Adoptive T-cell therapies engineered with T-cell receptors (TCRs) or TCR-like antibodies have shown considerable promise in cancer immunotherapy. However, identifying tumor antigen-specific TCR-like antibodies, particularly against human leukocyte antigen-presented neoantigens, remains challenging. Here, we present a function-based, rather than affinity-based, antibody screening platform utilizing Synthetic T-cell receptor and Antigen Receptor (STAR)-T cell libraries. We found that antigen engagement in STAR-T cells triggers synchronous receptor endocytosis and T-cell activation, and we integrated these paired processes into an Endocytosis-Activation (E-A) functional readout for antibody screening. Applying E-A functional screening, we rapidly identified multiple nanobodies targeting the cell-surface antigen CD22 as well as the intracellular neoantigen P53R175H. STAR-T cells engineered with these nanobodies mediated potent anti-tumor efficacy both in vitro and in vivo. Furthermore, this platform yielded nanobodies that can be directly reformatted into other therapeutic modalities, including chimeric antigen receptors and bispecific antibodies, while maintaining cytotoxic function. Overall, the E-A screening platform links antibody discovery directly to T-cell function, providing a robust approach for identifying therapeutic antibodies, especially neoantigen-specific nanobodies, for T cell-based cancer immunotherapy.
To determine the change in modified star excursion balance test scores in patients between 6 and 12 months following anterior cruciate ligament (ACL) reconstruction and the change in the proportion of patients who met recommended thresholds at each timepoint. A secondary aim was to assess whether change over time differed according to age, sex, graft type or activity level. Longitudinal cohort. Orthopaedic Clinic. 115 patients that completed 6- and 12-month testing after ACL reconstruction. Modified star excursion balance test scores (composite and individual reach scores). There was a significant increase of test scores for the surgical limb at 12-months compared to 6-months (p < 0.005) with no differences in limb symmetry index. Most patients (92%) reached threshold composite scores >90% equivalent to individual leg length at 6-months. This proportion did not change at 12-months. Patient-related factors had no effect on change in test scores across time. The modified star excursion balance test may be more applicable to assess dynamic balance performance 6-months post-surgery. Almost all patients met pre-determined criteria for dynamic balance performance by 6-months following surgery. There was no impact of patient age, sex, graft type or activity level on change in performance between 6- and 12-months.
Convectional heat transfer continues to receive extensive research attention due to its importance in vital applications such as solar energy collectors and cooling of nuclear reactors. This research investigates natural convection and associated entropy generation within a porous wavy enclosure with a star-shaped hot cylinder. The fluid saturated in the porous medium is assumed to be a water-Cu-Al2O3 hybrid nanofluid. The energy balance equation accounts for thermal non-equilibrium between the hybrid nanofluid and the local structure of the porous medium (LTNE). The effects of the number of lobes of the star-shaped cylinder (N), the Rayliegh number (Ra), the Darcy number (Da), the porosity of the medium (ε), and the volume fraction of the different nanoparticles (φAl2O3 and φCu) are inspected using numerical FEM analysis and optimized by the aid of artificial neural network (ANN). The results show that it is not possible to increase the Nusselt number without increasing entropy, and it is not possible to decrease entropy without decreasing heat transfer. The objective function (OBF), defined as the ratio of the Nusselt number to the total entropy generation, indicates that the best design is achieved with low obstacle waviness and low nanoparticle loading. The number of lobes and the nanoparticle volume fraction often increase entropy faster than they improve heat transfer. The maximum OBF = 3.877 (Nuavg = 38.705, ST = 9.9824) occurs at N = 1, φAl2O3 = 0, φCu = 0, Ra = 103, Da = 10-4, ε = 0.1. This study demonstrates the advantages of using artificial neural networks (ANN) to optimize the design of heat exchangers filled with nanofluids. This approach minimizes losses caused by thermal and flow irreversibility, thereby contributing to energy savings.
