Search Results (294)

Background/Objectives: Bile acid synthesis disorders (BASDs) represent a distinct category of progressive familiar cholestatic liver disease. A novel targeted mass spectrometry assay was developed for the accurate measurement of the major urinary atypical bile acids and bile alcohols that are biomarkers for HSD3B7, AKR1D1, CYP7B1 and CYP27A1 deficiencies, the four most common BASDs. Methods: Stable-isotope dilution UPLC tandem mass spectrometry was used for the simultaneous quantification of 12 key atypical bile acid biomarkers in urine from patients with BASD. Typical concentration ranges for these metabolites were established from urine samples from patients with biochemically and/or genetically confirmed BASD and compared with non-cholestatic and cholestatic controls. Results: The separation of major 3β-hydroxy-Δ⁵-bile acid sulfates, taurine- and glycine-conjugated 3-oxo-Δ⁴-bile acids, and bile alcohol glucuronides was achieved in a 20 min chromatographic run with intra- and inter-batch imprecisions of <15% for all metabolites. The mean ± SEM urinary concentration of total 3β-sulfated-Δ⁵-cholenoic acids in patients with HSD3B7 deficiency was 704 ± 204 µmol/L (n = 22), approximately 2000-fold higher than in cholestastic patients (n = 168) or non-cholestatic controls (n = 127). Similarly, the concentration of 5β-cholestane-3α,7α,12α,24,25-pentol-glucuronide, the major bile alcohol, in patients with CYP27A1 deficiency was 95 ± 17 µmol/L (n = 12). For CYP7B1 deficiency, two confirmed cases showed elevated levels (average, 7.5 µmol/L) of the glycine conjugate of 3β-sulfooxy-Δ⁵-bile acid. In AKR1D1 deficiency, total 3-oxo-Δ⁴-bile acids in urine were elevated (81 ± 16 µmol/L, n = 48), but concentrations showed overlap with cholestatic and non-cholestatic controls. Conclusions: A novel quantitative tandem mass spectrometry assay is described for the measurement of the major atypical metabolites and biomarkers in urine applicable to the accurate monitoring of treatment responses, and for the first time typical concentration ranges are established for each of these BASDs. Full article

Background/Objectives: Aldo–keto reductase isoforms AKR1C1–3 and 17β-hydroxysteroid dehydrogenase 1 and 2 (17β-HSD1 and 17β-HSD2) are key enzymes in steroid metabolism and validated targets in hormone-dependent cancers. Methods: In this study, Δ¹⁵- and 15β-substituted estrone derivatives were evaluated as inhibitors of AKR1C1–3 and 17β-HSD1 using enzymatic assays, cell viability assaysand computational modeling. Cellular uptake of the fluorescent estrone-based inhibitor was investigated using confocal microscopy. Results: The Δ¹⁵-estrone derivative showed potent and selective inhibition of 17β-HSD1 in the low nanomolar range, while 15β-O-propargyl and 15β-azide derivatives exhibited dual inhibitory activity against 17β-HSD1 and AKR1C2. The Δ¹⁵- and 15β-azide derivatives reduced cell viability in hormone-dependent breast, endometrial, and ovarian cancer cell lines in the sub- to low-micromolar range. A BODIPY-labeled 15β-O-propargyl analogue retained submicromolar inhibitory potency toward 17β-HSD1, representing the first fluorescent estrane-based inhibitor with preserved biological activity. Confocal microscopy confirmed efficient cellular uptake and predominant cytosolic localization in MCF-7 cells. Conclusions: These findings identify Δ¹⁵- and 15β-modified estrone derivatives as promising single- and dual-target inhibitors and introduce a fluorescent probe suitable for investigating intracellular steroid metabolism in hormone-dependent malignancies. Full article

