Gene Literature Dashboard

<h4>Aim</h4>To investigate whether early lactate dehydrogenase (LDH) and amplitude-integrated electroencephalography (aEEG) improve the prediction of outcome in outborn infants transported for therapeutic hypothermia.<h4>Method</h4>Secondary analysis of a randomised controlled trial (2016-2019) including 113 asphyxiated newborn infants randomised to transport with a phase-change material mattress or standard care. The analysis included 81 infants with available aEEG, of whom 50 had LDH measured on admission. Outcome at 18 months was categorised as good, defined as normal development or mild delay, or poor, defined as moderate or severe impairment or death.<h4>Result</h4>Mean (SD) rectal temperature on admission was lower in the PCM group (34.6 [1.1]°C vs. 35.2 [1.1]°C, p = 0.027), but the proportion within target temperature did not differ (39.5% vs. 25.6%, p = 0.235). Clinical characteristics, LDH, aEEG and outcomes were similar between groups. Poor outcome was predicted by LDH > 1000 U/L (89% sensitivity and 61% specificity), severely depressed aEEG within 12 h (87% sensitivity and 47% specificity) and the combination of both (91% sensitivity, 62% specificity and accuracy 81%).<h4>Conclusion</h4>Combined LDH and aEEG provide a strong early prediction of outcome in encephalopathic infants.

Neurocognitive disorders (NCDs) are major conditions which impair the cognitive abilities of older adults and other populations worldwide, with incidence rising steadily every year. They constitute a group of acquired disorders characterized by progressive cognitive decline, encompassing delirium, mild and major NCD. These conditions exert wide‑ranging and profound adverse effects, including diminished quality of life, increased risks of falls, malnutrition, infections, reduced treatment adherence, and greater family caregiving and societal burdens. The pathogenesis of NCDs involves multiple coexisting factors and complex pathway interactions. Such heterogeneity and interwoven mechanisms contribute to the limited efficacy of existing pharmacological therapies. Current drugs such as acetylcholinesterase inhibitors and N‑methyl‑D‑aspartate receptor antagonists can provide partial symptom relief but cannot fundamentally halt disease progression. Moreover, current drugs are limited by notable side effects and, importantly, no effective standard treatment strategies exist for mild cognitive impairment, postoperative cognitive dysfunction or delirium. Against this background, natural compounds with multi‑target mechanisms, diverse biological activities such as anti‑inflammatory, antioxidant, neurotrophic and synaptic regulatory activities, and relatively low toxicity, for example flavonoids, alkaloids, terpenoids and polyphenols, are emerging as promising therapeutic candidates and major research foci. The aim of the present review was to summarize the pathogenic mechanisms of NCDs and highlight recent advances in the study of natural products for treatment, thereby providing a theoretical and research foundation for future translational applications in clinical prevention and therapy.

FAT4 (FAT Atypical Cadherin 4), a member of the atypical cadherin family, represents the human ortholog of Drosophila Fat (Ft) and functions as a positive regulator of planar cell polarity in the Drosophila wing. It serves various functions during embryogenesis and adult tissue development, demonstrating calcium-dependent adhesive properties. FAT4 has been extensively studied regarding its physiological functions and tumor mechanisms, with numerous studies emphasizing its essential role in various biological processes. A systematic mechanistic analysis and review of its key functions across these various biological processes is lacking. FAT4, as a transmembrane adhesion receptor and tumor suppressor, is expressed in cells across multiple systems, diseases, and diverse cancer tissues. It performs essential functions in diverse biological processes by modulating cell proliferation, polarity, and the immune microenvironment through multiple signaling pathways. This review delineates the functions of FAT4 in tissue morphogenesis, autoimmune diseases, and malignancies. The inactivation of this gene (through mutation, methylation, and deletion) is closely associated with multisystem developmental anomalies, immune responses, and cancer progression, thereby establishing it as a potential prognostic marker and therapeutic target. This study aimed to enhance the understanding of FAT4 and evaluate its potential applications in clinical diagnosis and treatment.

Peripherally administered therapeutics for neurological indications are challenged with anatomical and physiological barriers that limit their ability to access their site of action in the central nervous system (CNS). This is particularly true for complex therapeutics such as antibodies, immunotherapeutics, and gene therapies. The blood-brain barrier is the specialized structure that functionally regulates the ability of blood constituents to access the CNS. Blood-brain barrier delivery technologies for protein therapeutics have been established in pre-clinical models and are beginning to be verified in clinical studies. Technologies reliant on the transcellular pathway across the blood-brain barrier utilize the receptor-mediated transcytosis mechanism. Research into the use of lipid nanoparticles (LNPs) to deliver complex therapeutics has tremendously expanded in recent years. Lipid nanoparticles represent a compelling alternative to viral vectors for the delivery of various gene therapy modalities, including messenger RNA, small interfering RNA, and antisense oligonucleotides. Functionalization of LNPs with blood-brain barrier-penetrant moieties is being explored as a means to enable CNS delivery of LNP-based therapeutics. The recent innovations and validation of LNP-based delivery systems have hastened the fulfillment of the promise of facile CNS-targeted gene therapies. This review focuses on functional aspects of the blood-brain barrier and how they relate to recent advances in LNP technologies for CNS delivery, as well as their potential impact on gene therapy.

Immune checkpoint inhibitors have transformed cancer therapy, yet many patients fail to achieve durable responses due to insufficient T cell reinvigoration. Cytokines offer promise for enhancing immunotherapy, but their clinical use is limited by toxicity and a narrow therapeutic index. Immunocytokines, engineered fusion proteins combining antibody specificity with cytokine activity, aim to overcome these challenges by targeting cytokine delivery to immune cells or the tumor microenvironment. We describe SAR445877 (SAR'877), a potentially novel PD-1-targeted immunocytokine that fuses a high-affinity anti-PD-1 antibody with a detuned IL-15/IL-15Rα sushi domain complex. SAR'877 blocks PD-1/PD-L1 and PD-1/PD-L2 interactions while selectively delivering IL-15 signals to PD-1+ T cells, enhancing proliferation and activation of antigen-experienced CD8+ and CD4+ T cells and NK cells, while minimizing systemic inflammation. Mechanistically, SAR'877 activates STAT5 signaling in PD-1+ lymphocytes and restores effector function in exhausted T cells. In preclinical models, a murine surrogate of SAR'877 accelerated viral clearance and induced robust antitumor immunity by expanding cytotoxic CD8+ T cells and promoting Th1 polarization. Notably, SAR'877 outperformed anti-PD-1 plus untargeted IL-15, highlighting the therapeutic potential of targeted IL-15 delivery. These findings position SAR'877 as a promising next-generation immunotherapy with enhanced efficacy and reduced cytokine-associated toxicities.

Osteoarthritis is a degenerative joint disease that disrupts the cellular homeostasis within cartilage tissues, promoting further disease progression that can lead to debilitating pain. Cartilage of the temporomandibular joint (TMJ) is unique among diarthrodial joints because they are of neural crest origin, rather than the mesoderm germ layer. TMJ cartilage also has different cellular architecture, where it is composed of a superficial fibrous layer, a proliferative layer, and a fibrocartilage layer overlying subchondral bone. Understanding of the cytopathological changes that occur during TMJ osteoarthritis (TMJOA) could potentiate therapies to mitigate disease progression and repair diseased tissues. Here, we analyzed the unique cell populations present in healthy and OA-induced condylar cartilage of adult rats through single-cell RNA-sequencing. TMJOA was established via our previous rat model to study the changes in the cellular composition of the condyle in response to OA. Several cell types could be uniquely identified, and the prominent matrix producing cells were fibroblasts and chondrocyte subsets. Our trajectory and pseudotime analysis revealed three cell fates stemming from a fibrochondrocyte-like population and two chondrocyte cell fates that stem from a shared progenitor population. We also found that Pleiotrophin is uniquely expressed in the proliferative zone by cells with a chondrocyte progenitor phenotype. In OA cartilage, differential gene expression in the fibroblast group revealed responses to inflammation, possibly through activation of chondrocyte differentiation. The chondrocyte group was highly metabolically active, indicative of rapid repair or remodeling. Cell-cell signaling analysis revealed that chondrocyte and chondrocyte progenitor communication became highly activated. Additionally, intracellular pathways that may contribute to cellular dysfunction and tissue remodeling were highly active, while pathways related to tissue catabolism appeared less active.

The disordered adapter protein linker for activation of T cells (LAT) propagates T cell receptor signaling. To interrogate how LAT coordinates multiple downstream pathways, we developed a single-cell screening approach, identifying widespread functional segments including protein interaction motifs and blocks of negative charge. Regardless of their position in LAT, individual segments generally conferred defects across all downstream signaling pathways. To understand the underlying mechanism, we used molecular biology, computational modeling, and imaging to demonstrate that disruption of LAT interaction with a single partner protein indirectly disrupts other partner interactions, likely through the dual roles of these proteins as effectors of downstream signaling and bridging factors between LAT molecules. Overall, we describe an extendable approach for interrogating sequence-function relationships for proteins with complex activities.

Haemochromatosis is an inherited disorder of iron metabolism affecting approximately 100 000 Australians. Iron overload may result in end organ dysfunction, most commonly manifesting as chronic liver disease, arthropathy and endocrinopathies. End organ complications can be prevented, minimised or reversed, with early diagnosis and initiation of treatment in the form of venesection. Our understanding of the pathogenesis and epidemiological characteristics of the disorder has evolved in recent decades, including identification of HFE gene variants and sex-specific differences in phenotypic expression of the disorder. This clinical perspective article summarises the up-to-date evidence regarding the pathophysiology, clinical manifestations and treatment recommendations for haemochromatosis.

Polygenic risk score values vary with genetic ancestry due to differences in population-specific allele frequencies and linkage disequilibrium patterns. We present a framework to calibrate polygenic risk scores based on ancestral makeup. We propose the "expected polygenic risk score" or ePRS, defined as the expected value of a polygenic risk score based on one's global or local admixture patterns. We further define the "residual polygenic risk score" or rPRS as measuring the deviation of the polygenic risk score from the ePRS. The ePRS reflects the baseline ancestry-driven component of genetic risk, whereas the rPRS isolates an ancestry-agnostic measure of genetic liability. Simulation studies confirm that it suffices to adjust for ePRS to obtain nearly unbiased estimates of the polygenic risk score-outcome association without further adjusting for principal components. Using the TOPMed and the All of Us datasets, effect size estimates for the rPRS (adjusted for ePRS) are similar to those obtained from polygenic risk scores adjusting for genetic principal components. The ePRS framework can protect from population stratification in association analysis and provide an equitable strategy to interpret genetic risk across diverse populations.

A consistent postmortem finding in schizophrenia (SCZ) is reduction in dendrites' size. However, neurons with larger dendritic trees have also been encountered. In vitro experiments with neurons and neuronal-like cells coming directly from patients with SCZ showed similar results, evidencing at times more extensions and at others less arborizations. The process of extending and retracting neuronal outgrowths depends on microtubules polymerization and depolymerization. The possibility that microtubule polymerization/depolymerization is related to defects in the neuronal structure comes from several microtubular anomalies reported in SCZ such as its abnormal distribution in the cytoplasm, irregular shape of microtubules and even absence of these cytoskeletal components in dendrites. Moreover, microtubules in olfactory neuroepithelial cells from patients with SCZ were resistant to depolymerization. But whether deficits in microtubules are associated with abnormalities in the neuronal structure has never been investigated in living cells coming directly from patients. Therefore, we studied dynamic neurostructural changes in Monocyte-Derived-Neuronal-like cells (MDNCs) from 12 controls and 13 patients with SCZ. First, we showed that human neuroprogenitor cells and MDNCs have comparable neurostructural plasticity. Then, we investigated whether colchicine, a microtubular stabilizing and depolymerizing agent, disrupts dynamic neurostructural changes. The lowest concentration of colchicine tested, stopped dynamic neurostructural changes in MDNCs from controls, while cells from patients with SCZ continued to extend and retract neuronal outgrowths. Following, we investigated if antipsychotics impact dynamic structural changes, but our results were inconclusive. Our data indicate that, under certain circumstances, neuronal-like cells from patients with SCZ evidenced hyperdynamic microtubule-based neurostructural changes and consequently, link deficits in microtubules with anomalies in the neuronal shape. While other potential confounders are unlikely to have influenced our results, the effects of medications cannot be excluded.

Alcohol consumption triggers neuroinflammation, potentially creating a feed-forward loop that increases drinking. Previous studies have shown that activation of the toll-like receptor 7 (TLR7) leads to escalated drinking. In this study, we aim to identify cell type-specific transcriptomic patterns underlying TLR7-induced neuroinflammation, potentially leading to escalated drinking. Therefore, male C57BL/6J mice were treated with the selective TLR7 agonist R848 every-other day for 20 days in total. After 10 treatment-free days, half of the cohort underwent two bottle-choice drinking; the other half was sacrificed and brains were collected for single-nucleus RNA-Sequencing (snRNA-Seq) in the medial prefrontal cortex (mPFC) and central amygdala (AMG). The AMG showed a greater number of differentially expressed genes (DEGs), primarily in inhibitory and excitatory neurons. Among glial cells, AMG astrocytes exhibited the greatest number of DEGs, which were involved in blood-brain barrier (BBB) regulation (e.g., Cldn5, Mecom, Nrg1), a finding supported by secondary validation using Xenium in situ spatial transcriptomics on AMG-containing sections. BBB-regulatory genes, including those in the Wnt signaling pathway (e.g., Notch3, Top2a, Aldoc), were altered across multiple cell types in both regions. Together with the alterations observed in neurons, these findings suggest that repeated TLR7 activation induces persistent BBB and neuronal dysregulation, potentially leading to TLR7-induced escalated drinking.

Dysregulated lipid metabolism has emerged as a defining hallmark of colorectal cancer (CRC) progression, particularly in metastatic disease, where metabolic adaptation and immune evasion are tightly interconnected, as demonstrated in both murine models and human studies. Increasing evidence has demonstrated that alterations in lipid synthesis, uptake, transport, and oxidation not only sustain tumor bioenergetics but also actively remodel the tumor immune microenvironment. Key lipid metabolic regulators-including FASN, SREBP signaling, CD36-mediated lipid uptake, cholesterol metabolism, and fatty acid oxidation-coordinate oncogenic signaling and promote immunosuppressive states characterized by T-cell exhaustion, macrophage polarization, and ferroptosis resistance, on the basis largely of correlative and preclinical evidence. Recent advances in multiomics technologies, including single-cell and spatial transcriptomics, metabolomics, and lipidomics, have enabled high-resolution mapping of lipid-dependent immune niches within metastatic CRC (mCRC) lesions. These approaches reveal lipid metabolism as a central organizer of tumor-immune interactions and identify previously unrecognized metabolic vulnerabilities. In this review, we integrate current knowledge on lipid metabolic reprogramming in CRC with emerging multiomics insights, highlighting the mechanisms linking lipid metabolism, ferroptosis, gut microbiota interactions, and immune remodeling. We further discuss therapeutic strategies targeting lipid metabolic pathways and their potential synergy with immunotherapy. Collectively, the results of this work suggest that understanding lipid metabolism is a unifying framework for understanding mCRC biology and developing metabolism-guided therapeutic interventions.

Antigen processing and presentation (APP) is essential for adaptive immunosurveillance. We uncover a mechanism whereby activated T cell-derived extracellular vesicles (AT_EVs) drive a positive feedback loop that enhances antigen presentation and immune responses in normal physiology and cancer. AT_EV-induced immunogenicity relies on extracellular vesicular double-stranded DNA (EV_DNA), which is notably abundant and primarily composed of genomic DNA enriched in immune-related genes, including those encoding APP machinery. Mechanistically, granzyme B (Gzmb) packaged by AT_EVs disrupts the nuclear envelope of recipient cells, facilitating intranuclear transfer and subsequent transient expression of EV_DNA encoding APP genes. DNase treatment removes most AT-EV_DNA, abrogating APP upregulation and thus T cell activation and recruitment to tumors. Notably, AT_EVs hold promise as an acellular immunotherapy, restoring APP and synergizing with checkpoint blockade in immunotherapy-refractory tumors. Collectively, our findings uncover a mechanism of transient, non-viral gene delivery by AT_EVs that boosts APP and anti-tumor immunity while limiting autoimmunity.

Gut microbiota dysbiosis is a driving factor in Alzheimer's disease (AD), yet the mechanisms behind remain elusive. Emerging evidence highlights that outer membrane vesicles (OMVs) are critical mediators of microbiota-host communication. Here, we observed a reduction in a gut probiotic Escherichia coli Nissle 1917 (EcN)-like strain in AD patients, and its levels are positively associated with cognitive ability. The EcN OMVs containing outer membrane protein A (OmpA) translocate to the brain, reshaping the dysregulated immune network. Specifically, EcN OMVs are internalized by glia and neurons, suppressing glial hyperactivation and restoring synaptic function, thereby reducing Aβ deposition and cognitive deficits. The results further show that OmpA plays an important role in vesicle trafficking and inflammatory pathways and may be the key regulator of inflammatory mediators in EcN OMVs, modulating astrocyte-microglia-neuron interactions and functionality. This work discloses the substantial therapeutic potential of the probiotic and its secreted OMVs in intervention and treatment of neurological disorders.

Spatial transcriptomics extends traditional transcriptomic methods by quantifying gene expression within intact tissues while preserving each cell's precise spatial context. This technology also captures gene expression under physiological conditions, including interactions with the surrounding microenvironment, thereby enhancing our understanding of cellular states in both health and disease. Rapid recent advances have improved throughput, transcript capture, accuracy, and overall data quality. In this review, we summarize the major spatial transcriptomics platforms and outline their strengths and limitations. We also highlight key applications in neuroscience, including brain cell-type identification, structure-function relationships, and developmental processes. Additionally, we examine spatial gene-expression patterns in psychiatric and neurodegenerative disorders such as Alzheimer's disease and depression. Finally, we discuss emerging directions, including spatial multi-omics integration and the potential for artificial intelligence to advance brain research. Collectively, this work provides a foundation for future studies in neuroscience and brain disorders.

Cancer stemness drives malignant progression and drug resistance in hepatocellular carcinoma (HCC). Although mitochondrial dynamics are known to influence HCC development, the precise mechanisms linking mitochondrial function to stemness remain largely elusive. Integrating bulk and single-cell transcriptomics, we identified Butyrophilin Subfamily 3 Member A3 (BTN3A3) as a novel oncogene driving HCC stemness. BTN3A3 depletion markedly reduced sphere formation, stemness-related gene expression, and the percentage of CD90⁺/EpCAM⁺ cancer stem cells. Rescue experiments confirmed that BTN3A3 promotes HCC cell proliferation, migration, and invasion. Furthermore, BTN3A3 depletion sensitized HCC cells to sorafenib by inducing ROS accumulation and apoptosis. Mechanistically, mass spectrometry and Co-IP identified TOMM22 as a key mitochondrial interactor of BTN3A3. Crucially, sorafenib stress actively promotes BTN3A3 mitochondrial translocation, where it shields TOMM22 from ubiquitin-proteasome-dependent degradation. BTN3A3 deficiency led to TOMM22 depletion, mitochondrial fragmentation, and impaired oxidative phosphorylation (OXPHOS) and ATP production. Importantly, silencing TOMM22 reversed BTN3A3-mediated stemness and sorafenib resistance. In vivo orthotopic xenograft models and patient-derived organoids (PDOs) further validated that BTN3A3 correlates with stemness and malignant tumor growth. Utilizing 5E08, a pan-BTN3 monoclonal antibody, markedly suppressed tumor growth and concurrently downregulated TOMM22 expression in vivo. In conclusion, our study unveils a previously unrecognized non-immunological role for BTN3A3 in mitochondrial reprogramming. We demonstrate that BTN3A3 drives HCC stemness and drug resistance by preventing TOMM22 ubiquitination to maintain mitochondrial homeostasis. These findings position BTN3A3 as a promising therapeutic target, with the pan-BTN3 monoclonal antibody 5E08 offering a potential strategy to overcome stemness-driven malignancy and resistance in HCC patients.

The development of safe and effective radioprotective agents with minimal side effects, particularly for high-dose exposure, remains a global priority. E0703, a novel steroidal compound structurally derived from estradiol, has shown promising radioprotective efficacy with limited estrogenic activity in prior pharmacodynamic studies. In this study, E0703 was found to significantly increase the abundance of <i>Akkermansia muciniphila</i> (AKK) in the intestines of both irradiated and non-irradiated mice. Co-administration of E0703 and AKK markedly improved the 7-day survival rate of mice exposed to a lethal 8.5 Gy dose of radiation. E0703 induced beneficial transcriptional changes in AKK, with enrichment in metabolic pathways such as amino acid biosynthesis, aminoacyl-tRNA biosynthesis, the tricarboxylic acid (TCA) cycle, and fatty acid biosynthesis. These alterations supported the production of glucosamine 6-phosphate (GlcN-6-P) by AKK, which contributed to intestinal tissue regeneration following irradiation. Single-cell transcriptomic analysis revealed that E0703 significantly increased the proportion of intestinal stem cells and goblet cells by Day 5 post irradiation. Mechanistically, E0703 modulated the oxidative phosphorylation pathway in these cell types, including regulation of Muc2 production. E0703 also enhanced AKK abundance in irradiated mice, particularly in the presence of mucin, thereby elevating the availability of GlcN-6-P-a critical substrate for intestinal organoid repair. These findings indicate that E0703 exerts direct effects on goblet cells and AKK, promoting host-microbe interactions that facilitate intestinal regeneration and improve survival following radiation exposure.

Hydrogen peroxide (H₂O₂) plays a dual role as both a signalling molecule and a mediator of oxidative stress. Although mitochondria are major producers of H₂O₂, the relative contributions of mitochondrial versus cytosolic antioxidant systems to mitochondrial H₂O₂ homeostasis in intact cells remain poorly defined. Here, we combined compartment-resolved live-cell imaging using HyPer7, inducible mitochondrial H₂O₂ generation (matrix-targeted d-amino acid oxidase), kinetic modelling, and a targeted CRISPR/Cas9 screen to dissect determinants of mitochondrial H₂O₂ dynamics in HEK293 cells. Unexpectedly, we found that the cytosolic peroxiredoxin PRDX1 is a dominant regulator of mitochondrial matrix H₂O₂ levels. Loss of cytosolic PRDXs markedly enhanced matrix Hyper7 signals under both exogenous and mitochondria-intrinsic H₂O₂ production, exceeding the effects of deleting mitochondrial peroxiredoxins. Modelling and transport experiments indicated a very high permeability of the mitochondrial inner membrane to H₂O₂ enabling rapid efflux and the establishment of steep concentration gradients. This permits the cytosol to function as a major sink to limit matrix H₂O₂ accumulation. PRDX1 deficiency sensitized cells to chronic mitochondrial oxidative stress. A targeted CRISPR screen identified the Rab7 GAP TBC1D5, linking mitophagy to cellular survival under these conditions. Consistently, PRDX1/2-deficient cells exhibited elevated mitophagic flux, indicating mitochondrial quality control as a compensatory response. Our study reveals that cytosolic PRDXs critically impact mitochondrial redox homeostasis and provides a systems-level framework for understanding compartmental redox control and stress adaptation.

The heme synthesis pathway consists of eight enzyme-catalyzed steps, and pathogenic variants in the genes encoding these enzymes cause porphyrias. Diagnosis of certain porphyrias is often significantly delayed, due to their episodic and nonspecific symptoms that mimic more common conditions. To improve genetic diagnostics, which are increasingly used as a first-line approach, the ClinGen General Inborn Errors of Metabolism Gene Curation Expert Panel identified the need to perform gene-disease curations for all genes in this pathway. During this process, we found that several genes (HMBS, UROD, CPOX, and PPOX) had multiple disease assertions, primarily differentiated by dominant versus recessive inheritance. For example, deficiency of HMBS is associated with both autosomal dominant acute intermittent porphyria (AIP) and autosomal recessive homozygous dominant AIP (HD-AIP). Yet, both conditions result from loss-of-function of the HMBS protein, and some recessive cases share identical causative variants with dominant cases. Therefore, our expert panel decided to include patients from both disease assertions (AIP and HD-AIP) as part of a single gene-disease curation rather than separating these groups of patients. This approach correctly links the variable affecting severity to allele dosage. This "lumping" process also required novel parent terms that encompass all disease subtypes and have gene-based nomenclature. Parent terms incorporating both monoallelic and biallelic subtypes were assigned a semidominant inheritance pattern, reflecting the overlapping variants and variable phenotypic severity based on residual enzyme activity. These gene-disease curations for the heme synthesis pathway pave the way for improved downstream variant curation which is a critical requirement to improve porphyria diagnostics. A key focus of this undertaking is to clarify and advance more appropriate nomenclature for these gene-disease relationships, enabling molecular laboratories to report variants in the heme synthesis pathway with greater precision.

Primary adrenal insufficiency is a rare disease characterized by deficient production of glucocorticoids with or without a deficiency of mineralocorticoids and adrenal androgens.1 This disease can present as a range of symptoms provoking a visit to the emergency department (ED). In this case, we present a 40-year-old female who reported tongue and gingival discoloration, unintentional weight loss, and skin hyperpigmentation. The patient was evaluated by an endocrinologist in the ED, received a dose of steroids in the ED, and discharged home with a prescription for steroids. This case report illustrates the distinctive tongue discoloration seen in primary adrenal insufficiency, highlighting the need for early recognition in the emergency setting to enable prompt and appropriate management.<h4>Topics</h4>Endocrine, Addison's disease, primary adrenal insufficiency, black tongue, steroids, skin hyperpigmentation.

<i>Background and aims</i>. Liver mechanical properties' (stiffness/viscoelasticity) evaluation is relevant for diagnosing/monitoring liver fibrosis. Due to limitations of the commonly used elastography, we propose the use of rheology and Low Field-Nuclear Magnetic Resonance (LF-NMR). <i>Methods.</i> In 30 liver samples from patients undergoing bariatric surgery and 18 control samples, we evaluated the shear modulus <i>G</i>/critical stress τc (elastic properties) and mean complex modulus Ga* (elastic/viscous properties) by rheology. LF-NMR was used to measure the spin-spin relaxation time (<i>T</i>_2m), reflecting iron content. The expression of iron-related proteins and of pro-fibrotic proteins were evaluated by qRT-PCR. Tissue histology was also determined. <i>Results</i>. Ga*/<i>G</i>/τ_c were higher in pathological samples, which also showed increased expression of pro-fibrotic proteins. Fibrosis determination displayed a correspondence of 4/30 samples for elastography/histology and 17/30 for rheology/histology. <i>T</i>_2m was significantly lower in pathological livers, indicating iron accumulation as confirmed by increased expression of iron-related proteins. <i>T</i>_2m was more effective than histology in detecting iron. An inverse correlation was observed between <i>T</i>_2m and Ga*/<i>G</i> showing that iron accumulation is associated with increased liver elasticity/viscoelasticity, i.e., fibrosis. Additionally, an inverse correlation of Ga*/<i>G</i> with transferrin, was observed. <i>Conclusion.</i> As our patients mostly have mild liver fibrosis, the combined use of rheology/LF-NMR can effectively detect early changes in liver mechanical properties, aiding in staging and diagnosis of fibrosis.

