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The cryopreservation of gametes and embryos has increased notably over the past 20 years and is now an essential part of assisted reproductive technologies (ARTs). However, because the cryopreservation process is un-physiological for human cells, gametes, and embryos, cryobiologists have suggested diverse methods to successfully cryopreserve human gametes and embryos in order to maintain their viability and assure successful pregnancy. During the first period of early development, major waves of epigenetic reprogramming-crucial for the fate of the embryo-occur. Recently, concerns relating to the increased incidence of epigenetic anomalies and genomic-imprinting disorders have been reported after ARTs and cryopreservation. Epigenetic reprogramming is particularly susceptible to environmental and un-physiological conditions such as ovarian stimulation, embryo culture, and cryopreservation that might collectively affect epigenetics dysregulation. Additionally, recent literature suggests that epigenetic and transcriptomic profiles are sensitive to the stress induced by vitrification, osmotic shock, oxidative stress, rapid temperature and pH changes, and cryoprotectants; it is therefore critical to have a more comprehensive understanding of the potential induced perturbations of epigenetic modifications that may be associated with vitrification. The aim of this paper is to present a critical evaluation of the association of gamete and embryo cryopreservation, use of cryoprotectants, and epigenetic dysregulations with potential long-term consequences for offspring health., (© 2024. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

Background: The usefulness of endometrium strain elastosonography (SE) for the evaluation of endometrial receptivity in women undergoing in vitro fertilization (IVF) remains controversial. The objective of this prospective, observational study was to evaluate the correlation between endometrial thickness (EMT) and its related strain (ESR) on the day of ovulation triggering (hCG-d) and in vitro fertilization outcomes. Additionally, 3D Power Doppler vascular indices (3DPDVI) were also analysed., Methods: We included all the patients undergoing fresh IVF-single blastocyst transfer cycle from January 2021 to August 2021 at our center. On hCG-d, after B-mode scanning was completed to measure the EMT, the mode was changed to elastosonography to evaluate the ESR (ratio between endometrial tissue and the myometrium below). At the end of examination, the Endometrial Volume (EV) and 3DPDVI (vascularization index [VI], flow index [FI] and vascularization flow index [VFI]), were assessed. Statistical analysis was completed using STATA MP16 software., Results: A total number of 57 women were included. Based on the EMT on hCG-d, women were divided into two groups, Group 1: <7 mm and Group 2 ≥ 7 mm. Women with EMT < 7 mm had a significantly higher ESR (p = 0.004) and lower pregnancy rate (p = 0.04). Additionally, low ESR values were correlated with high VFI values (rho = -0.8; 95% CI = -0.9- -0.6; p < 0.0001) and EMT ≥ 7 mm could be predicted by low ESR (OR = 0.01; 95% CI = 0.01-0.30; p = 0.008, area under the ROC curve: 0.70). After all, in multiple logistic regression analysis, low values of ESR (p = 0.050) and high values of EMT (p = 0.051) on hCG-d had borderline statistical effects on pregnancy rate., Conclusions: The ESR may be useful to improve the ultrasound evaluation of the endometrial quality in infertile women candidates to IVF/ICS. Given the small sample size of our study, the usefulness of strain elastosonography in this patients, needs further investigation., (© 2023. BioMed Central Ltd., part of Springer Nature.)