Cyclophosphamide (CTX)-induced premature ovarian failure (POF) is characterized by disruption of the follicular microenvironment, granulosa-cell loss, endocrine imbalance, and oxidative-inflammatory injury. Here, we evaluated two multi-bioactive formulations developed to enhance ovarian stress resilience: a base formulation containing coenzyme Q10, calcium L-5-methyltetrahydrofolate, and Vitex agnus-castus extract (Base), and a urolithin A-enriched formulation (Base + U). Using a CTX-induced female C57BL/6 mouse model, we integrated phenotypic, histological, endocrine, oxidative-inflammatory, and transcriptional readouts to assess efficacy and mechanistic consistency. CTX markedly reduced ovarian index, disrupted estrous cyclicity, shifted follicle development toward atresia, increased granulosa-cell apoptosis, and caused endocrine dysregulation, including decreased anti-Müllerian hormone and estradiol and increased GnRH, FSH, and LH. CoQ10, Base, and Base + U each partially alleviated these abnormalities, improving ovarian index and coat condition, showing a trend toward improved follicular distribution, and normalizing hormone profiles. CTX also induced an ovarian oxidative-inflammatory shift, as reflected by decreased GSH-Px, increased MDA, and elevated IL-1β, IL-6, and TNF-α, all of which were attenuated by the interventions. Notably, Base + U more effectively reduced lipid peroxidation and TNF-α than Base alone. Consistently, ovarian transcripts related to follicle responsiveness and steroid regulation, including Fshr, Esr1, and Hsd17b2, were restored, whereas hypothalamic qRT-PCR analysis did not reveal robust transcriptional alterations within the intervention window. These findings suggest that the urolithin A-enhanced formulation partially alleviates CTX-induced ovarian dysfunction by suppressing oxidative-inflammatory stress and preserving granulosa-cell and follicle fate. Full article

Background/Objective: Familial Mediterranean fever (FMF) is an autoinflammatory disease caused by missense MEFV mutations, leading to recurrent episodes of interleukin (IL)-1β-mediated inflammation, and represents a model of cytokine-induced growth hormone (GH) resistance. Chronic or relapsing inflammatory bouts may impair growth in FMF children through functional alterations of the GH-insulin-like growth factor 1 (IGF-1) axis; however, the impact and reversibility of growth deficit remain unclear. The aim of this review is to assess data related to linear growth in young patients with FMF. Methods: This scoping review was conducted following PRISMA guidelines, searching for studies evaluating growth outcomes in FMF via the PubMed database. Fourteen studies, including 1144 children, were analyzed, evaluating height, growth velocity, IGF-1 levels, and treatment effects of colchicine or IL-1–targeted biologics. Results: Growth was generally preserved in a considerable number of children with FMF. Longitudinal analyses showed improvement in height standard deviation scores (HSDS) along with earlier and higher cumulative doses of colchicine. FMF attack frequency and overall disease severity modestly seemed to influence growth, whereas inflammatory markers were inconsistently correlated with growth parameters. Biologic therapies targeting IL-1 (canakinumab and anakinra) also showed positive effects on HSDS. Children with specific MEFV variants (such as M694V) or higher disease activity scores were at risk of developing a subtle growth impairment. Conclusions: Data on final height, though limited, suggest the preservation of growth in most pediatric patients with FMF. The maintenance of a normal linear growth is related to regular treatment with colchicine, though IL-1 blockers also appear to be beneficial in refractory FMF cases. These data highlight the importance of periodic, proactive check-ups and regular growth monitoring in children with FMF. Full article

Caffeine (CAF) is a widely consumed psychostimulant known to modulate adenosine receptors and neurotransmitter systems, although its effects in invertebrates remain poorly understood. Environmentally relevant concentrations (5, 30, and 50 µg/L) are associated with altered behavior, including locomotion, exploration, and feeding, in the freshwater gastropod Physella acuta. This study examined molecular responses underlying these effects. Adult snails were exposed to CAF for 24 h and 7 days. Gene expression related to the nervous system and stress pathways was analyzed by RT-PCR, including A1AR, ADORA2B, AChE, GLRA2, DRD2, RYR, HSD11β, HSP70, SLC6A2, and SLC6A1. After 24 h, exposure to 50 µg/L CAF altered A1AR expression and caused downregulation of AChE, GLRA2, and DRD2, associated with observed behavioral changes. A1AR upregulation may indicate compensatory adjustment in adenosine signaling. After 7 days, A1AR remained upregulated, while genes linked to inhibitory neurotransmission showed partial recovery. Increased expression of genes involved in dopamine regulation and steroid metabolism suggested physiological adaptation. Overall, CAF induced dose- and time-dependent molecular responses in P. acuta, linking neurochemical disruption with behavioral changes and highlighting its ecological risk as an emerging freshwater contaminant. Full article