Huntington's disease (HD) is the best-known example of a neurodegenerative disorder caused by the expansion of a glutamine-encoding CAG repeat in the causative gene. Growing evidence indicates that somatic CAG expansions play a key role in disease progression, providing a strong rationale for therapeutic strategies directly targeting the repeat tract. However, achieving sufficient efficacy while maintaining allele selectivity and minimizing off-target effects remains a major challenge. Here, we developed allele-selective, CAG-targeting artificial microRNA (amiRNA) molecules that exhibit significantly reduced off-target risk. This was achieved by introducing specific substitutions at selected positions within the guide strand. These molecules effectively downregulated polyglutamine (polyQ) proteins in cellular models of HD, spinocerebellar ataxias types 1 and 3, and dentatorubral pallidoluysian atrophy. The most promising candidate, amiR136-13A, reduced mutant huntingtin levels in different brain regions of the HD mouse model and did not induce toxicity up to 28 weeks following a single administration of an AAV5 vector. Transcriptomic profiling of human HD neural stem cells treated with amiR136-13A revealed minor changes in gene expression. Moreover, amiR136-13A reduced the level of HTT1a, a short pathogenic isoform of huntingtin. Collectively, these findings identify amiR136-13A as a potent, selective, and safe therapeutic candidate for HD and potentially other polyQ disorders.

Ochratoxin A (OTA) is a common fungal toxin contaminant in feed that is primarily produced by fungi belonging to the <i>Aspergillus</i> and <i>Penicillium</i> genera. In animal husbandry, the long-term ingestion of OTA-contaminated feed by animals can lead to severe consequences, such as reduced production performance, immunosuppression, and kidney and liver damage. Therefore, the development of efficient OTA detection technology is crucial for ensuring animal health and enhancing farming efficiency. This study aimed to design and modify the molecular structure of OTA, compare the preparation and identification of two artificial antigens, and select the optimal artificial antigen. Based on the molecular structure of OTA, the OTA hapten was activated via the N-hydroxysuccinimide active ester (NHS) method to generate an intermediate product known as OTA-carboxylic acid ester. This intermediate was subsequently conjugated with bovine serum albumin (BSA) via the coupling agents N, N'-dicyclohexylcarbodiimide (DCC) or 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC) to prepare new and effective artificial antigens. Additionally, ovalbumin (OVA) was used to prepare detection antigens. The structures of the prepared artificial antigens were physically and chemically characterized via instrumental analysis techniques such as ultraviolet (UV) and infrared (IR) spectroscopy. Mice were immunized with artificial antigens to explore their immunogenicity. The results revealed successful conjugation of both artificial antigens with molecular binding ratios of OTA to BSA of 10.88:1 and 7.84:1. The immunized mice exhibited corresponding responses, and the titers of the mouse polyclonal antisera (pAbs) reached 1:3.2 × 10³, the OTA pAb that was prepared from two artificial antigens was able to recognize OTA with 100% accuracy. Among them, the mice immunized with the artificial antigen OTA-BSA (EDC) exhibited the best sensitivity, with a half-maximal inhibitory concentration (IC₅₀) of 11.40 ng/mL. Although the cross-reactivity rates with ochratoxin B (OTB) and ochratoxin C (OTC) were relatively high at 86.69% and 62.02%, respectively, the cross-reactivity rates with other compounds were less than 0.3%. In conclusion, this study successfully prepared two artificial antigens and selected the OTA-BSA (EDC) group as the optimal artificial antigen via identification. These results provide the antigenic foundation for the preparation of OTA monoclonal antibodies (mAbs) and the establishment of immunological analysis methods.

Pharmacogenetics has redefined the understanding of antidepressant response by demonstrating that genetic variability profoundly influences both drug efficacy and safety. This review synthesizes current evidence on the impact of pharmacokinetic and pharmacodynamic polymorphisms in guiding antidepressant therapy, focusing on the cytochrome P450 enzymes CYP2C19 and CYP2D6, as well as the serotonergic genes SLC6A4 and HTR2A. A comprehensive literature search in NCBI and Google Scholar (2017-2024) identified recent meta-analyses and clinical studies evaluating genotype-phenotype associations in patients treated with selective serotonin reuptake inhibitors (SSRIs). Findings indicate that <i>CYP2C19</i> and <i>CYP2D6</i> polymorphisms markedly affect plasma concentrations, therapeutic outcomes, and adverse-event risk-where poor metabolizers exhibit increased efficacy but greater toxicity, while ultrarapid metabolizers show reduced therapeutic response. Likewise, functional variants such as <i>SLC6A4</i> (5-HTTLPR) and <i>HTR2A</i> modulate serotonin transporter availability and receptor sensitivity, influencing clinical improvement and tolerability, especially in interaction with environmental stressors. The integration of these genetic markers into conceptual clinical frameworks enables more rational antidepressant selection, personalized dosing, and minimization of adverse reactions, demonstrating that both pharmacokinetic and pharmacodynamic polymorphisms jointly contribute to antidepressant efficacy within a stepwise precision-medicine approach.

Diabetes classified as category 3, usually referred to as type 3 diabetes, encompasses specific types due to other causes. These specific forms of diabetes are clinically and pathophysiologically very heterogeneous. Type 3 diabetes encompasses inherited forms of diabetes (monogenetic diabetes, neonatal diabetes, Down syndrome, Klinefelter syndrome, Turner syndrome), diabetes due to other endocrine disorders (acromegaly, Cushing's disease, glucagonoma) and drug-induced forms, e.g., caused by glucocorticoids, immune checkpoint inhibitors, calcineurin inhibitors, phosphatidylinositol 3 kinase inhibitors, highly active antiretroviral therapy (HAART) and antipsychotics. Additionally, this category includes pancreatogenic forms (after pancreatic surgery, after pancreatitis, pancreatic tumors, hemochromatosis, cystic fibrosis), rare infection-triggered (e.g., congenital rubella syndrome) and autoimmune forms other than type 1 diabetes (e.g., Stiffman syndrome, anti-insulin receptor antibodies, insulin autoimmune syndrome). An exact diagnosis is critical for correct and optimal treatment of affected patients. Exocrine pancreatic insufficiency is not only found in patients with pancreatogenic diabetes but also in patients with type 1 diabetes or long-standing type 2 diabetes.

<b>Background</b>: Carbonic Anhydrases (CAs) represent regulators of cell adaptation to hypoxia, pH regulation, and metabolic fitness. Among cancers, multiple myeloma (MM) is a plasma cell malignancy sustained by hypoxia-driven metabolic adaptation, extracellular acidification, and redox imbalance. Tight regulation of tumor extracellular pH, mediated by Carbonic Anhydrases IX and XII, is crucial for myeloma survival, progression, and stemness, making these isoforms attractive therapeutic targets. <b>Methods</b>: We designed and synthesized a library of terpenoid-based hybrids by derivatizing chlorothymol and 4-isopropyl-3-methylphenol with either the natural coumarin umbelliferon or the 2,2'-dipicolylamine (DPA) scaffold. This chemical strategy aimed to selectively inhibit tumor-associated CAs IX/XII through coumarin- or DPA-mediated recognition, while terpenoid fragments were introduced to enhance lipophilicity, membrane permeability, and potential redox-modulating properties. The compounds were tested by a Stopped-Flow assay for CA inhibition, in cell-based assays for antiproliferative properties and by means of several antioxidant assays. <b>Results</b>: The most active compounds, connecting the coumarin core to a terpenoid tail, inhibited the targeted CAs in the nanomolar range, showing up higher selectivity over off-target isoforms (I and II). In studies performed on MM cell lines, selected derivatives reduced viability (IC₅₀ = 15.8-85.4 µM) and displayed favorable selectivity over normal cells. In silico investigations suggested that the compounds were able to interact selectively with the target enzymes. <b>Conclusions</b>: Collectively, these results support a dual-targeting strategy in which selective inhibition of tumor-associated CAs, combined with redox modulation, interferes with adaptive mechanisms of MM cells, providing a rational framework for the development of multifunctional agents against metabolically resilient hematological malignancies.

Plastic products are extensively utilized in various industrial goods and consumer items. However, when these plastics fail to undergo complete degradation, they generate nanoplastic particles (NPs). As emerging environmental pollutants, such nanoplastics are highly likely to have widespread and adverse impacts on human health. Accumulating evidence indicates that NPs can penetrate biological barriers and exert toxic effects on multiple organs, including the nervous system. Although extensive studies have investigated the toxicity of NPs, the mechanisms underlying their long-term neurotoxic effects remain poorly understood. Here, we summarize the current understanding on the exposure pathways of NPs, their neurotoxic effects, and the molecular mechanisms involved in neurotoxicity. Emerging evidence suggests that NPs induce neurological damage through various mechanisms, including oxidative stress, neuroinflammation, ferroptosis, autophagy dysregulation, and gut-brain axis imbalance. A comprehensive understanding of these mechanisms will provide new insights into the potential impacts of environmental NPs exposure on the nervous system and contribute to more accurate health risk assessments.

The mammalian intestinal epithelium is a rapid self-renewal tissue in the body, serving as a critical interface between the host and the external environment. Maintaining the intestinal epithelium homeostasis requires precise coordination of cellular processes, including proliferation, migration, differentiation, autophagy, and cell-to-cell interaction. An increasing body of evidence has unveiled circular RNAs (circRNAs) as abundant and stable regulatory molecules that play pivotal roles in the intestinal epithelial biology and are intimately involved in many aspects of gut mucosal pathologies. Unlike linear RNAs, circRNAs form covalently closed loop structures through back-splicing events, conferring remarkable stability and resistance to exonucleolytic degradation. circRNAs regulate the growth of the intestinal mucosa, injury-induced epithelial regeneration, and gut barrier function via diverse mechanisms, including interactions with microRNAs and RNA-binding proteins. Deregulated circRNAs are implicated in the pathogenesis of various gut mucosal disorders such as inflammatory bowel disease and malignancies. In this review, we highlight pathobiological functions and mechanisms of intestinal epithelium-enriched circRNAs, particularly <i>circHIPK3</i>, <i>Cdrlas</i>, and <i>circPABPN1</i>, in the epithelium homeostasis and pathologies and also discuss potential clinical application of circRNAs as diagnostic biomarkers and therapeutic targets in patients with critical diseases.

<h4>Background</h4>Cuproptosis is a copper-induced mechanism of programmed cell death (PCD) that may influence the progression of papillary thyroid cancer (PTC). However, research linking this route to long non-coding RNAs (lncRNAs) in PTC is limited. This study assesses the correlation between cuproptosis-associated lncRNAs and immunotherapy sensitivity as well as clinical outcomes in PTC.<h4>Methods</h4>PTC transcriptomic profiles, tumor mutational burden (TMB), and clinical annotations were obtained from The Cancer Genome Atlas (TCGA). Cuproptosis-associated lncRNAs were identified via co-expression analysis with known cuproptosis genes. We employed least absolute shrinkage and selection operator (LASSO)-penalized Cox modeling to develop a lncRNA signature for predicting progression-free survival (PFS) and calculated a patient-level risk index. Subsequently, we evaluated the associations among this index, immune infiltration, predicted immunotherapy response, and clinical outcomes.<h4>Results</h4>Ten lncRNAs associated with cuproptosis-related risk were identified. Using the median value of risk score, we stratified individuals into higher- and lower-risk strata. Individuals in the higher-risk category exhibited poorer PFS, disease-specific survival (DSS), and overall survival (OS) rates. In multivariable Cox models, the lncRNA-derived score independently predicted PFS and outperformed traditional clinicopathologic factors as demonstrated by receiver operating characteristic (ROC) and C-index comparisons. A nomogram that incorporates the risk score alongside essential covariates exhibited well-calibrated estimates for PFS at 1, 3, and 5 years. The heightened risk was positively correlated with increased immune-cell infiltration, augmented immunological effector actions, and elevated expression of immune checkpoints. Model-based inference indicated that high-risk patients are more likely to get benefits from immune-checkpoint blocking, with enhanced results expected from anti-programmed cell death protein 1 (PD-1) or anti-cytotoxic T-lymphocyte-associated protein 4 (CTLA-4) monotherapy. The reverse transcription quantitative polymerase chain reaction (RT-qPCR) results demonstrated that LINC01545 overexpressed in PTC tissues while DLG3-AS1 downregulated in PTC tissues compared with the normal tissues, which is in consistent with the bioinformatic results.<h4>Conclusions</h4>A lncRNA signature associated with cuproptosis serves as an independent predictor of clinical outcomes and immunotherapy response in PTC, supporting the use of these transcripts as biomarkers for risk stratification and treatment selection guidance.

Gallbladder cancer (GBC) is a highly lethal disease which is usually diagnosed at advanced stage owing to unavailable screening tools and effective therapies. Persistent inflammation induced by various risk factors is the main cause of GBC, however, the molecular program remains elusive. Unveiling the molecular trajectory and events during gallbladder epithelium malignant transformation contribute to prevention and drug discovery for GBC. Single cell RNA sequencing was performed by using 4 gallbladder adenoma and cancer samples. Pseudotime trajectory analyses were employed to reconstruct epithelium transformation track in order to identify crucial genes which promoted GBC development. Functional and mechanism studies were performed to validate the regulatory network and cell behavior in vitro and in vivo. Three clusters of gallbladder epithelium were identified among GBC microenvironment which were characterized by distinct epithelial mesenchymal transition (EMT) and inflammation states. Cell adhesion molecular binding was the most significant GO term between EMT high and low states, in which OLFM4 was differentially expressed gene participated. Further studies implicated that OLFM4 could promote GBC metastasis and activate EMT through CEACAM6/AKT signaling cascade, and the CEACAM6 expression was regulated by TGF-β/Smad3 pathway. Interestingly, we disclosed that TGFβR1 was the functional receptor of OLFM4, through which the tumorigenic signaling of OLFM4 was transduced. These findings suggest the oncogenic role of OLFM4 during GBC carcinogenesis which can be a candidate biomarker of GBC, and OLFM4, TGFβR1 and downstream signaling elements are promising therapeutic targets for GBC.

<h4>Abstract</h4>Diabetic kidney disease (DKD) remains a leading cause of end-stage renal disease worldwide, characterized by increasing prevalence and limited therapeutic options. This review comprehensively synthesizes recent advances in the molecular mechanisms underlying DKD progression, focusing on key cellular events including podocyte injury, tubular damage, endothelial dysfunction, pericyte loss, and immune-inflammatory responses that drive glomerular and tubulointerstitial fibrosis. We further elaborate on critical pathways such as lipotoxicity, mitochondrial dysfunction and oxidative stress, aberrant autophagy, inflammasome activation, and profibrotic signaling. The review also highlights clinically approved agents including renin-angiotensin-aldosterone system inhibitor, sodium-glucose cotransporter 2 inhibitors, endothelin receptor antagonists, mineralocorticoid receptor antagonists, glucagon-like peptide-1 receptor agonists, and emerging therapeutic targets and agents currently under preclinical and clinical investigation, including anti-inflammatory, anti-fibrotic, and metabolic interventions. By integrating mechanistic insights with translational evidence, this work aims to provide a comprehensive review for the development of DKD.

Alzheimer's disease (AD) is a debilitating neurodegenerative disorder characterized by cognitive decline and memory loss. Current treatments offer limited efficacy, necessitating the development of innovative multitarget therapeutic strategies. Here, we present <i>N</i>³,<i>N</i>⁵-bis(2-(5-methoxy-1<i>H</i>-indol-3-yl)ethyl)-2,6-dimethyl-4-(2-nitrophenyl)pyridine-3,5-dicarboxamide (<b>HCM-01</b>), a novel compound developed to target multiple neurodegenerative pathways implicated in AD. In vitro assays included MTT-based cell viability analyses performed in two complementary experimental settings: primary neuronal cultures and astrocyte-based in vitro cell culture models exposed to glutamate. In primary hippocampal neuronal cultures, glutamate exposure induced a statistically significant reduction in cell viability compared with vehicle-treated controls, consistent with glutamate-induced excitotoxicity. Under these conditions, <b>HCM-01</b> treatment resulted in a statistically significant improvement in neuronal viability, showing a greater protective effect compared with donepezil and memantine. In contrast, in astrocyte-based in vitro cultures, the applied glutamate concentration did not induce overt cytotoxicity, in line with the intrinsic neuroprotective and glutamate-buffering role of astrocytes. Accordingly, astrocytic experiments were designed to assess functional modulation of glutamate-handling mechanisms rather than cell survival. Western blot analysis in C8-D1A astrocytic cells demonstrated increased expression of excitatory amino acid transporter 2 (EAAT2) following <b>HCM-01</b> treatment compared with control and reference drug-treated groups, suggesting modulation of astrocyte-mediated glutamate homeostasis. In parallel, redox analyses revealed that <b>HCM-01</b> improved oxidative/antioxidative balance, as evidenced by increased total antioxidant capacity (TAC) and reduced total oxidant status (TOS), supporting an indirect antioxidant contribution to its functional effects. In vivo behavioral assessment of <b>HCM-01</b> in a streptozotocin (STZ)-induced Alzheimer's model in female Sprague-Dawley rats demonstrated that administration of <b>HCM-01</b> at doses of 50 mg/kg orally (oral, P.O. and intraperitoneal, I.P.) and 100 mg/kg (P.O.), significantly improved cognitive and memory functions in the passive avoidance (PA), Morris water maze (MWM), and locomotor activity tests. Moreover, histopathological and immunohistochemical analyses of different hippocampal regions revealed reduced neuronal damage, attenuation of tau pathology, antiamyloidogenic effect, and restoration of cholinergic function. Complementary in silico studies, including molecular docking, molecular dynamics simulations (MDS), and free energy calculations, suggested potential interactions of <b>HCM-01</b> with the allosteric site of EAAT2. Taken together, these findings suggest that <b>HCM-01</b> exerts neuroprotective effects against glutamate-induced excitotoxicity in primary hippocampal neurons while additionally modulating glutamatergic homeostasis and redox balance through functional mechanisms in astrocyte-based models, supporting its relevance as a multitarget preclinical candidate for early stage AD mechanisms.

Microplastics (MiPs, ×5 mm in size) harbor complex biofilms that facilitate pathogen dissemination, yet standardized extraction protocols are lacking. Here, we developed and optimized a method for biofilm extraction from environmentally weathered MiPs. To reflect real-world conditions, the protocol was applied directly to bulk, heterogeneous, field-collected MiP mixtures (size range: 80 µm-5 mm) without prior sorting by polymer type or morphology. By optimizing extraction buffers, mechanical disruption, and MiP quantities (100-150 particles), we established an optimal protocol combining phosphate-buffered saline with 0.1% Tween 80, ultrasonication (40 kHz, 10 min), vortexing with glass beads, and a two-cycle extraction-disaggregation workflow. This approach involves an initial extraction followed by a repeated, exhaustive extraction step designed to maximize the recovery of recalcitrant biofilm residues. This protocol markedly enhanced recovery of viable, culturable cells, delivering a 2,950-fold enhancement in the recovery of viable, culturable cells (evaluated via CFU counts; 28,020 ± 11,034 CFU MiP⁻¹) vs. conventional PBS extraction (9.5 ± 3 CFU MiP⁻¹) and 102-fold vs. passive extraction (274 ± 59 CFU MiP⁻¹). The 10-min sonication empirically maximized viable cell recovery within the tested duration range. The two-step protocol with Tween 80-mediated disaggregation proved critical, increasing recovery 208-fold by disaggregating biofilm fragments. While DNA yields (26.5 ± 3.93 ng µL⁻¹) were sufficient for targeted PCR-based pathogen detection (<i>Aeromonas</i> spp., <i>Salmonella enterica</i>), the co-extraction of complex environmental matrices (<i>A</i>₂₆₀/<i>A</i>₂₈₀ ratio: 0.17-0.19) strictly requires an additional purification step prior to next-generation sequencing. Validation across contrasting aquatic environments confirmed the method's robustness. Comparative analysis demonstrates that conventional single-step approaches fail to recover the majority of viable cells trapped within weathered MiP biofilms. This optimized and validated protocol provides a critical methodological foundation for investigating plastisphere microbial ecology and pathogen transport dynamics, supporting evidence-based risk assessment of MiP contamination, especially public health risks associated with microplastic pollution.IMPORTANCEMicroplastic-associated biofilms (the "plastisphere") serve as vectors for waterborne pathogens and antibiotic resistance genes; however, the persistent use of inadequate extraction methods has systematically underestimated microbial abundance, presenting a critical barrier to global environmental risk assessment. By overcoming the limitations of conventional extractions-which fail to penetrate recalcitrant extracellular polymeric matrices on environmentally weathered microplastics-our standardized methodology liberates previously undetectable bacterial populations. The ability to accurately quantify these hidden communities, including key pathogens like <i>Aeromonas</i> spp. and <i>Salmonella enterica</i>, fundamentally transforms our understanding of microplastics as hidden biological reservoirs. Ultimately, this methodological advancement bridges a critical gap in microbial ecology, delivering the reliable, quantitative data strictly required by policymakers, environmental agencies, and public health officials to establish evidence-based guidelines mitigating the impacts of microplastic pollution on global water systems.

This study investigates the synthesis and evaluation of glycine-conjugated 5.0G polypropyleneimine dendrimers as an advanced nanocarrier system for the selective delivery of chloroquine. 5.0G PPI dendrimers were synthesized via a divergent method and subsequently conjugated with glycine. Comprehensive characterization confirmed the successful modification and an increase in nanocarrier size. Drug loading studies demonstrated a significantly enhanced entrapment of chloroquine (57.5% vs. 44.5%) in glycine-conjugated formulations, attributed to the potential complexation and sealing of dendritic branches. In vitro release profiles revealed a substantial reduction in chloroquine release from the glycine-coated dendrimers, indicating a sustained-release capability crucial for prolonged therapeutic action (only 32.07% release over 24 h compared to 88.80% from uncoated). Crucially, macrophage uptake studies indicated a four-fold reduction in phagocytic uptake of glycine-conjugated formulations, suggesting an effective bypass of macrophage recognition, thereby potentially minimizing non-specific clearance. In vivo pharmacokinetic analyses in rats showed a prolonged plasma concentration of chloroquine with the glycine-conjugated system, extending detectability up to 11 h. Furthermore, organ distribution studies highlighted a remarkable increase in liver accumulation (45.6% of the initial dose) compared to uncoated dendrimers or free drug, indicating highly effective liver targeting. These findings highlight the significant potential of glycine-conjugated PPI dendrimers to achieve highly efficient and sustained targeted drug delivery, particularly to the liver, while simultaneously reducing macrophage-mediated clearance.

Multiple spillovers of porcine deltacoronavirus (PDCoV) into humans in Haiti highlight its zoonotic potential and the need for targeted interventions. No approved vaccines or therapeutics are available for use in humans against any DCoVs. Here, we report the de novo design of PDCoV miniprotein inhibitors (aka minibinders, MBs) and show that one of them, MB11, binds with picomolar affinity to the PDCoV receptor-binding domain (RBD). MB11 potently inhibits PDCoV, outcompeting monoclonal antibodies, and cross-reacts with and broadly neutralizes a panel of distantly related DCoVs. We determined a cryoelectron microscopy structure of MB11 bound to the PDCoV RBD which reveals the molecular basis of broad DCoV neutralization through interference with host receptor engagement. Deep mutational scanning of the PDCoV RBD reveals that MB11 has a high barrier to viral escape with only few mutations mediating escape without dampening APN receptor binding. MB11 resists stringent biochemical stresses, including high temperature, low pH, and proteolysis, which may enable delivery to various tissues for viral inhibition. This work delineates a prime candidate for clinical evaluation against PDCoV infection and for pandemic preparedness.

In plants, long-distance transport and phloem-mediated signal distribution play crucial roles in regulating stress adaptation and development. Phloem sap contains various types of RNAs, including small RNAs (sRNAs), messenger RNAs (mRNAs), and long noncoding RNAs (lncRNAs). Recently, endogenous circular RNAs have been identified in phloem sap of apple trees. Some phloem RNAs have been shown to have long-distance signaling functions, but for most, no functions have yet been determined. Due to their stability, circRNAs are interesting candidates with potential functions in long-distance signaling. Therefore, we aimed to characterize the circRNA content in the phloem sap of the crop plant Brassica napus. To achieve this, we performed Illumina sequencing of rRNA-depleted, circRNA-enriched, and sRNA libraries. The analysis revealed 1,734 distinct circRNAs in the phloem sap of B. napus. Of these, we validated ten circRNAs by PCR amplification and Sanger sequencing of their back-splicing junctions (BSJs). Using circ-Panel Nanopore sequencing, we investigated the full-length sequences of 14 circRNAs from phloem sap and leaf samples, identifying seven high-confidence candidates that exhibit potential intron retention and isoform variation across the two tissues. The investigation of potential interaction partners from phloem circRNA identified miRNA target sites on multiple circRNAs, particularly for known phloem-mobile miRNAs like miR156, miR169, and miR395. With Microscale Thermophoresis (MST), we were able to show the ability of the abundant RNA-binding protein BnGRP7 to bind the phloem circRNA circBnaANL2(7,8) with a dissociation constant of around 1 µM, raising questions about the involvement of RBPs in circRNA transport, stabilization, and function in phloem sap.

Although unfractionated heparin (UFH) remains a vital agent for rapid anticoagulation, its reliance on animal-derived sources results in batch-to-batch variability and contamination risks, and its clinical application is further restricted by the possibility of heparin-induced thrombocytopenia. Dendritic polyglycerol sulfates were investigated as heparin analogues in 2004 due to their ability to mimic its charge, which is essential for its acting mechanism, and revealed an anticoagulant activity of 15-35% compared to UFH. In the current study, we found that the anticoagulant effect of polyglycerol sulfates further increases with their flexibility, resulting in a comparable activity of linear polyglycerol sulfate to UFH. Furthermore, we comprehensively analyzed the mechanism of action and discovered an antithrombin-independent, thrombin-selective mechanism. Moreover, we confirmed that FDA-approved protamine sulfate is a viable reversal agent for polyglycerol sulfate.