Background: In recent times, different novel GnRH-antagonist protocols with various combinations of gonadotropins and other molecules (e.g., aromatase inhibitors, selective estrogen receptor modulators) have been proposed for expected normal ovarian responders undergoing assisted reproductive treatments. The purpose of this study was to evaluate the effectiveness of a novel ovarian stimulation protocol based on the combination of corifollitropin-alfa plus five days of letrozole in E-NOR women undergoing IVF as compared with a daily recombinant-FSH regimen. Methods: We conducted a retrospective-controlled study on 182 couples undergoing their first IVF attempt. In Group A (experimental), letrozole (2.5 mg daily) was administered from day 2 (up to day 6 of the cycle), followed by corifollitropin-alfa on day 3 and daily recombinant FSH from day 10. In Group B, recombinant FSH from day 2 were administered (150 IU-225 IU daily). Statistical analysis was completed using SPSS Statistics. The primary outcome was the total number of MII oocytes retrieved. Results : Group A showed similar results compared to Group B in terms of MII oocytes, live birth, implantation, and clinical pregnancy rates ( p = ns). Nevertheless, the experimental group was associated with a trend towards a higher number of developing follicles, total oocytes, and embryos ( p < 0.05) with lower estradiol and progesterone values at ovulation induction compared to Group B, resulting in an increased chance of performing a fresh embryo transfer ( p < 0.05). Conclusions : The combination of CFα plus five days of letrozole was associated with a trend towards a higher number of developing follicles, total oocytes, and obtained embryos. Moreover, the experimental protocol resulted in lower estradiol and progesterone values at ovulation induction compared to daily rFSH, with an increased chance of performing a fresh embryo transfer (with no OHSS occurrence). Given the observational design of our study, further well-conducted RCTs are needed.

Chiral β-nitro alcohols are key intermediates in the synthesis of a wide range of active pharmaceutical ingredients. Despite their massive use for pharmaceutical applications, in-depth kinetics studies concerning their stability during formation and transformation reactions are scarce in the literature. In this study, the (1R,2S)-1-(m-benzyloxy)-2-nitro-1-propanol) (BNA), the precursor of the metaraminol, was selected as a molecular model and the retro-Henry reaction was explored by a multidisciplinary approach involving HPLC, electronic circular dichroism and theoretical methods. The enantio-, diastereo-, and chemo-selective high-performance liquid chromatographic method for determining the purity of β-nitro alcohol during its formation and degradation is based on the use of an amylose-derived chiral stationary phase under normal-phase eluent conditions. The influence of various factors (e.g. temperature, type of reaction solvent, basic and acid catalysts) on the degradation kinetics has been investigated. The retro-Henry reaction was found to be the major degradation of BNA, under spontaneous, solvent- and base-catalyzed conditions, resulting in the formation of its precursors 3-benzyloxybenzaldehyde and nitroethane., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier B.V. All rights reserved.)

A set of nine planar chiral 1,2-ferrocenes was analyzed by high-performance liquid chromatography (HPLC) on the amylose tris(3,5-dimethylphenylcarbamate) chiral stationary phase. The enantioseparations were carried out using neat methanol, ethanol, 1-propanol, and 2- propanol as well as mixtures of n-hexane-2-propanol as mobile phases. The differences in retention times between the second eluted (Rp)-enantiomers and the first eluted (Sp)-enantiomers were significantly influenced by elution modes and the steric hindrance of substituents at the aromatic rings of the ferrocene backbone. It has been demonstrated an ON/OFF switching of receptor-like chiral discrimination through the employment of different alcohols as mobile phases. In particular, the presence of pure 2-propanol triggers exceptional conditions of enantioselectivity that for some ferrocenes result in values of the enantioseparation factor higher than 80., (Copyright © 2022 Elsevier B.V. All rights reserved.)

Wingless/integrase-11 (WNT)/β-catenin pathway is a crucial upstream regulator of a huge array of cellular functions. Its dysregulation is correlated to neoplastic cellular transition and cancer proliferation. Members of the Dishevelled (DVL) family of proteins play an important role in the transduction of WNT signaling by contacting its cognate receptor, Frizzled, via a shared PDZ domain. Thus, negative modulators of DVL1 are able to impair the binding to Frizzled receptors, turning off the aberrant activation of the WNT pathway and leading to anti-cancer activity. Through structure-based virtual screening studies, we identified racemic compound RS4690 ( 1 ), which showed a promising selective DVL1 binding inhibition with an EC 50 of 0.74 ± 0.08 μM. Molecular dynamic simulations suggested a different binding mode for the enantiomers. In the in vitro assays, enantiomer ( S )- 1 showed better inhibition of DVL1 with an EC 50 of 0.49 ± 0.11 μM compared to the ( R )-enantiomer. Compound ( S )- 1 inhibited the growth of HCT116 cells expressing wild-type APC with an EC 50 of 7.1 ± 0.6 μM and caused a high level of ROS production. These results highlight ( S )- 1 as a lead compound for the development of new therapeutic agents against WNT-dependent colon cancer.

