factors to ensure appropriate guidance and intervention. Here, we review the available literature regarding the genetic and epigenetic changes associated with varicocele, and how these alterations are related to the different clinical manifestations of the disease.


Subject(s)
Epigenesis, Genetic , Infertility, Male/complications , Varicocele/genetics , DNA Fragmentation , DNA, Mitochondrial/chemistry , Humans , Infertility, Male/genetics , Male , Oxidative Stress , Semen Analysis , Varicocele/pathology , Varicocele/physiopathology
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