Progression of prostate cancer to castration-resistant disease is driven by early adaptive mechanisms that remain poorly defined. Increasing evidence suggests that senescence-associated programs and intratumoral steroidogenesis may contribute to therapeutic resistance, but their functional interplay in hormone-naïve disease is unclear. In this study, Hormone-naïve primary prostate cancer cultures (hnPCs) were established from diagnostic biopsies and characterized for proliferation, therapeutic response, senescence markers, and steroidogenic gene expression. Functional relevance of CDKN1A was assessed by knockdown experiments. Public transcriptomic datasets from ETS fusion-negative, AR signaling inhibitor-naïve metastatic castration-resistant prostate cancer (mCRPC) were interrogated for clinical validation. hnPCs exhibited low proliferative capacity yet intrinsic resistance to androgen deprivation therapy and docetaxel. Most cultures displayed coordinated overexpression of CDKN1A/p21 together with steroidogenic enzymes, particularly AKR1C3, alongside p16(INK4a) expression and SA-β-galactosidase activity, consistent with a partial senescence-like state. CDKN1A depletion selectively reduced AKR1C3 expression and increased proliferation in a subset of cultures, indicating that p21 maintains specific senescence-associated features while permitting adaptive plasticity. Analysis of ETS fusion-negative, ARSI-naïve mCRPC tumors revealed positive correlations between CDKN1A and steroidogenic genes, supporting the translational relevance of this program. These findings identify a previously underrecognized CDKN1A-associated partial senescence program that supports steroidogenic reprogramming, survival, and early therapeutic resistance in aggressive prostate cancer. This adaptive state may predispose tumors to progression toward castration resistance and represents a potential biomarker and therapeutic vulnerability in high-risk, ETS-negative disease. ### Competing Interest Statement The authors have declared no competing interest. Ministerio de Ciencia, Innovación y Universidades, https://ror.org/05r0vyz12, RTI2018-096055-B-I00

Background: Emergency Department (ED) viral hepatitis screening programs typically report short-term outcomes. We evaluated the medium-term impact of an ED-based screening for Hepatitis C (HCV), Hepatitis B (HBV), and Hepatitis D (HDV) viruses. Methods: Prospective opportunistic screening conducted between January 2020 and December 2024 at a referral hospital in Barcelona. Adults attending the ED were screened for HBV, HCV, and HDV. Positive cases were evaluated and linked to care when appropriate. To assess screening impact, HCV RNA and HBsAg prevalences were compared between 2020–2021 and 2023–2024 among participants aged ≤80 years. Results: Over five years, 39,532 individuals were screened: 220 (0.56%) had detectable HCV RNA, 234 (0.59%) HBsAg, and 4 HDV RNA. Of 110 patients with detectable HCV RNA eligible for linkage, 84 (76%) were treated and cured. Among 135 HBV-positive individuals eligible for linkage, 116 (86%) attended outpatient follow-up. HCV RNA prevalence significantly decreased (0.58% vs. 0.35%, p = 0.005), whereas HBsAg prevalence did not vary (0.7% vs. 0.58%, p = 0.224). Conclusions: Sustained ED-based screening enabled the identification and linkage to care of several individuals with HBV and HCV. This strategy contributed to a reduction in HCV prevalence among ED attendees and supports opportunistic ED screening in viral hepatitis elimination goals.

