This study aimed to develop an artificial intelligence-based scoring system to prioritize mosaic embryos according to live birth outcomes. This multicentre, observational, retrospective study included 264 transferred mosaic embryos from 2583 PGT-A (Preimplantation Genetic Testing for Aneuploidies) cycles performed between January 2017 and January 2023. Trophectoderm (TE) biopsies from day-5 (D5) or day-6 (D6) blastocysts were analysed using Next-Generation Sequencing (NGS) (VeriSeq, Illumina®, San Diego, CA, USA). Biopsied embryos were vitrified and subsequently transferred. Clinical, embryological, and laboratory variables were collected to build predictive machine learning models for live birth. Models excluding cohort-invariant variables were refined to derive the final scoring system. Among mosaic embryos, biochemical, clinical pregnancy, and live birth rates were 50.75%, 41.66%, and 36.36%, respectively. The best-performing model, validated through tenfold cross-validation, identified embryo quality and biopsy day as the most influential predictors (42% and 34% weights, respectively), while mosaicism-related factors such as monosomy/trisomy (18%) and mosaicism degree (6%) had lower influence. The resulting score suggests prioritizing high-quality embryos biopsied on D5, as the type and level of mosaicism play a minor role in gestational potential, except when comparing embryos of similar quality where lower mosaicism levels and absence of monosomy are advantageous. The AI-derived score highlights embryo quality as the primary determinant of success in mosaic embryo transfer, supporting prioritization of high-quality, D5-biopsied embryos to improve live birth outcomes in ART.

The goal of this study is to determine factors associated with fetal heartbeat and live birth rate for euploid frozen embryo transfers. This is a retrospective cohort study of IVF patients at a single private practice in Cincinnati, Ohio undergoing single frozen euploid embryo transfer from 1/1/2023 to 7/31/2025. Variables assessed are embryo transfer protocol, genetic testing laboratory, physician who performed the embryo transfer, and embryologist. The main outcomes of interest are fetal heartbeat and live birth rate per single euploid frozen embryo transfer. Fetal heartbeat rate was significantly associated with the frozen embryo transfer protocol (p < 0.01). Modified natural FET protocols without oral medication (such as letrozole) had the highest fetal heartbeat rate of 66% compared to the average fetal heartbeat rate of 52%. Fetal heartbeat rate was also associated with the genetic testing laboratory (p < 0.01) and the day of embryo biopsy (p < 0.01). Fetal heartbeat was not associated with any of the other modifiable measures evaluated including the embryologist performing the biopsy or the physician performing the embryo transfer. At our center, the highest ongoing pregnancy rate per euploid embryo is seen with modified natural FET protocols without oral medication (such as letrozole).

This study evaluated Preimplantation Genetic Testing for Structural Rearrangements (PGT‑SR) outcomes in carriers of complex chromosomal rearrangements (CCRs). Four CCR couples underwent PGT‑SR, and 20 karyotypically normal couples underwent Preimplantation Genetic Testing for Aneuploidy (PGT‑A) as controls. Embryo biopsies were analyzed by next‑generation sequencing (NGS), and Mapping Allele with Resolved Carrier Status (MaReCs) was used for breakpoint mapping. Literature data from 31 additional CCR carriers (222 embryos) were also summarized. Among the four CCR couples in this study, 14 blastocysts were biopsied. The high‑quality blastocyst rate (57.1%, 8/14) and euploidy rate (14.29%, 2/14) were significantly lower than in controls (71.5% and 42.37%, respectively; P < 0.05). In the literature cohort of 222 embryos, for CCR types A (three‑way), B (double two‑way), and C (exceptional), the high‑quality blastocyst rates (37.50%, 44.44%, and 32.91%) and chromosomal abnormality rates (86.25%, 92.06%, and 88.61%) showed no significant differences; the overall euploidy rate (11.26%) was significantly lower than that in the PGT‑A control group (42.37%, P < 0.001). In conclusion, CCR carriers show impaired embryonic potential and markedly lower euploidy rates. While Type B CCR embryos have good morphology, their genetic normality is extremely poor, ruling out morphology-only selection. Instead, PGT-SR combined with high-resolution breakpoint mapping is required to reliably identify viable embryos for healthy pregnancy.