<h4>Background</h4>Brain tumor is one of the most malignant diseases of the central nervous system, and early accurate detection is of great significance for improving patient survival rate. However, the heterogeneity of brain tumors in terms of morphology, size, and location on magnetic resonance imaging (MRI) image, as well as their similarity to surrounding normal brain tissue, poses significant challenges for tumor detection.<h4>Objective</h4>This study aims to develop a high-performance brain tumor detection framework that integrates feature enhancement, channel attention, and progressive pruning, achieving an optimal balance between detection accuracy, model efficiency, and interpretability for slice-level MRI tumor localization tasks.<h4>Methods</h4>This paper proposes a convolution Prewitt-and-pooling-based preprocessing (CSPP) approach, based on the "you only look once" version 11 (YOLOv11) framework, which highlights important structural detail more effectively than traditional statistics. A dynamic convolution-based C3k2 (DCC) module was integrated to more efficiently capture both local and global features. A channel prior convolutional attention (CPCA) module was introduced before the detection head, enabling the network to specifically focus on information-rich channels and key spatial regions. Through a progressive hybrid pruning strategy (PHPS), the model was optimized for efficient inference. Furthermore, Eigen-class activation mapping (Eigen-CAM) was used to interpret the prediction result, making them more transparent.<h4>Results</h4>Extensive experiments on 3 brain tumor MRI datasets demonstrated the superior performance of CDCP-YOLO (CSPP-DCC-CPCA-PHPS-YOLO). On Br35H, the mean average precision (mAP) at an intersection-over-union (IoU) threshold of 0.5 (mAP0.5) increased by 2.6%, average mAP over several IoU thresholds (0.50-0.95; mAP0.5:0.95) increased by 5.9%, and number of floating-point operations (×10⁹; GFLOPs) decreased by 47.7%. On Roboflow, mAP0.5 increased by 19.5%, mAP0.5:0.95 increased by 7.7%, and GFLOPs decreased by 47.7%. On Capstone, mAP0.5 increased by 6.9%, mAP0.5:0.95 increased by 5.8%, and GFLOPs decreased by 47.7%.<h4>Conclusions</h4>The proposed CDCP-YOLO framework achieves an optimal balance between accuracy, efficiency, and interpretability, providing a lightweight and reliable solution for slice-level brain tumor detection in MRI images.

<h4>Background</h4>Community-acquired pneumonia (CAP) remains a leading cause of hospitalization, morbidity, and mortality worldwide, particularly among older adults with multimorbidity and frailty. Despite advances in antimicrobial therapy, clinical outcomes have improved little, highlighting the need for safe, inexpensive adjunctive treatments. Vitamin C plays a critical role in immune function, redox homeostasis, and endothelial integrity, all disrupted during acute infection. Hypovitaminosis C is common in hospitalized patients with CAP and has been associated with increased disease severity, longer length of stay (LOS), and worse outcomes. However, prior randomized trials of vitamin C have produced inconsistent results, often focusing on critically ill patients with sepsis, using short treatment durations, and discontinuing therapy abruptly.<h4>Objective</h4>The Vitamin C in Community-Acquired Pneumonia (VitCAP) trial aims to evaluate whether high-dose oral vitamin C administered over an extended period improves clinical recovery and patient-centered outcomes in adults hospitalized with CAP.<h4>Methods</h4>VitCAP is a single-center, double-blind, placebo-controlled, parallel-group randomized clinical trial conducted at a tertiary hospital in Australia. Adults aged 18 years and older hospitalized with CAP will be randomized within 48 hours of admission in a 1:1 ratio to receive either oral sodium ascorbate (1 g 3 times daily for 7 days, followed by 500 mg twice daily for 30 days) or a matching placebo in addition to standard care. Randomization will be computer generated with allocation concealment via a centralized pharmacy service, and all participants, clinicians, investigators, and outcome assessors will remain blinded. The primary outcome is time to clinical stabilization, defined using standard physiological criteria. Secondary outcomes include early clinical response, symptom burden at 30 days, intensive care unit admission, need for ventilatory or vasopressor support, LOS, all-cause mortality at 30 days and 6 months, hospital readmission, health-related quality of life, and changes in inflammatory biomarkers (C-reactive protein and procalcitonin). Analyses will follow the intention-to-treat principle. The primary outcome will be analyzed using Cox proportional hazard regression adjusted for prespecified covariates, with sensitivity analyses including restricted mean survival time.<h4>Results</h4>The VitCAP trial received ethics approval from the Southern Adelaide Local Health Network Human Research Ethics Committee in 2025 and funding in September 2025. Recruitment is expected to commence in 2026 and continue for 18 to 24 months. A total of 124 participants will be enrolled to provide 80% power to detect a clinically meaningful difference in time to clinical stabilization while allowing for attrition. Data analysis will follow completion of follow-up, with primary results anticipated in 2028.<h4>Conclusions</h4>The VitCAP trial is designed to address important evidence gaps by evaluating sustained oral vitamin C supplementation in hospitalized patients with CAP using clinically meaningful patient-centered outcomes. If effective, vitamin C could represent a safe, low-cost, and scalable adjunct to standard CAP management.<h4>Trial registration</h4>Australian New Zealand Clinical Trials Registry (ANZCTR) ACTRN12625001361493; https://anzctr.org.au/Trial/Registration/TrialReview.aspx?ACTRN=12625001361493.<h4>International registered report identifier (irrid)</h4>PRR1-10.2196/91037.

The attachment of Post-Translational Modifications (PTMs) to proteins regulates their activities and stability. Here we utilized the nematode Caenorhabditis elegans to test whether UFMylation, a PTM which affects key biological functions, regulate aging and protein homeostasis (proteostasis). We find that lowering UFMylation extends lifespan and mitigates the toxicity of aggregation-prone proteins that underlie the development of neurodegenerative disorders in humans. Mass spectrometric analysis suggests that UFMylation of aging-regulating proteins, including of the nucleolar FIB-1-NOL-56 complex and the germline-resident proteins CAR-1 and CGH-1, governs proteostasis, probably across tissues. Functional analyses indicate that the proteostasis-regulating transcription factors DAF-16 and SKN-1 are crucial for the protective effects of reduced UFMylation. Counter-proteotoxic effect of reduced UFMylation are mediated by enhanced nascent protein quality control, reduced protein aggregation, and increased protein degradation by the ubiquitin-proteasome system. These insights highlight the important roles of PTMs in the regulation of proteostasis and point at research directions for the development of therapies for neurodegenerative disorders.

CXCL5 is an epithelial cell-derived neutrophil-activating peptide implicated in intestinal inflammation. We investigated the role of CXCL5 in inflammatory bowel disease (IBD), with a focus on its interaction with IL-17 signaling. CXCL5 expression in colonic tissues from patients with IBD and its association with clinical disease activity were analyzed. Publicly available single-cell RNA sequencing data were examined to identify CXCL5-expressing cell populations. Acute and chronic murine colitis models were used to assess the functional role of CXCL5 in vivo, and cytokine expression was evaluated. In vitro experiments were performed in human intestinal epithelial cells to examine CXCL5 regulation by inflammatory cytokines. CXCL5 expression positively correlated with fecal calprotectin levels in IBD patients and was significantly reduced in ulcerative colitis patients achieving endoscopic mucosal healing. Single-cell analysis revealed CXCL5 expression concentrated in a discrete epithelial cluster. IL17RA was broadly expressed across epithelial and immune cell populations, including the CXCL5-expressing epithelial subset, which may suggest the capacity of these cells to respond to IL-17A signaling. In acute DSS-induced colitis, CXCL5 deficiency markedly attenuated inflammation and neutrophil recruitment, accompanied by reduced IL-17A expression. Conversely, CXCL5 deficiency did not significantly alter disease severity in chronic colitis models. In vitro, TNF-α induced CXCL5 expression, which was further enhanced by IL-17A. CXCL5 contributes to neutrophil-driven intestinal inflammation in a phase-dependent manner and the CXCL5/IL-17 axis may represent a potential therapeutic target, warranting further mechanistic investigation.

Dioxin contamination represents a serious global environmental threat, particularly in developing and climate-vulnerable regions. This study evaluates the potential of Bacillus velezensis Bm3, isolated from a dioxin-contaminated site in central Syria, for the aerobic biodegradation of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD). Quantitative analyses demonstrated a strong biodegradative capacity, with 71% and 76% of the initial TCDD concentration removed after 5 and 6 weeks, respectively. Whole-genome sequencing revealed a 5.29 Mbp genome encoding 5411 predicted proteins, providing a solid genetic basis for this activity. A two-step degradation pathway is proposed, initiated by cytochrome P450-mediated hydroxylation of TCDD to dihydroxy-hydrodibenzo-p-dioxin, followed by conversion to the key intermediate catechol. This compound is subsequently cleaved by catechol 2,3-dioxygenase, yielding acetaldehyde and pyruvate. Gene expression and protein accumulation analyses supported this mechanism, showing strong induction of a specific cytochrome P450 isoform (CYPBM3_v1_ua3131) and cat-2,3-diox transcripts. These findings identify B. velezensis Bm3 as a promising candidate for TCDD bioremediation in warm, dry environments.

Diabetic kidney disease (DKD), the leading cause of kidney failure, is marked by clinical and molecular heterogeneity, making therapeutic development exceedingly difficult¹. Here we used Xenium and CosMx single-cell spatial transcriptomics, integrated with single-nucleus RNA sequencing, to build a cross-platform kidney atlas that makes tissue architecture computable for prognosis, non-invasive detection and patient selection. Using this atlas, we defined reproducible tissue niches and injury-linked microenvironments and uncovered a profibrotic context that expands with disease and tracks with worse kidney function. Within this architecture, we identified a B cell-predominant, tertiary lymphoid structure-like immune microenvironment that defines a distinct DKD subset with accelerated progression to renal end-points. We developed tissue biomarkers and a matched plasma protein panel that capture this biology, stratify patients in a population biobank and improve risk prediction beyond clinical models-supporting their potential for biomarker-guided selection in future B cell-targeted DKD trials.

<h4>Aims</h4>Metabolic dysfunction-associated steatohepatitis (MASH) is a leading cause of cirrhosis. NIS2+ is a recently approved serum-based test combining two biomarkers (miR-34a-5p and YKL-40) to identify at-risk MASH (i.e., MASH and significant fibrosis).<h4>Objective</h4>To assess the prevalence of at-risk MASH by NIS2+ in individuals from primary care or endocrinology clinics.<h4>Materials and methods</h4>798 participants recruited from outpatient clinics were risk-stratified by NIS2+ into low-risk, intermediate-risk or having at-risk MASH (NIS2+ score < 0.46, ≥ 0.46 and < 0.68 or ≥ 0.68, respectively). Presence of steatosis (CAP ≥ 288 dB/m) and clinically significant liver fibrosis (VCTE- ≥ 8.0 kPa) was established by transient elastography (FibroScan).<h4>Results</h4>At-risk MASH affected 29% of individuals with both obesity and T2D compared to 3% of those without either condition (p < 0.001). People with at-risk MASH, compared to those at low-risk, more often had steatosis (81% vs. 40%), clinically significant fibrosis (42% vs. 5%), AST or ALT ≥ 40 IU/L, hepatic insulin resistance (by HOMA-IR) and adipose tissue insulin resistance (by adipo-IR) (all p < 0.001). NIS2+ strongly correlated with diagnosis of at-risk MASH by FAST (r = 0.67; p < 0.001), as well as CK-18, ALT and AST and liver fibrosis by VCTE-LSM (all p < 0.001).<h4>Conclusion</h4>The prevalence of at-risk MASH is high in individuals with obesity and T2D attending outpatient primary care and endocrinology clinics. Their progression to cirrhosis may be prevented with early risk-stratification and timely intervention.

Mounting evidence implicates pathogen infections in the pathogenesis of Alzheimer's disease (AD), yet the cellular mechanisms underlying infection-induced neurodegeneration remain poorly understood. Central to this process is the dysfunction of astrocyte-neuron interactions, which are critical for maintaining neuroinflammatory balance and synaptic homeostasis. Here, we demonstrate that astrocytic protein tyrosine phosphatase 1B (PTP1B) acts as a key regulator of astrocyte reactivity during infection, leading to impaired neuroglial communications and cognitive decline. In a murine model of chronic Toxoplasma gondii (T. gondii) infection, elevated PTP1B levels in astrocytes were closely associated with neuroinflammation and cognitive impairments. Conditional deletion of astrocytic PTP1B or its pharmacological inhibition mitigated neuroinflammation, restored synaptic integrity, and rescued cognitive function. Mechanistically, astrocytic PTP1B induced the polarization of A1-like neurotoxic reactive astrocytes, enhanced glutamate-mediated excitotoxicity, and triggered neuronal senescence, collectively contributing to synaptic damage and cognitive deficits. Notably, elevated levels of PTP1B, GAFP and cellular senescence markers were observed in the serum samples from T. gondii IgG-seropositive individuals and in hippocampal transcriptomes from AD patients, underscoring the translational relevance. Together, our findings reveal that PTP1B-mediated disorder of astrocyte-neuron crosstalk represents a novel mechanism of pathogen-driven neurodegeneration.

Epithelial-mesenchymal transition (EMT) is a key cellular process that facilitates distant metastasis during tumor progression. Tumor cells lose their epithelial characteristics while obtaining mesenchymal features during EMT process. This process is associated with a complex interaction between tumor cells, microenvironment, and signaling pathways. Therefore, it is helpful to clarify the molecular mechanisms of EMT process to introduce novel diagnostic and therapeutic markers to target malignant tumor cells. C-MYC is a transcription factor that regulates cell proliferation, apoptosis, metabolism, and EMT process. C-MYC is an effector of various signaling pathways that regulates EMT process during tumor progression. Therefore, in the present review we discussed the role of signaling pathways in regulation of C-MYC mediated EMT process during tumor progression. It has been shown that WNT, MAPK, TGF-β, and PI3K/AKT pathways are the main regulators of C-MYC mediated EMT process in tumor cells. This review paves the way to introduce C-MYC as a reliable therapeutic target to reduce metastatic ability of tumor cells.

BACKGROUND: Liver fibrosis represents a critical stage in the progression of chronic liver diseases toward cirrhosis and hepatocellular carcinoma. However, there are no targeted therapies specifically designed for the treatment of liver fibrosis. This research aims to identify novel targets for anti-hepatic fibrosis and to provide new insights into therapeutic strategies for addressing this condition. METHODS: Dual-luciferase reporter assays and chromatin immunoprecipitation (ChIP) analyses were conducted to assess the modulation of UNC5B gene transcription. Co-immunoprecipitation experiments were performed to investigate the interactions between UNC5B and Src homology phosphotyrosyl phosphatase 2 (SHP2) or Focal adhesion kinase (FAK) proteins. Virtual screening was utilized to identify small-molecule inhibitors of UNC5B. The in vivo effects of Unc5b knockdown in liver fibrosis mouse models were evaluated using an Unc5b inhibitor (D359-0264) and an adeno-associated virus vector of serotype 6 (AAV6) containing short hairpin RNA targeting Unc5b. RESULTS: This study demonstrates that the pro-fibrotic pathway TGF-β1/SMAD3 transcriptionally regulates UNC5B expression. Unc5b expression was significantly upregulated in liver fibrosis mouse models. Knocking down Unc5b resulted in reduced hepatocyte damage and decreased fibrosis in these models. Additionally, UNC5B was shown to interact with the PTP domain of SHP2 and the Y397 site of FAK, indicating that UNC5B competitively binds to the Y397 site of FAK, thereby inhibiting FAK dephosphorylation by SHP2 and promoting liver fibrosis. CONCLUSION: This study identifies a novel regulatory axis in liver fibrosis models involving UNC5B, which is regulated by the TGF-β/SMAD signaling pathway. UNC5B facilitates liver fibrosis by competitively binding to the Y397 site of FAK with SHP2, thus enhancing its phosphorylation and activating FAK. Furthermore, the UNC5B inhibitor has demonstrated the therapeutic potential of targeting this molecule for the treatment of liver fibrosis.

<h4>Background</h4>MTHFD2, a critical enzyme in one-carbon metabolism, regulates cellular growth, redox balance, and energy homeostasis. Aberrant MTHFD2 expression has been implicated in the pathogenesis and progression of various diseases. MTHFD2 is a promising therapeutic target, however, its roles in diseases and upstream and downstream regulatory mechanisms have not been clarified.<h4>Methods</h4>This study reviewed literature from databases such as PubMed and Google Scholar, using keywords "MTHFD2", aiming to summarize current research on the functions and mechanisms of MTHFD2 on diseases and provide theoretical support for future investigation of related therapeutic strategies.<h4>Results</h4>High expression of MTHFD2 promotes cancer progression and MTHFD2 can be used as a sensitization target in cancer treatment. MTHFD2 influences tumor microenvironment through disturbing immune cell infiltration and immune checkpoints expression. Aberrant MTHFD2 expression can also participate in other diseases. Consequently, a range of MTHFD2 inhibitors has been developed, advancing to both preclinical and clinical evaluations.<h4>Conclusions</h4>This review examines the multifaceted roles of MTHFD2 in cancer biology and other pathological conditions, describing its influence on the tumor microenvironment and relevance in immunotherapeutic applications. Structural strategies for MTHFD2 inhibitor development are explored, alongside challenges and opportunities for leveraging MTHFD2 as a diagnostic, therapeutic, and prognostic target in disease management.

<h4>Background</h4>Healthcare workers (HCWs) remain at an elevated risk of SARS-CoV-2 infection due to occupational exposure and waning vaccine-induced immunity. We conducted a longitudinal study to estimate cumulative uptake, incidence, and factors associated with COVID-19 booster dose uptake among Bangladeshi HCWs.<h4>Methods</h4>Between March 2021 and December 2023, we followed 3099 HCWs recruited from 20 healthcare facilities across four divisions of Bangladesh. At enrollment, information was collected on sociodemographic and clinical characteristics, prior SARS-CoV-2 infection, and COVID-19 vaccination history. Booster uptake status was updated biweekly. Multivariable Cox proportional hazards regression models were used to estimate adjusted hazard ratios (aHRs) for predictors of booster uptake.<h4>Results</h4>During follow-up, 1964 (63.4%) HCWs received at least one COVID-19 booster dose. However, uptake of the 2nd booster dose was low (6%). The incidence of booster uptake was 54.5 per 100 person-years (95% CI 50.0-59.5). The hazard of booster uptake was higher among HCWs with prior SARS-CoV-2 infection (aHR = 1.13; 95% CI 1.01-1.26) and among those working in both COVID-19 and general wards (aHR = 1.36; 95% CI 1.22-1.53). In contrast, prior adverse events following the primary series (aHR = 0.36; 95% CI 0.32-0.41) were associated with a lower hazard of booster uptake.<h4>Conclusions</h4>Although more than half of HCWs received a booster, uptake declined with repeated dosing. Variations in the hazard of uptake by occupational exposure, prior infection, and adverse events reflect changing perceptions of risk and experiential factors in addition to access. Strengthening institutional vaccination policies and addressing behavioral barriers may thus support timely booster uptake among HCWs.

Endogenous ethanol (EtOH) production is a newly identified pathophysiological mechanism involved in metabolic dysfunction-associated steatohepatitis (MASH) and liver disease associated with hepatitis B virus (HBV). Therefore, the characterization of EtOH-producing species associated with liver disease could contribute to the development of gut microbiota-targeted approaches. We investigated EtOH production and tolerance, antimicrobial susceptibility and antimicrobial resistance gene(s) in 33 strains isolated in previous culturomics studies and belonging to species enriched in MASH, alcoholic hepatitis (AH) and HBV patients. <i>Enterocloster clostridioformis</i>, <i>Thomasclavelia ramosa</i> and <i>Peptinophilus grossensis</i> were identified as new EtOH-producing species associated with liver diseases. A strong association between EtOH tolerance and production was detected (<i>p</i> < 0.05). Yeast, <i>Enterocloster</i> species (strictly anaerobic bacteria) and <i>Limosilactobacillus fermentum</i> produced the highest concentrations of EtOH (0.8 to 3.3 g/L). The poorly absorbed drugs, amphotericin B, rifaximin and vancomycin together showed high <i>in vitro</i> susceptibility. Furthermore, <i>E. clostridioformis</i> EC38 harbored the <i>vanB</i> operon. New EtOH-producing species associated with liver diseases were identified thanks to culturomics. Notably, most of them are anaerobic bacteria. These findings underscore the need to further investigate anaerobic gut microbiota species enriched in liver diseases with the aim of developing gut microbiota-targeted therapies.

This work focused on the identification of angiotensin I-converting enzyme (ACE) inhibitory peptides from royal jelly (RJ) proteins and elucidated their inhibition patterns and mechanisms. RJ proteins were analyzed for ACE inhibition potential using in silico tools, and suitable enzymes were selected for peptide release. Hydrolysis conditions were optimized using response surface methodology (RSM), and the resulting peptides were fractionated and purified. Mass spectrometry identified 57 peptides, with seven selected for synthesis based on scoring. IDFDF, DVNFR, and SFHRL showed the highest ACE inhibition, with IC₅₀ values of 16.9 μM, 42.5 μM, and 242.6 μM, respectively. Lineweaver-Burk plots revealed IDFDF as a competitive inhibitor, DVNFR as a non-competitive inhibitor, and SFHRL as a mixed inhibitor. Molecular docking indicated that peptide-ACE interactions were primarily mediated through hydrogen bonds and Zn(II) coordination. This work promotes the sustainable utilization of RJ and the development of ACE inhibitory peptides derived from food sources.

Fetal skeletal muscle development involves coordinated interactions among myogenic, stromal, vascular, and immune compartments, yet the cellular and molecular programs guiding tissue maturation remain incompletely understood. To address this, we generated a high-resolution single-cell atlas of fetal female goat skeletal muscle and performed trajectory analysis, transcription factor activity profiling, and intercellular communication mapping. Unsupervised clustering identified RUNX2 mesenchymal progenitors, fibro-adipogenic progenitors (FAPs), myofibroblasts, endothelial cells, macrophages, differentiating myocytes, and mature skeletal muscle fibers, revealing a heterogeneous ecosystem in which stromal populations support myogenic progression and vascular and immune cells contribute to tissue organization. Pseudotime analysis traced a maturation continuum from differentiation-competent myocytes to contractile fibers, marked by sequential activation of extracellular matrix remodeling, cytoskeletal stabilization, and sarcomere assembly. KEGG and GO enrichment highlighted stage-specific engagement of ErbB, Hedgehog, and Hippo signaling, as well as cell cycle and ubiquitin-mediated proteolysis pathways, linking proliferation, differentiation, and structural maturation. Transcription factor profiling revealed early-stage proliferative and morphogenetically permissive states driven by E2F4/5, HMGA2, and HAND2, transitioning to late-stage differentiation, ECM remodeling, and tissue stabilization orchestrated by CEBPB, CREB3L1, ELK1, and E2F2. Cell-cell communication analysis showed a developmental redistribution of signaling authority, from ECM-driven, progenitor-centered networks to modular, structurally stabilized interactions. These findings define the cellular, transcriptional, and signaling framework orchestrating fetal skeletal muscle maturation.

Stress has been prevalent and has become an epidemic health burden, loaded with chronic disorders. The stress response is an adaptive mechanism that prepares an individual to respond to threats or other stressors in a fight-or-flight situation. The stress response involves the induction of neurological and hormonal networks and is usually resolved when stress subsides; however, persistent stress leads to permanent and detrimental impacts on health. With the rise of advanced single-cell analysis technologies, a wave of basic and translational research aimed at elucidating stress has shed light on the underlying mechanisms. Among 80 studies in this review, stressors are classified into acute/chronic physical, physiological, and psychological groups, whereas some studies have more than one stress source. Single-cell RNA-seq was the dominant technology utilized in these studies. This advanced technique systematically reveals cellular heterogeneity in gene expression patterns and the differential transcriptomic landscape of stress response in a wide array of tissues and organ systems, e.g., the nervous system, the endocrine system, the immune system, and others. Bioinformatics identified a single-cell atlas of stress-specific cell subtypes, cell-to-cell interactions, and enriched pathways, showing promise for stress syndrome biomarkers, attenuation, and targeted therapy. The limits of these stress studies were mainly focused on transcriptomics, so future studies using multi-omics approaches across multiple organ systems will yield insights into stress disorders and novel therapeutic strategies.

Neurodegenerative diseases, characterized by progressive neuronal loss and functional decline, impose a substantial global health burden. Autophagy, the principal intracellular degradative pathway for clearing misfolded proteins and damaged organelles, is vital for neuronal homeostasis, whereas maladaptive neuroinflammation is increasingly being recognized as a central driver of disease progression. A growing body of evidence indicates a bidirectional, tightly coupled relationship between autophagy and neuroinflammation: impaired autophagic flux promotes accumulation of damage-associated molecules that activate innate immune responses, while sustained inflammatory signaling further disrupts autophagy, together forming a self-reinforcing cycle that accelerates neurodegeneration. This interplay is regulated by diverse genetic, molecular, cellular, and environmental factors and manifests in cell-type-specific ways across microglia, astrocytes. Therapeutic strategies emerging from these insights include modulation of autophagic pathways (e.g., mTOR, AMPK, TFEB), targeted inhibition of inflammasome and pro-inflammatory mediators (notably NLRP3-related signaling), and delivery platforms for small molecules or nucleic acids, with increasing interest in multi-target and stage-specific interventions. This review integrates mechanistic evidence and translational advances, highlights gaps in cell-type and stage-specific understanding, and outlines priorities for developing safe, effective therapies that target the autophagy-neuroinflammation axis in neurodegenerative disorders.

This study focuses on the development and comprehensive evaluation of the physicochemical, mechanical, and biological properties of composites based on polyhydroxybutyrate (PHB), chitosan (Ch), and hydroxyapatite (HA) for biomedical applications. DSC and FTIR spectroscopy showed that the addition of hydroxyapatite did not significantly affect the structure of the materials, but AFM data revealed a change in the surface morphology. Variations in RMS roughness ranging from 13 to 150 nm were observed for chitosan and the composites. The density of the HA-containing samples was 0.06-0.067 g/cm³, which is higher than that of the unfilled composite (0.056 g/cm³). Optimal hydrophilic properties (contact angle 38.9°) and elasticity (damping factor 0.064) were recorded for the sample with 10% HA (PChHA10). The water absorption varied: the addition of chitosan increased the value to 7.5 g/g, compared to 2.7 g/g for pure PHB, while HA slowed the swelling kinetics (more than 180 min). A biodegradation study revealed that samples containing 10-20% HA exhibited the highest stability in an enzymatic environment, while further increases in HA content resulted in increased degradation rates. The PChHA10 is considered to offer the balanced combination of properties. The potential applications of this material in medicine include its use as a scaffold for the <i>in vitro</i> cultivation of osteoblasts and chondrocytes, as well as for implantation in models of bone and cartilage defects <i>in vivo</i>.

General pustular psoriasis (GPP) is a rare, potentially life-threatening neutrophilic dermatosis. Pediatric cases are uncommon and often misdiagnosed due to overlapping clinical and histopathological features with other pustular dermatoses. We present a case of an 11-year-old boy, initially diagnosed with Sneddon-Wilkinson syndrome, who presented with disseminated pustular eruptions, with no response to antibiotics, dapsone, and glucocorticosteroids. In histopathology, we observed subcorneal neutrophilic pustules. Due to atypical features and poor treatment response, the patient underwent genetic testing, which revealed a homozygous IL36RN gene c.338C>T (p.Ser113Leu) pathogenic variant, which enabled a definitive diagnosis of GPP. Treatment with acitretin led to clinical improvement. Pediatric GPP poses diagnostic and treatment challenges. Genetic testing for IL36RN pathogenic variants may aid in the diagnosis, especially in atypical cases. The presence of the biallelic IL36RN pathogenic variant supports the diagnosis of DITRA (Deficiency of the IL-36 Receptor Antagonist, ORPHA:404546)-a monogenic autoinflammatory form of GPP.