Biomineralized structures produced by living organisms are widely recognized for their exceptional mechanical performance, yet their potential optical roles are relatively less explored. Here, we demonstrate that within the calcitic ossicle-based skeleton of the sea star Protoreaster nodosus, where each ossicle represents a discrete skeletal element, one specialized ossicle, known as the terminal plate, contains a radially arranged array of light-guiding structures (LGSs). These LGSs exhibit an elongated, cone-like geometry (~250 μm in length) and are embedded within the porous stereom, a characteristic meshwork architecture of echinoderms analogous to open-cell cellular solids and composed of magnesium-containing single-crystalline calcite. Optical experiments demonstrate that, unlike other skeletal elements, the terminal plate can transmit and focus light into an internal cavity via the LGS array. Combined optical analyses using ray-tracing and finite-difference time-domain simulations reveal that each LGS transmits ca. 70% of incident light at normal incidence and concentrates it up to 2.8-fold at its exiting surface. Furthermore, when acting collectively as the LGS array within the terminal plate, the LGSs capture light over a broad field of view (~120°), resulting in an integrated transmitted intensity that is sixfold to eightfold greater than the incoming intensity perceived by a single LGS. Although the biological function of this optical capability remains uncertain, this natural porous structure demonstrates that cellular solids can integrate efficient light-guiding behavior while enhancing mechanical properties (i.e., threefold increase in stiffness compared with random stereom), offering design insights for lightweight, multifunctional structures.
Dark matter (DM) models with a conserved particle-antiparticle number, n_{χ}-n_{χ[over ˜]}, and the asymmetry in the cosmological abundance n_{χ}≠n_{χ[over ˜]}, are known to be challenged by the existence of old neutron stars (NSs), as the sufficient accumulation of DM will lead to the collapse of NSs into black holes. We demonstrate that the applicability of these constraints is much wider and covers models with symmetric populations of DM, n_{χ}=n_{χ[over ˜]}, as the process of DM capture regulated by a nucleon-DM scattering can be inherently asymmetric, σ_{χn}≠σ_{χ[over ˜]n}. The asymmetry is induced by the interference of different types of χ-n interactions, provided that their combination is odd under charge conjugation in the DM sector, C_{χ}, and even under combined parity P_{χ+n}. We provide a complete analysis of DM-nucleon bilinear χ-n interactions and find that this asymmetry is very generic. Using canonical NS parameters and local DM halo inputs, we exclude spin-averaged scattering cross sections down to σ_{nχ}≳10^{-46}  cm^{2} at DM mass m_{χ}≲10^{10}  GeV for the maximally asymmetric capture rate and show that the constraints persist down to very small values of the cross-section asymmetry, A=(σ_{χn}-σ_{χ[over ˜]n})/(σ_{χn}+σ_{χ[over ˜]n})≳10^{-5}.
