F&S science最新文献

Objective: To study the efficacy of mitochondrial activator BGP-15 to preserve sperm quality and competence against cellular damage.

Design: Spermatozoa from mice or humans were treated in vitro with BGP-15 and sperm quality markers assessed. Spermatozoa from young (8-12 weeks old) or reproductively old (>14 months old) mice were treated with BGP-15 for 1h and assessed for sperm quality and pre-implantation embryo development after in vitro fertilization (IVF). The safety of BGP-15 on offspring outcomes was assessed through embryo transfers. In parallel studies, spermatozoa from healthy (not infertile) men were incubated in hydrogen peroxide, to induce oxidative stress, plus increasing doses of BGP-15 , and sperm quality evaluated. Spermatozoa from patients undergoing assisted reproductive technology treatment (ART) were incubated in the optimized dose of BGP-15 for 30 min and sperm quality assessed.

Subjects and animals: C57BL/6 mice (N=4-15 per group) for sperm quality and embryo development. CBAF1 mice (n=6 per group) produced embryos for transfer. Human spermatozoa were from men with no infertility diagnosis (n=14-20), or men undergoing ART (n=33) at a local fertility clinic.

Exposure: Mouse spermatozoa were treated with 10μM BGP-15. Human spermatozoa were treated with BGP-15 at doses from 1μM to 100μM.

Main outcome measures: Sperm quality measures (mouse and human): motility, mitochondrial membrane potential (JC-1 dye), DNA fragmentation ('HALO' assay) and DNA oxidation (8-OHdG immunodetection). Mouse embryo and offspring measures: on-time development after IVF, morphokinetic analysis and blastocyst inner cell mass and trophectoderm cell number; growth and development from birth to 21 days post-natal.

Results: BGP-15 increased sperm motility, increased mitochondrial membrane potential and decreased DNA oxidation in old mice. BGP-15 improved on-time development of 2-cell and blastocyst embryos, and increased ICM blastomere number. Embryos from BGP-15-treated mouse spermatozoa produced normal offspring. In human spermatozoa subjected to in vitro oxidative stress, BGP-15 increased motility by 45% (p=0.03) and prevented DNA fragmentation (by 45%; p<0.0001) and oxidative damage (by 60%; p<0.0001). In spermatozoa from men attending a fertility clinic, BGP-15 increased motility by 12% (p=0.02), and reduced both DNA oxidation and fragmentation by >20% (p<0.05).

Conclusion: BGP-15 protects sperm against cellular damage and has the potential to improve ART outcomes.

Objective: To investigate whether endometrial receptivity is affected in patients with endometriosis using podocalyxin (PCX) as a functional biomarker; to study how endometriotic lesions display PCX and the potential pathological implications.

Design: We have previously reported that PCX, an anti-adhesion glycoprotein and barrier protector, is dynamically regulated in the endometrium and acts as a key negative regulator of epithelial receptivity. Early in the cycle both luminal epithelium (LE, lining the endometrial surface) and glandular epithelium (GE, residing within the tissue) strongly express PCX, but in the receptive window PCX is selectively down-regulated in LE, switching the endometrial surface to an adhesive state for embryo attachment/implantation; meanwhile PCX expression is maintained in GE until post-receptivity. Here, we immuno-stained PCX in endometrial tissues and ectopic lesions biopsied across the menstrual cycle from patients with endometriosis (EOS, n=41), and compared to endometrium of non-endometriosis controls (Non-EOS, n=55). We further investigated how PCX changes observed in ectopic lesions may influence their adhesive capacity.

Subjects: Women without and with endometriosis.

Exposure: N/A.

Main outcome measures: The window of endometrial receptivity might be shorter in patients with endometriosis; ectopic sites also down-regulate PCX cyclically, mirroring the eutopic endometrial cells in preparing for receptivity to increase their adhesion potential.

Results: Endometrial PCX levels were comparable between non-EOS and EOS early in the cycle, and in both groups PCX is down-regulated in LE during the expected window of receptivity, however, in EOS endometrium PCX is reduced earlier in GE as if the receptive window were shorter. In endometriotic lesions, PCX was detected in endometrial LE- and GE-like cells plus mesothelial cells enveloping peritoneal organs, but PCX was cyclically lost specifically in LE-like cells and reduced in GE-like cells as seen in the eutopic endometrium, which however may increase their adhesion potential to nearby organs (overlaid by mesothelial cells). This speculation was further corroborated in an in vitro model showing endometrial epithelial cells with lower PCX were indeed more adhesive to mesothelial cells.