Report of the PGDIS Task Group on the status of PGT-A.

Врожденная дисфункция коры надпочечников (ВДКН) — это группа заболеваний с аутосомно-рецессивным типом наследования, в основе которых лежит дефект ферментов стероидогенеза коры надпочечников. В зависимости от варианта ферментного блока спектр клинических проявлений ВДКН варьирует от малосимптомных до потенциально фатальных нарушений. В обзоре представлен детальный анализ шести основных форм ВДКН (липоидная гиперплазия, дефициты HSD3B2, CYP17A1, CYP21A2, CYP11B1, POR) с углубленным описанием их молекулярных основ, патогенеза и характерных клинико-лабораторных проявлений.Особое внимание уделено современным методам генетической диагностики ВДКН, включая анализ высокогомологичного локуса CYP21A2, пренатальную и преимплантационную диагностику. Детально описаны не только современные подходы к заместительной терапии, но и перспективные методы лечения: антагонисты рецепторов кортикотропин-рилизинг-гормона, генная терапия и клеточные технологии. Уникальность работы заключается в комплексном анализе заболевания от фундаментальных основ до прикладных аспектов ведения пациентов с учетом российских клинических реалий.

Preimplantation genetic diagnosis for monogenic disorders makes it possible to identify and select embryos free of specific genetic mutations prior to uterine transfer, using embryos produced via medically assisted reproductive technologies. Since 2015, the Italian legal system has allowed parents who are carriers of, or affected by, monogenic hereditary conditions, including autosomal dominant polycystic kidney disease (ADPKD), to access preimplantation genetic diagnosis. However, the experience remains limited, and its application in particular to a late onset disease like ADPKD raises clinical and ethical concerns. These include the potential health risks associated with assisted reproductive technologies for the woman, the risk of assisted reproductive technology failure, moral permissibility of embryo selection, psychosocial implications for the couple and child, and issues related to equitable access to these treatments. An effective preconception counseling strategy should be grounded in a shared decision-making framework involving the prospective parents and a multidisciplinary medical team, including specialists in nephrology, obstetrics, and human genetics. This review aims to discuss the principal bioethical considerations surrounding preimplantation genetic diagnosis for ADPKD, in order to support clinicians involved in genetic counseling and reproductive decision-making.

To our knowledge, only 29 individuals have been described in the literature with biallelic pathogenic variants in the valyl-tRNA synthetase 2 (VARS2) gene, responsible for changes in the mitochondrial respiratory chain complex. We report two siblings with a novel combination of biallelic variants in the VARS2 gene (c.1079C>T p.Ala360Val, likely pathogenic, and c.1258G>A p.Ala420Thr, likely pathogenic). Both presented early hypertrophic cardiomyopathy and lactic acidosis, with fatal outcomes within the first year of life. The first also presented severe fetal growth restriction and a ventricular septal defect; the second developed epilepsy, respiratory failure, and psychomotor delay. This genotype may be linked to a particularly severe cardiac phenotype. Our report broadens the clinical and genetic spectrum of VARS2-related mitochondrial disease, highlights the variability of phenotypic expression, and reinforces the importance of early molecular diagnosis in neonatal-onset cardiomyopathy. Genetic confirmation enables accurate genetic counselling and consideration of prenatal or preimplantation diagnosis in future pregnancies.