BACKGROUND:Accelerated biological ageing has been proposed as a key mechanism influencing disease progression in multiple sclerosis. Anti-Müllerian hormone (AMH), a marker of ovarian reserve, may reflect biological ageing in women and could be associated with disability progression. We aim to evaluate the prognostic implications of AMH levels measured at the first demyelinating event (FDE) suggestive of multiple sclerosis (MS). METHODS:This retrospective study analysed prospectively collected data from three Spanish neuroimmunology centres (since 1994). It included 365 females aged 20-45 years with serum AMH measured within 2 years of onset. According to AMH tertiles (n, median ng/mL (range)), participants were classified as follows: high (n=121; 4.6 (3.2-17.4)), medium (n=122; 2.1 (1.3-3.2)) and low (n=121; 0.6 (0.01-1.3)). A total of 145 healthy females served as controls. Age-adjusted multivariable linear regression models assessed associations between AMH and baseline clinical, radiological and serum biomarkers. Cox regression models evaluated the risk of Expanded Disability Status Scale 3.0, second relapse, progression independent of relapse activity and relapse-associated worsening. RESULTS:At FDE, median (range) age was 32.9 (22.2-44.9) years, follow-up 6.5 years (0.01-29.4) and AMH 2.2 (0.01-17.4); 68% received disease-modifying treatment. AMH levels did not differ between patients and controls and were inversely correlated with chronological age (β -1.21 (95% CI -1.42 to -1.00), p<0.001). After adjustment for chronological age, AMH showed no independent association with clinical, radiological or biomarker measures, or long-term outcomes. CONCLUSIONS:AMH levels at the time of an FDE suggestive of MS were not associated with baseline activity or long-term outcomes related to relapses or disability. Our findings suggest that the apparent associations between AMH and disease characteristics are largely driven by chronological age, supporting the concept that ovarian reserve primarily reflects biological ageing rather than an independent determinant of MS progression.

IMPORTANCE:The impact of bariatric surgery (BS) on the performance of the 1 mg dexamethasone suppression test (DST) is not well established. OBJECTIVE:(1) To evaluate the intraindividual results of the DST in a group of patients before and 2 years after BS, and (2) to assess plasma dexamethasone levels and other factors influencing the reliability of the DST. METHODS:We conducted a prospective longitudinal study, including 38 subjects evaluating DST before and 2 years after BS. We also compared DST results, plasma dexamethasone levels, and related factors across 3 groups: individuals of the previous cohort 2 years post-BS (n = 21), patients with severe obesity without BS (pwO; n = 10), and healthy controls (n = 7). RESULTS:Post-BS patients had higher cortisol levels after DST compared with prior (0.9 vs 0.7 µg/dL; P < .01). Four individuals post-BS had cortisol levels >1.8 µg/dL in the absence of autonomous cortisol secretion. Plasma dexamethasone levels were significantly lower in post-BS patients (1.9 ng/dL) compared with non-operated pwO (3.7 ng/dL) and healthy controls (4.0 ng/mL), P < .01. Multivariate analysis identified BS (β = -1.258, P = .01) and sex hormone-binding globulin levels (β = -.013, P = .04) as significant independent predictors of plasma dexamethasone concentrations. CONCLUSION:Post-BS subjects showed higher post-DST cortisol levels and reached lower plasma dexamethasone concentration compared with non-operated individuals, which may lead to false-positive results. These findings highlight the need to consider dexamethasone measurements to enhance DST interpretation in post-BS patients. Further studies are necessary to validate these findings in broader populations. The underlying mechanisms need to be explored.

BACKGROUND:Patients undergoing solid organ and hematopoietic stem cell transplantation are at risk of opportunistic pathogenic infections that increase morbidity and mortality. Universal antiviral prophylaxis improves the outcomes in this context. Therapeutic drug monitoring of antiviral drugs is not universally recommended but may be necessary in certain complex or polymorbid patients. The authors aimed to develop and validate a high-performance liquid chromatography-tandem mass spectrometry method to simultaneously quantify ganciclovir, acyclovir, and letermovir in human serum. METHODS:A stable isotopically labeled internal standard was used for each antiviral drug. Compounds were extracted by protein precipitation, evaporation, and reconstitution in an aqueous mobile phase. Samples were analyzed using reverse-phase chromatography with subsequent detection by electrospray ionization in the positive ion mode on a triple quadrupole mass spectrometer (run time: 6.5 minutes). RESULTS:Analytical curves for ganciclovir and acyclovir exhibited linearity within 0.1-25 mg/L (R 2 > 0.993), whereas for letermovir, the linear range was 0.01-2 mg/L (R 2 = 0.999). Matrix effects were not observed. Intraday and interday precision and accuracy were within ±15%. A therapeutic drug monitoring-guided strategy was explored to optimize preemptive antiviral drug therapy in 3 cohorts of transplant recipients. Seventy-nine samples from 35 patients were quantified, revealing median trough concentrations of 0.2 mg/L for ganciclovir (n = 21), 0.28 mg/L for acyclovir (n = 26), and 0.29 mg/L for letermovir (n = 32). CONCLUSIONS:This method has been successfully applied in clinical settings and allows reliable and accurate drug-level measurements.