Background: Sensorineural hearing loss represents a significant global health burden affecting over 1.5 billion individuals worldwide. Modern hearing aids, equipped with digital signal processing and smart connectivity features, constitute a cornerstone of neuro-sensory rehabilitation. However, the psychosocial impact of these assistive smart technologies on patient self-esteem remains incompletely characterized. Methods: A cross-sectional multivariate study was conducted with 245 participants, divided into three groups: normal-hearing controls (NH, n = 73), hearing-impaired patients using smart hearing aid technology (HA users, n = 86), and hearing-impaired patients not using hearing aid technology (HA non-users, n = 86). Self-esteem was measured using the Rosenberg Self-Esteem Scale (SES). Hearing disability and tinnitus severity were assessed with the Hearing Handicap Inventory for Adults (HHIA) and Tinnitus Handicap Inventory (THI), respectively. Data analysis included one-way ANOVA, Tukey’s HSD post hoc tests, Pearson correlations, and multivariate regression. Results: Hearing aid users showed significantly higher SES scores (35.41 ± 5.32) compared to non-users (22.99 ± 4.53; p < 0.001, Cohen’s d = 2.515). One-way ANOVA indicated highly significant differences among groups (F = 299.00, p < 0.001, η² = 0.712). SES was negatively correlated with HHIA (r = −0.573, p < 0.001) and THI (r = −0.443, p < 0.001), while HHIA and THI were strongly positively correlated (r = 0.729, p < 0.001). In multivariate analysis, HA use remained a strong independent predictor of self-esteem (β ≈ 11.9, p < 0.001), even after adjustment for age, sex, HHIA, and THI. Perceived hearing handicap was independently associated with lower self-esteem, whereas tinnitus severity was not a significant predictor in the fully adjusted model. The model explained approximately 65% of the variance in self-esteem scores. Conclusions: Smart hearing-aid use is strongly and independently associated with higher self-esteem in patients with sensorineural hearing loss. These results support the inclusion of modern audiological rehabilitation devices in comprehensive management strategies for long-term conditions and highlight psychosocial benefits that extend beyond hearing restoration. Full article

The placenta produces a variety of steroid hormones through the catalytic activity of steroidogenic enzymes, including cytochrome P450 (CYP) hydroxylases and hydroxysteroid dehydrogenases (HSD). Large amounts of progesterone produced by the placenta are essential for the maintenance of pregnancy. Although androgens and estrogens are also elevated in maternal circulation during gestation, there are conflicting reports on whether de novo synthesis of these steroids occurs in the human placenta. To address this issue, we performed a comprehensive analysis of steroidogenic gene expression in early and term placenta. While none of the genes examined showed binary expression changes, 17β-HSDs, including HSD17B1 and AKR1C3, were markedly upregulated in the term placenta. CYP19A1 and HSD11B2 genes were also markedly upregulated. In contrast, CYP17A1, CYP21A2, CYP11B1, CYP11B2, and HSD17B3 were almost undetectable. Consistent with these findings, the plasma ratios of active to precursor sex steroids (estradiol/estrone and testosterone/androstenedione) were higher in pregnant than in non-pregnant women, although concentrations of all steroids increased. In contrast, plasma levels and profiles of 11-oxygenated androgens were unchanged. These results indicate that the human placenta does not significantly contribute to circulating levels of either classical or novel classes of androgens. Therefore, this study provides new insights into the tissue of origin and the physiological significance of sex steroids during gestation. Full article

