Medicine and Pharmacology

Abstract: This study aimed to evaluate the effects of astaxanthin on ovarian reserve, histopathological injury, oocyte maturation, fertilization, and early embryo development in rat models of ovarian ischemia and ischemia–reperfusion. In this randomized controlled experimental study, 48 adult female Sprague–Dawley rats were equally allocated to six groups undergoing sham surgery, ischemia, or ischemia–reperfusion, with or without intraperitoneal astaxanthin administration. Two male rats were used for in vitro fertilization. Following ovarian hyperstimulation, serum anti-Müllerian hormone (AMH) concentrations, ovarian histopathology, Caspase-3 expression, oocyte maturation, fertilization, and embryo development were evaluated. Astaxanthin significantly increased total and metaphase II oocyte counts, fertilized oocytes, and two-cell embryos in both ischemia and ischemia–reperfusion groups, as well as Grade 1 embryos in the ischemia–reperfusion group. Astaxanthin also reduced hemorrhage, edema, necrotic cell damage, inflammatory cell infiltration, and Caspase-3 expression. Furthermore, astaxanthin attenuated the injury-associated decline in serum AMH concentrations. No significant reproductive or histopathological changes were observed in astaxanthin-treated sham-operated rats. Astaxanthin administered before torsion or detorsion improved reproductive outcomes and reduced ovarian tissue injury, suggesting a protective effect that warrants further investigation before clinical translation.

Abstract: Testicular torsion mainly affects children and adolescents, and the ischemia/reperfusion (I/R) injury that follows surgical detorsion can impair later fertility. Melatonin studies in this setting have used histological, hormonal, and semen endpoints, which cannot show whether the spermatozoa can still fertilize oocytes and support embryo development. Forty-eight male Wistar rats were assigned to sham, sustained ischemia (I), and I/R (720° torsion for 90 min, then detorsion) groups, each with or without a single intraperitoneal dose of melatonin (10 mg/kg; n = 8/group). After 20 days, testes were evaluated by Johnsen scoring and caspase-3 immunohistochemistry, serum testosterone was measured, and epididymal spermatozoa were analyzed and used for in vitro fertilization (IVF) of pooled oocytes from superovulated females. Both injuries lowered Johnsen scores, testosterone, and sperm concentration, motility, and normal morphology and increased caspase-3 immunoreactivity, with sustained ischemia causing greater damage. Fertilization fell from 55.0% in sham animals to 10.0% (I) and 15.0% (I/R), and blastocyst yield from 35.0% of inseminated oocytes to 2.5% and 7.5%, respectively. Melatonin improved all parameters, increasing fertilization to 30.0% (I+MLT) and 35.0% (I/R+MLT) and blastocyst yield to 17.5% and 25.0%, although most values remained below sham levels. A single melatonin dose therefore partially preserves the fertilizing and embryo-supporting capacity of spermatozoa after testicular ischemic injury.

Abstract: Anti-Müllerian hormone (AMH) and antral follicle count (AFC) predict ovarian response, while serial ultrasound documents the evolving antral cohort during controlled stimulation. Yet most visit-level summaries do not preserve which follicle is which across scans. We propose a dynamic ovarian recruitment phenotype, defined as a treatment-conditioned description of identity-linked antral follicle trajectories rather than a new measure of primordial reserve. Our primary, falsifiable hypothesis is that individual follicle identity contributes reproducible information about mature-oocyte yield beyond baseline predictors and the complete routine monitoring history available at the same early prediction landmark. A stringent test compares three nested models: baseline characteristics; baseline plus contemporaneous visit-level measurements, hormones and treatment exposure; and the same information augmented by identity-linked growth, persistence and synchrony, with a rank-matched pseudo-identity model as an intermediate comparator. Scans, prediction timing and modelling flexibility must be matched, with identity permutation as a negative control. A separate hypothesis asks whether any trajectory feature retains a patient-level component across comparable stimulation cycles. Spatial features are exploratory and are not evidence of primordial-follicle activation. Support requires reliable tracking, external validation, clinically relevant incremental prediction and, ultimately, evidence of benefit for a specific clinical decision. Equal performance of identity-free monitoring models, sensitivity to matching error or absent cross-cycle reproducibility would substantially weaken the proposed phenotype. This essay offers an operational definition and a validation pathway, not new clinical data or treatment recommendations.