Background: To evaluate the impact of applying alternative diagnostic criteria for gestational diabetes mellitus (GDM) during the COVID-19 pandemic on GDM prevalence, obstetrical and perinatal outcomes, and costs, as compared to the standard diagnostic method. Methods: A cohort of pregnant individuals undergoing GMD screening with the alternative GDM method, which uses plasma glucose (fasting or non-fasting) and HbA1c, was compared with a cohort of pregnant individuals undergoing the standard GDM screening method. Both cohorts were obtained from six hospitals across Catalonia, Spain, from April 2020 to April 2022. The primary outcome was large for gestational age rate at birth. The secondary outcomes were composite adverse outcomes, including pregnancy complications, delivery complications, and neonatal complications. The cost differences between screening methods were also evaluated. A similar analysis was performed in the subgroup diagnosed with GDM. Results: Data were collected from 1543 pregnant individuals in the standard screening group and 2197 in the alternative screening group. The standard screening group had a higher GDM diagnostic rate than the alternative screening group (10.8% vs. 6.9%, respectively; p < 0.0001). The primary outcome (large for gestational age rate) was similar between groups: 200/1543 (13.0%) vs. 303/2197 (13.8%). The adjusted OR for this outcome was 1.74 (95% CI: 0.74–4.10). An adjusted analysis showed no differences between groups in the composite adverse outcomes for pregnancy complications (OR: 1.11; 95% CI: 0.91–1.36), delivery complications (OR: 0.95; 95% CI: 0.75–1.19), and neonatal complications (OR: 1.28; 95% CI: 0.94–1.75). Among individuals diagnosed with GDM, the large for gestational age rate was similar between groups: 13/166 (7.8%) vs. 15/151 (9.9%). The OR adjusted for this outcome was 1.24 (95% CI: 0.51–3.09). An adjusted analysis showed no differences in the composite adverse outcomes for pregnancy complications (OR: 1.57; 95% CI: 0.84–2.98), delivery complications (OR: 1.21; 95% CI: 0.63–2.35), and neonatal complications (OR: 1.35; 95% CI: 0.61–3.04). The mean cost (which included expenses for consumables, equipment, and personnel) of the alternative screening method was 46.0 euros (22.3 SD), as compared to 85.6 euros (67.5 SD) for the standard screening method. Conclusions: In this Spanish population during the COVID-19 pandemic, GDM prevalence was lower in the alternative screening group than in the standard screening group. After adjusting for GDM risk factors, outcomes related to obstetrics, delivery, and neonatal complications were comparable between both groups. Finally, the alternative screening method was cheaper than the standard screening method.

Introduction:This case illustrates the challenges in the diagnosis of a rare disease with an intricate orientation. The definitive genetic diagnosis was carried out after establishing crucial correlations between the preliminary clinical indications and the laboratory findings. Case Presentation:The initial presentation was myoclonic jerks. This was a direct consequence of hyperinsulinaemic hypoglycaemia (HH), and not a phenotypic characteristic described in previous case reports. Linking this to glycaemia led to the evaluation of response to fasting, where inadequate insulin suppression resulted in hypoketotic hypoglycaemia. The diagnosis of chronic kidney dysfunction associated with atypical Fanconi renal tubulopathy syndrome type 4 (FRTS4) was indicated on the basis of a decreased estimated glomerular filtration rate, nephrocalcinosis, millimetric lithiasis, rickets, and complex proximal tubulopathy. This indicated atypical FRTS4 as associated with HH and necessitated further molecular genetic testing. Conclusion:The patient was identified as a carrier of the c.187C>T (p.Arg63Trp) variant in the HNF4A gene, which is a heterozygous missense variant classified as pathogenic. This entity is rare, and the published literature reporting HNF4A gene variants associated with atypical FRTS and HH is limited. It is therefore important to report such cases to contribute to the growing body of evidence and help identify pathogenic HNF4A variants and their implications.