Background: Diets composed of various components have been shown to influence inflammatory diseases such as asthma. While most studies have focused on fiber-rich diets to investigate their effects on the immune system and, consequently, on asthma, little is known about the impact of sugar-rich diets, particularly when such diets are consumed over short periods of time. Methods: To investigate the short-term effects of a sugar-rich diet on allergic airway inflammation, A/J mice were fed either a standard diet or a sugar-enriched diet and subsequently sensitized and challenged with ovalbumin or PBS. Airway inflammation was assessed by bronchoalveolar lavage (BAL) cell analysis, including eosinophil counts and cytokine levels (IL-4, TNF-α, IL-33), and by lung histology (H&E for inflammatory infiltrate and PAS for mucus). Serum IgE levels were also measured. In addition, glucose tolerance, visceral and subcutaneous adipose tissue mass, and inflammatory markers in visceral adipose tissue were evaluated. Results: Short-term consumption of a sugar-rich diet induced glucose intolerance and expansion of adipose tissue, particularly visceral fat, independent of ovalbumin sensitization. Gonadal adipose tissue analysis revealed a shift toward M1 macrophage polarization, characterized by elevated TNF-α, IL-6, and IL-1β, increased leptin levels, and reduced adiponectin. In OVA-sensitized mice, the sugar-rich diet significantly exacerbated eosinophil infiltration in BAL, increased IL-4, TNF-α, and IL-33, and enhanced PAS-positive mucus accumulation and inflammatory infiltrates in the lung. Moreover, total serum IgE was significantly higher in allergic mice fed the sugar-rich diet compared with allergic mice on the standard diet. Importantly, in non-sensitized mice fed the sugar-rich diet, no pulmonary inflammation was detected by BAL, demonstrating that HSD alone does not induce asthma but amplifies allergic responses when sensitization is present. Conclusions: Our findings demonstrate that short-term consumption of a sugar-rich diet is sufficient to exacerbate, but not initiate, allergic pulmonary inflammation. From a translational perspective, reducing dietary sugar intake may represent a valuable adjuvant strategy in the management of allergic asthma. Full article

The Hippo pathway effector Yes-associated protein (YAP), acting through TEA domain (TEAD) transcription factors, regulates transcriptional programs in ovarian tissues; however, its role in interferon tau (IFNT) signaling within bovine luteal cells has not been investigated. This study aimed to determine whether YAP-TEAD interaction is required for IFNT-induced interferon-stimulated gene (ISG) expression in primary bovine luteal cells and to perform an exploratory assessment of selected receptor genes (ITGB1, GRP78, VEGFR2). Primary luteal cells were treated with recombinant ovine IFNT (roIFNT; 1 ng/mL) in the presence or absence of verteporfin (VP; 0.1, 0.5, or 1.0 µM), a pharmacological YAP-TEAD inhibitor, and mRNA expression was quantified by RT-qPCR. VP dose-dependently suppressed YAP target genes (YAP1, CTGF, ANKRD1) and reduced roIFNT-induced expression of MX1, MX2, and OAS1, whereas ISG15 was unaffected. Steroidogenic gene expression (3β-HSD, P450scc, StAR) remained unchanged across treatments, indicating preserved cell viability. Among the exploratory receptor endpoints, VP decreased ITGB1 and increased GRP78 at the highest concentration, while VEGFR2 was unaffected. These findings indicate that YAP-TEAD activity contributes to IFNT-induced ISG responsiveness in bovine luteal cells, with preliminary evidence of effects on integrin-mediated signaling pathways. Full article

Severe winter nutritional deficiencies may impair reproductive performance in Mongolian mares, yet optimal protein requirements during late gestation remain undefined. This study aimed to determine the effects of varying protein levels in complementary feed on gestational performance, physiology, and gut health. Seventeen late-gestating mares were assigned to three isocaloric diets differing in crude protein (CP)—high (HCP, 13.25%), medium (MCP, 12.04%), and low (LCP, 10.85%)—for 40 days. Statistical analysis was conducted using one-way ANOVA followed by Tukey’s HSD post hoc test. Mares fed the MCP diet tended to show more favorable growth performance and nutrient digestibility relative to the HCP group (p < 0.05). Blood analysis suggested that MCP mares had comparatively lower serum creatinine and creatine kinase concentrations, along with higher antioxidant capacity (catalase) and interleukin-1β levels (p < 0.001). Fecal microbiota sequencing showed that MCP was associated with comparatively higher microbial diversity, while HCP was characterized by enrichment of Proteobacteria, and LCP by enrichment of Bacteroidetes. Metabolomics identified 533 differential metabolites linked to protein metabolism. The MCP diet may help balance immune function, antioxidant status, and microbial homeostasis. These findings suggest that a complementary feed containing 12.04% CP may be associated with favorable effects on maternal health-related indicators in late-gestating Mongolian mares during winter. Full article