Abstract: Polycystic ovary syndrome (PCOS) is a heterogeneous multisystem disorder characterized by reproductive, metabolic, and neuroendocrine abnormalities. Current management primarily targets individual manifestations through lifestyle modifications and pharmacological therapy, while growing evidence implicates altered autonomic and neuroendocrine regulation in PCOS pathophysiology. Electroceuticals (ECs) and neuromodulatory interventions may represent potential non-pharmacological approaches for modulating neural pathways involved in reproductive and metabolic regulation. This review examines the biological rationale, experimental evidence, and available clinical findings supporting the investigation of such interventions in PCOS. From a biomedical perspective, ECs may offer potential advantages in PCOS management; however, the limited evidence currently available precludes definitive conclusions regarding their short- and long-term therapeutic effects and potential adverse effects in humans. Therefore, further specialized preclinical and clinical studies are warranted to evaluate the safety and efficacy of this promising approach.

Abstract: Background: Quantification of levonorgestrel (LNG) remaining is crucial for understanding drug release, effectiveness, and potential for extended use beyond the labeled duration of contraceptive implants. The current study aimed to compare and characterize in vivo rate of LNG released from two types of implants, Femiplant (FP) and Levoplant (LP) users in Bangladesh. Methods: Between July and December 2025, explant sets were collected from reproductive age women who had an implant removed at four family planning settings and residual LNG was quantified using a validated reverse-phase high-performance liquid chromatography (RP-HPLC) method. Baseline LNG load was also determined from six unused implants of both FP and LP. Release kinetics were analyzed to identify the models that best fit the data. We determined Pregnancy rates using Pearl Index and recorded self-reported side-effects. Results: Mean baseline LNG load for FP and LP were 152.15 mg and 149.98 mg, respectively. Mean LNG residual per set of explants was 21.40 ± 3.05 (95% CI: 20.11–22.69) and 21.63 ± 2.70 (95% CI: 20.49–22.77), while LNG (%) of 76.0 ± 1.2 (95% CI: 75.5-76.8) and 74.3±1.5 (95% CI:73.5-75.1) were released from FP and LP, respectively. Additionally, f1 (difference factor) and f2 (similarity factor) estimated were 11.7 (95% CI: 9.5-17.8) and 62.5 (95% CI: 55.6-76.8) for fresh FP and LP, respectively. No pregnancy and serious adverse effect were observed, except implant related side effects such as irregular menstruation and bleeding pattern change. Overall satisfaction was 90.7% for FP, and 96.8% for LP and all participants continued implants for three years. Conclusion: FP could perform clinically similarly to LP as a long-acting reversible contraceptive implant demonstrating contraceptive effectiveness, safety, and acceptability with respective duration of action.

Abstract: Male factor infertility contributes to around half of all cases of couple infertility, and semen analysis remains the main first-line investigation of male fertility. However, conventional semen parameters do not always provide a complete picture of sperm quality, and there is growing interest in additional tests such as sperm DNA fragmentation analysis. A retrospective observational study was conducted using anonymised clinical data collected from men undergoing routine semen analysis between January 2024 and May 2026. Semen analyses were performed according to the World Health Organization laboratory manual for the examination and processing of human semen using computer-assisted semen analysis (CASA). Macroscopic and microscopic semen parameters were recorded alongside morphology, laboratory assessment and referral for sperm DNA fragmentation testing using the Halosperm sperm chromatin dispersion assay. We retrospectively analysed data from 387 men who underwent routine semen analysis at Marylebone Diagnostic Centre, London, between January 2024 and May 2026. Samples were assessed according to WHO 6th Edition recommendations using computer-assisted semen analysis (CASA). Macroscopic and microscopic semen parameters were recorded, together with clinical classification and subsequent sperm DNA fragmentation testing using the Halosperm® sperm chromatin dispersion assay where indicated. The mean age of the cohort was 36.6 years (range 21–66 years). Although average semen parameters were generally above WHO lower reference limits, approximately 39–43% of men fell below the reference limits for key measures including sperm concentration, total sperm number, motility and morphology. Following review by the attending doctor, 61.0% of samples were classified as good, 30.0% as poor and 9.0% as very poor. Among the 170 men with available DNA fragmentation results, 25.6% of those classified as good were positive for increased DNA fragmentation, compared with 64.6% of the poor group and 82.1% of the very poor group. These findings show that conventional semen analysis remains valuable for identifying abnormalities and guiding further investigation. Importantly, sperm DNA fragmentation was also identified in a proportion of men whose routine semen assessment was considered satisfactory, suggesting that selective DNA fragmentation testing may add useful information to the conventional diagnostic pathway.