Background:Exemestane, an aromatase inhibitor commonly used for breast cancer treatment, shares structural similarities with sex steroids analyzed in clinical laboratories. We aimed to investigate the influence of exemestane cross-reactivity in the measurement of sex steroids across various immunoassays. Methods:We conducted a multicenter study involving measurements of androstenedione, testosterone, estradiol, progesterone, and 17-hydroxyprogesterone in serum samples from women undergoing exemestane therapy (N=15; 25 mg/day). Measurements were performed using liquid chromatography-mass spectrometry (LC-MS) and various commercially available chemiluminescence immunoassays, ELISA, and radioimmunoassay. In-vitro cross-reactivity was assessed by adding exemestane and 17-hydroexemestane to serum samples. Results:Patients undergoing exemestane therapy had markedly falsely elevated androstenedione results in all immunoassays evaluated (N=4), which correlated with serum exemestane levels. In-vitro experiments confirmed this interference to be caused by cross-reactivity with exemestane. Additionally, one immunoassay yielded falsely elevated estradiol results in 20% of patients. However, in-vitro experiments did not confirm this to be caused by cross-reactivity with exemestane or 17-hydroexemestane. Conclusions:Exemestane cross-reacts with androstenedione immunoassays, causing falsely elevated results in treated patients. This analytical interference may raise unnecessary concerns, leading to expensive diagnostic workups.

OBJECTIVE:To identify distinct clinical phenotypes in acromegaly based on growth hormone (GH) assay standardization and unsupervised machine learning. METHODS:This was a multicenter cross-sectional analysis of 416 patients diagnosed with acromegaly from 2010 onward. Patients were stratified according to baseline serum GH levels standardized to the assay-specific upper limit of normal (GHxULN) using a binary classification (GH-B: <1.0×ULN vs ≥1.0×ULN) and a four-tier classification (GH-4: <0.25, 0.25-0.99, 1.0-9.9, ≥10×ULN). Unsupervised cluster analysis included age, GHxULN, insulin-like growth factor 1 (IGF-1)xULN, tumor diameter, and T2-weighted signal intensity. RESULTS:Overall, 36% of patients had GH levels within the normal reference range for their assay (GH-B <1.0×ULN). Microadenomas (23.1%) were more frequent in older patients and associated with lower GH/IGF-1 levels. Across GH-4 categories, significant gradients were observed for age (z = -5.34, P < .001), tumor size (z = 8.01, P < .001), IGF-1 (z = 9.00, P < .001), and symptom duration (z = 4.34, P < .001). Higher GH categories were associated with greater odds of arthropathy (odds ratio 3.5, P = .015 for 1.0-9.9×ULN and odds ratio 6.58, P = .002 for ≥10×ULN). Cluster analysis revealed 3 phenotypes: cluster 1 (49.0%) [older age, lower GH/IGF-1, intermediate tumor size]; cluster 2 (44.4%) [intermediate age, moderate biochemical activity, smaller tumors]; cluster 3 (6.6%) [younger age, markedly elevated GH/IGF-1, large aggressive tumors]. CONCLUSION:GH standardization to assay-specific ULN reveals clinically meaningful phenotypes in acromegaly that correlate with age, tumor characteristics, and disease severity (particularly arthropathy). GHxULN complements IGF-1 by capturing tumor secretory activity, and this stratification approach may support more individualized clinical decision-making.