Inflammation is a physiological and tightly regulated component of normal pregnancy, contributing to implantation, placental development, and the initiation of parturition. The placenta functions as an active immunological hub, coordinating innate and adaptive immune responses to maintain tolerance while protecting against infection. Preeclampsia and fetal growth restriction (FGR) are major causes of maternal and perinatal morbidity worldwide and represent central manifestations of placental disease. Increasing evidence indicates that these conditions share key pathophysiological mechanisms, including placental dysfunction and maladaptive maternal immune responses. When immune regulation at the maternal-fetal interface becomes disrupted, inflammatory pathways contribute to impaired placental development and vascular maladaptation. In this context, excessive immune activation-driven by inflammasome signaling, Th1/Th17 polarization, and altered natural killer and macrophage function-can compromise placental perfusion, promote antiangiogenic imbalance, and lead to systemic endothelial dysfunction. This review, therefore, focuses on how immune dysregulation contributes to placental dysfunction in preeclampsia and FGR, synthesizing current knowledge of the maternal-fetal immune interface and exploring therapeutic strategies that link pathogenic mechanisms to targeted interventions. A deeper understanding of placental immunology and inflammatory signaling is essential to develop precision therapies. Established therapies, including low-dose aspirin, low-molecular-weight heparin, and antenatal corticosteroids, aim to mitigate inflammation and optimize fetal outcomes, while adjunctive strategies target oxidative stress, nutritional deficits, and the maternal microbiome. Emerging approaches such as cytokine-targeted biologics, inflammasome inhibitors, and mesenchymal stem cell therapies show promise but require rigorous safety and efficacy evaluation. Future research should prioritize biomarker validation, pathway-specific interventions, and equitable implementation to reduce inflammation-driven pregnancy complications.

Emphysema is a well-recognized risk factor for lung cancer; however, its influence on the immunologic tumor microenvironment in lung adenocarcinoma remains poorly defined. In this pilot, hypothesis-generating study, immune-related gene expression profiling was performed using archival formalin-fixed paraffin-embedded tumor specimens from 12 patients with lung adenocarcinoma, including the Never-smoker group (never-smokers without emphysema; <i>n</i> = 4), the Smoker 1 group (smokers without emphysema; <i>n</i> = 3), and the Smoker 2 group (smokers with CT-defined emphysema; <i>n</i> = 5). Expression of 770 immune-related genes was analyzed using the nCounter PanCancer IO 360 Panel (NanoString Technologies, Seattle, WA, USA). Compared with the Never-smoker group, tumors from the Smoker 1 group showed marked upregulation of SFRP1, SERPINB5, and IL6, whereas tumors from the Smoker 2 group exhibited increased expression of KIR2DL3, BLK, and WNT2B. Relative to the Smoker 1 group, the Smoker 2 group demonstrated significant upregulation of MMP7, TDO2, and CCL18. Pathway enrichment analysis revealed cytokine-cytokine receptor interaction as the most prominently enriched pathway in both smoker groups, while the IL-17 signaling pathway was preferentially enriched in the Smoker 2 group. In addition, diffusing capacity for carbon monoxide showed significant correlations with immune-related genes including IL-6 and IL-6R. Collectively, these preliminary findings suggest that lung adenocarcinoma arising in emphysematous lungs may be characterized by a distinct pro-inflammatory immune microenvironment. Given the small sample size and potential confounders, these results should be regarded as hypothesis-generating. Emphysema-associated immune remodeling may nevertheless represent an important biological factor worthy of validation in larger, independent cohorts.

Colorectal cancer (CRC) remains a major cause of cancer-related death, and resistance to chemotherapy and radiotherapy continues to limit durable disease control. Ferroptosis, an iron-dependent form of cell death driven by lipid peroxidation, has therefore emerged as a potential therapeutic strategy. However, models focused solely on glutathione peroxidase 4 (GPX4) and solute carrier family 7 member 11 (SLC7A11) do not fully explain why CRC cells differ in their sensitivity to ferroptosis. In this review, we examine how ferroptosis in CRC is shaped by iron trafficking and selenium-dependent antioxidant defense. We first discuss the poly(rC)-binding proteins 1 and 2 (PCBP1/2)-nuclear receptor coactivator 4 (NCOA4) axis, which regulates iron storage, trafficking, and ferritinophagy. We then review the AlkB homolog 8 (ALKBH8)-directed selenoprotein network, which supports the detoxification of lipid peroxides and maintenance of redox homeostasis. We next consider how these two systems intersect and how their interplay influences ferroptosis sensitivity. We also discuss why concurrent disruption of iron handling and selenium-dependent defense mechanisms may enhance therapeutic efficacy. Finally, we outline potential clinical applications, including combination strategies and biomarker development.

Systemic lupus erythematosus (SLE) is a polygenic autoimmune disorder where genetic diversity drives significant clinical heterogeneity. This review summarizes the current understanding of the roles of genetic polymorphisms in immunological dysregulation, organ-specific manifestations and therapeutic response heterogeneity in individuals with SLE. The literature was obtained from PubMed, EBSCOhost, Web of Science and Scopus. The narrative review comprised 60 publications published within the last 12 years. The research consistently identifies the major histocompatibility complex (MHC) region as the most significant genetic risk factor for the onset of autoimmunity. Genetic variants in <i>STAT4</i> and <i>IRF5</i> exacerbate disease progression by facilitating chronic inflammation. These genetic factors are associated with various clinical outcomes, including renal and neuropsychiatric symptoms. Polymorphisms in HLA class II, <i>TLR7</i> and <i>FBN2</i> are notably linked to serious consequences, including lupus nephritis (LN). Progress in targeted therapy signifies a transition to personalized medicine with medications such as anifrolumab, litifilimab, iberdomide and Janus kinase (JAK) or Cyclin-Dependent Kinase (CDK) inhibitors, demonstrating potential for targeting pathways associated with the interferon gene signature and <i>STAT4</i> polymorphisms. Notwithstanding the problems presented by the heterogeneity of SLE, the identification of risk variations is anticipated to enhance predictive and therapeutic biomarkers, hence facilitating more precise and individualized disease management.

This study evaluated the effects of light spectral quality on shoot yield and essential oil of <i>Tagetes erecta</i> L. cultivated in controlled growth chambers under three LED lighting treatments with different red, blue, and white wavelength ratios and a constant 16 h photoperiod for up to 101 days. The F2 treatment (5 red:1 blue) produced yields of fresh shoots, early blooming flowers, and oils of 1586 ± 164 g/m², 569.77 ± 76.81 g/m², and 307 ± 31.7 mg/m², respectively. These values were significantly higher (<i>p</i> < 0.05) than those of the F1 treatment (white:red-phosphor), and represented increases of 1.37-, 1.26-, and 1.38-fold, respectively. Gas chromatography identified 30-31 compounds in the oil with three major constituents-(<i>E</i>)-<i>β</i>-ocimene (22.9-28.8%, highest under F3), (<i>E</i>)-myroxide (13.9-20.6%, highest under F1), and piperitone (7.3-9.6%, highest under F3). Essential oils inhibited from four to five of the seven tested microbial strains, with the notable activity against <i>Escherichia coli</i> and <i>Candida albicans</i> recorded in F2 and F1, respectively. These findings confirm that light spectral quality is a critical factor regulating flower, essential oil, and antimicrobial efficacy in <i>T. erecta</i>, demonstrating that optimized LED spectra offer a practical strategy to improve plant yield and phytochemical quality.

<h4>Introduction</h4>Neutrophils are proposed to contribute to inflammation at the onset of multiple sclerosis (MS). However, as neutrophils must be analysed rapidly following blood collection, their characterisation remains challenging, and the biology of neutrophils during episodes of MS is poorly understood. Neutrophils can comprise of several subpopulations and diverse phenotypes that are modified across health states and tissues. We hypothesised that neutrophil subpopulations would significantly differ in abundance between people with MS and controls.<h4>Method</h4>Our pilot study applied flow cytometry to analyse phenotypes of neutrophils present in the peripheral blood of 10 people with recently active MS or clinically isolated syndrome (CIS) and 12 control participants.<h4>Results</h4>Using both unsupervised and supervised analyses of flow cytometry data, we identified that CD10^low neutrophils were significantly enriched in the blood of people with CIS and MS compared with controls. These CD10^low neutrophils featured decreased CD16 and CD11b expression, with CD184 expression absent, suggesting they were an immature neutrophil population. In people with MS, the proportions of CD10^low neutrophils were non-significantly correlated with expanded disability status scores (p=0.06).<h4>Discussion</h4>These findings point to immature CD10^low blood neutrophils as a population of interest to active MS disease, whose functions should be studied in greater detail in context of MS pathology and biomarkers.

Periodontitis is a chronic inflammatory disease driven by dysbiotic biofilm and abnormal host adaptive immune responses, characterized by irreversible destruction of periodontal tissues. Recent studies indicate that ferroptosis-a form of iron-dependent lipid peroxidation-mediated cell death-is not a passive outcome of the disease process but rather an important active inducer and pathological amplifier in the development of periodontitis. This review synthesizes existing evidence to propose that ferroptosis acts as an important regulatory node within the pathological network of periodontitis, impairs epithelial barrier function, suppresses osteoblast differentiation and function, and amplifies pro-inflammatory signaling pathways through cell-type-specific mechanisms. Furthermore, ferroptosis exhibits complex cross-regulation with other cell death pathways (e.g., pyroptosis, apoptosis, autophagy), collectively forming a self-amplifying vicious cycle that persistently drives chronic inflammation and tissue destruction. Although iron death-targeting interventions, including iron chelators, GPX4 agonists, lipid peroxidation scavengers, natural bioactive compounds (e.g., curcumin, resveratrol), and nanodelivery systems, show promise in preclinical studies, the cell type specificity and context dependency of ferroptosis pose critical challenges for precise intervention. Therefore, this review positions ferroptosis as a pivotal node in the regulatory network of periodontal cell death, advocating for the development of stage-specific and cell-type-specific targeted strategies. This signifies a paradigm shift in periodontal therapy, transitioning from traditional antimicrobial approaches toward host-centered precision periodontal medicine.

<h4>Background</h4>ALK-positive histiocytosis is a rare form of cutaneous histiocytosis and has been explicitly designated as a novel histiocytic neoplasm in recent classification systems. This disease may involve multiple organs throughout the body. Its diagnosis primarily relies on pathological confirmation, and molecular evidence of <i>ALK</i> rearrangement is required to establish a definitive diagnosis. For patients with unresectable or disseminated ALK-positive histiocytosis, therapy with ALK inhibitors has demonstrated considerable efficacy.<h4>Case presentation</h4>An 8-month-old Chinese male infant was admitted with 1-day history of fever and newly diagnosed anemia of 0.5-day duration. Physical examination revealed chronic rashes on the bilateral lower extremities, including the plantar surfaces. Laboratory investigations demonstrated anemia (hemoglobin level: 63 g/L), elevated inflammatory markers, and an increased serum S-100 protein level (9.11 ug/L). Abdominal ultrasonography and contrast-enhanced computed tomography (CT) revealed hepatosplenomegaly and diffuse hypoechoic or cystic low-density lesions in the liver (largest approximately 7 × 8 mm), which were initially suspected to represent a liver abscess. Histopathological examination of a biopsy specimen from the lower extremity rash revealed dermal proliferation of histiocytes, foamy cells, and short spindle cells, accompanied by scattered multinucleated giant cells and lymphocytes. Immunohistochemical analysis demonstrated positivity for CD68, S100, and ALK, and negativity for CD1a, CD3, CD20, and other markers. Fluorescence <i>in situ</i> hybridization (FISH) confirmed <i>ALK</i> rearrangement, and next-generation sequencing (NGS) identified a novel <i>PTRH2::ALK</i> fusion. Positron emission tomography-computed tomography (PET-CT) subsequently revealed mediastinal space-occupying lesions, multiple hypermetabolic lesions in the liver, kidney, mesentery, and bones, as well as metabolically active systemic lymph nodes, findings consistent with ALK-positive histiocytosis. The patient remained clinically stable during follow-up after initiation of the ALK inhibitor crizotinib.<h4>Conclusion</h4>This study reports a rare case of infantile ALK-positive histiocytosis that was initially misdiagnosed as a liver abscess and identifies a novel <i>PTRH2::ALK</i> fusion gene. In addition, this study systematically summarizes, for the first time, the age, sex, lesion distribution, fusion genes, treatment strategies, and prognosis of all previously reported pediatric cases of ALK-positive histiocytosis.

<h4>Background</h4>Post-COVID-19 condition (PCC), commonly called long COVID, is a prevalent sequela of SARS-CoV-2 infection and can affect multiple organ systems. Cognitive dysfunction is one of the most common symptoms in PCC, with a prevalence of 22%. It can persist for years and significantly reduce patients' quality of life. The brain network is the neural basis underlying human cognitive processes. Diffusion tensor imaging (DTI) and functional magnetic resonance imaging (fMRI) studies have revealed that cognitive impairments across attention, memory, executive function, and language are associated with alterations in network characteristics for PCC. Currently, there is no accepted therapy for cognitive impairment in PCC. Acupuncture has the potential to improve cognitive deficits in PCC. This trial aims to investigate the effect of acupuncture on cognitive functions in patients with PCC, and to explore the underlying mechanism of its effects on cognition in this condition using DTI and fMRI.<h4>Methods</h4>In this three-armed, randomized controlled trial, 117 PCC patients with cognitive symptoms will be randomly assigned in a 1:1:1 ratio to verum acupuncture (VA), sham acupuncture (SA), or a waitlist control group. Participants in the VA and SA groups will receive three sessions of treatment per week for 8 weeks. The primary outcome measures are the changes in Addenbrooke's Cognitive Examination-III (ACE-III) total score and phonemic fluency test score at week 8. The secondary outcome measures include the Digit Span Test (DST), Symbol Digit Modality Test (SDMT), Trail Making Test (TMT), Rey's Auditory Verbal Learning Test (RAVLT), Rey-Osterrieth Complex Figure Test (RCFT), Stroop Color Word Test (SCWT), category fluency test, action fluency test, and Boston Naming Test (BNT-30), as well as global and regional topological measures of structural and functional brain networks constructed from DTI and fMRI data. Additionally, the Fatigue Severity Scale (FSS), the Generalized Anxiety Disorder-7 (GAD-7), the 24-item Hamilton Depression Scale (HAMD-24), and the MOS 36-item Short Form Health Survey (SF-36) will also be measured.<h4>Discussion</h4>The results of this study will reveal the effect of acupuncture treatment on cognitive functions for PCC and provide insights into the mechanisms by which acupuncture may improve cognition in PCC.<h4>Clinical trial registration</h4>ClinicalTrials.gov (www.clinicaltrials.gov), identifier: NCT07355751.

<h4>Introduction</h4>Bladder cancer exhibits sex-specific behavior, occurring more frequently in males but progressing to advanced stages more commonly in females. The activation of sex hormone receptors may explain these differences, but the exact genetic drivers remain poorly understood. Furthermore, current bladder cancer biomarkers have inconsistent sensitivities and specificities in practice, making early diagnosis a challenge.<h4>Methods</h4>This study approaches bladder cancer biomarker discovery through machine learning techniques on gender and disease-stratified RNA-seq data. Training sets limited to differentially expressed genes were subjected to four different feature selection methods: differential gene expression analysis adjusted p-value, recursive feature elimination with support vector machine, logistic regression, and an optimized random forest procedure. Gene panels were compared and aggregated across selection strategies and cross-validation folds to identify robust biomarkers for sex-specific bladder cancer development and progression.<h4>Results</h4>When applied to unseen datasets and limited to 50 genes or less, male and female-specific panels achieved areas under the receiver operating characteristic curve of 0.932 and 0.914, respectively, in distinguishing bladder cancer samples from non-tumor controls. In terms of enriched pathways, the male panel was associated with cell interactions and altered PI3K-AKT signaling, while the female panel was more closely connected to extracellular matrix reorganization. The panel differentiating male and female tumors had a poorer performance on external datasets compared to the sex-specific analyses, but still contained relevant genes.<h4>Discussion</h4>Genes such as PRAC1 and PCDH11Y were identified as high-impact predictors related to sex hormones or chromosomes for male tumor development. In the female-specific panel, genes related to aberrant androgen signaling across tumor types like androgen receptor, PLXNA1, USP54, and PMEPA1 were influential. These results offer potential targets for further in vivo/vitro experimentation and provide a framework for constructing high-performance gene panels related to sex-specific bladder cancer biology.

<h4>Introduction</h4>Phenolic acids are allelopathic substances that mediate plant-fungal interactions. However, the regulatory effects of phenolic acids on soil microbiota and crop phosphorus (P) uptake remain unclear. Thus, this study aimed to investigate the effects of continuous addition of cumalic acid (CA) on soil microbial communities and P uptake of rice.<h4>Methods</h4>We continuously added different concentrations of CA to the rhizosphere during the rice growth to investigate its effects on soil fungal communities and P uptake of rice.<h4>Results</h4>The addition of 5 and 10 μmol kg^-1 CA significantly increased P uptake in rice by 51.4% and 63.8%, respectively. In addition, the 10 μmol kg^-1 CA treatment significantly increased soil Olsen P content. Exogenous addition of CA altered the diversity and composition of soil microbial communities. The application of 5 and 10 μmol kg^-1 CA significantly elevated the relative abundance of Diversisporales from 46.3% to 68.7% and 67.1%, respectively; while suppressing Eurotiales from 15.8% to 2.6% and 5.7%, respectively. At the genus level, the treatment enhanced <i>Diversispora</i> (from 46.3% to 68.7% and 67.0%) and reduced <i>Penicillium</i> (from 9.0% to 1.2% and 2.2%). Notably, the relative abundance of <i>Diversispora</i> was significantly and positively correlated with P uptake of rice, whereas <i>Penicillium</i> was negatively correlated with P uptake.<h4>Conclusion</h4>CA stimulated the P uptake of rice by shaping keystone fungal taxa. Our results suggest that the indirect microbial-mediated mechanism dominated P uptake improvement of rice. This study provides critical insights into the potential application of allelopathic substances in agricultural production.

<h4>Purpose</h4>Obesity, characterized by abnormal fat accumulation with comorbidities, continues to increase dramatically, particularly in the pediatric population. Identifying the environmental and genetic causes underlying the development of obesity during early childhood is crucial for establishing preventive and protective treatments for this complex disease. We aimed to investigate genetic variants related to non-syndromic early-onset childhood obesity.<h4>Methods</h4>Whole-exome sequencing was performed in three independent consanguineous families with obesity, including three index cases and two additional affected siblings. Non-synonymous variants with minor allele frequency < 0.01 in all normal populations were filtered using the Genomize-SEQ Platform. Variant confirmations and familial segregations were analyzed by Sanger sequencing.<h4>Results</h4>WES revealed a shared <i>ATXN3</i> gene variant and two known variants of the <i>SH2B1</i> and <i>ADIPOQ</i> genes, which were reported to be associated with obesity. Additionally, five heterozygous novel gene variants of the <i>ANKK1</i>, <i>NEGR1</i>, <i>OGDH</i>, <i>ABCB1</i>, and <i>GSK3B</i> genes were identified, which are predicted to cause excessive fat accumulation and disruption of energy balance in individuals.<h4>Conclusions</h4>We suggest that the cumulative effects of all obesity-associated detected variants lead to the early-onset obesity phenotype observed in individuals. Hence, periodic follow-up and treatment opportunities are recommended for index cases, alongside the adoption of a more active lifestyle and healthy nutrition practices.

<b>Background:</b> Xinshubao tablet (XSB), a traditional Chinese medicine (TCM) formula composed of five medicinal herbs, is used clinically to alleviate cardiovascular diseases. This study aimed to investigate the coronary vasodilatory effects of XSB and its individual herbs, exploring its active constituents and the underlying mechanisms. <b>Methods:</b> The vasorelaxant effects of XSB and its individual herbal intestinal absorption solutions (IASs) were evaluated by ex vivo coronary artery ring assays. The chemical constituents of the best active herbal IAS were qualitatively identified using ultra-performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF-MS). Molecular docking and ex vivo assays were used to predict and validate the bioactive constituents and mechanisms responsible for coronary vasorelaxation. <b>Results:</b> Vasodilation experiments revealed that XSB-IAS and its individual herb IAS exhibited varying degrees of vasodilatory effects, in the range of 0.8-18 g raw materials/mL. At 6, 12, and 18 mg of raw materials/mL, <i>Crataegus pinnatifida</i> Bge (Shanzha) exhibited vasodilation rates of 26.45% ± 1.8%, 36.57% ± 3.5%, and 45.16% ± 6.3%, which were obviously higher than those of the other individual herbs. Fifty constituents in Shanzha IAS were identified by UPLC-Q-TOF-MS. Vasodilation-related protein-protein interaction (PPI) network revealed NOS3 as a core regulatory target. Molecular docking demonstrated that among the identified constituents, isochlorogenic acid B, betulin, etc., displayed binding affinity to NOS3. Isochlorogenic acid B was further validated to exhibit vasodilatory effects in the ranges of 0.05-2.5 mM. Mechanistic results showed that isochlorogenic acid B improved vasodilation by inhibiting Ca²⁺ influx through both voltage-dependent and receptor-operated Ca²⁺ channels, activating K⁺ channels, and exhibiting endothelium-dependent vasorelaxation. <b>Conclusions:</b> This study provides insights into the material basis and mechanisms underlying the vasorelaxant effects of XSB. Isochlorogenic acid B was firstly found to exert the coronary vasodilatory effect. This study can also contribute to the identification of efficacy-related quality markers in TCM.

Enteroviruses are positive-sense single-stranded RNA viruses and common pathogens that are responsible for diverse public health diseases. To facilitate the study of the virus biology and pathogenesis of enterovirus, we developed a rapid method for construction of the enteroviral cDNA clones including enterovirus A71 (EV-A71) and coxsackievirus B5 (CVB5). As described for EV-A71, the full-length cDNA of CVB5 was amplified by long-distance PCR and cloned into a T7 promoter-containing plasmid using directional seamless cloning technology. The virus was successfully rescued by single transfection into cells stably expressing T7 polymerase and exhibited characteristics similar to the parental virus. Next, through systematic construction and the optimization of the EV-A71 and CVB5 reporter viruses, we successfully generated two novel reporter virus panels with high virus titers, rapid replication, and relatively stable genetic inheritance across passages using the new fluorescence proteins mScarlet3-H and the smallest miRFP670nano3. Analysis of critical determinants for the reporter virus construction revealed that reporter gene sizes, genomic insertion sites, and the usage of protease recognition sites are crucial parameters. The EV-A71 and CVB5 reporter viruses enable antiviral drug evaluation, as demonstrated by our identification of gemcitabine as a broad-spectrum inhibitor of both viruses. These systems also facilitate the functional interrogation of host factors, exemplified by our discovery that METTL3 promotes EV-A71 and CVB5 replication. These reverse genetic tools, including infectious cDNA clones and reporter viruses, will advance basic enterovirus biology and accelerate antiviral drug discovery.

<b>Background:</b> Iron overload cardiomyopathy (IOC) is a major determinant of outcomes in hemochromatosis, and conventional echocardiography may miss early myocardial toxicity. Comparative data on primary (PH) versus secondary hemochromatosis (SH) using myocardial work (MW) indices are limited. <b>Methods:</b> We performed a retrospective cross-sectional study of 34 adults (16 PH and 18 SH patients) at a tertiary center. They all underwent echocardiography with speckle-tracking to obtain LV global longitudinal strain (GLS) and non-invasive MW indices from pressure-strain loops: global work index (GWI), global constructive work (GCW), global wasted work (GWW), and global work efficiency (GWE). Echocardiographic phenotypes were classified as a Normal, Dilated, Restrictive, or right ventricular/pulmonary hypertension (RVPH) phenotype. <b>Results:</b> SH patients showed higher iron burden and neurohormonal activation than PH patients (maximum ferritin 2954 vs. 444 ng/mL; BNP 93 vs. 13.5 pg/mL; both <i>p</i> < 0.001) and accounted for all deaths (33% vs. 0%) despite similar 3D LVEFs and GLSs. PH patients predominantly exhibited Normal phenotypes (81%), whereas SH patients more often showed advanced phenotypes, mainly RVPH and Dilated. GWI correlated inversely with ferritin (ρ ≈ -0.40), particularly ferritin at echocardiography in SH patients, while PH patients showed no significant correlations. GWW was higher in Dilated/RVPH compared to Normal phenotypes, and in SH patients, higher maximum ferritin was associated with impaired right ventricular free-wall strain. <b>Conclusions:</b> PH and SH patients exhibit distinct IOC phenotypes, with SH patients showing more advanced remodeling and worse outcomes. In this exploratory analysis, MW indices showed modest associations with iron burden markers, suggesting they may provide complementary information beyond LVEF and GLS. These preliminary findings require validation in larger, prospective studies.

The development of complex tissues relies on the precise assignment of cell identity. At the molecular scale, this process depends on the deposition of epigenetic modifications-such as methylation-that are regulated by complex biochemical networks and occur at specific regions on the DNA and chromatin. Here we show that despite the complexity of epigenetic regulation, dynamical scaling and self-similarity of DNA methylation marks emerge in embryonic development. Drawing on single-cell multi-omics experiments, super-resolution microscopy and statistical physics, we demonstrate that these phenomena originate in dynamical feedback between DNA methylation and the formation of nanoscale dynamic chromatin aggregates. These nanoscale processes lead to genome-wide increase in DNA methylation marks following a power law and self-similar correlation functions. Using this framework, we identify methylation patterns that precede gene expression changes in embryonic symmetry breaking. Our work identifies linear sequencing measurements as a laboratory to study mesoscopic biophysical processes in vivo.

<title>Abstract</title> <p> Cellulases are essential enzymes for hydrolyzing cellulose into simple sugars, with significant applications in bioethanol production, food processing, and agricultural waste management. This study investigated the efficacy of five agricultural residues—coconut fiber, rice straw, sugarcane bagasse, peanut shells, and rice bran—as substrates for endoglucanase (EG) production using <italic>Penicillium citrinum</italic> M5. Rice straw proved to be the most effective substrate, generating an endoglucanase (EG) activity of 39.69 U/gds, a yield 2.92 times greater than that of sugarcane bagasse. Preliminary one-factor-at-a-time experiments identified the optimal baseline parameters as pH 5.0, an inoculum of 10 ⁶ spores/mL, 1.5 g/L yeast extract, and 15 g/L maltose. Further refinement using a Response Surface Methodology-Box-Behnken Design (RSM-BBD) identified peak culture conditions: an inoculum density of 1.12 × 10⁶ spores/mL, 1.525 g/L yeast extract, and an initial pH of 5.05. This optimization boosted experimental EG activity to 88.74 U/gds, closely validating the model's predicted 89.33 U/gds. Additionally, scanning electron microscopy (SEM) was utilized to verify structural modifications on the rice straw's surface. The enzyme exhibited peak activity at a pH of 5.0 and a temperature of 50 °C. Ultimately, these results underscore the economic and biotechnological viability of pairing <italic>P. citrinum</italic> M5 with agricultural residues, particularly rice straw, for sustainable cellulase production. </p>

Although empirical evidence shows that antidepressants are effective and superior to placebo in treating depressive disorders, their mechanisms of action are still unclear. In this study, we examine the multidimensional molecular affinities of 18 commonly used antidepressants. Clustering analyses consistently generate three distinct clusters, providing a broader taxonomy that groups SSRIs and SNRIs together. Correlational analyses cautiously indicate a relationship between the affinity to metabotropic serotonin receptors and antidepressant efficacy. Next, we generate anatomical distribution profiles of drug action strengths by combining positron emission tomography-derived maps of cerebral neurotransmitter receptor and transporter densities from an open-access repository of healthy participants with the drugs' affinity profiles. We then relate these profiles to functional and structural neuroanatomical measures in health and disease. Our results reveal distinct, mechanistically interpretable differences between antidepressants with high 5-HTT affinity and atypical antidepressants. These differences could inform personalized drug selection and development.