Abstract: Background. Endometriosis is a chronic gynecological disorder affecting a substantial proportion of women of reproductive age and is associated with impaired quality of life, absenteeism, presenteeism, and reduced work participation. Despite its potential occupational implications, evidence on how occupational physicians (OPs) manage workers affected by endometriosis remains scarce. This study aimed to assess knowledge, attitudes, and practices (KAP) regarding endometriosis among Italian OPs. Methods: A cross-sectional web-based survey was conducted between 1 and 18 April 2026 among members of an Italian discussion group restricted to occupational physicians. The questionnaire investigated demographic characteristics, knowledge of endometriosis, risk perception, perceived barriers to workplace management, and previous experience in issuing conditioned fitness-for-work appraisals. Multivariable logistic regression was performed to identify factors independently associated with having previously issued work restrictions because of endometriosis. Results: A total of 155 occupational physicians participated in the survey. Most respondents considered themselves adequately informed about endometriosis (78.1%) and reported previous professional experience with affected workers (81.3%). However, objective knowledge revealed important gaps, particularly regarding disease prevalence (26.5% correct responses), familial predisposition (34.8%), and diagnostic pathways (31.6%). The mean General Knowledge Score was 64.1% (95%CI 62.2–66.1), while only 11.0% of respondents reported awareness of occupational recommendations specifically addressing endometriosis. Overall, 65.8% of participants reported having previously issued conditioned fitness-for-work appraisals, most frequently recommending increased work breaks, avoidance of night shifts, or limitation of shift work. In multivariable analysis, male gender (aOR 5.69, 95%CI 1.84–17.58), participation in continuing medical education activities (aOR 85.72, 95%CI 6.60–1114.21), information obtained from colleagues (aOR 4.67, 95%CI 1.23–17.77), and employment in healthcare settings (aOR 5.91, 95%CI 1.92–18.16) were independently associated with previous issuance of restricted fitness-for-work judgments, whereas relying exclusively on undergraduate medical education was negatively associated with this outcome (aOR 0.07, 95%CI 0.01–0.39). Conclusions: Italian occupational physicians generally recognized endometriosis as a clinically relevant condition and frequently encountered affected workers in daily practice. Nevertheless, important deficiencies persist regarding epidemiological knowledge, diagnostic issues, and awareness of occupational management strategies. Continuous professional education, interdisciplinary exchange, and practical experience appear to contribute more substantially to occupational decision-making than undergraduate training alone. These findings support the development of targeted educational interventions and shared occupational recommendations aimed at improving workplace management and promoting the inclusion and retention of women affected by endometriosis in the workforce.

Abstract: Background/Objectives: Fertility preservation is a key component of cancer care in women of reproductive age, and controlled ovarian stimulation (COS) followed by oo-cyte/embryo cryopreservation is considered the standard strategy. Although COS pro-tocols are generally standardized, systemic effects related to different malignancies may interfere with ovarian reserve assessment and influence stimulation planning and counselling. This study aimed to evaluate COS outcomes across different malignancy types in women undergoing fertility preservation. Methods: This retrospective study included 331 oncological patients who underwent COS for fertility preservation at a single referral center between May 2008 and May 2025. Patients were stratified into three groups according to malignancy type: breast cancer, hematological malignancies, and other cancers. Ovarian reserve was assessed using serum anti-Müllerian hormone (AMH) levels and antral follicle count (AFC). COS outcomes, including stimulation duration, total gonadotropin dose, number of retrieved and mature oocytes, follicular output rate (FORT), and follicle-to-oocyte index (FOI), were compared among groups. Results: Pa-tients with hematological malignancies were significantly younger and showed higher AFC values compared with the other groups, whereas AMH levels did not differ sig-nificantly. After adjustment for age and AFC, patients with hematological malignancies showed significantly lower AMH levels compared with breast cancer patients, supporting the presence of a relative AMH–AFC discordance. In unadjusted analyses, gonadotropin requirements were higher in breast cancer patients, whereas the number of retrieved and mature oocytes was higher in patients with hematological malignancies. However, after adjustment for age and AFC, malignancy type was not independently associated with retrieved oocytes, mature oocytes, total gonadotropin dose, or stimulation duration. FORT and FOI were comparable among groups. Conclusions: Malignancy type did not appear to independently impair ovarian responsiveness to stimulation. Patients with hemato-logical malignancies showed preserved stimulation outcomes despite AMH values not being proportionally higher than expected based on their younger age and higher AFC. These findings suggest that AMH should be interpreted with caution in systemic ma-lignancies and that AFC may be particularly useful for tailoring fertility preservation counselling and stimulation strategies when AMH and AFC are discordant.