Normal-phase and reversed-phase high-performance liquid chromatography methods for the separation of the active pharmaceutical ingredient escitalopram from its (R)-enantiomer impurity have been developed on the cellulose-based Chiralcel OJ-H chiral stationary phase. Both methods share two features: they use ethanol as a cosolvent and are able to give a complete enantioseparation without interference from other associated chiral impurities. With the green eluent mixture ethanol-water-diethylammine 70:30:0.1 (v/v/v), the resolution between escitalopram and (R)-enantiomer was 2.09 at 30°C. The limits of quantification for the (S) and (R) enantiomers were 4.5 and 3.8 μg mL -1 , respectively., (© 2022 Wiley-VCH GmbH.)

Intracytoplasmic sperm injection (ICSI) has allowed reproduction options through assisted reproductive technologies (ARTs) for men with no spermatozoa within the ejaculate (azoospermia). In men with non-obstructive azoospermia (NOA), the options for spermatozoa retrieval are testicular sperm extraction (TESE), testicular sperm aspiration (TESA), or micro-surgical sperm extraction (microTESE). At the initial time of spermatozoa removal from the testis, spermatozoa are immobile. Independent of the means of spermatozoa retrieval, the subsequent steps of removing spermatozoa from seminiferous tubules, determining spermatozoa viability, identifying enough spermatozoa for oocyte injections, and isolating viable spermatozoa for injection are currently performed manually by laboratory microscopic dissection and collection. These laboratory techniques are highly labor-intensive, with yield unknown, have an unpredictable efficiency and/or success rate, and are subject to inter-laboratory personnel and intra-laboratory variability. Here, we consider the potential utility, benefits, and shortcomings of developing technologies such as motility induction/stimulants, microfluidics, dielectrophoresis, and cell sorting as andrological laboratory add-ons to reduce the technical burdens and variabilities in viable spermatozoa isolation from testicular samples in men with NOA.

Purpose: Oocytes and embryos can be vitrified with and without dimethyl sulfoxide (DMSO). Objectives were to compare no vitrification (No-Vitr), vitrification with DMSO (Vitr + DMSO), and vitrification without DMSO (Vitr - DMSO) on fresh/warmed oocyte survival, induced parthenogenetic activation, parthenogenetic embryo development, and embryonic maternal imprinted gene expression., Methods: In this prospective controlled laboratory study, mature B6C3F1 female mouse metaphase II oocytes were treated as: i) No-Vitr, ii) Vitr + DMSO/warmed, and iii) Vitr - DMSO/warmed with subsequent parthenogenetic activation and culture to the blastocyst stage. Oocyte cryo-survival, parthenogenetic activation and embryo development, parthenogenetic embryo maternal imprinted gene expression were outcome measures., Results: Oocyte cryo-survival was significantly improved in Vitr + DMSO versus Vitr - DMSO at initial warming and 2 h after warming. Induced parthenogenetic activation was similar between all three intervention groups. While early preimplantation parthenogenetic embryo development was similar between control, Vitr + DMSO, Vitr - DMSO oocytes, the development to blastocysts was significantly inferior in the Vitr - DMSO oocytes group compared to the control and Vitr + DMSO oocyte groups. Finally, maternal imprinted gene expression was similar between intervention groups at both the 2-cell and blastocyst parthenogenetic embryo stage., Conclusion(s): Inclusion of DMSO in oocyte vitrification solutions improved cryo-survival and developmental potential of parthenogenetic embryos to the blastocyst stage without significantly altering maternal imprinted gene expression., (© 2021. Springer Science+Business Media, LLC, part of Springer Nature.)