Androgen receptor (AR) is a therapeutic target for prostate cancer. Despite effectively targeting its folded ligand-binding domain (LBD), resistance ultimately develops by mechanisms involving reactivation of AR signaling. These mechanisms include expression of constitutively active AR that lacks LBD and fueled the discovery of inhibitors that bind to AR's N-terminal intrinsically disordered transactivation domain (TAD). AR-TAD inhibitors (ARTADIs) are unique due to the paucity of small molecule inhibitors that bind directly to intrinsically disordered TADs, which have historically been considered undruggable. Leveraging our library of ARTADIs using cultured prostate cancer cells and multiple xenograft models, we reveal that small alterations in the chemical scaffold impact selectivity and potency within the AR-transcriptome; impacting signal transduction pathways involved in protumorigenic mechanisms. Mechanistically, these compounds differentially disrupt interactions between full-length AR or splice-variant AR-V7, and co-regulators, as revealed by rapid immunoprecipitation mass spectrometry of endogenous protein and the proximity ligation assay. Biophysically, several ARTADIs displayed exceptionally strong binding affinities that were better than, or were comparable to the LBD-inhibitor enzalutamide, with dissociation constants in the picomolar to low-nanomolar range as determined by surface plasmon resonance and microscale thermophoresis. MS/MS analysis revealed covalent binding to cysteine 129. In vivo, ARTADIs outperformed enzalutamide against prostate cancer xenografts in the presence of androgens, underscoring the therapeutic potential of targeting alternative AR domains. These findings support the feasibility - but also highlight the complexity - of developing drugs against an intrinsically disordered TAD impacted by multivalent binding interactions that may not occur in a stepwise fashion.

Dynamical systems exhibit transitions between ordered and disordered states, and a "critical state" occurs when the system lies at the borderline between these phases, where input is neither strongly damped nor excessively amplified. Systems maintaining critical dynamics are considered adaptive, and it is widely believed that brains operate at near-critical states where memory encoding and information processing are optimized. Impairments in brain function, such as dementia or epilepsy, may arise from failure of adaptive criticality, making deviation from criticality a potential biomarker for cognition-related neurological and psychiatric impairments. Wide-field calcium imaging using miniscopes is performed to record activity of hundreds of hippocampal CA1 neurons in freely behaving mice during rest, a novel object recognition (NOR) task, and NOR following scopolamine administration that severely impairs spatial memory. We find that while hippocampal networks exhibit some features of a near-critical system at rest, network activity shifts significantly closer to a critical state during NOR, and away from criticality when memory performance is impaired by the muscarinic antagonist scopolamine. These results support that hippocampal networks move toward criticality with increasing cognitive load, leveraging maximal dynamical range, information content, and transmission of critical regimes, while pathophysiological impairment alters criticality metrics reflecting network-level effects.

BACKGROUND: Telomerase reverse transcriptase (TERT) promoter mutation is among the most frequent genetic alterations in hepatocellular carcinoma (HCC), closely associated with tumor initiation, progression, and early recurrence; however, its prognostic role remains unclear. This study aimed to investigate the prevalence of TERT promoter mutations in HCC and evaluate their association with early recurrence, and to further explore whether its prognostic impact varies across clinically relevant subgroups. MATERIALS AND METHODS: We retrospectively analyzed 108 patients with histologically confirmed HCC who underwent curative hepatectomy. TERT promoter mutations were identified via Sanger sequencing. The risk of early recurrence, defined as tumor relapse within 24 months after surgery, was assessed using Cox proportional hazards models, including subgroup analyses. RESULTS: TERT promoter mutations were present in 45.4% of patients. While not predictive of early recurrence in the overall cohort (HR = 1.39; 95% CI: 0.76–2.56; p = 0.282), mutation status was significantly associated with reduced recurrence-free survival (RFS) in specific subgroups: patients < 60 years (HR = 3.45; 95% CI: 1.22–9.82; p = 0.020), males (HR = 2.03; 95% CI: 1.02–4.02; p = 0.043), and those with high mitotic activity (HR = 3.60; 95% CI: 1.01–12.80; p = 0.047). CONCLUSION: Although TERT promoter mutations did not predict early recurrence in the overall cohort, they were significantly associated with poorer RFS in defined clinical and pathological subgroups. These findings suggest the context-dependent prognostic value of TERT promoter mutations and may support their potential role in individualized risk stratification following curative resection in HCC.

Hydrogels are used for a wide range of biomedical applications. While mechanical characterization of hydrogels is frequently performed in isotonic saline, the chemical identity of these solutions may vary widely from the ionic environments encountered during their use. To explore this idea, we test the mechanical properties of a hydrogel cross-linked with dynamic covalent chemistry (DCC) in several physiologically relevant ionic solutions that mimic different biological conditions. Specifically, we evaluate rheological properties of a hydrazone-cross-linked hydrogel composed of recombinant, chemically modified hyaluronan and elastin-like protein (ELP). Our results show that the shear moduli and stress relaxation properties of DCC hydrogels can vary significantly in different ionic environments. We identify the thermoresponsive nature of ELP and changes in hydrazone bond kinetics as the primary reasons for the observed differences in mechanical properties. Taken together, this work elucidates mechanisms underpinning changes in hydrogel mechanics in different physiological solutions.

<h4>Main conclusion</h4>Almond trees prioritize C to yield rather than stem growth, and their overall lower NSC stores compared to forest trees may have consequences for resilience under environmental variability. Perennial tree crops store nonstructural carbohydrates (NSCs) as energy reserves that can be used to persist during both predictable periods of reduced activity like dormancy and more unpredictable periods associated with stress. For deciduous tree crops, which lose their leaves at the start of dormancy, the NSC reserves accrued by that time are critically important for fueling respiration but may also influence processes in the following growing season. To quantify the seasonal NSC fluctuation surrounding dormancy and its influence on downstream processes like growth and yield, we conducted a comparative study of four almond varieties in a commercial orchard (California, USA). Sugar and starch concentrations were quantified in branch, stem, and coarse root when entering and exiting dormancy. We then assessed the correlation between these NSC data and metrics of stem growth and yield in the following growing season. We further explored long-term trade-offs between stem growth and yield using historical data from 2017 to 2022. Overall, total NSC concentrations significantly decreased during the dormant season in all organs. We observed a significant positive correlation between branch total NSC concentration when entering dormancy and yield the next year. However, we unexpectedly found that stem total NSC concentrations when entering dormancy were negatively correlated with stem growth the next year, suggesting that stem reserves were primarily used to support wintertime respiration or translocated to other organs. A long-term trade-off between stem growth and yield was evident; as yield increased, basal area increment tended to decrease. Additionally, we found whole-tree NSC storage to be lower in almond trees compared to temperate forest trees, reflecting prioritization to yield over NSC storage. Overall, these findings advance our understanding of crop tree carbon physiology and provide insight into the resilience of different almond cultivars under changing environmental conditions.

Ewing sarcoma (ES) is an aggressive bone tumor in children and adolescents, and metastatic dissemination remains the major cause of treatment failure. Protocadherin 17 (PCDH17), a member of the cadherin superfamily, has been reported to exert context-dependent roles in solid tumors, but its significance in ES is unclear. Here, we analyzed Gene Expression Omnibus (GEO) datasets to evaluate PCDH17 expression, its association with overall survival and event-free survival, and the enrichment of biological pathways related to PCDH17 levels, and further examined PCDH17 expression in ES cell lines. The biological functions of PCDH17 were investigated using short hairpin RNA-mediated knockdown in A673 and RD-ES cells, followed by Cell counting kit-8 (CCK-8), colony formation, wound healing and transwell invasion assay, as well as western blotting (WB) and immunofluorescence (IF). PCDH17 was markedly upregulated in ES tissues and cell lines compared with normal counterparts, and high expression correlated with shorter survival. Gene set enrichment analysis (GSEA) indicated that high PCDH17 expression is associated with transcriptional programs driving cell cycle progression and oncogenic growth. Functionally, silencing PCDH17 significantly impairs proliferation, colony formation, migration and invasion of ES cells, and attenuates mesenchymal marker expression as well as associated morphological features. Collectively, our data indicate that PCDH17 is upregulated in ES and associated with worse survival, and that PCDH17 depletion suppresses malignant phenotypes in vitro. Together, these results nominate PCDH17 as a candidate prognostic biomarker and potential therapeutic target, which merits further validation.

Omics signatures for psychiatric conditions have been under-investigated. In this study, we used data from a representative observational cohort study of Hispanic/Latino adults to identify serum metabolites associated with symptoms of depression or anxiety and to develop and assess metabolite risk scores (MRSs) of these phenotypes. We conducted a metabolome-wide association study of 768 metabolites with depression and anxiety symptoms, using two separate assay batches measured from baseline samples collected between June 2008 and July 2011 for discovery (batch 1: n = 4002 adults) and replication (batch 2: n = 2178 adults). We estimated the associations via survey-based generalized linear regression, and applied Least Absolute Shrinkage and Selection Operator (LASSO) regression for metabolite selection and for obtaining MRS weights. In analyses adjusted for age, sex, field center and Hispanic/Latino background, we identified five metabolites of replicated associations with depression symptoms and six metabolites with anxiety symptoms. Key pathways related to the identified metabolites included arginine and proline, vitamin A, phospholipid, fatty acid and tyrosine metabolism. MRSs were associated with higher depression symptom score (1.07 points, 95% CI: 0.67 - 1.46, per 1 SD increase in depression MRS) and anxiety symptom score (1.05 points, 95% CI: 0.67 - 1.43, per 1 SD increase in anxiety MRS) in the replication dataset. Our study supports the associations between serum metabolites and symptoms of depression and anxiety in Hispanic/Latino adults. The developed MRSs may facilitate earlier and more objective screening of individuals for depression and anxiety apart from traditional risk factors.

Mechanochemical polymerization offers a solvent-free alternative to conventional solution-based methods. However, achieving controlled radical polymerization poses challenges due to the competing interplay between chain propagation and polymer degradation. Herein, we explore atom transfer radical polymerization (ATRP) under optimized mechanochemical conditions to enhance controlled polymerization and systematically analyze the overall reaction process, encompassing polymer synthesis, degradation, and depolymerization. By expanding the range of aromatic methacrylate monomers, we demonstrate that the size of pendant groups significantly impacts the kinetics of mechanochemical ATRP. Notably, the propagation rate constant in this process surpasses that of conventional ATRP, indicating significantly faster kinetics in solid-state reactions. Semi-quantitative MALDI-ToF analysis and computational simulations elucidated the contributions of main-chain, pendant-group, and end-group scissions with respect to the pendant group size, affirming the influence of steric effects on degradation pathways. Additionally, we observe the depolymerization process concurrent with polymer degradation, offering further insights into mechanochemical ATRP. This study provides valuable insights on efficient design and the mechanistic understanding of mechanochemical polymerization.

The rise of multi-drug resistant bacteria constitutes a global challenge with direct impact on millions of patients worldwide. The identification of bacterial species is essential to avoid unnecessary use of antibiotics and to minimize the emergence of additional resistance; however, there are few chemical strategies to identify resistant species in a rapid and unbiased manner. Cross-reactive arrays combine multiple sensor components to produce distinctive fingerprints for similar targets; herein, we present a cross-reactive sensing array built with long lifetime organic fluorophores. To the best of our knowledge, we synthesize the largest collection to date of bioconjugatable triangulenium fluorophores by including heteroatom and side-chain diversification for spectral and lifetime diversity, as well as water-soluble and orthogonal moieties for peptide tagging and compatibility with biological samples. After optimization, we evolve a four-fluorophore sensor array that correctly assigns all seven ESKAPEE bacterial species by combining their optical signatures. Lifetime chemical sensor arrays will open avenues for concentration-independent, reliable and sensitive optical detection without the need for prior knowledge of molecular targets.

The basal ganglia comprise interconnected subcortical nuclei essential for motor control, learning, emotion and cognition, yet how cell types relate to their circuit organization remains elusive. Here we show that spatial patterns of transcriptomic cell types in the basal ganglia and thalamic parafascicular nucleus relate to module-organized cortical inputs in the mouse. By co-registering genoarchitectural and connectivity datasets to the common coordinate framework, we delineate distinct three-dimensional subdivisions of these nuclei. Analyses of anterograde and retrograde viral tracing data and single-neuron reconstructions reveal that each subdivision receives convergent, modular and cell-type-specific cortical and subcortical inputs. Caudoputamen subdivisions primarily receive layer 5 intratelencephalic inputs from the entire isocortex, whereas smaller basal ganglia subdivisions receive ipsilateral layer 5 extratelencephalic inputs from limited cortical areas. The parafascicular nucleus subdivisions are connected with specific cortical modules and basal ganglia subdivisions. Our results suggest that combinatorial gene expression underlies a global map of parallel, modular and cell-type-specific basal ganglia and parafascicular nucleus subnetworks.

Colorectal cancer (CRC) is one of the most commonly diagnosed and globally spread malignant diseases. Cancer-associated fibroblasts (CAFs) are key architects of the tumor microenvironment, yet their origin, stability, and interconvertibility remain poorly understood. Using transcriptomic profiling of fibroblasts from colorectal cancer (CRC) patients, we identify highly expressed (HEX) markers that define fibroblast subpopulations and uncover mechanisms governing their plasticity. We find that ADH1B marks normal colon-associated fibroblasts (NAFs), which consist of PI16-NAFs and ADAMDEC1-NAFs. ITGA3 delineates the total CAF population, which comprises myofibroblastic CAFs (myCAFs), whose characterizing markers were associated with poor prognosis and proteolytic inflammatory CAFs (piCAFs), characterized by markers not associated with prognosis. An AGT/TGM2-expressing fibroblast subset is present in both healthy and tumor tissues, suggesting alternative trajectories to the classical NAF-to-CAF transition model. While PI16-NAFs, AGT/TGM2-fibroblasts, and myCAFs maintain stable identities in long-term culture, the ADAMDEC1-NAF and piCAF phenotypes are lost in vitro. ITGA3-CAFs demonstrate dynamic plasticity, with TGF-β stably inducing myCAF formation and TNF-α or inhibition of DNA methylation promoting transient piCAF emergence. These findings redefine fibroblast heterogeneity in CRC and reveal a coexisting stable and plastic fibroblast network that may be amenable to modulation and provides a framework for future functional and translational studies. We identified highly expressed markers (HEX markers) to distinguish CAFs, NAFs and corresponding subpopulations in CRC. ADH1B characterized NAFs, which consisted of stable (solid outline) PI16-NAFs and unstable (dashed outline) ADAMDEC1-NAFs. ITGA3 identified CAFs consisting of stable myCAFs associated with poor prognosis and unstable piCAFs not associated with prognosis. AGT/TGM2 fibroblasts did not express ADH1B or ITGA3, were stable in culture and could be detected in both healthy colon and CRC. Treatment of PI16-NAFs with LPS or IFN-γ induced ADAMDEC1-NAFs, TGF-β the formation of myCAFs, while treatment with TNF-α led to the formation of piCAFs. Reduced DNA methylation converted myCAFs and PI16-NAFs into piCAFs.

Doppler parameters used to predict the maintenance of sinus rhythm after cardioversion of supraventricular arrhythmia (SVA) may be influenced by heart rate. We included patients with septic shock who developed SVA, had normal to moderately reduced left ventricular systolic function (ejection fraction > 35%), and received continuous noradrenaline at doses < 1.0 µg/kg/min. Cardioversion was successful in 96% of the 209 patients with the echocardiographic data available for 187 patients post cardioversion. The mitral E wave velocity decreased from 87 (76;100) cm/s during SVA to 82 (70;98) cm/s at 4 h (p = 0.001), while the newly restored mitral A wave increased from 65 (52;79) cm/s at 1 h to 69 (56;83) cm/s at 4 h (p = 0.002). Similarly, the A wave velocity-time integral (Avti) increased from 7.00 (5.63;8.28) cm to 7.65 (6.10;9.40) cm, p < 0.001. A weak positive relationship between the heart rate (85 (77;95) bpm) and A wave (R² = 0.04, p < 0.001) was found. No significant relationship between the heart rate and the E wave (R² = 0.001, p = 0.56), and a heart rate related decrease in the E/A ratio (R² = 0.03, p = 0.002) were observed. With accelerating heart rate, the Avti (R² = 0.02, p = 0.016), Avti/LVOTvti ratio (R² = 0.03, p = 0.006) and Avti/stroke volume ratio (R² = 0.03, p = 0.004) all increased. In patients with septic shock, mitral Doppler parameters associated with mechanical sinus rhythm after cardioversion are not significantly affected by heart rate.

Targeted protein degradation (TPD) redefines therapeutic strategies by overcoming the key limitations of traditional occupancy-driven pharmacology. TPD introduces a paradigm shift by using the cell’s intrinsic degradation systems—primarily the ubiquitin–proteasome system (UPS), as well as lysosomal and autophagy pathways—to eliminate disease-causing proteins. Here, we first summarize the research progress of the most advanced TPD platform, proteolysis-targeting chimeras (PROTACs). While most TPD strategies currently rely on a few well-characterized E3 ligases, such as CRBN and VHL, ongoing research aims to expand the ligase repertoire to increase tissue specificity and overcome resistance. Therefore, we also characterized alternative TPD platforms, such as lysosome-targeting chimeras (LYTACs), autophagy-targeting chimeras (AUTACs), and ribonuclease-targeting chimeras (RIBOTACs), which exploit lysosomal and autophagy pathways, broadening the ability of TPD to reach extracellular and aggregated proteins. Moreover, the application of TPD in oncology, neurodegeneration, immunology, and infectious diseases, with several agents advancing into clinical trials, has also been concluded. On the other hand, the key challenges of TPD remain, including improving bioavailability, minimizing off-target effects, and understanding resistance mechanisms. Overall, as novel chemistries and delivery strategies continue to evolve, TPD represents a powerful and versatile modality with the potential to radically expand the druggable proteome and address unmet medical needs across a wide range of diseases.

OBJECTIVE: Clarify the shared genetic architecture between Alzheimer’s disease (AD) and major psychiatric disorders. METHODS: We integrated large GWAS summary datasets derived predominantly from individuals of European ancestry for AD and eight psychiatric disorders; estimated genome-wide genetic correlations with LDSC/HDL; mapped local genetic correlations with SUPERGNOVA; ran pairwise Multi-trait analysis of GWAS (MTAG (AD with ADHD, BIP, MDD, PTSD, SCZ)) followed by FUMA; performed cross-trait colocalization with HyPrColoc and trait–eQTL colocalization using GTEx v8 (49 tissues). RESULTS: HDL identified significant genome-wide correlations for five AD–psychiatric pairs, led by AD–MDD; LDSC was significant for AD–MDD. We mapped 18 locally correlated regions. MTAG yielded 33 AD-associated loci (118 SNPs), including 12 novel (e.g., RAB27B/rs12968702, PTCH1/rs3824488, EP300/rs12157997), and 336 psychiatric-trait signals across 265 loci. HyPrColoc detected 74 AD–psychiatric colocalized regions (40 with PP ≥ 0.8), with 13 driven by a single candidate causal variant. Trait–eQTL colocalization prioritized 25 genes in 122 associations; brain-tissue signals implicated P4HTM, GPX1, CCDC71, and—at an AD–SCZ locus—ADAM10. CONCLUSION: AD shares substantial pleiotropic architecture with psychiatric disorders, particularly MDD. Integrative multi-trait association, colocalization, and tissue-specific eQTL evidence highlights convergent mechanisms—exosome biology (RAB27B), astrocytic Ca²⁺ signaling (P4HTM), and non-amyloidogenic APP processing (ADAM10)—and nominates testable therapeutic targets.

<h4>Background</h4>Leukodystrophies are a clinically and genetically heterogeneous group of diseases characterized by white matter abnormalities on brain magnetic resonance imaging. Clinical, biochemical, molecular, and/or neuroimaging findings collectively support the diagnosis confirmation. The heterogeneous and overlapping clinical presentations of different leukodystrophies and non-diagnostic molecular testing pose a significant challenge to establishing a definitive diagnosis in these rare diseases. The Myelin Disorders Biorepository Project is an observational research program that aims to establish new tests to diagnose leukodystrophies and describe the natural history of these disorders. Ensuring an accurate diagnosis is critical to the goals of this project, and this paper aims to describe the rigorous diagnostic review and confirmation process which was developed.<h4>Results</h4>We present a diagnosis review process that contributes to an accurate diagnosis for participants enrolled in this study. Board-certified genetic counselors with expertise in these disorders audit medical records to carefully assess each enrolled participant's clinical, biochemical, and molecular features. A scale of diagnostic categories is assigned based on the record review, and a team of leukodystrophy physician experts consults for cases that require further characterization or clarification.<h4>Conclusions</h4>This robust review process has resulted in a database of individuals with verified diagnoses that may be easily queried for inclusion in appropriate natural history studies and/or treatment trials. This is a model framework that may be adapted and implemented by other rare disease groups.

<h4>Background</h4>MYCN non-amplified (MYCN-NA) neuroblastoma (NB) demonstrates considerable heterogeneity in both biological and clinical aspects, and its molecular and biological characteristics remain inadequately defined.<h4>Methods</h4>A comprehensive multi-omics analysis was performed on transcriptome and proteome data from 20 MYCN-NA NB tissues, including 11 low- and intermediate-risk neuroblastoma (LIR-NB) cases and 9 high-risk neuroblastoma (HR-NB) cases. Additionally, the expression levels and survival prognostic significance of key candidate genes were systematically assessed using public datasets (GSE85047 and TARGET-NB).<h4>Results</h4>Transcriptomic analysis revealed 1,955 differentially expressed genes (DEGs), with 899 upregulated and 1,056 downregulated in HR-NB (P < 0.05, |log₂FC| ≥ 1.5). Pathway enrichment analysis based on the Kyoto Encyclopedia of Genes and Genomes identified significant associations of these DEGs with glycolipid metabolism, signal transduction, and transcriptional regulation. Proteomic profiling identified 609 differentially expressed proteins (DEPs), with 24 upregulated and 585 downregulated in HR-NB. These DEPs were predominantly enriched in metabolic pathways, axon guidance, and Rho GTPase signaling. Integrated omics analysis revealed that both mRNA and protein expression of DPEP1 and FILIP1 were co-upregulated in HR-NB, while seven genes-ICA1L, KCNH2, LGI3, FAM184A, INPP5F, NELL2, and VKORC1L1-were co-downregulated. Genes exhibiting consistent changes in both protein and mRNA levels were primarily involved in glycerophospholipid metabolism and regulation of transport. Moreover, the top five genes (ICA1L, KCNH2, LGI3, FAM184A, and INPP5F) with the most significant concurrent changes were further validated for their expression levels and association with survival prognosis in GSE85047 and TARGET-NB datasets. Notably, increased expression of INPP5F and LGI3 correlated with an improvement in overall survival rate (P < 0.05).<h4>Conclusion</h4>MYCN-NA HR-NB demonstrates metabolic reprogramming characterized by disorders in glucose and lipid metabolism. Notably, the elevated expression of INPP5F and LGI3 is linked to improved overall survival and may serve as potential therapeutic targets.

No abstract available.

Kinesins, a class of microtubule (MT)-dependent molecular motors, regulate MT dynamics and MT-mediated transport. We previously identified Kif15 (kinesin-12) as a key player in axonal growth by modulating MT remodeling during neuronal development, and more recently, its involvement in protein localization. In this study, we observed that Kif15 knockout (Kif15 KO) mice exhibited accelerated functional recovery after sciatic nerve injury. To investigate the cellular responses underlying this enhanced recovery after axotomy, spinal cord tissues from the injured regions were collected for single-nucleus RNA sequencing (snRNA-seq). The snRNA-seq results revealed differential genes expression in neurons, indicating a neuroprotective shift in Kif15 KO mice, and in microglia, where a repair-promoting and synapse-modulating profile was observed. Notably, the CX3CL1-CX3CR1 signaling pathway, critical for neuronal-microglial communication, was downregulated in Kif15 KO mice compared to wild-type controls. Further molecular analysis indicated that Kif15 facilitated the expression and localization of neuronal CX3CL1, which, in turn, influenced microglial function via the receptor CX3CR1. Our findings highlight a novel role for Kif15 in regulating neuronal-microglial communication through modulation of CX3CL1 signaling.

Breast cancer is the most prevalent malignancy in women, and the limited effectiveness of current treatments highlights the need for novel immune regulatory mechanisms to improve long-term survival. This study investigated the role of Vim in PGI₂ synthesis and its impact on tumor immune regulation. Multiomics profiling revealed molecular alterations following Vim deletion, which were validated in murine breast cancer models using RT-qPCR, Western blot, ELISA, and flow cytometry, with rescue experiments involving exogenous PGI₂. The findings showed that Vim deletion downregulated arachidonic acid metabolism, reduced PTGIS expression, and significantly lowered PGI₂ levels. Functional assays demonstrated that Vim deficiency enhanced T cell-mediated antitumor immunity, evidenced by an increased proportion of CD8⁺ T cells, upregulation of cytotoxic genes (<i>Ifng</i>, <i>Gzmb</i>, <i>Tnf</i>, and <i>Klrd1</i>), and activation of inflammation-related signaling pathways, as indicated by enhanced phosphorylation of ERK1/2 and p65. Both exogenous PGI₂ supplementation and ozagrel treatment reversed these effects. In conclusion, the Vim-PGI₂ axis is identified as a key regulator of CD8⁺ T cell immunity in breast cancer, representing a potential therapeutic target and a critical consideration in anticoagulant management during cancer immunotherapy.

Several New World mammarenaviruses (NWMs), including Junín virus (JUNV), can cause a severe, potentially fatal hemorrhagic fever (HF) syndrome. In the absence of FDA-approved vaccines or antivirals, there is an urgent need to develop broadly active countermeasures against these viral infections. Cellular entry by all pathogenic NWMs is mediated by transferrin receptor 1 (TfR1). Here, we report on the pharmacokinetics (PK) and antiviral activity of a mouse/human chimeric antibody, targeting the apical domain of human TfR1 and lacking effector functions (ch128.1/IgG1 Fc-silent) in two transgenic mouse models of JUNV infection. Assessment of tolerability and PK in human TfR1 knock-in/mouse TfR1 knock-out mice (huTfR1 Tg mice) demonstrated that ch128.1/IgG1 Fc-silent (FcS) was well-tolerated at doses up to 800 μg, and that once-daily dosing may be the most effective regimen for achieving optimal therapeutic benefit. Prophylactic and post-infection intervention efficacy studies employing once-daily antibody treatments in huTfR1 Tg mice challenged with JUNV revealed significant protection against mortality, even when treatment was delayed until 5 days post-infection. In mice with TfR1 containing only the apical domain of the human receptor (huApTfR1 Tg mice), the ch128.1/IgG1 FcS treatment trended toward increased survival and significantly reduced weight loss following JUNV challenge. The antibody treatments also significantly decreased infectious JUNV titers in target organs (liver and spleen) in the more physiologically relevant huApTfR1 Tg mouse model. Taken together, effective treatment of advanced JUNV infection in two transgenic mouse models with ch128.1/IgG1 FcS antibody supports the development of a humanized version of this antibody for clinical application.

