F&S science最新文献

Objective: To investigate the association between Polycystic Ovary Syndrome (PCOS) and the rate of embryo development, using time-lapse monitoring systems (TLM), compared to a control group of women with mechanical (tubal) factor infertility.

Design: A retrospective case-control study conducted in a university affiliated IVF unit.

Patients: Women with PCOS undergoing in-vitro fertilization (IVF) treatments and those with non-PCOS controls with tubal factor infertility only. Development morphokinetic milestones were compared and analysis of covariance for time to distinct cell number as well as logistic mixed models to determine predictors for embryos over the 75^th percentile was performed.

Exposure: Not applicable.

Main outcome measures: Embryo development morphokinetic parameters in women with and without PCOS undergoing IVF treatments.

Results: The study included 791 embryos from 115 women, 364 embryos from 52 women with PCOS and 427 embryos from 63 women with non-PCOS controls with tubal factor infertility. The PCOS group was 4 years younger (30.07±6.03 vs. 34.08±4.84 years, p=0.002) and had higher number of oocytes retrieved (16.00 vs. 11.00, p<0.001), mature oocytes (11.00 vs. 7.00,<0.001) and fertilized oocytes (8.00 vs. 5.00, p<0.001). The PCOS and control groups demonstrated comparable clinical pregnancy rates (55.8% vs. 32.1%,p=0.053), miscarriage rate (12.5% vs. 11.8%, p=0.994) and live birth rate (48,8% vs. 31.2%, p=0.089). Morphokinetic parameters were comparable between the groups. While age was associated with later time to 5 and 8 discrete cells and start of blastulation (tSB), PCOS was only associated with later tSB, including tSB>75^th percentile.

Conclusion: This study demonstrated comparable IVF outcomes in women with PCOS and non-PCOS controls. An analysis of TLM data from these patients showed no evidence that PCOS negatively affects embryonic development rate in women undergoing IVF cycles.

Objective: To establish a murine model of chemotherapy-induced diminished ovarian reserve (DOR) and investigate residual fertility after chemotherapy exposure.

Design: Two different chemotherapy protocols were tested to establish a valid DOR model by comparing follicle densities in mice given either protocol versus physiological solution. An ovarian stimulation protocol was then selected from among different gonadotropins by counting the number of day-2 embryos obtained from normal mice. Finally, DOR mice were stimulated 5 and 8 weeks after chemotherapy with the chosen gonadotropin protocols and day-2 embryos were recovered after mating, as was ovarian tissue for further immunohistological analyses.

Subject: Seventy-two Naval Medical Research Institute (NMRI) mice.

Mean outcome measures: To compare day-2 embryo counts in both normal and chemotherapy-induced DOR mice. Ovarian histology and morphology were also investigated by follicle counting and classification, as was immunostaining for apoptosis (cleaved caspase-3), activation (phospho-Akt) and proliferation (Ki67).

Results: A dose of 12 mg/kg busulfan (Bu) + 120 mg/kg cyclophosphamide (Cy) was chosen to establish the DOR model, as it significantly reduced the ovarian reserve compared to both control mice (physiological solution) and the 1.2 mg/kg Bu + 12 mg/kg Cy protocol, without depleting it completely. When stimulated with 3.75 IU Menopur, normal mice produced significantly more embryos than DOR mice given 12 mg/kg Bu + 120 mg/kg Cy (41.40 ± 14.74 versus 23.67 ± 15.55 day-2 embryos). While the follicle count was statistically diminished after single-dose chemotherapy administration, the remaining follicles did not display any difference in terms of apoptosis, activation or proliferation rates.

Conclusion: We successfully established a chemotherapy-induced DOR model using 12 mg/kg Bu + 120 mg/kg Cy, as evidenced by lower, but not completely depleted, follicle numbers and fewer retrieved embryos. Histological study of ovarian tissue exposed to DOR-inducing chemotherapy revealed that surviving follicles were of the same quality as tissue not exposed to chemotherapy.

This study investigated whether luteinizing hormone receptor (LHR) expression varies in the granulosa cells of individual follicles according to the maturation stage of the oocytes harvested for assisted reproductive technology treatment. We observed minimal to no LHR messenger ribonucleic acid and protein expression in cumulus cells surrounding oocytes arrested in the germinal vesicle stage. Interestingly, their ability to mature was confirmed by rescue in vitro maturation, suggesting somatic cell LHR deficiency as a key factor for the retrieval of germinal vesicle oocytes in assisted reproductive technology procedures.

Objective: To determine if the oral gonadotropin-releasing hormone antagonist relugolix affects leiomyoma extracellular matrix production through the transforming growth factor-beta (TGF-β) pathway.

Design: Laboratory study.