Conclusion: Endometrial receptivity is subtly affected in patients with endometriosis with a shorter window. Cyclic down-regulation of PCX in ectopic sites may have pathological consequences.

Objective: To determine whether the bioactive sphingolipid sphingosine 1-phosphate (S1P) modulates cellular proliferation and synthesis of fibrotic proteins in leiomyoma differently than myometrial cells.

Design: A basic science study using human leiomyoma and myometrial cells.

Setting: Academic laboratory.

Exposure: Leiomyoma and myometrial cells were treated with S1P, as well as with selective antagonists for S1P specific G-protein coupled receptors (S1PR) and secondarily with inhibitors of extracellular-signal regulated kinase 1/2 (ERK1/2) and ezrin.

Main outcome measure(s): Cellular proliferation and fibrogenesis. Bromodeoxyuridine (BrdU) cell proliferation assay was employed to measure DNA synthesis and proliferation, while Western Blot analysis was used to assess the expression of the fibrotic markers N-cadherin, alpha Smooth Muscle Actin (αSMA), transgelin and Collagen type I alpha 1.

Result(s): S1P stimulates cellular proliferation of leiomyoma but not myometrial cells. The mitogenic effect elicited by S1P relies on the specific engagement of its specific receptor S1P₂ and is mediated by ERK1/2 and ezrin activation. Furthermore, S1P exerts a pro-fibrotic effect in a S1PR-dependent manner in leiomyoma but not myometrial cells.

Conclusion(s): These results, beside extending the knowledge on the molecular mechanism underlying uterine leiomyoma development and fibrosis, demonstrate the pathogenetic role of S1P in leiomyoma and support the rationale for targeting S1P signaling pathway as innovative potential treatment.

Objective: To investigate causal associations between blood-based inflammatory markers and female infertility using Mendelian randomization (MR).

Design: Mendelian randomization using genome-wide association study data.

Setting: Publicly available genome-wide association study data.

Patient(s): Large female-only cohorts of European ancestry.

Intervention(s): Blood-based inflammatory markers (C-reactive protein, interleukins, monocyte chemoattractant protein-1, tumor necrosis factor-α, interferon-γ).

Main outcomes measure(s): Anovulatory infertility (1,054 cases and 117,098 controls); female infertility of other/unspecified origin (5,667 cases and 117,098 controls); and medical treatment for female infertility (2,706 cases and 120,873 controls). Total causal effects were assessed using univariable two-sample methods including inverse variance weighted (IVW) as the primary analysis, as well as other secondary analyses (MR-Egger, weighted median, etc.), with relevant quality assessments.

Result(s): Interleukin-8 demonstrated a positive association with anovulatory infertility via IVW (odds ratio, 95% confidence interval; 1.51, 1.04-2.21) and weighted median (1.64, 1.05-2.57) methods. Monocyte chemoattractant protein-1 was associated with anovulatory infertility via MR-Egger (2.06, 1.13-3.77). Inverse associations were found for interleukins-12 and -18 via IVW, with higher interleukin-12 being associated with lower medical treatment for female infertility (0.75, 0.59-0.94), whereas higher interleukin-18 was associated with lower female infertility of other/unspecified origin (0.90, 0.83-0.97).

Conclusion(s): This is the first study to examine causal relationships between inflammation and female infertility using MR. Monocyte chemoattractant protein-1 and interleukin-8 are implicated in anovulatory infertility; however, only the relationship with interleukin-8 was evident in the primary analysis. Interleukins-12 and -18 demonstrated inverse associations with infertility outcomes. Further research is needed to uncover the mechanistic functions of these markers to confirm causality and examine their therapeutic potential for female infertility.

Objective: To investigate whether idiopathic nonobstructive azoospermia (iNOA) has its own transcriptomic signature.

