Background and Objectives: Sperm DNA fragmentation refers to any break in one or both of the strands of DNA in the head of a sperm. The most widely used methodologies for assessing sperm DNA fragmentation are the sperm chromatin structure assay (SCSA), the sperm chromatin dispersion assay (SCD), the single-cell gel electrophoresis assay (SCGE–comet), and the terminal-deoxynucleotidyl-transferase (TdT)-mediated dUTP nick end labelling (TUNEL) assay. The aim of this study was to compare the efficiency and sensitivity of the analysis of sperm DNA fragmentation using TUNEL via fluorescence microscopy, and flow cytometry. Materials and Methods: Semen samples were collected and analyzed for standard characteristics using light microscopy, and for sperm DNA fragmentation using both TUNEL via fluorescence microscopy, and flow cytometry. Results: There were no significant differences in the values of the sperm DNA fragmentation index (DFI) obtained when the analysis was performed using TUNEL or flow cytometry (p = 0.543). Spearman’s correlation analysis revealed a significant negative correlation between sperm motility (%) and sperm DNA fragmentation (p < 0.01), as well as between sperm concentration and sperm DNA fragmentation (p < 0.05). The Mann–Whitney U test showed no significant difference in the DFI among couples with repeated implantation failure (RIF) and miscarriages (p = 0.352). Conclusions: Both methods (TUNEL via fluorescence microscopy, and flow cytometry) have a high efficiency and sensitivity in accurately detecting sperm DNA fragmentation, and can be effectively used to assess male fertility.

Background and Objectives: Ankylosing spondylitis (AS) is a condition that affects 0.1% to 0.5% of the adult population. The aim of this case report was to investigate the possible effects of the drugs taken for treatment of AS as well as mRNA vaccination for COVID-19 on semen quality by performing a highly detailed analysis. Materials and Methods: Sperm characteristics were examined by light microscopy, DNA fragmentation (DFI) was analysed by flow cytometry and morphology was evaluated by transmission electron microscopy (TEM). Results: Semen analysis under therapy with (1) celecoxib and sulphasalazine showed: concentration 47 million/mL, 53% progressive motility, 7% normal morphology and 9.6% DFI, (2) Golimumab and before mRNA Vaccination showed: concentration 108 million/mL, 82% progressive motility, 1% normal morphology and 7.6% DFI, and (3) Golimumab and after 3 doses of mRNA Vaccination showed: concentration 142 million/mL, 85% progressive motility, 1% normal morphology and 6.8% DFI. TEM revealed head, neck and tail abnormalities, as well as the presence of cells with incomplete spermiogenesis white cells and phagocytes in the sample under therapy with celecoxib and sulphasalazine. Golimumab treatment lead to an increased incidence of elongated heads but in general reduced inflammation as no white cells were evident in TEM. Conclusion: The anti-inflamatory drugs celecoxib and sulphasalazine had no adverse effect on sperm quality as all parameters were within normal limits and the patient achieved under that treatment 2 pregnancies following natural conception that lead to the birth of a healthy boy and girl respectively. Anti-TNFa treatment with Golimumab exerted a negative effect on morphology but not on concentration, motility and DFI. After 3 doses of mRNA Vaccination, sperm concentration increased while motility, morphology and DFI remained similar to the values before vaccination suggesting no negative effect of the mRNA vaccine for COVID-19 on sperm quality.

Background/Objectives: Conventional Coagulation Tests (CCTs) fail to accurately reflect Preeclampsia’s (PE’s) coagulation status, disease progression, and hemostatic alterations. They do not differentiate between the normal hypercoagulability of healthy pregnancies and the pathological hypercoagulability associated with PE. Rotational Thromboelastometry (ROTEM) analyzes clot dynamics from initiation through amplification and propagation to termination and fibrinolysis. However, their application in PE, particularly in neonates born to women with PE, is limited. We aimed to identify the hemostatic alterations in pregnant women with PE using ROTEMs that remain undetected by CCTs and to assess PE’s impact on neonatal hemostasis at birth. Methods: This was a single-center observational study (March 2022–March 2024) including 31 women with PE (34 newborns) and 45 pregnancies without PE (47 newborns). Maternal blood was collected intrapartum before placental delivery. Neonatal arterial samples were obtained within the first hour of life before vitamin K administration. ROTEM (Intrinsic (INTEM), Extrinsic (EXTEM), Fibrinogen (FIBTEM), Aprotinin (APTEM)), and CCTs were performed. Subgroup analyses considered PE severity and onset. ROC analyses examined discrimination for persistent maternal thrombocytopenia within 7 days of delivery and association with maternal platelet transfusion. Results: In preeclamptic women, the INTEM and FIBTEM assays were more affected, with higher Actual Clot Firmness (ACF) (p = 0.03, p = 0.04, respectively) and a higher Clot Formation Rate (CFR) (p = 0.03, p = 0.02, respectively). Hyperfibrinolysis was present (CT-APTEM < CT-EXTEM, MCF-APTEM > MCF-EXTEM). Clot Formation Time CFT-EXTEM was an indicator of maternal platelet transfusion (AUC = 0.81). Across EXTEM, INTEM, and APTEM, A10 (Amplitude at 10 min) and CFT showed good discrimination capability for maternal persistent thrombocytopenia within 7 days of delivery (AUCs 0.82–0.95). Neonates of women with PE presented lower ACF across all assays (INTEM p = 0.003; EXTEM p = 0.001; FIBTEM p = 0.01; APTEM p < 0.001), consistent across severity/onset subgroups. Conclusions: In this cohort, ROTEM identified maternal hypercoagulability with hyperfibrinolysis and neonatal hypocoagulability during the first hour of life. Several alterations were not reflected in CCTs. Further prospective studies should evaluate the role and clinical utility of combining ROTEM with CCTs for hemostatic monitoring in women with PE and their neonates.