Blastocyst biopsy allows genomic selection of pre-implantation bovine embryos, reducing generation interval and enhancing the genetic gain. We aimed to evaluate the suitability of a simple biopsy procedure, performed by hands, using a splitting microblade and a stereoscope, on invitro-produced embryos. Crossbred Bos taurus x B. indicus blastocysts at Day 6 or 7 after invitro fertilization (IVF) were biopsied and cultured invitro for 48 h. Biopsy reduced (p < 0.01) embryo development as noted by the lower blastocoel formation/re-expansion rates in both Day 6 (33.3% vs. 71.4%) and Day 7 (66.6% vs. 89.8%) blastocysts, when compared to non-biopsied embryos. Biopsy on Day 7 decreased (p < 0.01) cell number (112.8 ± 6.3 vs. 149.9 ± 5.6) and increased (p < 0.01) apoptotic index (14.9 ± 1.4 vs. 6.2 ± 1.2). In a second experiment, the IVF blastocysts were biopsied in the farm, and cultured for 3 h. Re-expanded blastocysts, corresponding to 78.3% biopsied embryos, were transferred to synchronized recipients. Pregnancy (57.4% vs. 62%), delivery (51.0% vs. 48%), gestation length (285.5 ± 2.0 vs. 284.6 ± 1.2 days) and birthweight (30.5 ± 1.8 vs. 29.9 ± 1.2 kg) were similar (p > 0.05) between handmade biopsy and non-biopsied embryos. Whole genome amplification of biopsied samples resulted in 256.0 ± 155.9 ng/μL DNA. The mean call rate was 75.3% ± 4.3% and the imputed GEBV for 305-day milk yield was 839.21 ± 61.6 kg. In conclusion, handmade biopsy can be performed at small laboratories and at farm level, contributing to reduce costs for embryo genomic selection. Pregnancy, calf delivery and birthweight were not compromised, and samples are suitable for downstream applications such as genotyping.

Sperm vacuolization and mitochondrial activity in raw versus prepared semen: effect on embryological outcomes.

Clinical application progress and prospects of non-invasive preimplantation genetic testing (niPGT): A review.

Women and girls with inherited bleeding disorders (IBD) face distinct gynaecologic and obstetric challenges, largely due to increased bleeding risk during key reproductive milestones. Conditions such as heavy menstrual bleeding (HMB), which affects a significant proportion of women with IBD, require collaborative management utilizing hormonal therapies and antifibrinolytics. Pregnancy, labour and delivery, and the postpartum period are high-risk phases. While IBDs like von Willebrand disease and haemophilia carriers may not inherently impair fertility or increase miscarriage risk, other severe factor deficiencies (e.g., factor X deficiency, factor XIII deficiency, and fibrinogen disorders) are associated with higher rates of miscarriage and antenatal haemorrhage, often requiring prophylactic factor replacement. Advances in preconception genetic counselling and prenatal diagnosis, including non-invasive prenatal testing (NIPT) and preimplantation genetic diagnosis (PGD), are crucial for informed reproductive choices and delivery planning. Careful assessment of coagulation status is mandatory for procedures like neuraxial anaesthesia, and mode of delivery requires shared decision-making to minimize cranial bleeding risk in an affected foetus. All IBDs, notably von Willebrand disease and haemophilia carriers, elevate the risk of primary and secondary postpartum haemorrhage (PPH), necessitating a multidisciplinary team approach and individualized haemostatic support. Furthermore, overcoming the historical under-recognition of symptomatic female carriers requires systematic screening and education to ensure optimal, lifelong care and reduced maternal morbidity.

Preimplantation embryo genomic selection (eGS) enables selection prior to implantation and could accelerate genetic gain in cattle. A major hurdle is the limited DNA from embryo biopsies, requiring efficient whole-genome amplification (WGA) for accurate genomic analyses. However, alternative WGA methods and genotyping strategies have not been systematically compared in cattle. This study evaluated different methods for WGA (multiple displacement amplification (MDA) or multiple annealing and looping-based amplification cycles (MALBAC)) and for genotyping (single nucleotide polymorphism array (SNP-array), genotyping by targeted sequencing (GBTS), or whole-genome sequencing (WGS)) using 3-, 6-, and 9-cell bovine samples. MDA consistently outperformed MALBAC across various performance metrics, including amplification length, call rates, genome coverage (93.43-94.40% vs. 53.01-67.08%), and genotyping concordance (0.89-0.98 vs. 0.75-0.92). GBTS achieved the highest call rates, while SNP-array and GBTS showed excellent concordance and low error rates. WGS provided genome-wide data for precise aneuploidy detection. We further validated the workflow in trophectoderm biopsies and arrested embryos, generating reliable data for genomic evaluation, sex determination, and aneuploidy screening. MDA from ≥6 cells combined with GBTS or SNP-array showed a favorable balance of efficiency and accuracy for bovine eGS. This framework may facilitate the application of eGS in cattle breeding by enhancing selection intensity and accelerating genetic improvement.

Controlled Ovarian Stimulation Contributes to the Incidence of de Novo Chromosomal Abnormalities in Cleavage-Stage Embryos.