Abstract: Background: The SPARK (Symptoms of PCOS ameliorated by responses to Ke-to-adaptation) pilot study investigates whether nutritional ketosis can improve repro-ductive and metabolic outcomes in women with Polyendocrine Metabolic Ovarian Syndrome (PMOS). PMOS is a common endocrinopathy characterized by anovulation, hyperandrogenism, and metabolic dysfunction. Objective: To evaluate if a ketogenic diet (KD) or exogenous ketone supplementation (KS) with Qitone™ (bis-octanoyl-(R)-1,3-butanediol) can restore ovulation and menstrual activity. Methods: This ongoing 12-week randomized trial assigns women with PMOS to either a KD or KS intervention. Assessments include body composition by DXA, metabolic and reproduc-tive biomarkers (e.g., insulin, testosterone, SHBG), and cycle tracking via basal body temperature. Results: Preliminary data from seven participants (KD: n=3; KS: n=4) show that 100% demonstrated spontaneous menstrual activity within 2–7 weeks. Full menstrual reinstatement occurred in 86% of participants (KD=3, KS=3), including those with long-standing amenorrhea. Ovulation was predicted in 43% of the cohort. Key metabolic improvements included significant insulin reductions, favorable body composition changes (fat mass loss with lean mass preservation), and enhanced neurobehavioral well-being, including reduced anxiety and improved cognitive clarity. Conclusion: Preliminary findings suggest that both sustained and intermittent ketogenic interven-tions can restore menstrual and ovulatory function through multi-node endo-crine-metabolic recovery, independent of significant weight loss. These results highlight the potential of ketogenic therapies for PMOS but require confirmation in larger trials.

Abstract: This systematic review of 25 original studies examined the efficacy of density gradient centrifugation (DGC) combined with magnetic-activated cell sorting using Annexin V (MACS) for improving sperm quality in infertile men undergoing ICSI. The evidence consistently demonstrates that DGC-MACS significantly reduces sperm DNA fragmentation, with reported reductions ranging from 2.82% to 21.9% in absolute terms, and relative reductions of 39%-83%. The combination of DGC followed MACS achieved superior outcomes compared to either technique alone, reducing DNA fragmentation index (DFI) 4.1 to ± 1.3% compared with 8.1 ± 4.1% for DGC alone and 7.4 ± 3.9% for MACS alone. The treatment improved sperm motility, membrane integrity and overall spermatozoa health by reducing protamine deficiency and chromosomal abnormalities. Clinical results in ICSI cycles showed that although fertilization rates were similar between the treated and control groups, DGC followed by subsequent MACS treatment significantly improved embryo quality (72.5% vs. 51.47% top-quality day-3 embryos), blastocyst formation rate (69.69% vs. 48%), pregnancy rates (60.7% vs. 51.5%, p=0.014), and live birth rates (47.4% vs. 31.2%, p=0.001) with a reduced miscarriage rate (14.7% vs. 20.6%, p=0.034). The technique proved most beneficial in patients with high baseline DNA fragmentation (≥30%) and in those with asthenozoospermia or asthenoteratozoospermia. Studies suggest, and consistently supported by our study results, that performing DGC before MACS (DGC-MACS) may yield superior results compared to the reverse sequence. This is because DGC leads to a primary separation of sperm based on density, motility, and morphology and thus producing a high-density fractions enriched in morphologically normal sperms. MACS by specifically binding to phosphatidylserine (PS) residues on sperm membranes selectively removes PS‑positive/apoptotic sperm in a subsequent purification step. Overall, the literature strongly supports the use of combined DGC-MACS as an effective sperm preparation technique for ICSI in infertile men with elevated sperm DNA fragmentation.

