Background: Male infertility is a problem that affects 10–15% of men of reproductive age. In particular, gametogenesis is a complex process in which inflammation may play a central role through the secretion of cytokines and the expression of microRNAs. We assessed the potential role of proinflammatory cytokines (TNF-α, IL-6 and IL-1α) and microRNAs (miR-146a-5p, miR-34a-5p and miR-23a-3p) in the seminal plasma of infertile men compared to controls, evaluating their correlation with seminal and biochemical parameters. Methods and results: Expression of cytokines and microRNAs was analyzed by ELISA and q-PCR. Our data shows that IL-1α was significantly increased in the azoospermic group compared to controls, TNF-α mRNA was more expressed in the oligozoospermic group than controls. There were no significant differences in miRNAs expression among the three groups. The correlations between sperm parameters and inflammatory markers were evaluated, however no significance was highlighted. Conclusions: The determination of each inflammatory marker separately in the seminal plasma of subfertile men, despite some significant differences, does not have a diagnostic value in male infertility even if an assay of selective pro-inflammatory cytokines and microRNAs in the semen may improve the diagnosis of male infertility.

The abnormal production of matrix metalloproteinases (MMPs), especially MMP-9 and MMP-2, plays a pivotal role in hypertensive disorders of pregnancy, and as such, can influence the development of preeclampsia. These alterations may result from functional genetic polymorphisms in the promoter region of MMP-9 and MMP-2 genes, which modify MMP-9 and MMP-2 expression. We investigated the association of MMP-9 polymorphism rs3918242 (-1562 C>T) and MMP-2 polymorphism rs2285053 (-735 C>T) with the risk of preeclampsia. This case–control study was conducted on 345 women with preeclampsia and 281 age-matched women with normal pregnancies from Tunisian hospitals. Genomic DNA was extracted from whole blood collected at delivery. Genotypes for -1562 C>T and -735 C>T polymorphisms were performed using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). An increased frequency of heterozygous MMP-9 -1562 C/T genotype carriers was observed in women with preeclampsia compared to healthy controls (p = 0.03). In contrast, the MMP-2 -735 C>T polymorphism was not significantly different regarding frequency distribution of the allele and genotype between healthy pregnant women and women with preeclampsia. Our study suggests that the MMP-9 -1562 C/T variant, associated with high MMP-9 production, could be a genetic risk factor for preeclampsia in Tunisian women.

Study question: The aim of the study was to investigate risk factors for sperm DNA fragmentation to determine which factors influence significantly the sperm DNA fragmentation. Summary answer: Obesity seems to lead to higher risk of sperm DNA damage in Tunisian subfertile men. What is known already: A positive association between lifestyle conditions, varicocele, advanced age and exposure to toxicants and DNA fragmentation has been documented. Study design, size, duration: It’s a retrospective case control study. 61 Patients registered from October 2018 to December 2021 were divided into two groups: group “C” with good quality sperm DNA (n = 20) and group “F” with fragmented sperm DNA (n = 41). Participants/materials, setting, methods: DNA fragmentation was measured using terminal deoxynucleotidyltransferase dUTP nick and labeling assay. The Odds Ratio (OR) and their 95% CI were calculated using univariate logistic regression in order to quantify the association between the variable of interest “Sperm DNA fragmentation” and the various risk factors: lifestyle conditions (e.g. tobacco smoking, alcohol consumption, and obesity), age, professional exposure to high temperature and toxic products and urological history (e.g. varicocele, testicular hypotrophy or hypertrophy). Main results and the role of chance: Our results showed that tobacco, professional exposure to high temperature and toxic products and urological history are risk factors that may alter sperm DNA with an odds ratio (OR) of 2.1, 1.9 and 1.5, respectively. Interestingly, we demonstrated that obesity seems to be the most significant risk factor of DNA fragmentation with an OR of 4.2 (p = 0.002). Limitations, reasons for caution: Further studies are needed to investigate the mechanism by which obesity occur sperm DNA damage. Wider implications of the findings: Our finding are in accordance with several retrospective studies demonstrating that obesity is liable to sperm DNA damage Trial registration number: not applicable

In vitro manipulation of human spermatozoa during Assisted Reproductive Technology (ART) can induce several damages to sperm structure and functions. This study investigated the protective effects of Raphanus sativus seed extract and its active compounds on several sperm parameters during in vitro incubation and cryopreservation. Extracts from five seed-batches were characterized by HPLC-DAD-MS and ¹H-NMR, identifying sinapine and sinipic glycosides as the main characteristic compounds. Sperm DNA fragmentation (sDF) was detected by the Sperm Chromatin Dispersion test and LiveTUNEL. Excessive reactive oxygen species (ROS) production was detected by MitoSOX Red in viable spermatozoa. Caspase activity was detected by FLICA. Cryopreservation was conducted with two alternative freezing media. In vitro incubation with the extract protected against the loss of motility and reduced the induction of sDF, sperm ROS production, and caspase activity. Similarly, during cryopreservation, it allowed much better recoveries of sperm viability, motility, and DNA integrity by decreasing sperm ROS production with both freezing media. Sinapine and sinapic acid completely mimicked the protective effects of the whole extract during both in vitro incubation and cryopreservation, suggesting that they are included among the active principles. These findings support Raphanus sativus seed extract and its active compounds as candidates for inclusion in handling and freezing media for human spermatozoa in ART.