PURPOSE: To characterize the clinical and genetic features of Chinese FHH type 1 (FHH1) patients and evaluate biochemical discriminators between FHH1 and sporadic primary hyperparathyroidism (s-PHPT). METHODS: This retrospective study included 11 genetically confirmed FHH1 and 55s-PHPT patients. Their clinical, biochemical, and imaging data were compared. Genetic testing was conducted via targeted next-generation or whole-exome sequencing. The key discriminators were assessed by receiver operating characteristic (ROC) analysis. RESULTS: Among the 11 FHH1 patients, 63.6% were females and 63.6% had a family history. Incidentally detected hypercalcemia was the most common cause (72.7%). Compared with s-PHPT, FHH1 were younger at diagnosis [38.1 ± 17.4 years vs. 53.6 ± 14.0, P = 0.002], had lower parathyroid hormone level [41.80 (28.89, 124.00) pg/mL vs. 233.90 (136.6, 521.6) pg/mL, P < 0.001], higher serum phosphate (1.06 ± 0.28 mmol/L vs. 0.77 ± 0.17 mmol/L, P = 0.008), and lower 24-hour urinary calcium excretion (24hUCa) (P < 0.001). Bone turnover markers (BTMs) were lower (P ≤ 0.001), low bone mass was less frequent (P = 0.005), and parathyroid imaging results were frequently negative in FHH1. Eight CASR variants were identified, including 4 novel variants (c.197G > C, c.488C >T, c.1042del, c.2386A >G). ROC analysis indicated that 24hUCa < 3.71 mmol/24h optimally distinguished FHH1 from s-PHPT. CONCLUSION: The relatively large single-center Chinese FHH1 cohort presented younger with similar serum calcium, milder PTH elevation, relatively normal BTMs, and less target-organ involvement than s-PHPT. A new cut-off value of 24hUCa may serve as a practical pre-genetic screening tool.

Objective: This study aimed to investigate the families with different GJB2 gene mutations undergoing preimplantation genetic testing (PGT), in order to provide reference for clinical practice and implementation strategies. Methods: Five families undergoing PGT for GJB2 gene mutations at Chinese People's Liberation Army (PLA) General Hospital from 2015 to 2024 were selected. All families received genetic counseling, and the audiological data of the patients and their relatives were collected. Blood samples from the probands and their related family members were taken for genetic sequencing. After entering the PGT process, oocyte retrieval, in vitro fertilization, embryo biopsy, and genetic testing were performed. Healthy embryos were selected for transfer, and the pregnant women were followed up. Results: The 5 families included in the study were as follows: one family with both parents being carriers of GJB2 c.235delC, one family with both parents being carriers of GJB2 c.109G>A, one family with one parent affected by autosomal dominant inheritance of GJB2, and in two other families, the probands were married to each other, and both GJB2 and SLC26A4 gene mutations were involved. Through different PGT strategies, the five families underwent a total of four assisted reproductive cycles, all of which resulted in clinical pregnancies. Four couples had delivered four healthy offsprings, all of whom passed hearing screening. Conclusions: GJB2 gene mutations, due to their involvement in diverse phenotypes, varying disease severity, and different inheritance patterns, require careful consideration when performing PGT. Different linkage analysis strategies should be chosen for families with varying genetic inheritance modes. For GJB2 c.109G>A with milder phenotypes, it is not recommended to proactively suggest prenatal or preimplantation interventions during genetic counseling, unless the couple strongly requests it, in which case, thorough genetic counseling and informed consent, should be obtained. Additionally, PGT may be performed for GJB2 c.109G>A concurrently if the couple is involved in other high-risk genetic diseases.

Genetically engineered offspring are produced by genetic modification to affect the characteristics of the offspring, or from an embryo chosen by pre-implantation genetic diagnosis (PGD). The main goal of designing offspring is to prevent genetically-based disorders that are caused by mutations in DNA. It is now possible to produce precisely modified designer babies—as evidenced by the advancements in mitochondrial DNA transfer and genome editing techniques. But there isn't enough information available about the risks associated with these editing tools, particularly when the changes are inherited. In addition, there are moral questions about whether or not we should use these technologies to produce designer offspring. that further study is needed to refine the technique and determine its long-term safety, and that these germ line genetic editing techniques should still be regarded as experimental operations. created and put into use a teaching module for both an advanced genetics course and a genetics course that introduces students to CRISPR-cas9 technology. The design, strategy, conceptual modelling, and implementation of CRISPR-cas9 technology are based on the current study findings about CCR5 gene editing in twin girls.