Chimeric antigen receptor (CAR) T cells offer a promising strategy for the treatment of autoimmune diseases. However, clinical translation is limited by the high cost, complexity, and poor scalability of current manufacturing, restricting broad patient access and persisting safety concerns including insertional mutagenesis risk from integrating vectors and uncontrolled long-term CAR T cell persistence. <i>In-vitro</i>-transcribed (IVT) mRNA enables transient, non-integrating CAR expression with improved safety and scalability, making it particularly suited for non-malignant indications where prolonged persistence may not be required. Here, we systematically compare two IVT mRNA delivery platforms, electroporation and lipid nanoparticles (LNPs), for transient CAR T cell engineering in primary human T cells using single-cell transcriptomics and functional cell assays. We show that electroporation yields higher transfection efficiency and more sustained CAR surface expression, whereas LNP delivery reduces stress- and senescence-related transcriptional signatures as well as exhaustion marker expression, while enhancing antigen-driven activation, chemotactic responses, and cytotoxic function. Our comparative analysis highlights that the mode of mRNA delivery is associated with distinct transcriptional signatures and functional properties of CAR T cells, providing a framework to guide future development of mRNA-based approaches. These insights support LNP-mediated delivery as a functionally favorable strategy for transient CAR T cell engineering in autoimmune disease and beyond.

<b>Background:</b> High-grade serous ovarian carcinoma (HG-SOC) remains the most lethal gynecological malignancy, largely due to intrinsic or acquired resistance to platinum-based chemotherapy. Although large-scale sequencing studies have delineated the genomic landscape of HG-SOC, clinically actionable biomarkers predictive of platinum response and outcome are still lacking. This study aimed to identify genomic alterations associated with platinum sensitivity, resistance, or refractoriness, and to assess their prognostic relevance. <b>Methods:</b> Tumor DNA from 24 HG-SOC patients with optimal cytoreductive resection, classified as platinum-sensitive (<i>n</i> = 9), platinum-resistant (<i>n</i> = 8), or platinum-refractory (<i>n</i> = 7) underwent targeted next-generation sequencing of 409 cancer-associated genes. Somatic variants were filtered and classified for oncogenicity using established criteria incorporating predicted functional impact, REVEL scores, and population allele frequencies. Associations between mutational profiles, platinum response, and overall survival (OS) were evaluated using Kaplan-Meier and Cox regression analyses. Key findings were validated in the TCGA ovarian serous carcinoma (TCGA-OV) dataset using survival analyses. <b>Results:</b> A total of 1367 protein-altering somatic variants across 301 genes were identified. While <i>TP53</i> mutations were ubiquitous, platinum-resistant and platinum-refractory tumors showed enrichment of pathogenic alterations affecting DNA repair, transcriptional regulation, epigenetic modification, and oncogenic signaling, including <i>FANCA</i>, <i>ATF1</i>, <i>MAF</i>, <i>NCOA2</i>, <i>PIK3CA</i>, and <i>TET1</i>. Mutations in these genes were associated with reduced overall survival in exploratory analyses (median 2.5-9 months vs. 27.5-45 months). Multivariate analysis identified <i>FANCA</i> and <i>ATF1</i> as potential independent predictors in exploratory modeling. In the TCGA-OV cohort, patients harboring pathogenic variants in a multi-gene panel derived from this study (excluding <i>BRCA1/2</i>) exhibited significantly worse survival compared with both <i>BRCA1/2</i>-mutated cases and the overall cohort. <b>Conclusions:</b> This exploratory study identifies a set of genomic alterations converging on transcriptional and epigenetic regulation, DNA repair, and oncogenic signaling that are associated with platinum resistance and adverse prognosis in HG-SOC. Independent validation in TCGA supports the potential clinical relevance of this mutational signature. These findings warrant further validation in larger prospective cohorts and functional studies to clarify their role as biomarkers of aggressive disease and therapeutic vulnerability.

Non-glycosylated liver-derived acute-phase amyloid A1 and A2 proteins (SAA1 and SAA2, 104 amino acids), generated by two different genes in humans (<i>SAA1</i>/<i>2</i>) and other mammalian species, are considered the prime acute-phase reactants following inflammatory conditions during host defense in cells, tissues, and the circulation. While human <i>SAA3</i> has been identified as a pseudogene, <i>Saa3</i> genes in other mammalian species are coding for primarily extrahepatically expressed Saa3 proteins that also may act as suitable inflammatory markers. The discovery of SAA4 (112 amino acids, carrying an octapeptide insert) in humans and mice has paved a new avenue for the exploration of different functions of this so far unknown member of the SAA superfamily. SAA4 has originally been termed a "constitutively" expressed SAA protein, apparently due to its nature not to act as an inflammatory marker. The present overview aimed to cover possible functions-so far identified-for human SAA4 (following its expression in various diseases on mRNA and protein level) and to work out whether SAA4 might be considered-at least in part-an acute-phase protein. Alternatively, we are raising the question whether SAA4 may solely act as a bystander or even underdog within the whole SAA family, where SAA1 and SAA2 proteins (commonly termed acute-phase SAA) hold undoubtedly an eminent status during inflammatory conditions, not only as host defense reactants but also as long-lasting markers for chronic diseases and malignancies in humans.

Tumor pathology is undergoing a profound transformation, shifting from pure morphological description toward multidimensional functional network analysis. This Mini Review focuses on the tumor microenvironment (TME) as a central concept driving tumor initiation, progression, and therapeutic resistance. We first outline the limitations of traditional pathological classification and elaborate on how the dynamic co-evolution of cellular components (such as cancer-associated fibroblasts (CAFs) and immune cells) along with the extracellular matrix (ECM) constitutes a functional unit. Key controversies are discussed, including the translational hurdles of TME-directed therapies and the challenge of spatiotemporally assessing tumor heterogeneity. We further identify critical research gaps, particularly the mechanistic understanding of the tumor-host interface across scales. Finally, we envision that the integration of artificial intelligence-driven spatial pathology, single-cell multi-omics, and <i>in vivo</i> imaging will usher in a new era of "functional pathology," merging morphology, molecular profiling, and dynamic insights.

Small G proteins are molecular switches in the realm of cell biology, switching from the GTP-bound active form to the GDP-bound inactive form. They participate in controlling various essential cellular functions, one of which is cell migration. Cell migration is a sine <i>qua non</i> for the proper functioning of the cell, as it aids in a litany of cellular processes, including embryonic development, morphogenesis, organogenesis, wound healing, and pathogenic states such as cancer metastasis. Amongst the five families of small G proteins (with over 150 members), Rho GTPase family is known to have significant roles in cell migration. Rho GTPases (primarily RhoA, Rac1, and Cdc42) translate extracellular cues into the spatiotemporal coordination of the actin cytoskeleton, allowing cells to polarize, migrate and navigate their environment. They integrate biochemical and biophysical signals through guanine nucleotide exchange factors (GEFs) and GTPase-activating proteins (GAPs). Rac1 and Cdc42 are known to act at the front end of a migrating cell and aid in cell polarisation, formation of protrusions and adhesive structures, while RhoA, though having overlapping roles, majorly functions at the cell rear, where it regulates cell body contraction and tail detachment, all of which we look into elaborately in this review. We also discuss the roles played by the other Rho GTPases in cell migration and have touched on the physiological and pathological impacts that Rho GTPase-regulated cell migration has on human health.

<h4>Background</h4>Although patients with papillary thyroid cancer (PTC) generally have a favourable prognosis, they still face the possibility of recurrence after surgery. Identifying biomarkers related to PTC initiation and progression is crucial. The role and regulatory mechanisms of tripartite motif containing 35 (TRIM35) in PTC remain unclear. This study aimed to investigate the expression, biological functions, and underlying mechanisms of TRIM35 in PTC.<h4>Methods</h4>The expression of TRIM35 was evaluated via immunohistochemistry, and its association with patient clinicopathological features was examined with Fisher's exact test. In PTC cell lines, TRIM35 was silenced using small interfering RNA (siRNA). A series of functional assays were then performed. Cell viability was measured with the Cell Counting Kit-8. Cell motility was assessed via wound healing assays. Cell migration and invasion were evaluated using Transwell chambers, with invasion assays specifically employing a Matrigel coating. Commercial assay kits were employed to quantify lactate production along with the expression of key glycolytic enzymes-hexokinase (HK) and lactate dehydrogenase A (LDHA). Transcriptomic sequencing was conducted to identify differentially expressed genes (DEGs) in TRIM35-knockdown K1 cells. Finally, protein levels of peroxisome proliferator-activated receptor gamma (PPARγ) and cluster of differentiation 36 (CD36) were detected by western blot analysis.<h4>Results</h4>TRIM35 expression was elevated in cancerous tissues, and its levels were closely linked to tumour size and the presence of lymphovascular invasion. Knockdown of TRIM35 notably suppressed cellular growth and metastasis of PTC cells. Interestingly, PTC cells with reduced TRIM35 levels produced significantly lower amounts of lactate. Moreover, we detected a notable decrease in the levels of HK and LDHA, in TRIM35-knockdown K1 and TPC-1 cells. Seventy-five downregulated and 103 upregulated DEGs were identified in TRIM35-knockdown cells. PPAR signalling pathway was enriched in the top 20 enriched Kyoto Encyclopedia of Genes and Genomes pathways. TRIM35 knockdown significantly suppressed the expression of both PPAR γ and CD36.<h4>Conclusions</h4>Elevated TRIM35 expression was associated with larger tumour size and the presence of lymphovascular invasion. Functionally, TRIM35 drives malignant phenotypes and glycolytic metabolism in PTC cells, likely via the PPAR pathway. Our present study indicates that TRIM35 may serve as a promising biomarker indicative of PTC progression.

Spinal commissural neurons play a fundamental role in motor control and sensory perception. Robo3, a receptor expressed on pre-crossing commissural axons, is required for midline crossing. Its downregulation in post-crossing axons is essential for forming synapses with contralateral CNS targets. We demonstrate that, at the transcriptional level, the dynamic expression of <i>Robo3</i> in pre-crossing neurons is controlled by incoherent feedforward loops (iFFLs) and negative feedback loops (NFLs) that precisely regulate its transience and thereby determine midline crossing. Lhx2 and Lhx9 activate <i>Robo3</i> while also inducing its repressor, Barhl2. Additionally, NFLs fine-tune the transience of <i>Robo3</i> expression by adjusting the relative strength of the activation and repression modules. Our genomic analysis reveals that this regulatory circuitry converges on an enhancer element of the <i>Robo3</i> gene. These findings imply that diverse iFFLs, together with NFLs, are essential for regulating the extent of midline crossing across different spinal commissural subtypes.

Feeding behavior is a key factor influencing growth performance and feed efficiency in poultry, yet its complex phenotypes remain difficult to quantify accurately, limiting the application of large-scale behavioral data in genetic and molecular analyses. In this study, feeding behavior and feed efficiency-related phenotypes were recorded in 829 Tianfu Nonghua ducks using an automated feeding monitoring system integrated with radio frequency identification (RFID) technology. Genetic parameter estimation indicated that the heritability of feeding behavior traits ranged from 0.25 to 0.39. Individuals were classified into high-frequency feeding (HFF) and low-frequency feeding (LFF) groups. Phenotypic analysis showed that ducks in the HFF group had significantly higher residual feed intake (RFI) and feed conversion ratio (FCR) than those in the LFF group. Based on this grouping, transcriptome sequencing was performed on the hypothalamus, pituitary, and liver tissues. A total of 266, 181, and 94 differentially expressed genes (DEGs) were identified in the three tissues, respectively. Enrichment analysis highlighted tissue-specific pathways, including neuroactive ligand-receptor interaction and calcium signaling in the hypothalamus, the Apelin signaling pathway in the pituitary, and steroid hormone biosynthesis in the liver. Appetite-related genes in the hypothalamus (PMCH, HCRT, and CCKAR) and endocrine and metabolic genes in the pituitary and liver (TRH and NEGR1) showed differential expression between feeding strategies. Weighted gene co-expression network analysis further identified hub genes (ITPKB, BAMBI, and BCKDHB) associated with total feeding bouts (TFB). These findings provide new insights into the genetic architecture of feeding behavior traits and the molecular differences associated with divergent feeding patterns in ducks.

<title>Abstract</title> <p> Schizophrenia (SCZ) and bipolar disorder (BD) are complex psychiatric disorders with partially overlapping genetic architectures and shared features with neurodegenerative diseases. Short tandem repeats (STRs), particularly CAG expansions in <italic>HTT</italic> , <italic>ATXN1</italic> and <italic>ATXN2</italic> , are established causes of neurodegenerative disorders, yet their role as genetic modifiers in major mental disorders remains poorly understood. We analysed CAG repeat sizes in a cohort of 1,604 individuals, including 234 SCZ patients, 329 BD patients and 1,041 healthy controls. Repeat lengths were determined by fluorescent PCR and capillary electrophoresis, and associations with disease risk and clinical phenotypes were evaluated using non-parametric tests, multinomial logistic regression and survival analyses. Intermediate <italic>HTT</italic> alleles were more frequent in BD type I compared with controls (8.8% vs 4.4%, p = 0.033), whereas <italic>ATXN1</italic> intermediate alleles were less frequent in SCZ than in BD (4.7% vs 11.2%, p = 0.02). Although overall differences in repeat size were modest, <italic>ATXN2</italic> CAG length showed consistent shifts across diagnostic groups, with slightly larger repeats in BD and SCZ than in controls. Multinomial models identified the <italic>ATXN2</italic> long allele as associated with disease risk with a non-linear effect. Intermediate <italic>ATXN2</italic> alleles were also associated with shorter disease duration in BD (p = 0.00048). Functional enrichment analysis revealed convergence of these genes with SCZ- and BD-related networks involved in synaptic function and RNA metabolism. These findings support a role for CAG repeat variation as a genetic modifier influencing susceptibility and clinical trajectories in psychiatric disorders. </p>

The corpus callosum (CC), the largest interhemispheric white-matter tract, plays a central role in higher-order cognitive functions and is frequently altered in neurodevelopmental and psychiatric conditions. However, most diffusion MRI studies rely on tract-averaged measures, which obscure spatially specific microstructural variation along the tract that may hold biologically and functionally meaningful information. This study aimed to provide a fine-grained characterisation of spatial variation in callosal structure and to determine its developmental, genetic, and cognitive correlates. This study leveraged multimodal data from the Philadelphia Neurodevelopmental Cohort (PNC), comprising 1342 participants aged 8–21 years. Diffusion MRI and tractography-based segmentation were used to extract fine-grained spatial variation in diffusion metrics along seven callosal tracts. A candidate gene approach targeted four genes ( DCC , CDH2 , AKT3 , and GLI3 ) previously linked to the neurobiological mechanisms underlying CC formation. Five cognitive factors were derived from the Penn Computerized Neurocognitive Battery using factor analysis. A two-stage functional data analysis approach to examine global and local associations between along-tract diffusion metrics and age, candidate genetic variants, and behavioural outcomes. Results revealed distinct midline-to-cortical variations of age-related diffusion metrics change across callosal subdivisions. Frontal and parietal heteromodal callosal pathways showed pronounced distal-segment maturation (F = 13 – 23, p ≤ 2.8×10e-16), whereas posterior sensorimotor and occipital callosal tracts exhibited more stable age associations along their lengths (F = 4.2 – 4.3, p = 10e-3). Genetic variations in callosal axon-guidance genes (ROBO1, IQCJ-SCHIP1, NRP1 and DCC) were associated with spatial variation in callosal diffusion metrics, particularly in anterior (rostrum) and posterior callosal subdivisions (isthmus and splenium) (F = 4.29 – 18.61, p-values = 2.2e-16 – 1.4e-04). These regions correspond to early-forming callosal compartments, suggesting that prenatal axon- guidance mechanisms leave enduring spatial patterns on callosal organisation. Finally, spatial variation in callosal microstructure was significantly associated with behavioural performance (F = 2.9 – 21.3, P = 2.8×10e-16 – 0.04), with the strongest and most spatially heterogeneous effects observed for complex cognition and executive functioning. Across all analyses, functional data models generally outperformed tract-averaged linear models, supporting the value of explicitly preserving spatial variation along callosal tracts. Our findings converge on the CC as a spatially differentiated structure in which early genetic modulators and developmental constraints shape region- and segment-specific microstructural architecture that is behaviourally relevant through childhood and adolescence.

The limited durability of vaccine protection and the rapid emergence of SARS-CoV-2 variants highlight the need for antiviral strategies that extend beyond vaccination and conventional small-molecule inhibitors. Here, we explored a dual approach combining structure-guided peptides predicted to interfere with the viral replication complex with lethal mutagenesis to limit SARS-CoV-2 replication. Using the crystallographic interfaces of nsp10 with nsp14 and nsp16, we designed short inhibitory peptides predicted to interact with the viral proofreading and RNA-capping machinery. In parallel, the mutagenic analog 5-fluorouracil was evaluated to determine its effect on SARS-CoV-2 in Vero E6 cells. Peptides P1 and P6 exhibited potent antiviral activity with minimal cytotoxicity, whereas 5-FU reduced specific infectivity without impairing genome replication. Combined treatment with 5-FU and peptide P1 resulted in >10⁴-fold reduction in infectious virus, achieving near-complete loss of infectivity at non-cytotoxic concentrations. Next-generation sequencing revealed that dual treatment increased mutation frequency, altered mutant spectra, and decreased genome stability, consistent with progression toward error catastrophe. Principal component analysis confirmed that combined treatment generated mutant spectra distinct from either monotherapy. These findings are consistent with a dual antiviral strategy in which structure-guided peptides designed to interact with components of the SARS-CoV-2 replication complex act in combination with lethal mutagenesis to produce a synergistic interaction between these two complementary processes. This integrated approach suggests a potential broad-spectrum antiviral strategy with applicability to other coronaviruses.

Hepatocellular carcinoma (HCC) is an extremely aggressive type of cancer with a poor prognosis and is often associated with amplification of chromosome 8q. Tonsoku-like DNA repair protein (TONSL) gene, located on 8q, plays a role in repairing DNA double-strand breaks. However, its functional relevance in HCC is not yet fully understood. In this study, we investigated the biological significance of TONSL in HCC progression. Analyses of public datasets and clinical specimens revealed that TONSL expression was markedly elevated in tumor tissues. TONSL levels showed a positive correlation with copy number variation and high levels were significantly associated with unfavorable patient outcomes. Functional assays demonstrated that TONSL knockout substantially reduced cell growth and increased apoptosis. Mechanistically, loss of TONSL impaired the recruitment of RAD51 to DNA damage sites, resulting in defective homologous recombination repair and enhanced apoptotic activity. Furthermore, TONSL-knockout HCC cells showed increased sensitivity to poly (ADP-ribose) polymerase (PARP) inhibitor. In a xenograft model, tumors generated from TONSL-knockout cells exhibited significantly reduced growth, underscoring the importance of TONSL in promoting HCC progression. Collectively, these findings indicate that TONSL facilitates HCC development by supporting RAD51-mediated DNA repair and suppressing apoptosis, and its elevated expression serves as a marker of poor prognosis. Thus, TONSL represents a promising biomarker of tumor aggressiveness and a potential therapeutic target in HCC.

Kinases represent one of the main targets of interest in recent years, completely revolutionizing small molecule anticancer therapy. Particularly, several medicinal chemistry strategies (e.g., covalent and allosteric inhibitors) allowed overcoming of the challenging selectivity issue, thus enabling the massive clinical translation of kinase inhibitors. However, the same success has not been detected yet for tackling neurodegenerative diseases, despite plenty of experimental evidence regarding kinases' pivotal roles in onset and development of neurodegenerative processes. In this perspective we highlight the therapeutic potential of allosteric kinase modulators in this respect, by showcasing the developmental processes and neuroprotective properties of CNS-directed allosteric kinases modulators developed so far. Moreover, we present additional kinases whose modulation is related to neuroprotection and featured by validated allosteric modulators not yet exploited in this context. This, along with a critical discussion on main drawbacks and future directions, may foster the development of allosteric kinase modulators in neurodegenerative diseases.

Myeloid/lymphoid neoplasms with eosinophilia (MLN-eo) associated with PDGFRB rearrangements represent a rare but clinically significant category of hematologic malignancies, characterized by clonal eosinophilia and exceptional sensitivity to tyrosine kinase inhibitors. Among these, the MPRIP::PDGFRB fusion is exceptionally uncommon, with only three patients previously reported. Here, we report the first United States patient with MPRIP::PDGFRB fusion in a 34-year-old man presenting with leukocytosis, marked eosinophilia, anemia, thrombocytopenia, mucocutaneous lesions, hepatosplenomegaly, chronic gastrointestinal symptoms, and tree-in-bud pulmonary nodularity. Bone marrow evaluation revealed a markedly hypercellular marrow with increased eosinophils, grade 1 reticulin fibrosis, and dysmegakaryopoiesis. Fluorescence in situ hybridization (FISH) demonstrated a PDGFRB rearrangement in 76% of cells, and chromosome analysis revealed a balanced translocation t(5;17)(q32;p11.2). Next-generation sequencing RNA-based myeloid fusion panel analysis identified an MPRIP::PDGFRB fusion formed by an in-frame junction of MPRIP exon 20 and PDGFRB exon 12. The patient was initiated on imatinib therapy and achieved rapid remission. To contextualize this patient, we compared the clinical and molecular features of all previously published patients with MPRIP::PDGFRB fusion, noting shared findings of marked eosinophilia and excellent imatinib sensitivity, but notable heterogeneity in symptom burden, degree of marrow fibrosis, and associated immune-related manifestations. Additionally, we provide an overview of 45 reported PDGFRB fusion partner genes, summarizing their cytogenetic characteristics, and associated diseases. This case report expands the clinical spectrum of MPRIP::PDGFRB positive MLN-eo and underscores the essential role of cytogenetic and molecular testing in the diagnostic evaluation of eosinophilia, given the significant therapeutic implications of identifying PDGFRB fusions.

PURPOSE OF REVIEW: Folate is a key regulator of one-carbon metabolism (OCM), which supports essential physiological processes, including DNA synthesis, repair, methylation, amino acid homeostasis, and redox balance. It is also crucial for brain health throughout life, from neural tube formation during early development to neurotransmitter synthesis, myelination, neuronal development, synaptic plasticity and cognitive function during later stages of life. Disruption of folate-mediated OCM (FOCM) can adversely affect brain health and contribute to neurodegeneration. In this review, we summarize current evidence linking FOCM dysregulation to neurodegenerative diseases, emphasizing disease-specific mechanisms and the therapeutic potential of modulating folate metabolism, as evidenced by experimental and clinical studies. RECENT FINDINGS: Disruption of FOCM can lead to oxidative stress, impaired methylation, excitotoxicity, and neuroinflammation, thereby contributing to neurodegenerative diseases. In Alzheimer’s disease, impaired FOCM promotes amyloid-β accumulation, tau pathology, cognitive decline, and vascular dysfunction, consistent with low folate and elevated homocysteine observed clinically, though supplementation outcomes remain mixed. In Parkinson’s disease, folate deficiency and hyperhomocysteinemia exacerbate motor deficits and dopaminergic neurodegeneration via oxidative stress, mitochondrial dysfunction, and NLRP3-mediated inflammation and combined folate and vitamin B12 supplementation may reduce levodopa-associated risks. Elevated homocysteine and aberrant FOCM have also been reported in Amyotrophic Lateral Sclerosis, Multiple Sclerosis, and Huntington’s disease and are associated with neuroinflammation, demyelination, neuronal loss, and severe disease phenotypes in these conditions. Overall, maintaining optimal folate levels may be a promising strategy to support brain health and reduce the risk of neurodegenerative disorders.

<h4>Background</h4>Aquaculture is the fastest growing farmed food sector and is a key part of global food security. Crustacean aquaculture, while being one of the most profitable sectors, is threatened by pathogenic diseases such as white spot disease (WSD), which results in severe stock losses and threatens animal health and welfare. In this study, we used novel techniques to study the impact of white spot syndrome virus (WSSV), the causative agent of WSD, on Pacific whiteleg shrimp (Litopenaeus vannamei), and uncover additional information that may be used in creating WSSV-resistant shrimp stocks.<h4>Results</h4>We successfully developed a novel nuclei isolation protocol optimized for shrimp tissues to prepare our samples for single nuclei RNA-sequencing following a pathogen challenge comprising 32 adult whiteleg shrimp infected with WSSV either through their feed or by injection. We constructed the first penaeid shrimp lymphoid organ cell atlas to improve our understanding of the characteristics of this immune organ, constructed UMAPs and identified marker genes that define distinct cell clusters to reveal the biological functions of different cell types within this organ.<h4>Conclusion</h4>By comparing gene expression between the control and WSSV-infected samples we uncovered multiple genes of interest that have the potential to be used as targets for CRISPR gene editing for WSSV-resistance in Pacific whiteleg shrimp. These genes have the potential to be valuable assets for future gene editing studies in commercially important penaeid shrimp species.

Retinal ganglion cell (RGC) axons form the optic nerve (ON). Numerous age-related ON diseases, including glaucoma, the second most common cause of worldwide blindness, result from multiple RGC stressors. Nearly all ON astrocytes in the optic nerve head (ONH): the junctional region between the ON and the retina in young-adult rodents expresses the homeodomain only (Hopx) protein. Hopx(+) ONH astrocytes are depleted during aging. ONH primary cultures which include Hopx(+) astrocytes secrete extracellular vesicles (ONH-EVs) which selectively enhance RGC survival and neurite extension in culture, while extracellular vesicles (EVs) secreted from distal ON cultures lacking Hopx(+) astrocytes do not. ONH-EVs also enhance RGC survival in vivo in a rodent model of glaucoma. Combining rat ONH single-cell (scRNA-seq) sequencing with EV proteomic analysis, we identified ONH-Hopx(+) astrocyte secreted factors. We interrogated the online Broad institute scRNA-seq database for rat RGC gene expression in control animals and following rodent ON crush, an RGC stress model, to correlate ONH-astrocyte secreted factors with RGC gene expression changes. Following stress, RGCs upregulate the complementary pathways involving Hopx(+) astrocytic-associated factors, suggesting reciprocal communication. Using a highly selective transgenic Hopx-cre ONH knockdown strategy, we demonstrate that eliminating Hopx(+) astrocytes also results in upregulation of RGC stress responses. Our results implicate age-related loss of young ONH-astrocytes as a crucial factor in the development of age-related optic nerve diseases, and discuss replacing ONH associated factors as a paradigm shift for ON disease treatment.