Background: Letrozole is widely employed as ovulation induction agent in women with PCOS, but its use in mild stimulation (MS) protocols for IVF is limited. Aim of the present study was to evaluate the feasibility of a MS protocol with letrozole plus hMG in non-obese PCOS women undergoing IVF after a metformin pre-treatment., Methods: We retrospectively evaluated the data of 125 non-obese PCOS undergoing MS with letrozole plus hMG, 150 IU as starting dose, (group 1, N = 80) compared to those undergoing a conventional IVF stimulation protocols (CS) (group 2, N = 45) prior to IVF. All patients had received metformin extended release 1200-2000 mg daily for three to six months before IVF. GnRH antagonist was administered in both groups when the leading follicles reached 14 mm., Results: Both groups were comparable for age, BMI and ovarian reserve markers. Both groups showed lower than expected AFC and AMH values as a consequence of metformin pre-treatment. Letrozole-treated patients required a significantly lower amount of gonadotropins units (p < 0.0001), and showed significantly lower day 5, day 8 and hCG day E2 levels compared to patients undergoing the CS protocol (p < 0.0001, p < 0.0001 and p = 0.001 respectively). The oocyte yield, in terms of total (6, IQR 3, vs 6, IQR 4 respectively,) and MII oocytes (5, IQR 3, vs 5, IQR 3, respectively) number, did not differ among groups; the number of total (3, IQR 2, vs 3, IQR 1 respectively) and good quality embryos (2, IQR1 vs 2, IQR 1,5 respectively) obtained was comparable as well in the two groups. The number of fresh transfers was significantly higher in group 1 compared to group 2 (80% vs 60%, p = 0.016). A trend for higher cumulative clinical pregnancy rate was found in women undergoing MS compared to CS (42.5%vs 24,4%, p = 0.044), but the study was not powered to detect this difference., Conclusions: The present study suggests that the use of letrozole as adjuvant treatment to MS protocols for IVF may be an effective alternative to CS protocols for non-obese PCOS patients pre-treated with metformin, as it provides comparable IVF outcome without requiring high FSH dose, and avoiding supraphysiological estradiol levels.

Background: Ultrasound elastography is a non-invasive medical imaging technique able to quantitatively characterize the stiffness of a given tissue. It has been shown to predict the risk for cervical insufficiency and preterm delivery, and to allow differentiation of malignancy from normal tissue. The present study sought to evaluate whether cervical tissue dishomogeneity, as assessed by cervical ultrasound elastography, may predict the embryo transfer (ET) ease in infertile women undergoing IVF/ICSI., Methods: We evaluated 154 infertile patients with no history of previous ET or intrauterine insemination. Cervical stiffness was evaluated in six regions of interest (ROI), compared two by two to obtain strain ratio (SR) values. Since a SR value of 1 was suggestive of tissue homogeneity, we computed 1-SR/SR-1 values to obtain a measure of the degree of cervical tissue dishomogeneity that we named "dishomogeneity index" (DI). Ultrasound-guided ET was performed by an expert operator blinded to the results of cervical elastography. The prediction ability of elastography on ET ease was evaluated by binary logistic regression, and the predictive accuracy of the independent variables was quantified with area under the curve (AUC) estimates derived from receiver operating characteristic (ROC) curve., Results: ET resulted to be easy in 99 out of 154 patients (64,2%), difficult in 54 patients (35%), and impossible in one. DI values in cervical medial lips region correctly classified 86.9% of patients, according to binary logistic regression, with a sensitivity of 81.4% and a specificity of 89,9%, positive likelihood ratio (LR) 8.07 and negative LR of 0.21. A DI cut-off value of 0.29 predicted a difficulty of ET with a sensitivity of 88,9% and a specificity of 85%., Conclusions: Cervical ultrasound elastography, by allowing the identification of cervical tissue dishomogeneity, may be of help in predicting the ET ease in infertile women candidates to IVF/ICSI.