Investigating DNA contamination affecting embryo biopsy specimens taken for PGT-A: Incidence and impact on the accuracy of chromosomal analysis.

Public understanding of assisted reproductive technology (ART) is poorly documented in many countries. This study aims to explore the awareness about ART among the general Jordanian population and its association with sociodemographic and reproductive health education factors. A cross-sectional online survey was conducted among Jordanian adults aged between 20 and 65 years. In addition to the demographic data, the questionnaire assessed knowledge about in vitro fertilization (IVF), intrauterine insemination (IUI), and ovulation induction (OI). From the 902 participants, about half could neither define IVF nor distinguish between IVF from ICSI. Most couldn’t define IUI (70.73%), or ovarian induction (OI) (63.75%). The majority were unaware embryo gluing (89.14%) or assisted hatching (88.47%). Most were unaware of the success rates and didn’t know if using frozen eggs, sperms, or frozen embryos, affected the success rate. Most didn’t know the possible side effects of ovarian stimulation. According to their opinions, IVF success factors were the wife’s age, utilizing preimplantation genetic screening (PGS), and luteal support injections. Significant gaps in Jordanian community’s knowledge of ART were found by this study. Most participants were unaware of IVF, ICSI, IUI, OI, and their success rates and complex procedures. The need for focused educational interventions to enhance reproductive health literacy.

Hypophosphatemia during pregnancy poses unique clinical challenges due to physiological changes in mineral metabolism, with potential impacts on maternal and fetal health, but limited evidence to guide treatment. We describe the case of a woman with X-linked hypophosphatemia, previously treated with burosumab for lower limb pseudofractures. Due to her desire for pregnancy and the absence of safety data on burosumab during gestation, the drug was discontinued and treatment with phosphate supplements and calcitriol was initiated. Following assisted reproduction with preimplantation genetic diagnosis, the patient successfully carried the pregnancy to term and delivered a healthy newborn. Monthly biochemical monitoring allowed for safe adjustment of therapy, maintaining serum phosphate levels within or near the normal range without maternal or fetal complications. We also review the available literature on hypophosphatemia in pregnancy, emphasizing the need for individualized management and close monitoring.

To maintain market competitiveness, raw milk production must reconcile rigorous safety protocols with operational efficiency. This review provides a comprehensive taxonomy of biotechnological solutions that support the precision-driven optimization of milk quality at the point of origin. Milk production cycle consists of four interconnected technological modules. 1) Modern genetic and reproductive technologies include genomic selection for loci CSN3, DGAT1, BoLA-DRB3, as well as the OPU-IVP-TE complex with preimplantation diagnosis. They enable the targeted development and rapid dissemination of superior genotypes characterized by enhanced productivity, optimized milk composition, and robust disease resistance. 2) Nutritional biotechnology and microbiome engineering facilitate the precision modulation of ruminal fermentation through the strategic application of probiotics and enzymes. In addition, microbial inoculants and adsorbents in feed storage make it possible to plan nutritional value of animal diet and minimize chemical contamination (mycotoxins). 3) Biological methods aimed at udder health control and microbiological purity of raw materials. Probiotics, bacteriophages, immunomodulators, and enzymes offer a safe alternative to antibiotics. This group also includes technologies for biological conditioning of equipment. 4) Biosensorics and digital platforms provide continuous predictive monitoring, e.g., in-line sensors for composition, somatic cell count, pathogens, inhibitory substances; the digital twin herd, etc. Implemented as an integrated, synergistic framework, these solutions mitigate the impact of empirical variables on raw milk quality, ensuring a more consistent and predictable production profile. Biotechnologies are an economically justified strategy that guarantees compliance of raw materials with regulatory standards by providing raw milk with reproducible functional and technological properties. They facilitate deep processing and encourage import substitution, thus strengthening the export potential of the domestic dairy industry.