Abstract: The evaluation of peripheral pulmonary lesions (PPLs) represents a persistent clinical challenge, particularly in the context of rapidly expanding lung cancer screening programs generating unprecedented volumes of screen-detected nodules requiring tissue evaluation. Conventional bronchoscopic techniques and earlier navigation platforms, including electromagnetic navigation bronchoscopy (ENB), improved access to peripheral airways but demonstrated variable and generally suboptimal diagnostic performance—historically plateauing at 60–70%—attributable principally to CT-to-body divergence and the diagnostic drop-off phenomenon. Robotic-assisted bronchoscopy (RAB) has emerged as a significant advancement integrating robotic catheter control, shape-sensing or electromagnetic navigation, and adjunctive intraprocedural imaging—principally cone-beam computed tomography (CBCT)—to improve lesion localization and procedural stability. Available evidence suggests that RAB achieves improved diagnostic yields compared with conventional bronchoscopic techniques, particularly when combined with real-time CBCT-guided tool-in-lesion confirmation and multimodal biopsy strategies including transbronchial cryobiopsy. Reported diagnostic yields generally range from 70% to 87% across pooled meta-analytic datasets, with higher yields described in centers employing CBCT and cryobiopsy. A favorable safety profile—with pooled pneumothorax rates of approximately 2%, substantially below the 20–25% associated with CT-guided transthoracic needle biopsy—represents a clinically important differentiator, particularly in patients with emphysema, coagulopathy, or contralateral lung compromise [1,3]. RAB additionally enables procedural capabilities beyond peripheral lesion biopsy, including simultaneous mediastinal lymph node staging, bilateral same-session lesion sampling, fiducial marker placement for stereotactic body radiotherapy, and investigational therapeutic applications. However, current evidence is largely derived from observational studies and single-arm prospective cohorts, with limited randomized data directly comparing RAB with transthoracic biopsy approaches. Outcomes are meaningfully influenced by lesion characteristics, operator experience, institutional volume, and availability of advanced imaging infrastructure. This review summarizes current evidence on RAB platforms, procedural optimization, diagnostic performance, safety, molecular tissue adequacy, and emerging applications, while explicitly addressing limitations, controversies, and areas requiring further investigation. This review was conducted and reported in accordance with PRISMA 2020 guidelines adapted for narrative reviews (SANRA framework).

Abstract: Background: In recent years, time-lapse incubators (TLs) with the ability to maintain a stable culture environment have been developed for in vitro fertilization treatment. Methods: In this retrospective cross-sectional study, data from 1,200 women who visited the Creation and Love fertility center, between April 2021 and October 2023 were reviewed. Among them, electronic medical records of 400 women aged 22-45 who underwent in vitro fertilization treatment were selected and divided into 2 groups according to the type of incubator used: 100 in the TL group and 300 in the CI group to evaluate differences in embryo development. Out of these participants, pregnancy outcomes were compared in 150 who underwent fresh-embryo transfer. Controlled ovarian stimulation was performed by standard antagonist protocol. The primary outcome was live birth rate, and the secondary outcome was good embryo rate. Results: The mean age of the study population was 34.6±5.4. Groups had a similar good embryo rate (67.7% vs. 65.5%, p=0.84), clinical pregnancy rate (49.0% vs. 52.0%, p=0.86) and live birth rate (35.0% vs. 42.0%, p=0.76). Furthermore, subgroup analyses of the live birth rate in women aged >35 was higher in the TL group than the CI group (38.0% vs. 23.4%, p=0.03). Conclusions: Based on the comparative analysis of live birth rates between women cultured using TLs and those using CI during 3-day fresh embryo transfer cycles, our study demonstrated a statistically significant increase in live birth rates among women >35 yr of age in the TL group.