Design: Testicular tissue biopsies were retrieved, processed, and prepared for ribonucleic acid (RNA) extraction from 26 consented patients diagnosed with iNOA. Samples were grouped into four pools based on the presence of testicular spermatozoa: two replicate pools for "No presence" (Null-spz-1 and Null-spz-2 pools), one for "High presence" (High-spz pool), and one for "Rare presence" (Rare-spz pool). A second set of replicate pools (CF-1 and CF-2) were used from patients with obstructive azoospermia (OA) and served as controls. RNA sequencing (RNA-seq) and comparative transcriptomics analysis were performed, followed by differential gene expression analysis focused on protein-coding genes only. Differentially expressed genes (DEGs) exclusively upregulated or downregulated were further analyzed using the Gene Ontology (GO), STRING, and Kyoto Encyclopedia of Genes and Genome bioinformatic platforms.

Setting: Private Fertility Clinic and Public University.

Patients: Males in whom iNOA was diagnosed.

Exposure: Testicular biopsies from men in whom iNOA was diagnosed.

Main outcome measures: Protein-coding DEGs.

Results: A significantly altered transcriptomic profile of protein-coding genes was identified in the testicular tissues from men with iNOA. A total of 3,858 genes exhibited dysregulated expression, with 1,994 genes being exclusively downregulated and 1,734 upregulated. Biological processes such as male gamete generation (GO:0048232) and meiotic cycle (GO:0051321) were significantly enriched by the downregulated DEGs whereas the upregulated DEGs enriched BPs such as regulation of cell death (GO:0010941), regulation of cell adhesion (GO:0030155), and defense response (GO:0006952). Interactome analysis identified hub genes among the downregulated DEGs, including PCNA, PLK1, MCM4, CDK1, CCNB1, AURKA, CCNA2, and CDC6, and among the upregulated DEGs, including EGFR, RELA, CTNNB1, MYC, JUN, SMAD3, STAT3 NFKB1, TGFB1, and ACTB. In addition, Kyoto Encyclopedia of Genes and Genome analysis demonstrated that pathways such as cell cycle (hsa04110) and oocyte meiosis (hsa04114) are primarily affected by the downregulated genes, whereas the upregulated genes mainly affected pathways such as the focal adhesion (hsa04510) and the PI3-Akt signaling pathway (hsa04151).

Conclusion: A distinct messenger RNA expression profile and altered transcriptomic activity were identified in the testicular tissues of men with iNOA.

Clinical trial registration number: University of Thessaly 1, 15.04.2016 and the Greek National Authority 701/15.9.2017.

Objective: To investigate the follicular fluid (FF) phthalate levels in adolescents undergoing fertility preservation compared with oocyte donors and explore its association with ovarian reserve and cumulus cell (CC) gene expression.

Design: Retrospective study and molecular analysis of CCs and FF.

Setting: Not applicable.

Patient(s): Adolescents (n = 20, 16.7 ± 0.6 years) undergoing fertility preservation and oocyte donors (n = 24, 26.2 ± 0.4 years).

Intervention(s): Not applicable.

Main outcome measure(s): Patient demographics, ovarian stimulation, and oocyte retrieval outcomes were analyzed for each group. The FF levels of 9 phthalate metabolites were assessed individually and as molar sums representative of common compounds (all phthalates, ƩPhthalates; di(2-ethylhexyl) phthalate (DEHP)-associated phthalate metabolites, ƩDEHP), exposure sources (plastics, ƩPlastic; personal care products, ƩPCP), and modes of action (antiandrogenic, ƩAA) and compared between the 2 groups. The transcriptome of CC associated with mature oocytes was compared between adolescents and oocyte donors using bulk ribonucleic acid sequencing.

Result(s): The FF ƩPlastic and ƩPCP levels were significantly higher in adolescents than in oocyte donors. The FF ƩDEHP, ƩPlastic, ƩPCP, ƩAA, and ƩPhthalates levels were positively associated with antral follicle count in oocyte donors when adjusted for age, body mass index, and race/ethnicity. Ribonucleic acid sequencing analysis revealed 248 differentially expressed genes in CCs of adolescents within the top quartile (n = 4) of the FF ƩPhthalates levels compared with those of the adolescents within the bottom half (n = 9). Genes enriched in pathways involved in cell motility and development were significantly down-regulated.

Conclusion(s): Adolescents undergoing fertility preservation cycles demonstrate higher levels of phthalate metabolites in their FF than oocyte donors. Higher phthalate levels are associated with alterations in cumulus cells transcriptome in adolescents. The phthalate metabolite levels in FF are associated with higher antral follicle count levels in oocyte donors.