Dehydroepiandrosterone modulates the PTEN/PI3K/AKT signaling pathway to alleviate 4-vinylcyclohexene diepoxide-induced premature ovarian insufficiency in rats.

Dehydroepiandrosterone (DHEA) is frequently integrated as an adjuvant in over a quarter of controlled ovarian hyperstimulation (COH) protocols, despite the ongoing debate regarding its impact. This study aimed to evaluate the efficacy and mechanism of action of DHEA on ovarian follicular development and ovarian response in rats with varying ovarian reserves. The study involved 75 rats categorized into 15 distinct groups. The ovarian tissues of rats in both the normal ovarian reserve group and the premature ovarian insufficiency (POI) group, induced by 4-vinylcyclohexene diepoxide (VCD) injection, were subjected to histomorphological and biochemical analyses following the administration of DHEA, either alone or in combination with COH. Follicle counting was performed on histological sections obtained from various tissues. Serum concentrations of AMH and the quantification of specific proteins in ovarian tissue, including PTEN, PI3K, AKT, COX-2, caspase-3, as well as assessments of total antioxidant status and total oxidant status, were conducted employing the Enzyme-Linked Immunosorbent Assay (ELISA) method. The impact of DHEA exhibited variability based on ovarian reserve. In the POI model, DHEA augmented follicular development and ovarian response to the COH protocol by upregulating the PTEN/PI3K/pAKT signaling pathway, mitigating apoptosis, inflammation, and oxidative stress, contrary to its effects in the normal ovarian reserve group. In conclusion, it has been determined that DHEA may exert beneficial effects on ovarian stimulation response by enhancing the initiation of primordial follicles and supporting antral follicle populations.

Do we overlook predictive factors in Poseidon 1 patients? A retrospective analysis co-evaluating antral follicle counts & diameters.

BACKGROUND: An unexpected impaired ovarian response pertains to an insufficient reaction to controlled ovarian hyperstimulation. This deficient reaction is identified by a reduced count of mature follicles and retrieved oocytes during an IVF cycle, potentially diminishing the likelihood of a successful pregnancy. This research seeks to examine whether the characteristics of antral follicles can serve as predictive indicators for the unexpected impaired ovarian response to controlled ovarian stimulation (COS). METHODS: This retrospective cohort study was conducted at a tertiary university hospital. The electronic database of the ART (assisted reproductive technologies) center was screened between the years 2012-2022. Infertile women under 35 years, with normal ovarian reserve [anti-Müllerian hormone (AMH) > 1.2 ng/ml, antral follicle count (AFC) > 5] who underwent their first controlled ovarian stimulation (COS) cycle were selected. Women with < 9 oocytes retrieved (group 1 of the Poseidon classification) constituted the group A, whereas those with ≥ 9 oocytes severed as control (normo-responders) one (group B). Demographic, anthropometric and hormonal variables together with COS parameters of the two groups were compared. RESULTS: The number of patients with < 9 oocytes (group A) was 404, and those with ≥ 9 oocytes were 602 (group B). The mean age of the group A was significantly higher (30.1 + 2.9 vs. 29.4 + 2.9, p = 0.01). Group A displayed lower AMH and AFC [with interquartile ranges (IQR); AMH 1.6 ng/ml (1-2.6) vs. 3.5 ng/ml (2.2-5.4) p < 0.01, AFC 8 (6-12) vs. 12 (9-17), p < 0.01]. The number of small antral follicles (2-5 mm) of the group A was significantly lower [6 (4-8) vs. 8 (6-12) p < 0.01), while the larger follicles (5-10 mm) remained similar [3 (1-5) vs. 3(1-6) p = 0.3] between the groups. CONCLUSION: The propensity of low ovarian reserve and higher age are the main risk factors for the impaired ovarian response. The proportion of the small antral follicles may be a predictive factor for ovarian response to prevent unexpected poor results.

Reliability of hypo-osmotic swelling test on fresh and frozen-thawed ejaculated or testicular immotile sperm: A sibling oocyte study.