The World Health Organization has identified health literacy as a key pillar for resilient health systems in its current global strategy for 2025-2028. In this editorial, we argue that effectively addressing health literacy requires its integration into key strategic frameworks at both the global and national levels, as this represents a fundamental precondition for a more coordinated and systematic approach to the issue. Slovenia has followed these global directions by adopting the National Health Literacy Strategy 2025-2035 in 2025, establishing a ten-year strategic framework to strengthen health literacy. The country is also adhering to recommendations for ongoing research in this field; in 2026, the second national health literacy survey will be conducted. Looking ahead, the focus should be on developing and implementing practical public health interventions, and on strengthening coordination with existing health promotion and prevention programmes in Slovenia that are already delivering measurable impact. A key challenge will be to strengthen collaboration between researchers, policy-makers, and practitioners to help create a supportive, health-literate environment in Slovenia. Svetovna zdravstvena organizacija je v svoji aktualni globalni strategiji za obdobje 2025—2028 poudarila, da je zdravstvena pismenost eden ključnih stebrov odpornosti zdravstvenih sistemov. V tem uvodniku zato razpravljamo, da je za učinkovito naslavljanje zdravstvene pismenosti nujno njeno umeščanje v ključne strateške dokumente tako na globalni kot nacionalni ravni, saj to predstavlja temeljni pogoj za bolj usklajeno in sistematično obravnavo tega področja. Slovenija tem globalnim usmeritvam sledi, saj je leta 2025 sprejela Nacionalno strategijo zdravstvene pismenosti 2025—2035, s čimer je vzpostavila dolgoročni strateški okvir za razvoj in krepitev zdravstvene pismenosti. Hkrati sledimo usmeritvam glede kontinuiranega raziskovanja na tem področju – v letu 2026 poteka druga nacionalna raziskava zdravstvene pismenosti, ki bo omogočila vpogled v trenutno stanje in spremljanje trendov skozi čas. V prihodnje bo pomemben poudarek na razvoju in implementaciji konkretnih javnozdravstvenih intervencij ter na boljši koordinaciji z že obstoječimi promocijskimi in preventivnimi programi v Sloveniji, ki izkazujejo pomembne javnozdravstvene učinke. Ključni izziv bo predvsem krepitev povezovanja med raziskovalci, odločevalci in strokovnjaki iz prakse, saj bo le s takšnim sodelovanjem mogoče soustvarjati podporno zdravstveno pismeno okolje v Sloveniji.
A new study examines how distinct neuronal ensembles underlie the encoding and retrieval of complex spatial memories. The paper also explores how distinct memory representations interact with one another to drive appropriate behavior in a rodent foraging task.
Whether obstructive sleep apnea (OSA) severity is independently associated with sarcopenia, beyond the effects of age, obesity and sex, has not been established in a single-centre cohort using standardised ultrasound-based assessment. We examined sarcopenia prevalence and its components across OSA severity strata in a Kuwaiti cohort using the ISarcoPRM sarcopenia algorithm. Cross-sectional within-cohort analysis of 110 adults aged 50 years or older with confirmed OSA (apnea-hypopnea index [AHI] 5 or more events/h by Level 3 portable monitoring; SomnoTouch, Somnomedics, Germany), stratified as mild (AHI 5-14.99, n = 28), moderate (AHI 15-29.99, n = 39) or severe (AHI 30 or more, n = 43). Sarcopenia was assessed using the ISarcoPRM algorithm: quadriceps muscle thickness by ultrasound, Sonographic Thigh Adjustment Ratio (STAR), handgrip strength (Jamar dynamometer) and chair stand test (CST). Demographic and comorbidity profiles were balanced across severity groups (all p > 0.05). Quadriceps muscle thickness, STAR and handgrip strength did not differ significantly across severity strata (all Kruskal-Wallis p > 0.05). CST time showed a significant gradient across severity strata (Kruskal-Wallis p = 0.047), and both AHI and ODI correlated modestly with CST time (r = +0.209, p = 0.029 and r = +0.203, p = 0.034, respectively). Sarcopenia prevalence was 21.4%, 30.8% and 34.9% in mild, moderate and severe OSA, respectively, with no significant trend (Cochran-Armitage p = 0.237). Age (OR 1.12 per year, 95%CI 1.05-1.19, p < 0.001) and BMI (OR 1.10 per kg/m2, 95%CI 1.02-1.18, p = 0.009) were the independent predictors of sarcopenia; OSA severity was not (adjusted OR 1.19, 95%CI 0.65-2.18, p = 0.577). Low STAR prevalence was 83.6%, driven by the high-obesity burden in this cohort and the origin of STAR cut-offs in a lower BMI Turkish reference population. In this Kuwaiti OSA cohort, age and BMI are the dominant determinants of sarcopenia, with no independent contribution from OSA severity. A modest association between OSA severity indices and CST time suggests that physical function may be more sensitive to OSA-related changes than muscle mass per se. The near-universal low STAR prevalence points to the need for population-specific normative data in high-obesity cohorts.