Background/Objectives: Platelet transfusions are administered to preterm neonates with thrombocytopenia prophylactically to decrease their bleeding risk. The amplitude difference between the extrinsic rotational thromboelastometry (EXTEM) and the fibrinogen rotational thromboelastometry (FIBTEM) assays is considered an index of platelet contribution to clot strength, guiding transfusion management. The difference in maximum clot elasticity (MCE) (namely the platelet contribution to clot elasticity—MCEplatelet) is considered highly accurate. Limited data exist to specify the contribution of platelets and fibrinogen in clot formation during sepsis in neonates with thrombocytopenia. We investigated the potential of MCFplatelet (platelet contribution to clot firmness) and MCEplatelet in reflecting platelet count and function in septic preterm neonates. We simultaneously assessed the contribution of both platelets and fibrinogen to clot strength during sepsis. Methods: We compared 28 preterm neonates with sepsis born (gestational age 24⁺¹-34⁺³) with 30 healthy counterparts by using rotational thromboelastometry (ROTEM) and platelet flow cytometry. Results: MCEplatelet showed a higher association with platelet count in the sepsis group than MCFplatelet (R² = 47.66% vs. R² = 18.79%). MCEplatelet (AUC = 0.81) had better discrimination capability than MCFplatelet (AUC = 0.78) in platelet count <100 × 10³/L. MCEplatelet was poorly associated with platelet function. The contribution of platelets was significantly lower (MCEplatelet = 84.03 vs. 89.21; p < 0.001) compared with fibrinogen (36.9 vs. 25.92; p < 0.001) in the sepsis group. Conclusions: MCEplatelet has a better predictive value than MCFplatelet. In clinical practice, the elasticity difference between EXTEM and FIBTEM may replace the amplitude difference. The higher contribution of fibrinogen in clot strength during neonatal sepsis results in higher MCF, even in neonates with thrombocytopenia.

Background/Objectives: Preeclampsia (PE) may be associated with maternal endothelial activation and altered early neonatal inflammation. The primary objective was to compare maternal and early neonatal serum interleukin 2 (IL2), IL6, IL8, tumor necrosis factor α (TNFα), and endothelin 1 (ET1) concentrations between PE and control mother–neonate pairs. Methods: This secondary report from a single-center observational cohort included 31 women with PE (34 neonates) and 45 control women (47 neonates). Biomarker concentrations were log-transformed, and PE–control differences were estimated using linear models, with results expressed as geometric mean ratios (GMRs). Neonatal models used pregnancy-clustered standard errors to account for twins. The Benjamini–Hochberg procedure was applied to control the false discovery rate (FDR). Additional neonatal models adjusted for gestational age at birth served as sensitivity analyses. Selected biomarker–outcome associations were explored post hoc using univariable Firth bias-reduced logistic regression. Results: Maternal ET1 was higher in PE (GMR = 1.79, 95% confidence interval—CI—1.53–2.09; q < 0.001). Neonatal IL2 (GMR = 1.37, 95% CI 1.11–1.71; q = 0.016) and TNFα (GMR = 1.49, 95% CI 1.22–1.82; q < 0.001) were higher in PE-exposed neonates. After gestational age at birth adjustment, the TNFα difference persisted (GMR = 1.33, 95% CI 1.14–1.55; q = 0.003), whereas IL2 was attenuated (GMR = 1.20, 95% CI 0.99–1.45; q = 0.164). None of the exploratory Firth associations remained significant after FDR correction. Thrombocytopenia at birth occurred in 5/33 PE-exposed and 1/47 control neonates; intraventricular hemorrhage occurred in 2/34 and 2/47, respectively. Conclusions: PE was associated with between-group differences in maternal endothelial and early neonatal inflammatory biomarkers, with maternal ET1 and neonatal TNFα as the most robust signals. Complication-related findings remain hypothesis-generating and do not support predictive cut-offs, risk stratification, or causal inference.