Abstract: Background/Objectives: Follitropin delta is an approved recombinant follicle stimulating hormone (rFSH) expressed in a human cell line. The FSH protein is a heterodimer containing α and β subunits. The FSH α-subunit (FSHα) contains five intramolecular disulfide bonds: C7‑C31, C10‑C60, C28‑C82, C32‑C84 and C59‑C87. The α subunit has four closely spaced cysteine residues that form disulfide bonds. These clustered sites resist protease cleavage, making it difficult to accurately characterize the disulfide bond pairing in this type of protein therapeutic. Methods: The study uses an X-ray crystallography method to determine a high-resolution three-dimensional model structure of the recombinant FSH α-subunit. Results: The crystal structure of FSHα at 2.29 Å resolution, containing the disulfide bonds, is presented. This high-resolution structure provides definitive structural evidence that the disulfide bonds in rFSH are consistent with the expected native structural conformation. Conclusions: The rFSH structure validates the expected cysteine connectivity, overcoming limitations of prior mass spectrometry-based disulfide mapping attempts.

Abstract: Ultrasound is the primary imaging modality for evaluation of the scrotum and male reproductive tract. While high-frequency linear probes allow detailed assessment of testicular parenchyma, comprehensive visualization of epididymal anatomy and its spatial relationship to the testis may be limited using conventional two-dimensional imaging alone . This limitation is clinically relevant in the evaluation of male infertility, where epididymal abnormalities may contribute to obstructive processes We present a clinical image demonstrating a novel external scrotal application of a conventional three-dimensional (3D) transvaginal ultrasound probe. The images were obtained in an adult male undergoing infertility evaluation. With generous coupling gel and minimal probe pressure, the transvaginal probe was applied externally over the scrotum. High-resolution 2D images were first obtained, followed by volumetric 3D acquisition. The acquired dataset was analyzed using multiplanar reconstruction and volume-rendering techniques. This approach enabled clear visualization of the epididymal head, body, and tail, along with their anatomical continuity and relationship to the adjacent testis. The volume-rendered images provided an intuitive 3D depiction of epididymal curvature and spatial orientation, features that can be challenging to appreciate using standard linear scrotal ultrasound alone. Importantly, no patient discomfort, adverse effects, or technical complications were observed during the examination. This clinical image highlights the feasibility of repurposing a 3D transvaginal probe for external scrotal imaging to improve anatomical depiction of epididymal structures. While the technique is not intended to replace conventional scrotal ultra-sound, it may offer additional anatomical insight in selected infertility cases. This re-port serves as a hypothesis-generating illustration and supports further evaluation of its diagnostic utility in larger studies.

Abstract: Backgound/Objective: Breast cancer (BC) management has traditionally relied on static clinicopathologic and immunohistochemical biomarkers (hormone receptor status, HER2 expression, and proliferative activity assessed at diagnosis. However, these biomarkers are typically evaluated at a single time point and may not reflect therapy-induced mo-lecular evolution. This study evaluates whether longitudinal molecular profiling before and after treatment better characterizes tumor dynamics and provides clinically ac-tionable insights into treatment response, resistance, and prognosis. Methods: Thirty-two patients with invasive breast carcinoma were analyzed using his-topathology, immunohistochemistry, tissue-based next-generation sequencing, and plasma circulating tumor DNA (ctDNA) analysis. Paired tumor tissue and plasma sam-ples were collected before and after treatment when available. Changes in biomarker expression, molecular subtype, and genomic alterations were assessed to characterize molecular plasticity under therapeutic pressure. Results: The cohort had a median age of 54 years (range 29–86), predominantly invasive ductal carcinoma (>85%) and high-grade disease. Hormone receptor–positive tumors accounted for 78.1%. Molecular subtypes were Luminal A (34.4%), Luminal B HER2− (40.6%), Luminal B HER2+ (6.3%), HER2-enriched (6.3%), and triple-negative breast cancer (12.5%). Initial tissue sequencing identified PI3K/AKT pathway alterations in 28.1% of cases. Post-treatment analyses revealed substantial molecular discordance, including progesterone receptor loss (33.3%), HER2 status changes (33.3%), and Ki67 variability (77.8%). Plasma ctDNA analysis was informative in 53.1% of patients and identified additional clinically relevant alterations, including FGFR1 amplification and BRCA1/2 variants not detected in tissue. Conclusion: BC molecular profiles are dynamic and frequently altered by therapy. Longitudinal molecular assessment reveals clinically actionable changes overlooked by static subtyping, supporting a dynamic model of molecular classification, highlighting the potential value of adaptive molecular subtyping to improve treatment stratification and resistance monitoring.