This study reviews the main candidate genes involved in the pathophysiology of Polycystic Ovary Syndrome (PCOS). PCOS is a common endocrine–metabolic disorder in women of reproductive age, characterized by menstrual irregularity, hyperandrogenism, and polycystic ovarian morphology. It is associated with increased metabolic and cardiovascular risk and is a leading cause of infertility. Although its pathophysiology is not fully understood, alterations in the hypothalamic–pituitary–ovarian axis, insulin metabolism, and steroidogenesis have been described. Polymorphisms in genes encoding hormones, enzymes, and receptors in these pathways contribute to clinical variability and ethnic differences, offering potential for early diagnosis and personalized medicine. This review summarizes key candidate genes related to insulin metabolism (INS, INSR, IRS-1), the hypothalamic–pituitary–ovarian axis (LHβ, LHCGR, FSHR, GnRHR, AMH, AMHR2, KISS1, CAPN10), steroidogenesis (CYP11A, CYP17A1, CYP19A1, CYP21, 17β-HSD, SHBG, AR, STAR), and other clinically relevant mechanisms such as obesity, lipid metabolism (PPARG, VDR, FTO), and follicular development (ACE). Full article

Endothelial dysfunction, chronic inflammation, immune dysregulation, oxidative stress, mitochondrial dysfunction, and metabolic disturbances collectively contribute to cardiovascular diseases (CVDs) associated with blood stasis patterns. Xuefu Zhuyu Decoction (XFZYD) is widely used clinically for the management of CVDs. Based on serum-exposed prototype profiling in rats, two pharmacology-driven core component sets of XFZYD were defined as the core set for the promotion of blood circulation and the elimination of blood stasis (CPBEB; HSYA, GRo, FA, β-ECD, AMY, ALB, PF) and the core set for the regulation of qi and the relief of pain (CRQRP; LIQ, NR, NAR, ROF, HSD, NHP, LTG, NRG, ISL, FNT, NOB, PD, SSa). CPBEB primarily targets vascular pathology by regulating endothelial dysfunction with dyslipidemia-driven arterial lipid deposition. Mechanistically, CPBEB is associated with improved endothelial function, reduced plaque instability, attenuated chronic inflammation and oxidative stress, normalized lipid and bile acid metabolism, and decreased thrombosis. CRQRP primarily modulates vascular tone and systemic energy metabolism. These effects are linked to enhanced AMPK/SIRT1-driven antioxidant defenses and mitochondrial homeostasis, increased NO/cGMP signaling, coordinated crosstalk among the TLR4/NF-κB, JAK/STAT, NLRP3, and PPAR pathways, and remodeling of the gut microbiota–immune network. In summary, this review integrates modern analytical approaches with network pharmacology and the literature evidence to clarify the material basis underlying XFZYD’s therapeutic effects in CVDs, thereby supporting the modernization and internationalization of traditional Chinese medicine. Full article

The hypothalamic–pituitary–adrenal (HPA) axis coordinates metabolic, immune, and behavioral responses to a changing environment. Its molecular effectors are the nuclear receptors for glucocorticoids and mineralocorticoids (the GRs/MRs), encoded by nr3c1/nr3c2. The MR serves as the high-affinity sensor of basal hormone concentrations, whereas the GR amplifies the stress response and mediates negative feedback. Despite their shared domain architecture, the receptors have diverged functionally: isoform composition, post-translational modifications, and the complement of co-regulators together determine which genes are activated or repressed in a given tissue at a given time. The regulation of the HPA axis activity is a major determinant of embryonic development. Pregnancy adds a placental control layer that meters maternal signals: 11β-hydroxysteroid dehydrogenase type 2 (11β-HSD2) in the syncytiotrophoblast inactivates cortisol, whereas 11β-hydroxysteroid dehydrogenase type 1 (11β-HSD1) can regenerate it, and systemic buffering by transcortin (cortisol-binding globulin, CBG) limits the free hormone fraction. Under stress, inflammation, or hypoxia, this barrier weakens, exposing the fetus to stronger glucocorticoid pulses during windows of heightened vulnerability for brain and immune development. Such overexposure not only reshapes ongoing transcription but is also epigenetically inscribed: the methylation of alternative nr3c1 promoters, the remodeling of histones, and the shifts in ncRNA profiles recalibrate the axis sensitivity for the long term. At the phenotypic level, this manifests as variability in stress reactivity, cognitive and affective trajectories, and an immune and metabolic risk across later ontogeny. In this review, we integrate evidence on the structure and functions of the GR, the mechanisms of its post-translational and epigenetic regulation, and the role of the placenta, to provide a coherent framework for understanding the multifaceted consequences of prenatal stress and to identify potential targets for early prevention. Full article