NR5A1 encodes a transcription factor essential for adrenal and gonadal development. Gene variants are a known cause of heterogeneous 46,XY disorders of sex development (DSD), but the mechanisms underlying the phenotypic variability remain unclear. We investigated how different NR5A1 variants affect downstream gene regulation and contribute to DSD pathogenesis. We analyzed four naturally occurring NR5A1 variants identified in patients with 46,XY DSD-two novel (p.Cys65Ser, p.His310Arg) and two previously reported (p.Cys30Ser, p.Gln329*). We performed protein and transcriptomic analyses to characterize variant effects and identify dysregulated and candidate target genes, validated by qPCR and luciferase assays. Transcriptomic and CUT&Tag analyses focused on the p.Gln329* truncating variant. All four variants occurred at conserved residues and resulted in reduced NR5A1 protein expression and impaired nuclear localization upon transfection in HEK293T cells. Transcriptomic analysis using the p.Gln329* variant revealed broad downregulation of genes involved in steroidogenesis, including CYP11A1, STAR, and CYP17A1. Notably, AMHR2 and STARD8 were significantly downregulated and showed reduced CUT&Tag signal in variant-transfected cells. Promoter assays confirmed that all variants diminished CYP11A1 and AMHR2 promoter activity. Only the p.Gln329* variant affected STARD8 promoter activity. These findings indicate that NR5A1 variants impair protein expression and localization, leading to transcriptional dysregulation of genes involved in steroid hormone biosynthesis and sexual development. Based on analysis of the p.Gln329* truncating variant, AMHR2 and STARD8 are strong candidate novel downstream targets of NR5A1, offering further insight into the mechanisms driving 46,XY DSD. Differences in sex development can occur when the genetic instructions for building male reproductive organs are disrupted before birth. This study focuses on a key gene called NR5A1, which acts like a master switch to turn on other genes essential for testis development and hormone production. We investigated four variants in this gene found in children with 46,XY differences in sex development. Using cell models, we showed that these variants cause the NR5A1 protein to be produced at lower levels and prevent it from reaching the cell’s nucleus, where it normally works. As a result, the variant proteins fail to properly switch on several crucial target genes involved in making male hormones. We identified two new candidate genes controlled by NR5A1, called AMHR2 and STARD8, which may play important roles in this process. Our research helps explain how changes in a single gene can lead to a broad range of developmental outcomes and provides a clearer picture of the molecular steps involved in human sex development.
Community-level sociodemographic factors and hospital quality are associated with access to cancer care and outcomes. Using Medicare data (2016-2018), we evaluated the association between social determinants of health (SDoH) and hospital quality with 30-day mortality and readmission after selected elective cancer surgery. Separate multivariable logistic regression models sequentially adjusted for comorbidities and SDoH factors (Social Vulnerability Index (SVI) and Distressed Communities Index (DCI)) and then Hospital Star Rating. Among 16,869 patients, the "At Risk" DCI group and higher SVI were associated with higher 30-day mortality, and higher SVI was associated with higher odds of 30-day readmission and mortality. Subsequent models demonstrated that higher Hospital Star Rating was associated with lower odds of 30-day mortality and readmission and SDoH factors lost significance after adjusting for Hospital Star Rating. Hospital quality may have a greater impact on short-term outcomes than SDoH factors.
Adaptive optics aims to restore diffraction limited resolution in an imaging system in the presence of optical aberrations. To this end, the wavefront error needs to be measured prior to its compensation. Traditional wavefront sensors typically measure the local gradient of the wavefront emitted from a guide star (or another known point source) using a microlens array in front of a camera. They are typically dedicated devices that need to be integrated into an imaging system. Here we have tested the concept of estimating the wavefront error directly from an image of a guide star by training a machine learning model. We produce a two-photon laser spot in a water fluorescein media, and introduce random, but known aberrations using a deformable mirror to form a large training set. After validation, we integrated the machine learning wavefront sensor in an AO feedback loop with the deformable mirror. We demonstrate proof of principle compensation of sample-introduced optical aberrations with this system.
