Follicular Fluid Components in Reduced Ovarian Reserve, Endometriosis, and Idiopathic Infertility.

Follicular fluid (FF) molecules, and their increase or decrease, can contribute to appropriate follicular growth and oocyte maturation, thus being related to female infertility conditions. In this paper, we studied the changes and the relationships of some biochemical components, hormones, antioxidant enzymes, F2-Isoprostanes (F2-IsoPs), and resolvin (Rv) D1 in the FF of infertile women with different reproductive conditions such as endometriosis, reduced ovarian reserve, and idiopathic infertility during assisted reproductive techniques (ART). In the whole population, positive correlations between albumin (ALB)/iron (Fe), ALB/beta-2-microglobulin (B2MG), and F2-IsoPs/RvD1 were detected in the FF. In FF from aged women, increased levels of follicle stimulating hormone (FSH) and reduced anti-Müllerian hormone (AMH) levels were associated with a worse oocyte quality. The negative ART outcome was influenced by patient age and AMH, B2MG, and FSH levels. Moreover, the reduced ovarian reserve condition was characterised by a significant decrease in oocyte number and quality, AMH amount, and lactate dehydrogenase (LDH) activity, as well as by an increase in age and FSH levels. In the presence of endometriosis, high levels of MDA and RvD1 were detected in FF, with a decrease in luteinising hormone (LH). Finally, among the molecules examined, none characterised the condition of idiopathic infertility. These data could support the identification of new FF markers in different reproductive disorders, suggesting the need for personalised therapeutic approaches and optimised ART outcomes. In particular, the evaluation of resolvins and lipid mediators in FF could be a promising field of investigation with which to understand the entity of oxidative stress and inflammation in some female infertility conditions.

Evaluation of Known Markers of Ferroptosis in Semen of Patients with Different Reproductive Pathologies and Fertile Men.

This study aims to investigate the role of ferroptosis, an iron-dependent form of regulated cell death, in male infertility. The motivation behind this research stems from the increasing recognition of oxidative stress and iron metabolism dysregulation as critical factors in male reproductive health. In this study, 28 infertile patients (grouped by the presence of urogenital infections or varicocele) and 19 fertile men were selected. Spermiograms were performed by light microscopy (WHO, 2021). Testosterone, ferritin, transferrin-bound iron, transferrin, and F2-isoprostanes (F2-IsoPs) were detected in seminal plasma. Glutathione peroxidase 4 (GPX4) and acyl coenzyme A synthetase long chain family member 4 (ACSL4) were also assessed in sperm cells using enzyme-linked immunosorbent assays (ELISA). All the variables were correlated (statistically significant Spearman's rank correlations) in the whole population, and then the comparison between variables of the different groups of men were carried out. Seminal ferritin and transferrin positively correlated with seminal F2-IsoPs, which had positive correlations with ACSL4 detected in sperm cells. Ferritin and ACSL4 negatively correlated with the seminal parameters. No correlation was detected for GPX4. Comparing the variables in the three examined groups, elevated levels of ACSL4 were observed in infertile patients with urogenital infections and varicocele; GPX4 levels were similar in the three groups. These results suggested a mechanism of ferroptosis, identified by increased ACSL4 levels and the occurrence of lipid peroxidation. Such events appear to be GPX4-independent in reproductive pathologies such as varicocele and urogenital infections.

Semen Biochemical Components in Varicocele, Leukocytospermia, and Idiopathic Infertility.

The evaluation of the seminal plasma plays a relevant role in the definition of male infertility and in assisted reproduction outcomes; for this reason, it would be recommended to find biochemical markers able to characterize sperm pathology. In this study, 53 infertile patients (grouped by the presence leukocytospermia, idiopathic infertility, or varicocele) and 10 fertile men were selected. Spermiogram was performed by light microscopy, and sperm ultrastructure was evaluated by transmission electron microscopy (TEM) mathematically elaborated. Testosterone (TESTO), estradiol (E2), ferritin (FERR), iron (Fe), transferrin (TRSF), triglycerides (TRG), cholesterol (CHOL), and isoprostanes (F2-IsoPs) were detected in seminal plasma. Sperm characteristics and biochemical components were correlated by Spearman's rank correlation coefficient in the whole population and in each group. The levels of TESTO and E2 were positively correlated with sperm quality in particular, and E2 was correlated with fertility index expressing the number of sperm free of ultrastructural defects evaluated by TEM. On the contrary, the indices of iron metabolism (FERR, Fe, and TRSF) were positively associated with low sperm quality and sperm necrosis, particularly in leukocytospermia and varicocele groups, pathologies in which an inflammatory status and oxidative stress condition are present. The study of the seminal plasma composition deserves attention because the levels of the various components seem to be associated with specific reproductive pathologies.