Aim:  A common anatomical consequence of low-segment cesarean section is the presence of a pouch on the anterior uterine wall that can be detected by sonography or hysteroscopy. Different suturing techniques have been compared (single vs double layer) and showed no substantial differences. This prospective longitudinal study was aimed at evaluating the outcome of the cesarean scar, comparing two different types of single-layer sutures by transvaginal ultrasound and hysteroscopy., Material and Methods: The study sample consisted of two groups of 30 singleton primiparae at term who delivered by elective low segment cesarean section. In the first group, uterine closure was done with locked continuous single-layer sutures and in the second group, with single-layer interrupted sutures. Patients were assessed by transvaginal ultrasound and hysteroscopy, between the 6th and the 12th month after delivery, and again at the 24th month. Ultrasound measurements were made of the pouch area, if present., Results: A bell-shaped uterine wall defect was seen at ultrasound in 36 (85.71%) of 42 patients who completed the follow up at the 24th month. It was larger in the group of patients with closure by continuous sutures (6.2 [2.1-14.7] mm2) as compared to interrupted sutures (4.6 [1.9-8.2] mm2, P = 0.03). Hysteroscopy confirmed the presence of the wall defect in all 36 cases, but different hysteroscopic outcomes were observed., Conclusion: Locked continuous sutures seem to cause a larger defect as compared to interrupted sutures, probably due to a greater ischemic effect exerted on the uterine tissue., (© 2012 The Authors. Journal of Obstetrics and Gynaecology Research © 2012 Japan Society of Obstetrics and Gynecology.)

Objective: Aim of this study was to evaluate the semen quality and the serum concentration of follicle-stimulating hormone (FSH) and Testosterone (T) in infertile patients with and without varicocele., Material and Methods: 365 infertile patients undergoing Assisted Reproduction Tecnique (ART) were retrospectively included in the study. All subject were evaluated by history, physical examination, semen analysis, semen culture, mixed anti-immunoglobulin reaction test (MAR) for demonstration of sperm agglutination antibodies IgG and IgA, serum FSH and T determination., Results: We observed 97 (26.6%) patients affected by varicocele compared to 268 (73.4%) without varicocele. A significant reduced percentage of motile spermatozoa (24.58 +/- 21.68 vs 21.01 +/- 12.62, p < 0.001) and lower sperm concentration (15.50 +/- 23.30 vs 16.50 +/- 15.22, p < 0.001) were observed in patients with varicocele compared to patients without varicocele. No significant differences were observed in sperm vitality between the two population of men with and without varicocele. Serum FSH (10.42 +/- 10.84 vs 9.11 +/- 18.81, p < 0.001) and Testosterone (5.73 +/- 5.97 vs 5.21 +/- 2.43, p < 0.001) levels were significantly higher in patients with varicocele compared to patients without varicocele. Detection of IgG and IgA sperm antibodies were negative in both man with and without varicocele., Conclusion: The direct connection between varicocele and infertility is not clear. The data of the present study suggest that the presence of a clinical varicocele rule out fertility in men affecting the hypothalamic pituitary-gonadal axis.

The authors present a rare form of orbital cavitary rhabdomyosarcoma in which lymphangioma was mistakenly diagnosed on the basis of echography and MRI. Rhabdomyosarcoma can usually be differentiated from lymphangioma by echographic and MR imaging, because cavitation is very rare in orbital rhabdomyosarcoma, but rhabdomyosarcoma should be suspected whenever the clinical presentation of a rapidly progressive unilateral exophthalmos is observed in a child.