In the USA, the government certifies nursing homes to receive government funding and uses a 5-star rating system to share information about the quality of care in these nursing homes with the public on the CMS website. However, a star rating is not posted for the poorest performing nursing homes, the "Special Focus Facilities." We interviewed older adults to get their perspectives on the information posted online about SFFs. Respondents reported the term "Special Focus Facility" is misleading and confusing and the posted information incomplete and inadequate. Their suggestions for communication improvement about SFFs are included.
To test whether baseline depressive symptom profiles are associated with remission in Major Depressive Disorder (MDD) and to replicate previous findings from GSRD and STAR∗D in the PANDORA trial. We performed a secondary analysis of 215 adults with MDD enrolled in PANDORA and starting a new antidepressant. Baseline severity was assessed with the 17-item Hamilton Depression Rating Scale (HAM-D). Remission at week 12 was defined as HAM-D ≤ 7. Logistic regression models examined associations between baseline HAM-D total and item scores and remission overall and by sex, adjusting for age and sex and other covariates; sensitivity analyses further adjusted for treatment allocation (treatment as usual vs genetically guided therapy). Remission was achieved by 53% of patients (n = 114). Higher baseline HAM-D total score and higher scores on depressed mood, suicide, psychomotor retardation and hypochondriasis were associated with lower odds of remission in the total sample. In women, higher baseline severity, suicide and psychomotor retardation were associated with non-remission, whereas in men hypochondriasis was the only significant prognostic marker. These associations remained significant after adjustment for the treatment arm. Baseline depressive severity and selected symptom dimensions, particularly suicidal ideation and worry-related symptoms, relate to remission in MDD and may inform sex-sensitive, symptom-guided treatment strategies.
A self-fitting scaffold could enable a regenerative engineering approach to treat irregular craniomaxillofacial (CMF) bone defects. We have previously reported conformally fitting, shape memory polymer (SMP) scaffolds based on poly(ε-caprolactone) (PCL). The fitting temperature (i.e., melt temperature, T m ) was tuned based on PCL architecture: linear-PCL-diacrylate (linear-PCL-DA; T m = ~55 °C) or star-PCL-tetraacrylate (star-PCL-TA, T m = ~45 °C). Scaffolds were also formed as semi-interpenetrating networks (semi-IPNs) by incorporating thermoplastic poly(L-lactic acid) (PLLA). The inclusion of a polydimethylsiloxane-dimethacrylate (PDMS-DMA) macromer and 45S5 Bioglass® (BG) were independently shown to promote hydroxyapatite (HAp) mineralization, as well as to accelerate degradation. In this study, PDMS-containing, composite SMP scaffolds were prepared with varying macromer compositions and BG concentrations. PCL/PDMS co-matrix scaffolds were formed with either linear-PCL-DA or star-PCL-TA, and PDMS-DMA (75:25 wt%). PCL/PLLA/PDMS (75:12.5:12.5 wt%) co-matrix-semi-IPNs were also formed. BG was included at relatively low concentrations (5 and 10 wt%). Composite scaffolds were fabricated to concentrate BG on the pore walls via a modified solvent-cast particulate leaching (SCPL) approach with a fused salt/BG template. PDMS-containing scaffolds preserved shape memory behavior and were non-brittle. In vitro degradation rates were accelerated for PDMS-containing composite scaffolds, owing to a combination of phase separation of polymer components and hydrophilicity imparted by the BG. Additionally, robust bioactivity was observed for PDMS-containing composite scaffolds with HAp mineralization commencing in just 1 day (1X simulated body fluid; SBF).