Hydroxyapatite (HAp) is a key bioceramic for biomedical applications, but conventional pressureless sintering (PS) requires high temperatures that can promote phase degradation. Here, we compare PS (1100 °C/180 min) and cold sintering process (CSP) (150 °C/450 MPa/30 min) for pure HAp and an HAp composite containing 4 wt.% niobium–phosphate bioglass (BG), using a 2 M H₃PO₄ transient liquid (10 wt.%). CSP increased relative density from 73.10% to 79.92% for HAp and from 68.43% to 83.54% for HAp/BG, representing up to a 22.1% gain compared with PS. One-way ANOVA confirmed a significant effect of processing route/composition on relative density (F(3,24) = 919.69, p < 0.05), and Tukey HSD indicated that all groups differed statistically. SEM revealed a markedly more consolidated and homogeneous microstructure for CSP, particularly for HAp/BG, consistent with enhanced dissolution–reprecipitation and pore filling. XRD showed that PS at 1100 °C led to partial HAp degradation with β-TCP formation, whereas CSP preserved the HAp phase with broader peaks, smaller crystallite size, and higher specific surface area. These results demonstrate CSP as an efficient low-temperature alternative for densifying HAp-based bioceramics, with BG addition further improving consolidation. Full article

Pregnanolone glutamate (PG) is a synthetic neurosteroid analog showing promise for treating ischemic brain injury, yet its blood–brain barrier (BBB) transport and metabolic fate remain unclear. We investigated the pharmacokinetics of PG in postnatal day 12 rats of both sexes subjected to endothelin-1 (ET-1)-induced focal hippocampal ischemia. Animals received PG (1 mg/kg intraperitoneal (i.p.)) or vehicle; serum and hippocampal steroidomes were profiled 60 min post-administration using gas chromatography-tandem mass spectrometry (GC-MS/MS) (hippocampus: n = 16 PG+, n = 27 PG−; multi-tissue subset: n = 6 PG+, n = 21 PG−). Our data revealed a “dual-fate” mechanism: PG undergoes systemic hydrolysis as a prodrug, as suggested by the tissue distribution pattern at 60 min post-administration, but also crosses the BBB intact, with significant parent conjugate accumulation in the hippocampus (42.3 pmol/g). The brain functioned as a “metabolic sink”, passively accumulating metabolites generated in peripheral organs—such as 17-hydroxypregnanolone—despite local absence of synthesizing enzymes (e.g., CYP17A1). Crucially, PG induced “metabolic segregation” within the central nervous system (CNS): the pharmacological 5β-pathway was saturated (~170-fold pregnanolone increase), while endogenous neuroprotective 5α-pathway (allopregnanolone) homeostasis remained preserved, contrasting with peripheral metabolic saturation. Preferential hippocampal accumulation of 3-oxo and 3β-isomers suggests autonomous regulatory buffering via oxidative 17β-hydroxysteroid dehydrogenase (HSD17B) enzymes, protecting against excessive GABAergic inhibition. This unique pharmacokinetic profile—combining metabolic segregation with active central buffering—defines PG as a dual-mechanism delivery system that generates central neuroactive metabolites—several with previously established GABAergic and neuroprotective activity—without disrupting endogenous neurosteroidogenesis, positioning it as a promising neurotherapeutic candidate minimizing physiological steroid homeostasis disruption. Importantly, the present study characterizes the pharmacokinetic and metabolic fate of PG; the neuroprotective efficacy of PG was demonstrated in our prior functional studies using the same model. Full article