The viability of sperm is a crucial factor for achieving a successful pregnancy in intracytoplasmic sperm injection (ICSI) cycles. The aim of this study was to evaluate the accuracy of the hypo-osmotic swelling test (HOST) in fresh and frozen-thawed sperm samples of different origins (ejaculated/testicular). A retrospective analysis was conducted on the outcomes of 2167 oocytes subjected to ICSI using motile and immotile-HOST-positive sperm from 2011 to 2023. We evaluated embryonic development, as well as clinical, obstetric, and neonatal outcomes in four groups based on different sperm origins (ejaculated/testicular) and processing (fresh/frozen). When comparing the results of ICSI between motile and immotile-HOST-positive sperm within each group, it was observed that there were no significant differences in the outcomes for fresh samples. However, for frozen-thawed samples, fertilization rates and blastocyst development rates were significantly lower when ICSI was performed with immotile-HOST-positive sperm compared to motile sperm. Of note, clinical, obstetric, and neonatal outcomes were statistically similar across all groups. Our findings indicate that HOST is more reliable in fresh samples than in those subjected to the freeze-thaw process. Nonetheless, HOST is considered a safe method for selecting viable sperm in all subgroups.

Comparison of semen parameters in the same patients before and after diagnosis of COVID-19.

Clinical and histopathological evidence suggest that the male reproductive system may be negatively impacted in patients with coronavirus disease (COVID-19). The objective of this study is to investigate the effect of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) on semen parameters by comparing semen analyses before and after COVID-19 diagnosis in the same patient. We retrospectively analyzed 342 semen analyses by reviewing medical records. The study included two groups of patients: (i) those who underwent two consecutive semen analyses within 6 months, one before (n = 114) and one after (n = 114) COVID-19 diagnosis, and (ii) a control group (n = 114) that was age-matched and did not receive a diagnosis of COVID-19. The study results indicated a significant decrease in semen volume, total sperm count per ejaculate, progressive motile sperm count, total motile sperm count, and normal sperm morphology after SARS-CoV-2 infection in comparison to their respective values before the infection. Subgroup analyses showed that the duration of COVID-19 diagnosis (short-term vs. long-term) did not impact the changes in semen parameters. However, fever during the COVID-19 process had a negative effect on semen parameters, particularly sperm concentration, unlike in patients without fever. In conclusion, our findings suggest that SARS-CoV-2 infection is associated with a decline in semen quality, which may potentially impact male fertility. Furthermore, it's important to note that the negative effects on semen parameters may persist in the long-term. Our results also indicate that fever during active infection could be a significant risk factor that negatively affects spermatogenesis.

Low-Pressure Deuterium Storage on Palladium-Coated Titanium Nanofilms: A Versatile Model System for Tritium-Based Betavoltaic Battery Applications.

Deuterium (D2(g)) storage of Pd-coated Ti ultra-thin films at relatively low pressures is fine-tuned by systematically controlling the thicknesses of the catalytic Pd overlayer, underlying Ti ultra-thin film domain, D2(g) pressure (PD2), duration of D2(g) exposure, and the thin film temperature. Structural properties of the Ti/Pd nanofilms are investigated via XRD, XPS, AFM, SEM, and TPD to explore new structure-functionality relationships. Ti/Pd thin film systems are deuterated to obtain a D/Ti ratio of up to 1.53 forming crystallographically ordered titanium deuteride (TiDx) phases with strong Tix+-Dy- electronic interactions and high thermal stability, where >90% of the stored D resides in the Ti component, thermally desorbing at >460 °C in the form of D2(g). Electronic interaction between Pd and D is weak, yielding metallic (Pd0) states where D storage occurs mostly on the Pd film surface (i.e., without forming ordered bulk PdDx phases) leading to the thermal desorption of primarily DOH(g) and D2O(g) at <265 °C. D-storage typically increases with increasing Ti film thickness, PD2, T, and t, whereas D-storage is found to be sensitive to the thickness and the surface roughness of the catalytic Pd overlayer. Optimum Pd film thickness is determined to be 10 nm providing sufficient surface coverage for adequate wetting of the underlying Ti film while offering an appropriate number of surface defects (roughness) for D immobilization and a relatively short transport pathlength for efficient D diffusion from Pd to Ti. The currently used D-storage optimization strategy is also extended to a realistic tritium-based betavoltaic battery (BVB) device producing promising ß-particle emission yields of 164 mCi/cm2, an open circuit potential (VOC) of 2.04 V, and a short circuit current (ISC) of 7.2 nA.

Low Molecular Weight Heparin-Aspirin-Prednisolone Combination Does Not Increase the Live Birth Rate in Recurrent Implantation Failure: A Retrospective Cohort Study.