Abstract: Background/Objectives: Fertility preservation (FP) spans oncological and non‑oncological indications, including gonadotoxic therapies, benign haematological/metabolic disorders, conditions at risk of premature ovarian insufficiency, severe andrological disease, and elective plans to defer childbearing. We provide a sex‑integrated overview of established and emerging FP strategies and their real‑world effectiveness [1–10]. Methods: Narrative synthesis of guidelines and high‑quality studies (to 2025), prioritising ASCO/ESHRE guidance, systematic reviews, and multicentre/registry data covering oocyte/embryo cryopreservation, ovarian tissue cryopreservation (OTC), GnRH agonists, ovarian transposition, in vitro maturation (IVM), sperm banking, surgical sperm retrieval (SSR), testicular tissue cryopreservation (TTC), and spermatogonial stem‑cell (SSC) approaches [1–10]. Results: For women, oocyte vitrification is the first‑line option when time allows; OTC is established when stimulation is infeasible or in prepubertal girls, with endocrine recovery common and increasing pregnancy/live‑birth reports. For men, sperm cryopreservation before therapy is standard; SSR supports selected cases. Paediatric TTC is feasible but remains experimental, whereas paediatric OTC is clinically implemented. Random‑start COS and letrozole/tamoxifen protocols minimise treatment delay and hormonal exposure; GnRHa co‑treatment preserves ovarian function as an adjunct. Utilisation of banked gametes/tissue remains modest, underscoring the need for pathway optimisation [1–10]. Conclusions: FP should be embedded across oncology and non‑oncology pathways with timely referral, clear counselling on probabilities of live birth, and robust follow‑up. Standardisation, registry‑based evidence and long‑term offspring safety data are priorities to bridge the gap between laboratory potential and clinical effectiveness [1–10].

Abstract: Background: Optimizing intrauterine insemination (IUI) outcomes while minimizing gonadotropin exposure, treatment cost, and the risk of ovarian hyperstimulation syndrome (OHSS) remains a central challenge in ovulation induction, particularly among women with polycystic ovary syndrome (PCOS) or high antral follicle counts (AFC). Sequential stimulation protocols incorporating early letrozole followed by delayed recombinant follicle-stimulating hormone (rFSH) have been proposed as a strategy to improve clinical efficiency while preserving safety and pregnancy outcomes. Objective To compare pregnancy outcomes, gonadotropin consumption, and safety profiles of a letrozole plus late-onset rFSH protocol versus conventional rFSH stimulation in IUI cycles, with particular emphasis on patients with PCOS and high ovarian reserve. Methods: This retrospective comparative cohort study included 764 IUI cycles performed between January 2022 and December 2025 at a tertiary assisted reproductive technologies center. Cycles were stimulated either with early letrozole followed by late-onset rFSH (n = 392) or with conventional rFSH alone (n = 372). The primary outcome was pregnancy per cycle, defined by a positive serum β-hCG test. Secondary outcomes included total gonadotropin dose, endometrial thickness, cycle cancellation, OHSS incidence, and obstetric outcomes. Multivariable logistic regression, propensity score matching (PSM), inverse probability of treatment weighting (IPTW), mediation analysis, and doubly robust methods were applied to account for baseline imbalances and confounding. Results: In unadjusted analyses, different stimulation protocols influenced pregnancy rates, but the letrozole plus late-rFSH group used significantly lower gonadotropin doses. After accounting for factors like female age and ovarian reserve, stimulation protocol did not independently predict pregnancy outcomes; female age was the main predictor. The rates of cycle cancellation and OHSS were low and similar across protocols, including in women with PCOS and high AFC. Propensity score analyses verified that the letrozole-based protocol produced pregnancy outcomes comparable to standard rFSH stimulation while decreasing gonadotropin use. Conclusions: Sequential stimulation with letrozole plus late-onset rFSH provides pregnancy outcomes comparable to conventional rFSH stimulation while significantly reducing gonadotropin requirements and maintaining favorable safety profiles, even in high-risk populations such as PCOS and high AFC patients. These findings support individualized ovarian stimulation strategies that prioritize both clinical effectiveness and treatment efficiency in IUI cycles.