ABSTRACT Prostate cancer (PCa) is the most common cancer in men and the third leading cause of cancer death in Europe. The TP53 gene, the most frequently mutated gene in human cancer, is a tumor suppressor gene with crucial functions in preventing tumor development. The single nucleotide polymorphism rs1042522, characterized by the substitution of a proline (PRO) for an arginine (ARG) at the position 72 of the p53 protein (P72R SNP), was studied in 12 primary tumor cultures from prostate biopsies of untreated hormone-naïve patients (hnPCs) with aggressive-metastatic cancer (Gleason ≥8), 11 radical prostatectomies, and a cohort of 94 serums from patient with aggressive prostate cancer using DNA sequencing and melting curve analysis. The results identified a high frequency of the P72R SNP in prostate cancer samples compared to the general European (non-cancer) population, suggesting a very significant association (p<0.0001) between this SNP and the risk of prostate cancer with an odds ratio of 7.937 (IC 95% 5.37-11.00). The G allele (R72) was more frequent in patients with high Gleason scores (≥8) suggesting its association to more undifferentiated-malignant PCa lesions.

Background: Bariatric surgery (BS) is effective for achieving significant weight loss. However, weight regain (WR) is an emerging problem. Objective: To assess the prognostic value of morning serum cortisol, a 1 mg dexamethasone suppression test (DST), 24 h urinary free cortisol (UFC) and late-night salivary cortisol (LNSC) in a cohort of patients with severe obesity (pwSO) undergoing BS in terms of weight loss and WR. Methods: Patients scheduled for BS underwent the following procedures at baseline, 12 months and 24 months after BS: medical history, anthropometric data, blood analysis and cortisol tests. We evaluated total weight loss (TWL) ≥ 30% at 1 year and WR after 2 years as an increase of ≥10% of the maximum weight lost. Results: In total, 142 subjects were included; 101 (71.1%) were females and the mean age was 45.9 ± 9.2 years. Up to 76.8% of subjects achieved ≥30% TWL, without statistically significant differences in DST results or morning serum cortisol, UFC or LNSC levels. However, a higher pre-surgery morning serum cortisol level was a significant predictor of a WR ≥ 10% (cortisol 17.8 [IQR 13.1–18.5] vs. 12.0 [IQR 8.8–15.8] μg/dL; p < 0.01); OR of 1.216 (95% CI 1.069–1.384); AUC [0.761, CI: (0.616–0.906); p < 0.01]. A cut-off value of cortisol > 13.0 μg/dL was predictive of a WR ≥ 10% (sensitivity 0.71; specificity 0.63). Conclusions: No cortisol test was useful in predicting weight loss; however, the pre-surgery morning serum cortisol level was able to predict a WR ≥ 10% in a cohort of pwSO 2 years after BS. A cut-off value of cortisol > 13 μg/dL might be an easy tool to identify patients at higher risk of WR, enabling healthcare providers to implement tailored, long-term strategies to minimize this outcome.

Mutations in GBA1, encoding glucocerebrosidase beta 1 (GCase), are the most common genetic risk factor for Parkinson's disease (PD). GCase dysfunction leads to an accumulation of glucosylceramide (GluCer) substrates in different organs and fluids. Despite the challenges in quantifying GluCer isoforms in biological samples, their potential clinical interest as PD biomarkers justifies the development of robust assays. An extensively evaluated high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) method for quantifying 14 GluCer and galactosylceramide (GalCer) isoforms in human cerebrospinal fluid (CSF) samples is presented. Sample pretreatment, HPLC, and MS/MS parameters were optimized. Evaluation was performed according to the recommendations of the Clinical and Laboratory Standards Institute and European Medicines Agency guidelines. Four 7-point calibration curves were generated, with a linearity interval from 2.5 to 200 nM (R2 ≥ 0.995). The limit of quantification was set at 5 nM. Between-run precision and accuracy were up to 12.5 and 9%, respectively. After method validation, we measured the levels of GluCer and GalCer isoforms in CSF human samples, including 6 healthy controls (HC), 22 idiopathic GBA1 wild-type PD (iPD) patients, and 5 GBA1-associated PD (PD-GBA) patients. GluCer/GalCer median ratios were found to be higher in the CSF of PD-GBA patients, particularly in severe GBA1 mutations, than those in iPD and HC. The observed trends in GluCer/GalCer ratios among groups provide novel information for the comprehensive analysis of sphingolipids as potential biomarkers of PD.