This study investigates the triple combination of adjuvants (low molecular weight heparin (LMWH)-aspirin-prednisolone) whether it improves the live birth rates of IVF&ICSI patients with previous implantation failure. This retrospective study included 1095 patients with >2 failed either fresh or frozen single embryo transfer cycles between 2014 Jan and 2021 Jan. Patients were divided into two subgroups. Group A consisted of patients with only vaginal progesterone for luteal phase support. Group B consisted of patients with triple (daily subcutaneous LMWH, daily 150 mg aspirin, and daily 16 mg prednisolone) luteal phase supplementation to vaginal progesterone. Demographic parameters, cycle characteristics, embryology, and pregnancy outcomes were compared, and the study's primary outcome was the live birth rate. Demographic parameters were similar between the groups. Positive b-hCG, miscarriage, and live birth rates were similar between groups as Group A vs. Group B, positive b-hCG 30.8% (190/617) vs. 35.4% (169/478), miscarriage rates 4.4% (27/617) vs. 6.7% (32/478), and live birth rates 20.4% (126/617) vs. 23.8% (114/478), respectively. When patients were stratified according to previous failures, live birth rates were still similar. Pregnancy outcomes were significantly improved in only patients with diminished ovarian reserve (Group A vs. Group B, positive b-hCG 24.2% vs. 34.3%, live birth rate 12.1% vs. 21.9%, p < 0.01). Whether the embryo transfer was fresh or frozen-thawed did not affect the results. A combined supplementation of LMWH, aspirin, and prednisolone in the luteal phase does not improve live birth rates of IVF&ICSI patients with previous implantation failure except potentially for patients with diminished ovarian reserve.

The Kisspeptin and Kisspeptin receptor in follicular microenvironment: is that really necessary for oocyte maturation and fertilisation?

The aim of this study was to determine whether Kisspeptin and Kisspeptin receptor in the follicular microenvironment is necessary for human oocyte maturation and fertilisation. The cumulus cell (CC) and follicle fluids (FF) obtained from the first aspirated follicles (n = 52) from 32 patients were divided into three groups considering nuclear maturation and fertilisation results of oocytes: (1) Metaphase I or germinal vesicle stage oocytes (incomplete nuclear maturation, n = 10), (2) unfertilised metaphase II oocytes (incomplete cytoplasmic maturation, n = 16), and (3) fertilised metaphase II oocytes (completed nuclear-cytoplasmic maturation, n = 26). The gene expression levels were assessed by RT-PCR. The levels of Kisspeptin (KISS1) and Kisspeptin receptor (KISS1R) were measured by ELISA. There were no significant efficacy KISS1 and KISS1R gene expressions in cumulus cells in terms of oocyte nuclear maturation stage (Group 1, vs Group 2 + Group 3) (respectively p = .49; p = .45). In terms of the cytoplasmic maturation stage (Group 2, vs Group 3); KISS1 and KISS1R expressions in CCs were comparable (respectively p = .07; p = .08). In FFs, KISS1 and KISS1R concentrations were similar between all groups (respectively p = .86; p = .26). In conclusion, the relative KISS1 and KISS1R expressions in CC and also KISS1 and KISS1R level of FF were independent of oocytes nuclear and/or cytoplasmic maturation. Impact statementWhat is already known on this subject? It has been demonstrated that Kisspeptin is an essential regulator of reproductive function and plays a key role in the modulation of GnRH secretion and gonadotropin release. Still, no information is available about the link between gene expression or concentration in the follicular microenvironment and oocyte development.What do the results of this study add? The study has shown that the relative Kisspeptin (KISS1) and Kisspeptin receptor (KISS1R) and expressions in cumulus cell (CC) and also KISS1 and KISS1R levels of follicle fluids (FF) were independent of oocytes nuclear and/or cytoplasmic maturation.What are the implications of these findings for clinical practice and/or further research? Based on the findings, it is difficult to establish a concept that kisspeptin can directly induce oocyte maturation. Nevertheless, to confirm these findings, further studies with a larger sample size are needed.

Which is more predictive ovarian sensitivity marker if there is discordance between serum anti-Müllerian hormone levels and antral follicle count? A retrospective analysis.