To assess the knowledge, attitudes, and practices (KAP) regarding herpes zoster (HZ) and its vaccination among rheumatologists in China. A nationwide cross-sectional survey was conducted between April 1 and May 31, 2025, using the Questionnaire Star online platform. The questionnaire assessed sociodemographic characteristics, knowledge of HZ and its vaccine (via a 13-item scale), attitudes and behaviors regarding vaccination, and willingness to disseminate information. Ordinal logistic regression was applied to identify associated factors of knowledge scores, while subgroup analyses were conducted to evaluate attitude-related factors. A total of 1,772 rheumatologists from a wide range of provinces participated (mean age: 41.4 y; 67.3% female). Regarding professional background, most held a master's degree (44.1%), followed by a bachelor's (38.9%) and doctoral degree (17.0%). Clinical titles were distributed as associate chief (31.3%), chief (26.4%), attending (26.2%), and junior physicians (16.0%). The median knowledge score was 7 (IQR: 6-9). Multivariable analysis revealed that higher regional GDP per capita (β = 0.020, p = .040), advanced education (β = 0.234, p < .001), and greater clinical seniority (β = 0.296, p < .001) were independently associated with higher knowledge scores. Clinically, physicians expressed significant concern primarily in three areas: HZ's potential to interfere with the underlying autoimmune inflammatory rheumatic diseases (AIIRDs) (89.0%), HZ-related complications (81.5%), and deterioration of patient quality of life (73.0%). Notably, 65.6% of respondents were concerned about all three aspects simultaneously. Regarding vaccination attitudes, 96.4% of respondents reported conditional support for vaccinating patients with AIIRDs, contingent upon pre-vaccination risk assessment and adherence to recommended protocols. Additionally, 80.0% favored prioritizing recombinant vaccines. Subgroup analyses indicated that higher clinical seniority was significantly associated with more supportive attitudes (p < .001). In clinical practice, 76.5% of physicians reported actively recommending HZ vaccination, although several barriers remained. The main reasons for not recommending included limited consultation time (52.2%), perception of low HZ incidence (42.8%), and lack of patient concern (31.3%). Targeted education and strengthened rheumatologist-led counseling may help address existing barriers, particularly among junior physicians in under-resourced settings. Such measures could contribute to improving HZ vaccination uptake and protection in patients with AIIRDs.
To establish a non-invasive predictive model for microdissection testicular sperm extraction (micro-TESE) outcomes in FSH-normal non-obstructive azoospermia (NOA) patients by integrating gut microbiota profiling with serum biomarkers. We conducted a retrospective clinical analysis of 58 men and established a busulfan-induced FSH-normal NOA mouse model. Serum hormone levels (FSH, INH-B, AMH, testosterone) were measured by ELISA, and gut microbiota was analyzed via 16S rRNA sequencing. Testicular histology and ultrastructure were assessed by H&E staining and TEM, while protein expression was evaluated by IHC, IF, and Western blot. Receiver operating characteristic (ROC) curve analysis was used to evaluate the predictive efficacy of the serum AMH/INH-B ratio for sperm retrieval outcomes. In both patients and model mice, serum INH-B, AMH, and the AMH/INH-B ratio were significantly decreased (P < 0.01), correlating with severe spermatogenic impairment. Mice exhibited a marked reduction in the abundance of Bacteroidales and Muribaculaceae. Fecal microbiota transplantation (FMT) restored these microbial populations, improved testicular function, and upregulated key proteins involved in proliferation (PCNA, PGK2), blood-testis barrier integrity (ZO-1, Claudin11), and steroidogenesis (StAR, CYP17A1) (P < 0.05). Mechanistically, FMT increased serum short-chain fatty acid (SCFA) levels, which served as the chemical messengers correlating directly with the recovery of BTB proteins and steroidogenic enzymes. Clinically, the serum AMH/INH-B ratio showed strong predictive efficacy for micro-TESE outcomes, with an area under the ROC curve (AUC) of 0.92 (95% CI: 0.86-0.98), optimal cut-off value of 0.65, sensitivity of 88.2%, and specificity of 85.7%. The gut Bacteroidales abundance (from mouse data) was mechanistically linked to spermatogenic function, suggesting its potential as a future clinical biomarker pending validation. Our findings elucidate an SCFA-mediated gut-testis axis, highlighting the therapeutic potential of microbiota modulation and providing a novel tool to guide clinical decision-making, potentially reducing unnecessary surgeries in FSH-normal NOA.Additionally, the serum AMH/INH-B ratio serves as a robust non-invasive biomarker for predicting micro-TESE outcomes in FSH-normal NOA, while gut Bacteroidales abundance may represent a complementary mechanistic target for future clinical investigation.