<h4>Primary objective</h4>To evaluate histopathological renal response in re-biopsy after induction therapy and to correlate with the Target Renal Response (TRR) and secondary objective: to correlate serum and urinary biomarkers with creatinine, 24-hour proteinuria, histological class, activity index and chronicity index.<h4>Methods</h4>Open, longitudinal, multicentre study in lupus nephritis (LN) confirmed through renal biopsy. Two kidney biopsies (T1 and T2) were performed. Laboratory evaluations included urinary sediment examination, renal function, 24-hour proteinuria, serum C3/C4, anti-dsDNA, antinucleosome, anticardiolipin immunoglobulin M/immunoglobulin G, other serum (anti-C1q, monocyte chemoattractant protein 1 (MCP-1), lipocalin-associated neutrophil gelatinase (NGAL) and kidney injury molecule 1 (KIM-1)) and urinary biomarkers (tumour necrosis factor-related weak inducer of apoptosis (TWEAK), anti-interleukin 16, activated leucocyte cell adhesion molecule, MCP-1, NGAL, KIM-1, adiponectin, haemopexin and ceruloplasmin).<h4>Results</h4>24 patients (mean age 32.5±8.2 years) were studied. After induction therapy, 18 patients achieved TRR. Among six patients who did not reach TRR, three failed to meet the 24-hour proteinuria criterion, two to the creatinine criteria and one to both criteria. Histological response was achieved by 14 patients; among these, four patients did not reach the TRR. We found 12 discordances between clinical and histological responses. Eight patients achieved TRR without histological response and four patients achieved histological response without TRR. Serum NGAL and serum KIM-1 showed correlation with urea and with creatinine and higher levels of urinary TWEAK were found in patients who remained with abnormal haematuria at T2. However, no correlation of these new biomarkers was found with activity score or histological class in this study.<h4>Conclusion</h4>Discordance between clinical and histological response was confirmed, more patients achieved clinical response than histological response and, even in patients with histological response, not everyone presented clinical response. In a short interval of time, we observed early reduction in proteinuria and change in histological class after induction therapy. The lack of correlation between biomarkers and baseline histological parameters, which is inconsistent with prior studies, renders the biomarker findings inconclusive.

FAM60A (also known as SINHCAF) is a subunit of the Sin3/HDAC histone deacetylase complex with established roles in chromatin remodeling, yet its broader cellular functions remain largely undefined. Using immunological, biochemical, CRISPR/Cas9, genomic, and proteomic approaches, we mapped the FAM60A interaction network and its functional impact. We reveal that FAM60A binds directly to HDAC1 to recruit Sin3/HDAC, while a dual-domain architecture mediates additional associations with RNA and DNA-binding proteins. CRISPR/Cas9-mediated HDAC1 knockout abolishes the FAM60A-SIN3A interaction, confirming this dependency. Loss of FAM60A triggers widespread transcriptional rewiring, including downregulation of WWC3-a scaffold for LATS1/2 activation. Consequently, YAP1 dephosphorylation and nuclear accumulation shifted cell-cycle dynamics toward G₁ enrichment and conferred resistance to metabolic stress. Restoration of FAM60A or exogenous WWC3 reactivated Hippo "off" signaling, normalized cell-cycle distribution, and reversed stress resistance. These findings establish FAM60A as a pivotal epigenetic tuner linking histone deacetylation to Hippo pathway regulation and nominate the FAM60A-HDAC1-WWC3 axis as a potential therapeutic target to restore growth control in YAP-driven cancers.

Detergent-free immunolabeling has been proven feasible for volumetric correlated light and electron microscopy (vCLEM) with the application of small immunoprobes, providing a reliable method to superimpose essential molecular information of cells onto their high-resolution electron microscopic images. However, the application of this method is restricted by the availability of suitable affinity reagents. Here we introduce CAptVE, a method using slow off-rate modified aptamers for cell fluorescence labeling on ultrastructurally reconstructable electron micrographs. CAptVE provides labeling for a wide range of biomarkers, offering a pathway to integrate molecular analysis into volumetric electron microscopic studies of tissues, including neural circuit investigations via connectomics.

Chemical exposures significantly affect individual and public health, yet there is no unifying framework describing how diverse chemical compounds may interfere with biological processes and contribute to disease risk. Here, we use a network-based approach to construct a comprehensive map linking 9887 exposures through their shared genetic effects. This map can be used to define classes of exposures that affect common biomolecular processes, even when they are chemically distinct. We find that exposures target specific modules in the human interactome of protein-protein interactions, and that exposure harmfulness relates to interactome connectivity. Systematically comparing exposure modules with disease modules suggests that their interactome proximity predicts exposure-disease relationships. We validate these predictions by integrating nationwide disease prevalence data with reported environmental exposures, finding higher disease incidence where exposure and disease modules overlap. Together, our study provides a blueprint for the systematic investigation of the pathobiological impact of chemical exposures ranging from the molecular to the population level.

RNA-binding proteins (RBPs) play critical roles in the regulation of gene expression. Recent studies have begun to detail the RNA recognition mechanisms of diverse RBPs. However, given the array of RBPs studied so far, it is implausible to experimentally profile RBP-binding peaks for hundreds of RBPs in multiple non-model organisms. Here, we introduce MuSIC (Multi-Species RBP-RNA Interactions using Conservation), a deep learning-based framework for predicting cross-species RBP-RNA interactions by leveraging label smoothing and evolutionary conservation of RBPs across 11 phylogenetically diverse species ranging from human to yeast. MuSIC outperforms state-of-the-art computational methods, and achieves highly accurate prediction of RBP-binding peaks across species. The prediction confidence is higher in the metazoan species, partially reflecting differences in RBP conservation patterns. Finally, the effects of homologous genetic variants on RBP binding can be computationally quantified across species, followed by experimental validations. The target transcripts with disrupted binding events are enriched in the ubiquitination-associated pathways. To summarize, MuSIC provides a useful computational framework for predicting RBP-RNA interactions cross-species and quantifying the effects of genetic variants on RBP binding, offering insights into the RBP-mediated regulatory mechanisms implicated in human diseases.

Hemodialysis (HD) patients suffer from chronic inflammation which is a serious complication and contributes significantly to morbidity and mortality. Dialysis technology has advanced over many years but despite advances, its underlying molecular mechanism remains poorly understood. We have designed an exploratory study and investigated how hemodialysis influences epigenetic changes by understanding the DNA methylation patterns which are associated with inflammation. By using the same polysulfone (PS) membrane dialyzer, we have collected the blood samples from the patients before and after dialysis. For DNA methylation analysis, we have extracted genomic DNA using the Monarch® Genomic DNA Purification Kit. Bisulfite conversion was done using the QIAGEN Epitect Bisulfite Kit. Genome Studio, which is a computational software, was used to identify the DNA methylation profiles. These profiles revealed significant epigenetic changes across the hemodialysis patients’ samples. From these DNA methylation profiles, we identified the differential methylated regions (DMRs) and focused on the genes that are associated with these DMRs with the change greater than 80% to confirm robustness. We identified the chromosomal mapping of DMR-associated genes on CpG islands and shore localization, promotor or enhancer regions, to evaluate the functional impact of methylation changes. One of the bioinformatics tools, named Discovery Annotation, Visualization, Integration Database (DAVID) was used to perform functional enrichment analysis to identify the affected biological pathways involved in neuronal signaling, immune regulation, inflammation, and metabolism. Protein- Protein Interaction network was constructed using another bioinformatics tool, named String Database, and It analysis was performed using a bioinformatics desktop-based tool called Cytoscape, which helped in identifying critical hub genes including TWIST1, SHANK3, FGF20, MEF2C, RUNX2, CAV1, CDKN2A, WNT3A, and CXCL12, which play crucial roles in immune system compromise and inflammatory regulation. A total of 93 genes, across the samples of HD patients, showed significant methylation alterations. Of which, 41 genes showed changes exceeding 100% and mostly mapped on Promoter/enhancer regions at CpG islands and shores. In HD patients, these functional consequences suggest changes in their role in impaired vascular function, cognitive decline and immune dysregulation. Fibrinogen (FB) behaviour during dialysis, particularly its adsorption onto the dialysis membranes, was understood using Synchrotron Imaging at the Canadian Light Source (CLS) which provided us a high-resolution visualization. To confirm significant FB fouling between initial and intermediate membrane layers, these targeted regions were identified using raw CT scan images and their magnified views. Similarly, we identified that the level of inflammatory biomarkers, which were elevated in post-dialysis samples of HD patients including vWF, CRP, Serpin C1, Properdin, and PF4 indicating an amplified inflammatory response. Particularly, the critical hub genes that are associated with vascular regulation including CAV1, TWIST1, CDKN2A, and CXCL1, were reported as hypermethylated after dialysis, meaning their expression may reduce which affects their contribution in endothelial dysfunction. These findings suggest a self-perpetuating inflammatory loop, where FB adsorption initiates molecular and immunological responses that reinforce epigenetic dysregulation. One of the important limiting factors is sample size, which influenced the reliability of findings across the molecular and clinical levels, and this underlines that further investigation is needed. This study helped us designed the foundational insights to target the anti-inflammatory strategies that may lead us to the development of more hemocompatible dialysis membranes to reduce long-term complications in HD patients.

We investigated the pathogenicity of a homozygous intronic variant in CDK5RAP3, a key UFMylation adapter, in three individuals from two unrelated families with a lethal neurodevelopmental disorder. CDK5RAP3 variants have not been linked to human disorders to date; however, murine Cdk5rap3 knockout is embryonic lethal and variants in five other UFMylation components cause severe neurodevelopmental conditions. A segregating homozygous variant, chr17(GRCh38):g.47974691G > A, CDK5RAP3 NM_176096.3:c.334 + 243G > A, was identified by trio whole-genome and proband RNA sequencing in Family A and by trio whole-exome sequencing data reanalysis in Family B. Variant pathogenicity investigations included RT-PCR, Western blot, co-immunoprecipitation and (phospho)proteomics to assess transcript, protein and UFMylation complex effects. Antisense oligonucleotide-mediated rescue of CDK5RAP3 expression combined with proteomics and phosphoproteomics defined the mechanistic impact of CDK5RAP3 deficiency and rescue in amniocytes from an affected individual. All three affected individuals showed foetal growth restriction, foetal akinesia, pontocerebellar hypoplasia, arthrogryposis and hepatic pathology. CDK5RAP3 c.334 + 243G > A activates a cryptic donor splice-site causing pseudoexon/intron inclusion triggering nonsense-mediated decay and deficiency of full-length CDK5RAP3 (NP_788276.1), while potentially allowing retained expression of C-terminal alternative isoforms. Co-immunoprecipitation revealed only full-length CDK5RAP3 binds UFL1, whereas C-terminal isoforms cannot. Primary amniocytes showed CDK5RAP3 deficiency was associated with impaired UFMylation of known substrates, RPL26 and UFBP1. Proteomic and phosphoproteomic analyses revealed dysregulation of extracellular matrix organisation, cell adhesion, mitotic/genome stability pathways, cytoskeletal networks and neuronal guidance, which were reversed by restoration of canonical CDK5RAP3 expression via splice-correcting antisense oligonucleotides. Phosphoproteomic data implicate CDK5RAP3 as an upstream regulator of UFL1 S462 phosphorylation, known to be regulated by Ataxia-telangiectasia mutated (ATM) signalling. Our findings provide strong evidence linking deficiency of full-length CDK5RAP3 to severe neurodevelopmental, liver and muscle dysfunction. This study further highlights the therapeutic potential of ASO-based deep-intronic splicing defect correction.

Considering that the circulating proteome represents a primary source of candidate biomarkers and therapeutic targets, we conducted a large-scale Mendelian randomization (MR) study to identify plasma proteins potentially involved in the pathogenesis and treatment of common urological cancers (UCs), including bladder cancer (BC), prostate cancer (PC), renal cell carcinoma (RCC), and testicular cancer (TC). Cis-protein quantitative trait loci (cis-pQTLs) were derived from two large-scale genome-wide association studies (GWASs) of plasma proteomes. GWAS for UCs were obtained from FinnGen and pan-UKBB meta-analysis, FinnGen and UK Biobank. Colocalization analysis and summary data-based MR (SMR) were performed to evaluate the robustness of the associations. Further evaluations involved Bulk RNA-seq differential expression and single cell-type expression analysis, protein–protein interaction, and druggability evaluation. For BC, we identified four protein markers: one associated with increased risk (PSCA) and three with decreased risk (GSTM1, GSTM3, GSTM4), which are mainly expressed in pericyte, urothelial, and NK cells in bladder tumors. Regarding PC, we found 15 protein markers: seven linked to increased risk (AGER, ALAD, CHMP2B, PEX14, ZG16B, PPP1R14A, SERPINA3) and eight to decreased risk (BTN2A1, CEACAM21, DNAJB9, MSMB, PYGL, HLA-E, SOD2, TOR1AIP1), enriched in epithelial cells, monocytes/macrophages in prostate tumors. The strongest evidence from MR and colocalization analyses supported GSTM4 (BC), SOD2 and CHMP2B (PC) as causal markers. Notably, seven identified proteins have been previously targeted by drugs for other cancers and immune disorders, indicating their potential therapeutic relevance in UCs. This study highlights several proteins with predictive value for BC and PC risk and supports their utility in biomarker discovery and drug development.

The sustainable valorization of poultry by-products into functional materials offers an eco-friendly approach for wastewater remediation and waste management. In this work, chicken bone waste (CBW) was converted into biochar (CBWB) via controlled pyrolysis for 2 h at 750 °C and examined as a low-cost adsorbent for the uptake of methyl red (MR) dye from aqueous solutions. The physicochemical characteristics of CBW and CBWB were evaluated using Fourier transform infrared spectroscopy (FTIR), Brunauer-Emmett-Teller (BET) surface area analysis, scanning electron microscopy (SEM), X-ray diffraction (XRD), and thermogravimetric analysis (TGA). Pyrolysis greatly improved porosity and surface area, increasing the BET surface area from 37.3 m²/g for CBW to 76.4 m²/g for CBWB. Batch sorption studies examined the impacts of solution pH, initial dye concentration, adsorbent dosage, contact time, and temperature on MR elimination. Adsorption kinetics followed the pseudo-first-order model for CBW and the pseudo-second-order model for CBWB, signifying stronger surface interactions after pyrolysis. Equilibrium data were best explained by the Langmuir isotherm for CBW and the Freundlich isotherm for CBWB, with maximum adsorption capacities of 84.84 and 138.58 mg/g, respectively. Thermodynamic assessment confirmed that the adsorption process was spontaneous and endothermic. Regeneration studies showed that CBWB reserved approximately 69% of its initial adsorption efficiency after five cycles of operation. These findings revealed that chicken bone waste-derived biochar is reusable, efficient, and sustainable adsorbent for dye-contaminated wastewater treatment.

The healthy donor effect (HDE) refers to the lower mortality observed among blood donors compared to the general population. While HDE arises due to healthier individuals being more likely to donate, the extent to which it is influenced by genetic differences remains unclear. To elucidate the genetic basis of HDE, we conducted a genome-wide association study (GWAS) involving 53,688 active blood donors with extensive donation histories and 228,060 controls from biobank cohorts within the FinnGen project. We identified 46 fine-mapped genome-wide significant loci associated with several health-related endpoints, plasma protein levels and laboratory measurements. Genetic correlation analyses across FinnGen endpoints revealed that blood donors are genetically protected against several diseases beyond those affecting donation eligibility. Using the correlated endpoints as exposures in multivariable Mendelian randomization (MVMR) to inform priors for Bayesian GWAS, we found that 25 of the fine-mapped loci exert a direct effect on blood donorship (BD) rather than acting through disease mediation, suggesting a genetic contribution to maintaining a health state conducive to long-term donation. We also performed MVMR analyses of laboratory traits. The results indicated that normal liver function, blood glucose, and low inflammation independently increase the likelihood of becoming a blood donor, while iron levels showed no causal relationship. Functional enrichments among the proteins regulated by the 46 fine-mapped variants included mainly red cell antigen-related cell adhesion processes. In conclusion, our findings demonstrate that HDE is partly explained by genetic factors, involving both direct health-promoting effects and indirect eligibility selection.

BACKGROUND: Drought intensity and frequency are increasing under global warming in the boreal forests, and breeding for drought resistance will facilitate adaptation of new planting material to changing climate conditions. We used a tree-ring dataset of 559 individuals to study Scots pine genetic variation and the efficiency of genomic selection of drought-response traits (drought resistance, recovery and resilience), for the first time. From genotyping-by-sequencing (GBS), 31,101 SNPs were generated and used for the study. RESULTS: Significant genetic variation was detected for drought-response and other growth, wood-anatomy and wood density traits. Heritability estimates for wood-anatomical traits were higher than those for drought-response and growth traits. Genetic correlations between drought-response and wood-anatomical traits were generally high but mostly nonsignificant. In contrast, drought resistance and recovery showed positive and significant correlations with basal area increment and height. We found that the predictive ability and accuracy for drought-response traits were lower than those for wood-anatomical traits, and were comparable between GBLUP and ABLUP. Greater genetic gain per year can be achieved through genomic selection relative to pedigree-based selection if the generation interval is reduced. CONCLUSIONS: The positive genetic correlation between drought-response and growth traits will enable simultaneous selection for improved growth and increased drought resistant trees in Scots pine breeding through either pedigreed-based and genomic selection.

<h4>Background</h4>Phosphatidylinositol transfer protein-1 (pitp-1) is involved in the phosphoinositide (PIP) cycle. The role of pitp-1 in promoting healthy longevity remains unknown. Our previous work showed that the PIP cycle-related genes diacylglycerol lipase-1 (dagl-1) and diacylglycerol kinase-5 (dgk-5) regulate lifespan, as overexpression of dagl-1 or knockdown of dgk-5 prolongs lifespan and enhances oxidative stress resistance through target of rapamycin (TOR) signaling. As pitp-1 is a key component of this pathway, we investigated its role in lifespan regulation and the underlying mechanisms, aiming to clarify whether it represents a critical regulator of healthy longevity and how it coordinates conserved signaling pathways to regulate aging.<h4>Methods</h4>Caenorhabditis elegans (C. elegans) mutants, RNAi-mediated knockdown, and transgenic overexpression were applied to assess lifespan, motility, and stress resistance. Temporal and tissue-specific RNAi were applied to identify critical time window and tissue for pitp-1-mediated lifespan regulation. TOR signaling was measured by phosphorylated S6 kinase (p-S6K) and puromycin incorporation, and transcriptomic analysis identified affected pathways.<h4>Results</h4>pitp-1 negatively regulated lifespan and healthspan in C. elegans. Genetic deletion or RNAi-mediated knockdown of pitp-1 extended lifespan, attenuated age-related motility decline, and increased oxidative stress resistance. Temporal and spatial analyses revealed that suppression of pitp-1 in neurons during early adulthood was sufficient to promote healthy longevity. Mechanistically, these beneficial effects upon pitp-1 reduction were associated with suppression of TOR signaling. Conversely, pitp-1 overexpression shortened lifespan and impaired healthspan via TOR activation. Moreover, pitp-1 was transcriptionally repressed by DAF-16 downstream of insulin/IGF-1 signaling (IIS), and contributed to IIS-mediated longevity regulation. Furthermore, pitp-1 reduction also improved organismal proteostasis, as evidenced by decreased polyglutamine (polyQ) aggregation and enhanced motility in a neuronal proteotoxicity model.<h4>Conclusions</h4>These findings identify pitp-1 as a novel regulator of healthy aging, suggesting a role in coordinating IIS and TOR signaling and providing new insights into conserved mechanisms of longevity regulation.

Gastric cancer (GC) remains a major cause of cancer incidence and mortality worldwide. Alternative splicing (AS), a post-transcriptional mechanism that expands transcript diversity from a single pre-mRNA, plays critical regulatory roles in GC. This review summarizes the current knowledge of AS in GC. Accumulating evidence shows that aberrant AS contributes to the oncogenesis, progression, invasion, metastasis and drug resistance of GC. In addition, AS-related biomarkers like RBM4 and prognostic models for subtype prediction have been reported. However, most existing studies have focused on basic research, while patient-level validation remains limited and standardized detection and analytical workflows are still lacking. Future work should prioritize prospective cohorts and standardized pipelines.

BACKGROUND: Ovarian cancer (OC) is the most deadly gynecological malignancy, mainly due to insufficient studies on its mechanisms and lack of effective therapies. Alternative splicing (AS) dysregulation has deepened understanding of cancer pathogenesis. As a constitutive molecular consisting spliceosome, the pathogenesis and therapeutic potential of SF3a1 in solid tumors are rarely studied. METHODS: The expressions and prognostic values of SF3a1 were accessed using bioinformatics. The functions of SF3a1 were evaluated using gain- and loss-of-function strategies. Whole transcriptome sequencing and mass spectrometry analysis were conducted to capture signaling axis of SF3a1. Semi-qPCR, qRT–PCR, WB, IHC and rescue experiments were exploited to validate the sequencing results. Targeted antisense oligonucleotide (ASO) was designed and its treatment efficacy was verified. Virtual screening was used to identify small molecule inhibitor of SF3a1, whose therapeutic potential on OC was evaluated via in vitro and in vivo assays. RESULTS: This study found SF3a1 played an indispensable role in OC pathogenesis. SF3a1 knockdown impaired malignant behaviors and reprogrammed splicing landscape of OC cells. Skipped exon (SE) event of MDM4-exon6 (exon7 in mouse) increased, leading to formation of MDM4-short (MDM4-s) isoform, which was subsequently degraded. Total MDM4 level was therefore downregulated, leading to P53 pathway activation and OC inhibition. Additionally, ASO targeting MDM4 pre-mRNA sequence bound by SF3a1 mimicked the alternative regulation caused by SF3a1 knockdown, and showed significant inhibitory effect on OC proliferation. Rifaximin, identified as an inhibitor targeting SF3a1, effectively killed OC cells and enhanced cisplatin’s efficacy in combination therapy. Moreover, CDK6 was found to phosphorylate SF3a1 at S359, which decreased its protein half-life and influenced downstream MDM4 splicing. CONCLUSIONS: Our findings uncover the oncogenic impact of SF3a1 in OC and highlight the prospects to target it by rifaximin and downstream MDM4 AS by ASO, synergistically improving the therapeutic effectiveness of cisplatin in OC.

Intestinal organoids (IOs) are versatile, physiologically relevant models of the human gut. In cystic fibrosis research, IOs help evaluate CFTR correctors, potentiators, and translational readthrough compounds. Here, we present a protocol for generating induced pluripotent stem cell (iPSC)-derived IOs with high phenotypic similarity to primary material. We describe steps for establishing iPSC culture, generating intestinal progenitors via directed differentiation, and maturing these intestinal progenitors using three-dimensional culture. We then detail procedures for establishing intestinal organoid cultures from these progenitors using dissection-based techniques.

Bladder cancer (BCa) is a histologically and molecularly heterogeneous disease and is one of the leading causes of cancer death globally. The main risk factors are sex (with incidence 3 to 4 times higher in men), tobacco usage, occupational exposure to carcinogens, and persistent infections, such as those caused by Schistosoma haematobium. Urine and the bladder were recently confirmed to be non-sterile, prompting investigations into the urinary and intratumoral microbiomes and their roles in tumor stage, prognosis, and therapy response. In this context, the role of the urinary and intratumoral microbiome in bladder carcinoma is among the most promising areas in translational uro-oncology. Recent evidence demonstrates the presence and diversity of microbial communities in both urine and bladder cancer tissue, with patterns associated with tumor stage and prognosis. Chronic inflammation, genotoxin production, altered carcinogen metabolism, and modulation of the immune microenvironment are biological processes that provide a rationale for the functional role of these microorganisms in the bladder. Furthermore, microbial profiles have been correlated with responses to intravesical therapies (such as BCG - Bacillus Calmette-Guérin) and, potentially, with systemic immunotherapies. The microbiome can help identify predictors of treatment response and potential adjuvant interventions, and offers a non-invasive, translational pathway for diagnosis and surveillance. This review summarizes current evidence on the microbiome in bladder cancer patients and its prognostic and therapeutic potential.

Tandem repeats (TRs) are dynamic genomic elements characterized by exceptionally high mutation rates, reversibility, and the capacity to generate graduated phenotypic effects. These properties enable TRs to modulate gene regulation at multiple levels-from transcription and chromatin organization to splicing and protein function. In the human lineage, TR expansions have been recurrently associated with brain evolution, with human-specific repeats enriched near neurodevelopmental genes and linked to neural-specific regulatory elements. However, this same mutational plasticity that facilitated cognitive innovation also introduced genomic instability, rendering the nervous system particularly vulnerable to repeat-associated dysfunction. The brain's unique cellular and molecular properties create an environment particularly permissive to repeat-mediated pathology. Recent studies have revealed diverse pathogenic mechanisms beyond classical models, highlighting the multifaceted ways in which repeat instability can disrupt neural function. Understanding TRs as integral components of neural regulatory architecture, rather than isolated pathogenic loci, provides a framework for interpreting the genetic basis of neurological and psychiatric disorders.

Neurodegenerative diseases (NDs) are characterized by progressive neuronal loss and proteostatic failure, driven by impaired clearance of misfolded proteins via the ubiquitin-proteasome system (UPS) and autophagy. In UPS, E3 ubiquitin ligases are crucial for regulating protein ubiquitination and degradation. Mutations in E3 ligases, along with dysfunctions of specific ligases such as Parkin, the C-terminus of HSC70-interacting protein (CHIP), and tripartite motif-containing proteins, have been identified as key factors in the buildup of amyloid-β, α-synuclein, tau, trans-active response DNA-binding protein 43, and mutant huntingtin. These accumulations are associated with NDs like Parkinson's disease, Alzheimer's disease, Huntington's disease, and amyotrophic lateral sclerosis. Therapeutic strategies targeting E3 ligases, particularly proteolysis-targeting chimeras (PROTACs), are being developed for ND treatment and are currently in clinical trials. These approaches aim to enhance E3 ligase activity and promote selective protein degradation. Here, we examine how individual E3 ligases influence cell-fate decisions in NDs, showing that their substrate selection determines whether neurons survive or die. Building on this knowledge, we present an innovative therapeutic pipeline that includes ligase activators, PROTAC degraders, and miRNA switches, which are molecules designed to transition from research to clinical application.