Searchable abstracts of presentations at key conferences in endocrinology ISSN 1470-3947 (print) | ISSN 1479-6848 (online)

BACKGROUND AND PURPOSE:Vitamin D is considered to play a role in multiple sclerosis (MS) etiopathogenesis. A polymorphism in the CYP24A1 gene, rs2762943, was recently identified that was associated with an increased MS risk. CYP24A1 encodes a protein involved in the catabolism of the active form of vitamin D. The immunological effects of carrying the rs2762943 risk allele were investigated, as well as its role as genetic modifier.METHODS:Serum levels of 25-hydroxyvitamin D and 1,25-dihydroxyvitamin D (1,25(OH)2 D) were measured in a cohort of 167 MS patients. In a subgroup of patients, expression levels of major histocompatibility complex class II and co-stimulatory molecules were determined by flow cytometry, and serum levels of pro-inflammatory (interferon gamma, granulocyte macrophage colony-stimulating factor, C-X-C motif chemokine ligand 13) and anti-inflammatory (interleukin 10) cytokines and neurofilament light chain were measured by single-molecule array assays. The effect of the rs2762943 polymorphism on disease activity and disability measures was evaluated in 340 MS patients.RESULTS:Compared to non-carriers, carriers of the rs2762943 risk allele were characterized by reduced levels of 1,25(OH)2 D (p = 0.0001) and elevated levels of interferon gamma (p = 0.03) and granulocyte macrophage colony-stimulating factor (p = 0.008), whereas no significant differences were observed for the other markers. The presence of the rs2762943 risk allele had no significant impact on disease activity and disability outcomes during follow-up. However, risk allele carriers were younger at disease onset (p = 0.04).CONCLUSIONS:These findings suggest that the CYP24A1 rs2762943 polymorphism plays a more important role in MS susceptibility than in disease prognosis and is associated with lower 1,25(OH)2 D levels and a heightened pro-inflammatory environment in MS patients.

BACKGROUND Gestational hypothyroidism has been shown to be associated with adverse pregnancy outcomes as well as adverse outcomes for the child. Thyroid hormones concentrations change in gestation, especially within the first trimester, so the results of thyroid function test often are outside non-pregnant reference ranges. The objective of this study was to establish the first trimester reference ranges for thyroid stimulating hormone (TSH) and free thyroxine (FT4) for pregnant women in Barcelona (Spain). METHODS It was a prospective study in which 673 women were recruited during their first trimester of gestation (8-13 weeks). Serum TSH, FT4 and antithyroid peroxidase antibodies (TPOAb) were measured with Atellica® IM 1600 (Siemens Healthineers). After excluding 418 women, the reference ranges for TSH and FT4 were calculated by the 2.5th and 97.5th percentiles. Potential variables examined in this study were age, body mass index (BMI), ethnicity, iodine supplementation and smoking habit. RESULTS The reference ranges established on the Atellica® IM 1600 for the first trimester pregnancy in our population were 0.111 to 4.291 mIU/L for TSH and 11.45 to 17.76 pmol/L for FT4. No significant differences were found in thyroid hormones concentrations regarding maternal age (≤30 years versus >30 years) (p=0.117), iodine supplementation (p=0.683) and smoking habit (p=0.363). The prevalence of TPOAb was estimated at 10.0%. CONCLUSIONS We found that in our local population the optimal TSH upper reference limit in the first trimester of gestation was 4.3 mIU/L, similar to that proposed by de ATA-2017 guideline (4.0 mIU/L).