Abstract: Levonorgestrel (LNG) used in contraceptive methods, necessitates accurate pharmacokinetic (PK) profiling to ensure efficacy and safety. The objective of this study was to develop and validate a highly sensitive LC-MS/MS method for quantification of LNG in human serum and its application in a pharmacokinetic study. 500 µL sample was processed using a liquid-liquid extraction (LLE) protocol optimized for LNG recovery that yielded consistent analyte recovery while minimizing matrix interferences. Chromatographic separation was performed on Waters ACQUITY UPLC BEH C18 reverse-phase analytical column (2.1× 100 mm, 1.7 µm) using mobile phase consisting of acetonitrile and water with 0.1% formic acid (70:30, v/v), delivered under a gradient elution at 300 µL/min flow rate. These conditions provided sharp peak resolution and minimized run times. Mass spectrometric analysis was run with Waters XEVO TQ ABSOLUTE triple quadrupole instrument operating in positive electrospray ionization mode and monitored LNG using multiple reaction monitoring with precursor ion at m/z 312.98>81.468, 90.921, 108.963 amu. Optimized fragmentation parameters yielded high signal-to-noise ratios, enabling trace-level detection in biological matrices. The method demonstrated linearity over calibration ranges of 32.5–2000 pg/mL, with correlation coefficients (R²) exceeding 0.997. Precision and accuracy met bioanalytical method validation guidelines, confirming the robustness of assay for PKs. The validated method was successfully employed to measure serum LNG in a PK study among women implanted in Bangladesh. The method’s sensitivity facilitated LNG detection supporting assessment of drug-release performance in clinical settings. The method proved highly reliable, sensitive, and specific for quantifying LNG in human serum, supporting its use in both clinical monitoring and research applications.

Abstract: This study evaluated whether a gonadotropin-releasing hormone (GnRH) agonist can be effectively used for luteal phase support following a dual trigger in IVF. A total of 284 patients undergoing fresh embryo transfer were included: 116 received a dual trigger and 168 an hCG trigger. All patients received luteal support with Nafarelin nasal spray twice daily for two weeks, starting on the day of oocyte retrieval. Women in the dual trigger group were older (36.2 vs. 32.3 years, p <  0.01), had lower antral follicle counts (7.0 vs. 17.5, p = 0.03), and received higher total gonadotropin doses (3000 vs. 2041 IU, p <  0.01). There were no significant differences between groups in retrieved and mature oocytes, fertilization rate, or blastulation rate. Positive pregnancy rate (β-hCG ≥ 25 mIU/mL) was 46.6% in the dual trigger group and 41.1% in the hCG group, without a significant difference, with similar live birth and miscarriage rates. Logistic regression identified maternal age as a predictor of positive pregnancy (OR = 0.95; 95% CI: 0.90–0.99; p = 0.024), while trigger type was not. Propensity score matching confirmed comparable IVF and pregnancy outcomes between groups. These findings indicate that a dual trigger followed by luteal-phase GnRH agonist support yields IVF and pregnancy outcomes similar to an hCG trigger.

Abstract: Pre-birth gene correction offers a precise and potentially transformative strategy to prevent severe genetic diseases by repairing pathogenic variants in embryos or gametes. Unlike selection-based preimplantation genetic testing (PGT), which depends on the availability of unaffected embryos, pre-birth gene correction directly addresses the underlying mutation, expanding the number of viable embryos available for transfer. We modeled the prospective impact of this technology in the United States across four exemplar monogenic diseases (sickle cell disease, cystic fibrosis, Marfan syndrome, and Huntington’s disease) under scenarios reflecting current and expanded access to assisted reproductive technologies. Depending on accessibility and implementation, pre-birth gene correction could correct hundreds to thousands of affected embryos each year, offering a viable path to parenthood for families who currently lack unaffected embryos through IVF and PGT alone. While translation will require rigorous evaluation of safety, efficacy, and ethical governance, these findings underscore this technology’s potential to broaden reproductive autonomy and meaningfully reduce the burden of severe genetic disease.