This retrospective study aims to determine the more predictive ovarian reserve marker when there is discordance between anti-Müllerian hormone (AMH) and antral follicle count (AFC) in patients with diminished ovarian reserve (DOR). Patients who underwent ICSI because of DOR were divided into three groups. Group 1: patients with low AMH (<1.1 ng/ml) and AFC (n < 7), group 2: patients with low AMH (<1.1 ng/ml) and normal AFC (n ≥ 7) and group 3: patients with normal AMH (≥1.1 ng/dl) and low AFC (n < 7). Demographic values, follicle output rate (FORT) score and follicle to oocyte index (FOI) score of the groups were compared. Totally, 662 cycles were enrolled in the study. There were 418 cycles in group 1, 167 cycles in group 2 and 77 cycles in group 3. As the primary result, FORT and FOI scores were higher in group 3 than the other two groups. Median FORT Score with quartiles: group 1: 100 (66-150), group 2: 71 (57-100), group 3: 136 (96-200), p<.01 - median FOI score with quartiles: group 1: 83 (50-140), group 2: 71 (40-100), group 3: 116 (66-216), p<.01. In conclusion, serum AMH level has more predictive value for stimulation success if there is discordance with AFC.Impact StatementWhat is already known on this subject? Female age, serum Anti-Müllerian Hormone (AMH) levels, and antral follicle count (AFC) are commonly used to assess ovarian reserve and predict response to ovarian stimulation. AMH and AFC are both positively correlated with ovarian reserve.What do the results of this study add? If there is discordance between AFC and AMH in patients with diminished ovarian reserve (DOR), the ovarian response is better in patients with high AMH and low AFC than the patients with low AMH and high AFC.What are the implications of these findings for clinical practice and/or further research? It is important to assess both AFC and AMH before controlled ovarian hyperstimulation, to predict ovarian response in DOR patients, rather than assessing AFC or AMH alone.

Fertilisation and early embryonic development of immature and rescue in vitro-matured sibling oocytes.

The objective of this study was to assess the effect of rescue in vitro maturation and immediate intracytoplasmic sperm injection (ICSI) application on fertilisation success and early embryonic development of metaphase I (MI) oocytes. This was a retrospective cohort study including 2425 sibling oocytes in 259 ICSI cycles. ICSI was performed on 104 GV (germinal vesicle) oocytes which had reached the metaphase II (MII) stage (Group 1) and 231 MI oocytes which had reached the MII stage (Group 2) following IVM (in vitro maturation). Immediate ICSI was applied following oocyte aspiration on 292 MI stage (Group 3) and 1798 MII stage oocytes (Group 4). Normal fertilisation rates in Groups 1, 2, 3 and 4 were 51.9%, 39%, 30.1% and 59.5%, respectively. The rates of blastocyst development per oocyte and per zygote were calculated as 3.8%, 3.0%, 6.8%, 14.1% and 7.4%, 7.7%, 22.7%, 23.6% for Groups 1, 2, 3 and 4, respectively. The blastocyst development rate was significantly higher in the MI-ICSI group compared with other immature oocytes. Even though performing ICSI on the oocytes at the MI stage on the day of oocyte aspiration resulted in lower fertilisation rates, it was associated with significantly higher rates of blastocyst development.

Supernumerary embryos, do they show the cycle success in a fresh embryo transfer? A retrospective analysis.

OBJECTIVE: To determine the predictive value of cryopreserved embryos during single fresh blastocyst transfer. METHODS: This retrospective cohort study was conducted at a tertiary university hospital ART center. Patients who underwent a single fresh blastocyst transfer were selected from the electronic database. Patients with diminished ovarian reserve, uterine pathology, advanced age (>40 years) were excluded from the study. Patients were divided into two groups. Group A consisted of patients with only one available embryo for the fresh transfer. Group B consisted of patients with cryopreserved embryos apart from the fresh transferred embryo. Demographic and embryology parameters, pregnancy results were compared. RESULTS: Demographic values, ovarian reserve parameters were comparable between groups. The number of the picked-up oocyte, metaphase-2 oocyte, and two-pronucleus embryo was lower in Group A. Positive pregnancy rates were two-fold higher in Group B (23.4% vs. 50.9% p < .01). Pregnancy results did not change depending on the number of cryopreserved embryos in Group B. CONCLUSION: According to our results, the presence of cryopreserved embryos results in higher pregnancy rates in patients with fresh blastocyst embryo transfer independent from the woman's age and ovarian reserve. Thus, these results may help us to predict the implantation rate before embryo transfer.

Luteal oestradiol for patients with serum oestradiol levels lower than expected per oocyte.