Duchenne muscular dystrophy (DMD) is an X-linked recessive disorder characterized by progressive and severe muscle degeneration. Motor function tests are commonly used to evaluate treatment efficacy in clinical trials. However, they are subject to interobserver variability and may lack sensitivity for detecting early changes in disease progression. These limitations highlight the need for blood-based biomarkers to monitor disease status and progression. In this study, we used tandem mass tag-based mass spectrometry to quantify proteins in longitudinal serum samples from patients with DMD and to identify proteins associated with motor function performance. Serum samples collected at three time points (baseline, 12, and 24 months) were obtained from participants in the FOR-DMD trial (NCT01603407) and processed for multiplexed analysis using TMT 6-plex isobaric tags and LC-MS/MS. Protein intensities were log2-transformed and analyzed using linear mixed-effects models to assess their associations with age and repeated functional outcome measurements, such as the North Star Ambulatory Assessment (NSAA) score, 6-minute walk test (6MWT), rise from supine velocity (RSV), and 10-meter run/walk velocity (10mRWV). P-values were adjusted for multiple comparisons, with FDR < 0.05 considered statistically significant. Mixed-model analysis identified 22 proteins associated with age and 77 proteins associated with at least 1 functional outcome, including 26 associated with 2 clinical outcomes after FDR correction. Most associations were observed with NSAA (73 proteins), followed by the 6MWT (28 proteins) and RSV (3 proteins). These proteins spanned multiple disease-relevant categories, including muscle-associated proteins, extracellular matrix (ECM), complement and inflammatory pathways, coagulation/hemostasis, carrier proteins, proteolysis, and cell adhesion. Using longitudinal serum proteome profiles and clinical outcome data, we identified proteins that associate with age and functional outcomes, particularly NSAA and 6MWT, highlighting key molecular pathways in DMD disease progression. The FOR-DMD clinical trial was registered on ClinicalTrials.gov (registration no. NCT01603407). First submission: 03/04/2012.
Mitochondria play crucial roles in energy metabolism and steroidogenesis. Heat stress suppresses steroidogenesis in ovarian granulosa cells (GCs) and impairs poultry reproduction. Our previous study revealed that mitophagy protects duck GCs from acute heat exposure. Notably, numerous studies have demonstrated that the AMP-activated protein kinase/Unc-51-like kinase 1 (AMPK/ULK1) pathway is a key regulatory pathway for mitophagy. However, how AMPK coordinates mitophagy and steroidogenesis under heat stress remains unclear. This study aimed to elucidate the role of the AMPK/ULK1 axis in heat-induced mitophagy and steroidogenesis. Using a duck heat-treatment model and in vitro GC culture models, we combined Western blotting, targeted metabolomics, siRNA-mediated gene knockdown, and mitochondrial function assays to assess the impacts of the AMPK/ULK1 axis on mitophagy and steroidogenesis. We showed that heat treatment induced mitochondrial dysfunction, activated the AMPK/ULK1 pathway, triggered mitophagy, and suppressed steroidogenesis in duck ovarian tissue. In vitro, GCs exhibited similar mitochondrial impairment and AMPK/ULK1-dependent mitophagic activation under heat treatment. The LC-MS/MS analysis also confirmed elevated intracellular AMP levels in heat treatment GCs. Furthermore, activation of AMPK by 5'-aminoimidazole-4´-carboxamide ribonucleotide (AICAR) triggered mitophagy and enhanced steroidogenesis in GCs under heat treatment by upregulating the expression of steroidogenic acute regulatory protein (StAR), cytochrome P450 11A1 (CYP11A1), and cytochrome P450 19A1 (CYP19A1), whereas inhibition of AMPK by compound C exerted the opposite effect. Additionally, genetic knockdown of AMPK or ULK1 via siRNA reduced mitophagy and steroidogenesis under heat treatment. Taken together, the AMPK/ULK1 axis mediates heat-induced mitophagy and enhances GC steroidogenesis, positioning AMPK as a therapeutic target to improve heat-stressed poultry reproduction.