Subjects: None.

Exposure: Exposure of human leiomyoma cells to TGF-β and/or relugolix.

Main outcome measures: Production of TGF-β, pSMAD2/3, SMAD2/3, collagen 1A1 (COL1A1), fibronectin (FN1), and versican (VCAN) in treated and untreated leiomyoma cells.

Results: Transforming growth factor-beta 3 production decreased at 24 hours with relugolix 10 nM (0.80 ± 0.09-fold) and 100 nM (0.86 ± 0.06-fold) and at 48 hours with relugolix 1 nM (0.86 ± 0.05-fold) and 100 nM (0.86 ± 0.06-fold). pSMAD2/3 production decreased at 24 hours with relugolix 1 nM (0.71 ± 0.01-fold), 10 nM (0.68 ± 0.01-fold), and 100 nM (0.41 ± 0.10-fold). Compared with relugolix treatment alone at the same concentration, combination treatment at 24 hours resulted in significantly increased COL1A1, FN1, and VCAN production with relugolix 1 nM, 10 nM, and 100 nM. At 48 hours, combination treatment resulted in significantly increased COL1A1, FN1, and VCAN production with relugolix 10 nM and 100 nM.

Conclusion: Relugolix regulated leiomyoma size by decreasing COL1A1, FN1, and VCAN production. This effect is at least partly through the TGF-β pathway.

Objective: To introduce an innovative non-contact method for denudation process of cumulus-oocyte complexes (COCs) for intracytoplasmic sperm injection (ICSI).

Design: We designed and fabricated novel acousto-hydrodynamic tweezers (AHT) to perform contactless denudation and tested them in mouse model. Cumulus removal efficiency, preimplantation development and live birth were assessed and compared to conventional manual pipetting denudation.

Subjects: Fourteen female B6D2F1/J mice (∼4 weeks of age), 9 male B6D2F1/J mice (6-12 weeks of age), and 28 CD-1 female mice (∼6 weeks of age) were used for experiment INTERVENTION AND METHODS: We designed a contactless platform for oocyte denudation based on the principle of focalized acoustic waves. We first investigate the acoustic intensity and thermal variability by measuring the surface displacement and temperature with a thermal camera to ensure a safe operation. COCs were denuded by conventional manual pipetting (MP) with 40 IU/ml hyaluronidase serving as control, or by AHTs with reduced amount of hyaluronidase (15 IU/ml). Piezo-ICSI was performed on both experimental and control groups. A triplicate of denudation and insemination experiments were performed. All embryos were monitored in time-lapse incubator. Embryo developmental rates were compared by chi-square test. Embryo morphokinetic timing as a continuous variable was compared by one-way ANOVA. Embryo transfers were performed on some blastocysts.

Main outcome measures: The procedural time for each denudation method was measured and compared. Fertilization, embryo development and morphokinetics, pregnancy and live birth rate were compared between control and experimental cohorts.

Results: Facile non-contact denudation is achieved without any damage to oocyte. Acoustic induced fluidic shear is the main contributor to COC denudation. The average denudation time per oocyte is reduced by 46% (15 seconds per oocyte for control versus 8 seconds per oocyte for AHT) while using a lower concentration of hyaluronidase. Piezo-ICSI on oocytes processed by MP and AHTs resulted in comparable survival (86.1% vs 85.3%, P=0.80), fertilization (96.7% vs 94.1%, P=0.09) and blastocyst rates (88.0% vs 81.3%, P=0.06). Embryo morphokinetics for both experimental and control cohorts were comparable, showing no impact of sound waves on the embryo development. Eventual delivery rates were also comparable between the MP and AHT cohort (51.3% vs 55.4%).

Conclusion: Acousto-hydrodynamic tweezers (AHTs) are used for contactless removal of the cumulus cells from the COCs prior to ICSI in an expedited, safe, and reliable manner. Embryo development outcomes confirm their safety and validate their potential for a comprehensive ICSI-on-chip device.

Objective: Hereditary hemochromatosis (HH) is a common genetic disorder characterized by iron overload, which, if undiagnosed, can lead to severe organ damage. There are 4 types of HH. Type 1 HH, the most common form, is primarily caused by a common variant in Western Europe (p.Cys282Tyr, C282Y, or c.845 G>A). It is generally preventable during in vitro fertilization if proper genetic testing is performed before implantation. Here, we demonstrated a direct detection and cost-effective approach using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) in preimplantation genetic testing (PGT) settings.

Design: We began by validating the assay with genomic deoxyribonucleic acid (DNA) from Coriell cell lines of known HFE C282Y genotypes, followed by testing patients' genomic DNA samples. After establishing the assay on genomic DNA, we extended the assay to whole-genome amplified DNA from embryo biopsies.