Although the efficiency of progesterone in providing luteal phase support has been established, the role of oestradiol supplementation during the luteal phase remains controversial. We evaluated pregnancy outcomes of patients who had a ratio of serum E2 levels on the hCG day to the number of oocytes retrieved (oestradiol/oocyte ratio - EOR) levels of <100 pg/ml by supporting them with additional oestradiol during the luteal phase. In total, 150 patients with an EOR < 100 pg/ml of oestradiol undergoing antagonist intracytoplasmic sperm injection (ICSI) cycles were randomly assigned into two groups to receive either oral oestradiol (4 mg/d) plus vaginal progesterone (90 mg, 2 × 1/day) (group 1) or vaginal progesterone (90 mg, 2 × 1/d) alone (group 2). Implantation rate following transfer of a single embryo did not differ between the two groups (group 1 = 33.3%; group 2 = 34.9%; p = 0.85). Similarly, both groups gave comparable clinical pregnancy rates per embryo transfer with 31.7% in group 1 compared with 28.6% in group 2 (p = 0.69). In conclusion the study suggests that the addition of 4 mg oral E2 to progesterone does not increase the probability of pregnancy.

Glutamatergic Activation of Neuronostatin Neurons in the Periventricular Nucleus of the Hypothalamus.

Neuronostatin, a newly identified anorexigenic peptide, is present in the central nervous system. We tested the hypothesis that neuronostatin neurons are activated by feeding as a peripheral factor and that the glutamatergic system has regulatory influences on neuronostatin neurons. The first set of experiments analyzed the activation of neuronostatin neurons by refeeding as a physiological stimulus and the effectiveness of the glutamatergic system on this physiological stimulation. The subjects were randomly divided into three groups: the fasting group, refeeding group, and 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX)+refeeding group. We found that refeeding increased the phosphorylated signal transducers and transcription activator-5 (pSTAT5) expression in neuronostatin-positive neurons and that the CNQX injection significantly suppressed the number of pSTAT5-expressing neuronostatin neurons. The second set of experiments analyzed the activation pathways of neuronostatin neurons and the regulating effects of the glutamatergic system on neuronostatin neurons. The animals received intraperitoneal injections of glutamate receptor agonists (kainic acid, α-amino-3-hydroxy-5methyl-4-isoazepropionic acid (AMPA), and N-methyl-D-aspartate (NMDA)) or 0.9% NaCl. The number of c-Fos-expressing neuronostatin neurons significantly increased following the AMPA and NMDA injections. In conclusion, we found that the neuronostatin neurons were activated by peripheral or central signals, including food intake and/or glutamatergic innervation, and that the glutamate receptors played an important role in this activation.

What is the effect of embryo morphology on serum ß-hCG levels?

OBJECTIVE: To determine the effect of embryonic factors on serum beta human chorionic gonadotropin (ß-hCG) levels in pregnancy and live birth resulting after a single fresh cleavage embryo and blastocyst transfer. STUDY DESIGN: This was a retrospective cohort study conducted at a tertiary care hospital. All fresh single embryo transfers (sETs) between September 2011 and December 2016 were included. The correlation analysis was performed to determine the association of embryo morphological parameters on mean serum ß-hCG levels on day 12 after the transfer of a fresh single cleavage embryo and a fresh single blastocyst embryo. RESULTS: Out of a total of 455 fresh sETs, 60 positive ß-hCG results after the transfer of a single fresh cleavage-stage embryo and 82 after the transfer of a single fresh blastocyst. The mean ß-hCG level resulting from a single fresh blastocyst ET was 371.7 ± 52.7 IU/L, which was similar to the mean ß-hCG level resulting from a cleavage ET (314.5 ± 36.9 IU/L) (p = .70). Interestingly, serum ß-hCG levels resulting from a single fresh blastocyst ET showed a correlation with day 5 blastocoele expansion, trophectoderm cell number and blastocyst quality score in ongoing pregnancy (r = .33, p = .02; r = .29, p = .04; and r = .31, p = .03, respectively). Moreover, day 5 blastocoele expansion and blastocyst quality score showed a correlation with the serum ß-hCG levels resulting from a single fresh blastocyst ET in live birth (r = .36, p = .02; r = .31, p = .04, respectively). CONCLUSION: Our study suggests that serum ß-hCG levels resulting from a single fresh blastocyst ET showed a correlation with day 5 blastocoele expansion and blastocyst quality score in both ongoing pregnancy and live birth.