Atopic dermatitis (AD) is the most common inflammatory skin disease and carries the highest disability-adjusted life-years burden, ranking 15th among all non-fatal diseases globally. It is characterized by intensely itchy skin and is associated with multiple comorbidities, such as food allergy, asthma, allergic rhinitis and eosinophilic oesophagitis, which are mainly driven by type 2 immune responses. Other comorbidities include mental health disorders, disordered bone health, and cutaneous and extracutaneous infections. AD is also associated with other immune-mediated inflammatory diseases, including alopecia areata, vitiligo and inflammatory bowel disease. AD most often starts in the first 2 years of life but can occur at any life stage and onset at >60 years of age is increasingly common. The twenty-first century has brought greater insights into disease pathology, with an understanding of the complex interplay between the skin barrier, cutaneous and systemic immune pathways, cutaneous microbiome and neural networks. This improved mechanistic understanding has enabled rational drug design and a shift from non-specific broad immunomodulation to targeted biologic therapies and small molecules for severe disease and from topical corticosteroids to next-generation therapies for mild and moderate disease. Yet, considerable global inequity remains in access to these novel therapeutics.
Ovarian stimulation using exogenous follicle-stimulating hormone may enhance follicular recruitment; however, in the context of artificial insemination, it does not substantially improve pregnancy rates. Moreover, such physiological stimulation may impair oocyte developmental competence. To investigate the underlying metabolic mechanisms, the present study employed targeted metabolomics to compare the amino acid profiles in the follicular fluid of dairy cows during the natural estrous cycle (control group) and following ovarian stimulation (treated group). In total, 21 differentially abundant amino acids were identified: 7 were significantly upregulated, and 14 were significantly downregulated in the treatment group. Among the downregulated amino acids, L-serine was identified as a candidate hub metabolite via network analysis. Kyoto Encyclopedia of Genes and Genomes pathway enrichment analysis revealed that these differentially abundant amino acids were primarily enriched in the metabolic pathways of glycine, serine, and threonine, as well as in monocyclic lactam and arginine biosynthesis. Supplementation with 25-μM L-serine in the in vitro maturation medium significantly improved the oocyte maturation rate, cleavage rate, and blastocyst rate of oocytes from dairy cows undergoing ovarian stimulation. Additionally, supplementation promoted cumulus cell expansion and reduced the apoptosis rate of the formed blastocyst cells. The in vitro culture of bovine granulosa cells further demonstrated that L-serine could significantly promote cell proliferation and increase the estradiol to progesterone ratio (E2/P4). Real-time quantitative PCR analysis revealed that L-serine significantly upregulated the mRNA expression of key functional genes, including aromatase (CYP19A1), steroidogenic acute regulatory protein (STAR), and proliferating cell nuclear antigen (PCNA). These findings suggest a potential role for L-serine in supporting oocyte and embryo development, offering metabolic insights for further optimization of ovarian stimulation protocols.