Effect of hydrostatic pressure of various magnitudes on osteoarthritic chondrocytes exposed to IL-1beta.

BACKGROUND & OBJECTIVES: Several in vitro studies have shown the importance of mechanical compression or hydrostatic pressure (HP) as a modulator of cartilage metabolism. The present study was undertaken to evaluate the in vitro effects of cyclical low HP (1-5 MPa) and continuous high HP (24 MPa) applied in the presence or absence of interleukin (IL)-1beta on human osteoarthritis (OA) chondrocytes. METHODS: Chondrocytes obtained from OA cartilage were cultivated for 48 h and then exposed to pressurization in the presence or absence of IL-1beta. After pressurization, the culture medium was collected to detect the amount of proteoglycans (PG) and nitric oxide (NO) and the chondrocytes were immediately fixed for transmission electron microscopy (TEM) and processed for immunocytochemistry to localize the inducible nitric oxide synthase (iNOS). RESULTS: A significant increase in the level of PG and a small, non-significant, decrease in NO production were observed upon exposure to cyclical low HP. On the other hand, exposure to continuous high HP resulted in a significant decrease in the PG levels and a significant increase in NO production. The presence of IL-1beta led to a significant decrease in PG levels as well as a significant increase in NO production. The cyclical low HP did not increase the PG levels significantly but caused a statistically significant decrease in NO production in cultures damaged with IL-1beta. The continuous high HP in chondrocyte cultures stimulated with IL-1beta did not significantly decrease PG production, but significantly increased NO production. The results concerning metabolic production were further confirmed by morphological findings obtained by TEM and immunocytochemical studies. INTERPRETATION & CONCLUSION: The findings of this study confirmed that the response of chondrocytes varies with magnitude and frequency of HP. These findings are important to understand aetiopathogenetic mechanisms of OA and to find out which type of physical activity may be best suited for the prevention and therapy of OA.

Associations between biochemical components of human semen with seminal conditions.

The aim of the study was to assess whether abnormal levels of seminal biochemical components could be associated with semen alterations and infertility. In this study, 92 human ejaculates from selected men were analyzed. Albumin, estradiol, ferritin, total proteins (TP), folic acid (FA), vitamin B12, alkaline phosphatase (ALP), creatine kinase (CK), gamma-glutamyl transpeptidase (GGT), lactate dehydrogenase were evaluated. Semen parameters and biochemical components of the 92 samples were correlated bySpearman's rho coefficient. Albumin showed a negative correlation with sperm progressive motility and vitality (P < 0.05), CK with sperm concentration and vitality (P < 0.05), ferritin with sperm morphology (P < 0.05). FA negatively correlated with sperm concentration (P < 0.05) and GGT with sperm motility (P < 0.05). The values of biochemical components were compared for each semen parameters (concentration, motility, morphology, vitality) in samples ≤5th percentile with those >5th percentile and in patients with/without leukocytospermia, presence/absence of germ cells, increased/normal viscosity by Mann Whitney U test. The albumin (P < 0.001) and TP (P < 0.05) levels and the GGT activity (P < 0.001) were significantly higher in patients with sperm motility ≤5th percentile. Patients with sperm vitality ≤5th percentile showed increased albumin concentration (P < 0.01) and the CK activity (P < 0.001). The presence of germ cells in semen was concomitant with high values of ferritin (P < 0.01); the ALP activity (P < 0.01) and FA level (P < 0.001) were decreased in hyperviscous semen. The FA and estradiol levels were significantly decreased in the smoker group compared to those measured in the non-smoker group. Subjects were grouped in infertile patients and men with unknown reproductive potential. Infertile patients albumin and ferritin were significantly increased (P < 0.05). This study suggests that some biochemical components may be associated with human seminal pathological conditions. Abbreviations: ALP: alkaline phosphatase; LDH: lactate dehydrogenase; GGT: γ-glutamyl transferase; CK: creatine kinase; ACP: acid phosphatase; ALB: albumin; TP: total proteins; FERR: ferritin, E: estradiol; FOL: folic acid; B12: vitamin B12; FSH: follicle stimulating hormone; LH: luteinizing hormone; T: testosterone; BMI: body mass index; WHO: World Health Organization.