Subjects: The subjects include cell line samples and human specimens and human embryo biopsies.

Exposure: Patients and embryos either carried or did not carry the HFE C282Y variant in their genome. No intervention was applied.

Main outcome measures: The readout included the genotype of samples at the HFE C282Y locus and accuracy of PCR-RFLP results.

Results: An accuracy of >99% was achieved across 80 cell line samples, 38 patient samples, and 81 embryo biopsies.

Conclusion: In this study, we demonstrated the feasibility of using the PCR-RFLP approach to PGT. Specifically, we validated the assay for the HFE C282Y variant, the primary cause of type 1 hemochromatosis. The assay was tested on genomic DNA and DNA resulting from whole-genome amplification, achieving >99% accuracy, sensitivity, precision, and specificity. These results also suggest the possibility for extending the PCR-RFLP approach to cover a broader range of conditions, such as spinal muscular atrophy, to benefit more patients currently ineligible for testing at PGT laboratories.

Objective: To evaluate the uterine microbiome among women with endometrial polyps and submucosal fibroids and to compare results between endometrial sampling techniques.

Design: Patients with polyps or fibroids were prospectively recruited prior to hysteroscopy, while patients undergoing retrieval for planned oocyte cryopreservation were recruited prospectively as controls. Three specimen types obtained for each patient were the distal 5 mm of an embryo catheter passed to the uterine fundus (C), endometrial tissue from an endometrial biopsy (T), and formalin fixed paraffin embedded endometrial tissue from the same endometrial biopsy (FFPE). 16S rRNAgene amplicon sequencing was performed to analyze the structure of the endometrial microbiome.

Subjects: Thirty-seven participants including 28 women with polyps and/or fibroids and 9 controls.

Intervention/exposure: None MAIN OUTCOME MEASURE(S): Microbial taxonomic alpha and beta diversity; differential abundance of taxa RESULTS: Across all sample types, participants with polyps had higher microbial alpha diversity compared to controls (4.3 vs 5.1, q = 0.049), and microbial communities were significantly different (pairwise PERMANOVA pseudo-F = 2.1, q = 0.003). These differences were observed when examining C specimens alone (5.4 vs 6.4, q = 0.001; pairwise PERMANOVA pseudo-F = 2.5, q = 0.003), though did not reach significance when examining either T or FFPE specimens alone. Participants with fibroids had similar alpha diversity yet significant differences in beta diversity compared to controls in analyses combining all specimens (Pairwise PERMANOVA pseudo-F = 1.475, q = 0.030); however, these differences did not achieve significance when analyzing C, T, or FFPE specimens alone. When comparing C and T specimens vs. FFPE specimens overall, alpha diversity was significantly higher (q<0.001 and <0.001, respectively) and there were significant differences in beta diversity (q<0.003 and q<0.003, respectively). Analyses of C specimens generated a larger number of significantly differentially abundant taxa compared to other sampling methods. Although not statistically significant, relative abundance of putative pathogens was higher in participants with polyps compared to controls regardless of sampling technique.

Conclusion: Results of this exploratory study suggest significant microbial differences exist among patients with endometrial polyps vs. healthy controls. However, results varied by sampling technique, highlighting a need to identify optimal sampling methods prior to validating findings in larger prospective cohort studies.

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 were assessed. Spermatozoa from young (8-12 weeks old) or reproductively old (>14 months old) mice were treated with BGP-15 for 1 hour and assessed for sperm quality and preimplantation embryo development after in vitro fertilization. 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 was evaluated. Spermatozoa from patients undergoing assisted reproductive technology (ART) treatment were incubated in the optimized dose of BGP-15 for 30 minutes, and sperm quality was 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 10-μM BGP-15. Human spermatozoa were treated with BGP-15 at doses from 1 to 100 μM.

Main outcome measures: Sperm quality measures (mouse and human) included motility, mitochondrial membrane potential (JC-1 dye), deoxyribonucleic acid (DNA) fragmentation ("HALO" assay), and DNA oxidation (8-oxoguanine immunodetection). Mouse embryo and offspring measures included on-time development after in vitro fertilization, morphokinetic analysis, and blastocyst inner cell mass and trophectoderm cell number, and growth and development from birth to 21 days postnatally.

Results: BGP-15 increased sperm motility and mitochondrial membrane potential and decreased DNA oxidation in old mice. BGP-15 improved on-time development of 2-cell and blastocyst embryos and increased the inner cell mass 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% and prevented DNA fragmentation (by 45%) and oxidative damage (by 60%). In spermatozoa from men attending a fertility clinic, BGP-15 increased motility by 12% and reduced both DNA oxidation and fragmentation by >20%.

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