<h4>Background</h4>Regional citrate anticoagulation (RCA) is suggested as the preferred anticoagulation strategy during continuous renal replacement therapy (CRRT), as it prolongs circuit lifespan while minimizing bleeding complications. However, evidence on its use in CRRT circuits during extracorporeal membrane oxygenation (ECMO) is limited. In patients receiving ECMO, systemic anticoagulation with unfractionated heparin (UFH) is routinely administered to maintain circuit patency and is often relied upon to anticoagulate the CRRT circuit, limiting the use of regional citrate anticoagulation. The aim of this study is to evaluate whether adding regional citrate anticoagulation (RCA) to systemic unfractionated heparin (UFH) reduces CRRT circuits clotting in patients undergoing venovenous extracorporeal membrane oxygenation (VV ECMO).<h4>Results</h4>Patients were randomized to receive alternating CRRT circuits anticoagulated with either systemic UFH alone or regional citrate anticoagulation added to systemic UFH (RCA + UFH), according to a predefined crossover sequence. Each circuit was maintained for up to 72 h or until clotting occurred. Coagulation parameters, CRRT pressures, and thromboelastography (TEG) data were collected.A total of 66 CRRT circuits from 18 patients were analyzed (33 RCA + UFH; 33 UFH). No clotting events occurred in the RCA + UFH circuits, whereas 6 events were observed with UFH alone (0% vs 19%; <i>p</i> < 0.001). Cox proportional hazards analysis showed significantly longer circuit survival with RCA + UFH compared to UFH alone (p = 0.029). Platelet counts increased during RCA + UFH but declined during UFH alone (median change +6 vs -7 ×10³/μL; <i>p</i> < 0.001), with a significantly more favorable overall trend under RCA + UFH (effect estimate +13 × 10³/μL, 95% CI 8-19). D-dimer levels increased significantly during UFH alone, whereas a lower increase was observed with RCA + UFH (effect estimate -782 μg/L, 95% CI -1525 to -39).Thromboelastography performed at the circuit level showed significantly prolonged R-times with RCA + UFH compared with UFH alone (median R-time 90 <i>vs.</i> 30 min; <i>p</i> < 0.001). No clinically relevant RCA-related metabolic complications were observed, including no episodes of severe hypernatremia, metabolic alkalosis, or citrate accumulation.<h4>Conclusions</h4>In patients undergoing VV ECMO, adding regional citrate anticoagulation to systemic unfractionated heparin reduced CRRT circuit clotting, prevented platelet consumption. This technique was feasible, safe, and may improve CRRT efficiency in this high-risk population.<h4>Clinical trial</h4>ClinicalTrials.gov Identifier NCT05148026.

Acute myocardial infarction (AMI), a life-threatening event caused by cardiomyocyte death due to oxygen deprivation, drives cardiac dysfunction through ferroptosis-an iron-dependent cell death mechanism involving lipid peroxidation. By delivering multifunctional cargoes with low immunogenicity, extracellular vesicles (EVs) hold the therapeutic potential to inhibit cardiomyocyte ferroptosis through the regulation of iron metabolism and the mitigation of oxidative damage. Their dual role as targeted drug carriers and natural imaging probes enhances precision in AMI management. EVs enable the non-invasive tracking of biodistribution and therapeutic responses in real time when integrated with molecular imaging technology, offering insights into cardiac repair mechanisms. This synergy between EV-based therapy and advanced imaging presents a novel strategy for AMI diagnosis and targeted intervention.

<b>Background</b>: Tumor invasion is the central barrier to effective immunotherapy in lung adenocarcinoma (LUAD) and glioblastoma. Cell adhesion signaling critically shapes tumor-microenvironment interactions, yet the upstream regulators coordinating these invasive programs at single-cell resolution remain incompletely understood. Methyltransferase Like protein 7B (METTL7B) has recently emerged as a candidate oncogenic regulator, but its lineage-specific functions and the potential downstream effectors are unclear. <b>Methods</b>: We integrated publicly available single-cell RNA sequencing datasets from LUAD and glioblastoma with The Cancer Genome Atlas (TCGA) transcriptomic analyses to resolve <i>METTL7B</i>-associated malignant cell states and microenvironmental interactions. Functional enrichment analyses identified invasion- and focal adhesion pathways linked to <i>METTL7B</i> expression. Gain- and loss-of-function experiments were conducted in LUAD and glioblastoma cell lines to validate downstream cell adhesion effectors. Spatial expression patterns were examined using immunofluorescence, and transwell assays were used to assess migratory and invasive phenotypes. <b>Results</b>: Single-cell analyses revealed that <i>METTL7B</i> was selectively enriched in malignant epithelial cells in LUAD and glioblastoma and defined a transcriptional program characterized by cell adhesion signaling. Integrin Alpha 3 (<i>ITGA3</i>) emerged as a conserved downstream effector of <i>METTL7B</i>, with progressive upregulation from minimally invasive to invasive LUAD and glioblastoma. Functional perturbation confirmed that METTL7B enhances tumor cell migration and invasion through integrin-associated pathways. <b>Conclusions</b>: METTL7B acts as a potential lineage-enriched regulator of invasive tumor states by activating cell adhesion signaling in LUAD and glioblastoma. These findings position METTL7B as a putative prognostic factor for strategies aimed at limiting invasion in lung cancer and glioblastoma.

Pancreatic ductal adenocarcinoma (PDAC) remains among the deadliest cancers, with a 5-year survival rate of 13% and broad resistance to therapy. It is driven by severe tumor hypoxia from desmoplasia, aberrant vasculature, and high interstitial pressure. Hypoxia stabilizes hypoxia-inducible factors (HIFs), reshaping the tumor microenvironment (TME) into a nutrient-poor, acidic milieu that fosters immune exclusion and suppression. While immune checkpoint inhibitors (ICIs) have revolutionized treatment, PDAC responses have been negligible. As hypoxia centrally drives PDAC's ICI-refractory TME, targeted alleviation could offer synergy with ICIs; however, no such combination is being applied in the clinic. One impediment could be the one-size-fits-all approach when investigating hypoxia-modifying therapy. Indeed, using hypoxia gene signatures, we and others have shown that PDAC tumors are not equally hypoxic, with patients having more hypoxic tumors experiencing worse survival and immunosuppressed TME. This review dissects hypoxia's mechanistic role in PDAC immune evasion and gives an update on the therapeutic advances that directly or indirectly target hypoxia, such as the inhibition of HIFs, hypoxia-activated prodrugs, and vascular and oxygen delivery approaches, with emphasis on their potential to enhance responses to ICIs. It further evaluates the need for hypoxia biomarkers and proposes gene signatures as detection tools to enable precision hypoxia modulation, potentially converting immune-cold PDAC into an ICI-responsive disease.

<h4>Background</h4>Esophageal squamous cell carcinoma (ESCC) exhibits heterogeneous responses to neoadjuvant chemoradiotherapy (nCRT) and understanding the molecular features of ESCC could improve patient management. This study aimed to investigate the differences in somatic mutations and gene expression profiles among esophageal cancer patients exhibiting varying responses to nCRT.<h4>Methods</h4>A prospective cohort of 30 patients with ESCC undergoing nCRT was enrolled. Tumor and blood samples were collected prior to the initiation of nCRT. Following transcriptome profiling and identification of differentially expressed genes, the singscore algorithm was applied to construct a response prediction model, and its predictive performance was assessed.<h4>Results</h4>The singscore method generated a unique score for each case. The score range of patients with Tumor Regression Score (TRS) 0-1 was -0.59 to 1. The area under the receiver operating characteristic (ROC) curve was 0.888 [95% confidence interval (CI): 0.773-1], with singscore =0 identified as the optimal cutoff value of the score (sensitivity: 80.0%, specificity: 78.9%, positive predictive value: 88.2%, negative predictive value: 69.2%). Singscore score was positively correlated with better overall survival (hazard ratio: 0.14, 95% CI: 0-0.374, P=0.01).<h4>Conclusions</h4>In this exploratory study, the singscore model demonstrated promising predictive performance for nCRT response in ESCC, with singscore =0 as the optimal cutoff in our cohort. However, these findings require validation in larger, multi-center studies before clinical application can be considered.

Sox9, a pivotal transcription factor belonging to the Sox family, orchestrates critical processes throughout embryonic development, maintenance and differentiation, and exerts a profound influence on organogenesis. Its regulatory versatility stems from precise binding to defined DNA regions, often in collaboration with tissue-specific partners. The dysregulation of Sox9 during chondrogenesis leads to a skeletal malformation termed campomelic dysplasia and has emerged as a significant factor in various other human diseases, including cancer. A point mutation at position 76 (alanine to glutamic acid, A76E) of Sox9 is recognized as one of the causes of campomelic dysplasia. We have used a combination of biophysical, structural and computational techniques to characterize the Sox9 A76E mutant and compare it with the wild-type (WT) Sox9. WT and A76E Sox9 assemble as homodimers, but form predominantly monomeric complexes in the presence of Sox-specific DNA. A CD analysis shows that the A76E mutant preserves the folding as well as the overall secondary structure of Sox9. Both A76E and WT Sox9 behave similarly in the presence of Sox-specific DNA. Perturbation, with increased temperature, displays a lower melting point for A76E, relative to WT Sox9, indicating decreased stability that may arise due to the long and charged side chain of glutamic acid compared to the small hydrophobic alanine, making unfavorable intra-molecular interactions. The destabilizing effect of the A76E mutant may disturb the formation of a stable higher-order complex that is a prerequisite for normal gene expression.

Idiopathic inflammatory myopathies constitute a group of immune-mediated disorders primarily affecting skeletal muscle, but they may also lead to significant involvement of internal organs. These conditions are highly heterogeneous, encompassing diverse clinical manifestations and multiple underlying pathophysiological mechanisms. A unifying feature across this disease spectrum is an autoimmune response characterized by the production of highly specific autoantibodies, which are detected in the majority of patients. Genetic studies have identified the principal susceptibility background as the 8.1 ancestral haplotype within the HLA region on chromosome 6. However, genetic predisposition extends beyond HLA loci and includes numerous genes encoding key molecules involved in cytokine production, the regulation of immune signaling pathways, and metabolic processes. In this paper, we review the currently identified genetic loci associated with inflammatory myopathies, with particular emphasis on the HLA system, as well as non-HLA genes and newly identified candidates.

<h4>Background</h4>Cancer remains a leading cause of death worldwide, constrained by limitations of current therapies, such as systemic toxicity, narrow therapeutic windows, and acquired drug resistance, highlighting the urgent need for novel agents. This study aimed to evaluate the antitumor efficacy of parimifasor and elucidate its molecular mechanism through transcriptomic profiling.<h4>Methods</h4>This study evaluated the efficacy of parimifasor <i>in vitro</i> and <i>in vivo</i>. Transcriptomic profiling [RNA sequencing (RNA-seq)] of treated cells was performed to identify differentially expressed genes (DEGs), followed by functional enrichment, validation in The Cancer Genome Atlas (TCGA) database, and quantitative polymerase chain reaction (qPCR) confirmation.<h4>Results</h4>This study presents the first mechanistic exploration of the small-molecule immunomodulator parimifasor (CAS: 1796641-10-5) as a broad-spectrum antitumor agent. <i>In vitro</i> cytotoxicity assays demonstrated potent suppression of 12 human tumor cell lines, with 50% inhibitory concentration (IC₅₀) values <1 μM for nine cell lines. In Bagg Albino mouse (BALB)/c nude xenograft models, intraperitoneal administration of parimifasor (2 and 4 mg/kg) significantly inhibited tumor growth in esophageal squamous carcinoma (KYSE140) and glioblastoma (LN229) in a dose-dependent manner (P<0.05). RNA sequencing of parimifasor-treated cells revealed 81 common DEGs enriched in the RIG-I-like, nucleotide-binding oligomerization domain (NOD)-like, and Toll-like receptor signaling pathways. Integration with TCGA data identified 10 oncogenic genes (<i>PARP10</i>, <i>OAS1</i>, <i>OAS3</i>, <i>PRR4</i>, <i>ZNFX1</i>, <i>HELZ2</i>, <i>SP110</i>, <i>CSRNP1</i>, <i>DDX3L</i>, and <i>PARP14</i>) dysregulated across 20+ cancer types; qPCR confirmed their consistent downregulation posttreatment in both cell lines (P<0.05).<h4>Conclusions</h4>This study establishes parimifasor's antitumor efficacy via the suppression of immune pathway-associated oncogenes, supporting its repurposing as a cancer therapeutic agent and providing biomarker candidates for clinical development.

<title>Abstract</title> <p>Anti-IgLON5 disease is a rare neurological disorder at the interface of autoimmunity and neurodegeneration. It is characterized by autoantibodies against the neuronal adhesion molecule IgLON5 and is associated with profound brain dysfunction and tau pathology. Despite its severe clinical manifestations, the molecular basis of antibody recognition and its contribution to disease pathogenesis remain unclear. Here, we profile the B cell receptor repertoire of a patient with anti-IgLON5 disease and identify a highly polyclonal response lacking dominant clonal expansion. We isolate a human monoclonal IgG4 antibody that binds IgLON5 with high affinity and determine its structure in complex with IgLON5 by cryo–electron microscopy. Biochemical and structural analyses show that antibody binding preserves IgLON5 adhesion interfaces while promoting higher-order clustering of IgLON5 dimers. These findings provide mechanistic insight into autoantibody recognition of neuronal surface proteins and establish a framework for understanding antibody-mediated neurodegeneration in anti-IgLON5 disease.</p>

Post-stroke depressive symptoms (PSDS) are a frequent and disabling consequence of stroke. While lesion-network studies implicate disruption of large-scale affective circuits in PSDS, the neurobiological factors determining why certain network disruptions confer vulnerability to PSDS remain insufficiently understood. We analyzed data from two independent stroke cohorts (total n = 435). Acute lesion masks were embedded within normative structural connectomes, weighted by positron-emission tomography-derived maps of 19 neurotransmitter receptors and transporters, to quantify neurotransmitter (NT)-informed network damage. Partial least squares regression with variable importance measures was used to identify NT-specific damage scores that were informative for PSDS, as quantified by the Hospital Anxiety and Depression Scale at follow-up. Informative NT-systems were subsequently evaluated in multivariable logistic regression models adjusted for age, sex, lesion volume, and neurological deficit. Across cohorts, multivariate analyses converged on a neurochemical signature involving serotonergic, cholinergic, dopaminergic, and GABAergic networks. Damage to networks related to the serotonin transporter (5-HTT) and the vesicular acetylcholine transporter (VAChT) was independently associated with increased odds of PSDS in covariate-adjusted models and improved model fit beyond clinical and lesion-based predictors. In contrast, associations with other NT systems, including dopaminergic networks, were not consistently implicated across cohorts. These findings identify the serotonergic and cholinergic network architecture as a key neurochemical substrate that modulates vulnerability to PSDS. By integrating structural disconnection mapping with NT-informed connectomics, this study provides a mechanistic framework that links stroke-induced network disruption to PSDS and highlights serotonergic and cholinergic systems as central pathways for hypothesis-driven risk stratification and future multimodal investigations.

BACKGROUND: TMEM132A has recently been identified as a potential contributor to tumorigenesis. Nevertheless, a thorough examination of TMEM132A in human tumors across various cancer types has not been conducted to date. METHODS: The expression of TMEM132A was analyzed in 33 different types of human tumors by utilizing data from the TCGA and GTEx databases. RESULTS: TMEM132A was found to be significantly increased in 27 different types of cancer. High TMEM132A expression was linked to worse overall survival in KIRC, MESO, SARC and UVM, as well as unfavorable disease-free survival in KIRC and PRAD. The S530 locus of TMEM132A had lower phosphorylation in GBM but higher in HNSC, PAAD, LUAD and LUSC tumors. The S914 locus had higher phosphorylation in HNSC, KIRC, PAAD and LUSC tumors. TMEM132A genetic changes were thoroughly studied in different types of tumors using TCGA data. SKCM had the highest TMEM132A alteration frequency at 6.53%, predominantly characterized by “Mutation.” TMEM132A was significantly correlated with m1A, m5C and m6A genes in STAD, UCEC, GBM and DLBC tumors. TMEM132A was also associated with CAFs in various TCGA tumors. TMEM132A was positively correlated with immune genes in most cancers, but negatively correlated in CHOL and LIHC. We found a strong correlation between TMEM132A and TMB, MSI and HRD in various types of cancer. TMEM132A was positively correlated with APBA2, B4GALNT4, CDK5, CLSTN1 and PRPF19 genes. The heatmap confirms this correlation across different tumors. An analysis of the overlap between these groups identified ENOPH1 as a shared member. Finaly, GO enrichment and KEGG pathway were used to understand the functional role of TMEM132A in tumor development. GO enrichment highlighted terms like “endomembrane system” and “nervous system development” as important for TMEM132A’s impact. KEGG pathway analysis identified pathways such as “Axon guidance” and “Wnt signaling pathway” as relevant to TMEM132A’s influence on tumor pathogenesis. CONCLUSION: This is the first pan-cancer study to deeply understand the role of TMEM132A in tumor development across various types of cancers.

BACKGROUND: The efficacy of current bone-targeting agents, notably bisphosphonates, in the treatment of bone metastases remains limited by their systemic toxicity and excessively long half-life. This study aims to develop bone-targeting agents inspired by osteotropic peptides involved in the bone mineralization process. These agents are intended to provide an innovative alternative to bisphosphonates for precision bone targeting. RESULTS: Osteotropic peptides and phosphopeptides were obtained by solid-phase synthesis and conjugated to DOTA. The peptides were radiolabeled with gallium-68 or lutetium-177, and their binding affinity to bone was tested in vivo. Osteopontin and matrix extracellular phosphoglycoprotein (MEPE) derived peptides did not show strong binding to bone. Systematic variations in oligoglutamic acid chain length, as well as the positioning and clustering of phosphorylated serine residues, enabled the identification of an optimized phosphopeptide. Clustering phosphorylated sites within the peptide sequence provided significant advantages over phosphorylated moieties scattered throughout the peptide sequence. DOTA-pS4E8 showed the strongest affinity for bone, comparable to the clinically used bone targeting agent methylene bisphosphonate (MBP). CONCLUSIONS: The novel phosphopeptides match the outstanding bone-targeting capabilities of bisphosphonates and show comparable pharmacokinetics. Owing to their peptidic nature and the consequently anticipated favorable toxicological profile, these agents warrant further investigation as versatile bone-targeting vectors.

Epidemiological evidence indicates that HP-I increases the risk of CRC, yet the underlying pathological mechanisms remain unclear. This study aimed to identify DEGs associated with HP-I and CRC by analyzing public RNA sequencing data and to investigate potential molecular pathways. Gene expression profiles of HP-I and CRC were obtained from the GEO and TCGA databases. Shared DEGs were identified, followed by functional annotation, PPI network construction, hub gene identification, and validation of hub gene expression and diagnostic and prognostic value in CRC. Immune infiltration analysis, TFs prediction for hub genes, and potential small-molecule compound screening were also performed. A total of 112 shared DEGs were identified, including 74 up-regulated genes and 38 down-regulated genes. Enrichment analysis indicated that these DEGs were predominantly involved in immune and inflammation-related pathways. Eight hub genes (AGT, CCL20, CXCL1, CXCL2, CXCL5, CXCL9, CXCL10, and MMP9) were identified as central players contributing to the development of comorbid HP-I and CRC. Notably, these genes demonstrated good diagnostic performance in CRC, with AUC values ranging from 0.682 to 0.933. CXCL1 and CXCL9 were identified as prognostic markers for CRC. Immune infiltration analysis revealed a significant dysregulation of CD4⁺ T cell subsets in the context of HP-I and CRC. Finally, DY-131 was identified as a potential therapeutic candidate through the CMap database. This study reveals the potential common molecular mechanisms between HP-I and CRC, identifies shared hub genes, and highlights candidate therapeutic compounds.

Viral myocarditis (VM) is a cardiac inflammatory condition caused by viral infection and serves as a critical precursor to life-threatening complications, such as dilated cardiomyopathy and heart failure. Coxsackievirus B3 (CVB3), a predominant etiological agent of VM, lacks targeted therapeutic interventions despite ongoing antiviral development. Mitophagy is a selective mitochondrial quality control mechanism mediated by PINK1. It has two key roles: maintaining mitochondrial homeostasis and regulating innate antiviral immunity. Here, we employed single-cell RNA sequencing to reveal a significant correlation between impaired mitophagy and cardiomyocyte pathology in CVB3-induced myocarditis. We demonstrated that CVB3 infection suppresses PINK1-dependent mitophagy, while the attenuation of PINK1 reciprocally enhances CVB3 replication. Mechanistically, CVB3 non-structural protein 3C promotes the degradation of mitochondrial antiviral signaling protein (MAVS). MAVS interacts with PINK1 to form a regulatory loop: PINK1 deficiency boosts MAVS reduction, which further promotes viral replication and worsens myocardial injury. Furthermore, we identify the transcription factor FOSL1 as a novel negative regulator of PINK1 transcription through direct promoter binding. Collectively, these findings show that the 3C/FOSL1/PINK1/MAVS signaling axis is a key mechanism in CVB3 pathogenesis. We propose innovative therapeutic targets for viral myocarditis through restoration of mitochondrial homeostasis and modulation of host-virus interactions.

Tamoxifen (TAM) resistance remains a significant challenge in treating estrogen receptor (ER)-positive breast cancer. Overexpression of epidermal growth factor receptor (EGFR), variations in TAM metabolism, and increased expression of drug-resistant proteins (P-glycoprotein, P-gp) all contribute to the development of TAM-resistant breast cancer. The lack of specific drug distribution to tumors further complicates the treatment of TAM-resistant breast cancer. Simultaneously addressing these four factors—EGFR overexpression, TAM metabolic variations, P-gp-mediated drug efflux, and poor tumor-specific delivery—can significantly enhance the treatment of TAM-resistant breast cancer. This study aimed to design a multifunctional and dual-targeting polymeric prodrug nanomicelle platform. The platform was self-assembled using the amphipathic precursor drug material [hyaluronic acid (HA)-4-hydroxytamoxifen (4-OH-TAM) ester (esterified by HA and 4-OH-TAM)] and D-α-tocopheryl polyethylene glycol succinate (TPGS), loaded with dasatinib (DAS). The nanosystem could target breast cancer mediated by HA and 4-OH-TAM, releasing DAS, 4-OH-TAM, and TPGS in tumor microenvironment. 4-OH-TAM, an active metabolite of TAM, substantially impacted TAM-resistant breast cancer by avoiding the metabolic differences in TAM. TPGS inhibited P-gp-mediated drug efflux, thereby increasing intracellular accumulation of DAS and enhancing its inhibitory effect on TAM-resistant breast cancer cells. The results showed that the nanomicelles released drugs in a pH-sensitive manner. The cumulative release rates of 4-OH-TAM and DAS were (72.73 ± 3.99)% and (78.39 ± 3.09)%, respectively, within 48 h in a pH 5.0 solution, significantly higher than those in pH 6.0 and 7.4 solutions. The findings regarding cellular uptake and biodistribution indicated that the nanomicelles exhibited effective targeting capabilities toward TAM-resistant breast cancer. Both in vitro and in vivo studies demonstrated a marked effect of nanomicelles against TAM-resistant breast cancer through the targeting characteristic of nanoparticles and the actions of drugs (4-OH-TAM, DAS, and TPGS). The study provides fresh perspectives and suggestions for the clinical treatment of TAM-resistant breast cancer.

<h4>Aim</h4>There is a need for updated reference intervals for iron status biomarkers during infancy. This study aimed to investigate reference interval trends and diurnal variation of iron, transferrin, and transferrin saturation in infants subjected to delayed cord clamping at birth (DCC).<h4>Methods</h4>Data analysis from population-based Swedish studies, including 362 term-born infants subjected to DCC ≥ 60 s. The 2.5th and 97.5th percentiles were calculated from serum samples: In cord blood, at 48-118 h, four months and 12 months. We used Spearman's rho to test for associations and Student's t-test to compare groups.<h4>Results</h4>Iron concentrations in cord blood were initially high, 14-41 μmol/L, and had decreased by 48-118 h to 6-16 μmol/L, remaining mainly constant thereafter. Conversely, early transferrin concentrations were low. Transferrin slowly increased; concentrations at 4 months were positively associated with the average weight gained per day, rho = 0.46, p < 0.001. A small diurnal difference in iron was observed at 48-118 h: Samples collected between 10:30-11:59 were, on average, 1.4 μmol/L (95% CI -2.8 to -0.0) lower compared with samples collected between 15:30-19:59.<h4>Conclusion</h4>Developmental trends in iron, transferrin, and transferrin saturation in the first year of life need to be considered in the interpretation of test results from infants.

<h4>Background</h4>Drug-induced liver injury (DILI) can lead to potentially fatal complications, such as acute liver failure and even death. In clinical practice, glucocorticoids have been considered in some cases of DILI, especially those with hyperbilirubinemia. However, the available evidence remains controversial and its quality is also very limited.<h4>Objectives</h4>To explore the efficacy and safety of glucocorticoids in patients with acute DILI and hyperbilirubinemia.<h4>Design</h4>An open-label, multicenter, randomized controlled trial (RCT).<h4>Methods and analysis</h4>Overall, 232 patients with acute DILI with hyperbilirubinemia will be enrolled, and then, randomly assigned at a ratio of 1:1 to the conventional treatment alone or combined with glucocorticoids groups. The primary endpoint is the improvement of DILI, which is defined as total bilirubin level decreased by 50% after treatment, at the 2nd week. Secondary endpoints include the improvement of DILI at the 4th week and rates of progressive liver injury, liver failure, liver transplantation, survival, and adverse events. As for exploratory endpoints, we will also identify the beneficial population and assess the changes of inflammatory factors following glucocorticoids treatment.<h4>Ethics</h4>The study has been approved by the Medical Ethical Committee of the General Hospital of Northern Theater Command (ethical approval number Y2024-226) as well as 11 other participating centers.<h4>Discussion</h4>This RCT will provide solid evidence to clarify the benefits of glucocorticoids in patients with acute DILI and hyperbilirubinemia.<h4>Trial registration</h4>ClinicalTrials.gov identifier: NCT06922669.

Baicalin is a natural compound sourced from <i>Scutellaria baicalensis</i> which possesses various biological activities. To date, a large amount of research has been conducted on the antibacterial activity and related mechanisms of baicalin, making it a promising candidate for new broad-spectrum antibacterial drugs. However, the solubility of baicalin is limited. To improve its solubility and overcome the clinical application bottleneck, researchers have developed various solubilization techniques. Therefore, this article introduces the biological characteristics of baicalin; explores its effects as an antibacterial agent on bacterial biofilms, quorum sensing, virulence factors, inflammatory responses, and the immune system; and discusses the applications of nano-carrier loading technology, cyclodextrin inclusion technology, metal ion coordination and organometallic complexation technology, and dynamic covalent hydrogel assembly technology in improving the solubility of baicalin, thereby enhancing its antibacterial activity.

<h4>Background</h4>Lung adenocarcinoma (LUAD), the most prevalent and clinically heterogeneous form of lung cancer, lacks robust biomarkers for prognosis and immunotherapy response prediction. This study aimed to identify key prognostic genes and develop a predictive framework to guide personalized treatment strategies.<h4>Methods</h4>We integrated summary data-based Mendelian randomization (SMR) with machine learning to identify LUAD-associated genes. RNA-sequencing and clinical data from The Cancer Genome Atlas (TCGA) and Gene Expression Omnibus (GEO) were analyzed. A prognostic model was constructed with 101 combinations of 10 machine learning algorithms and validated via receiver operating characteristic (ROC) curve and concordance index analyses. Single-cell RNA-sequencing and spatial transcriptomic data were further examined to clarify the therapeutic implications of the identified genes.<h4>Results</h4>SMR identified 316 LUAD-associated genes, among which 33 were associated with prognosis according to univariate Cox regression. The optimal model combining random survival forest and ridge regression achieved superior predictive accuracy, with the area under the curves for 1-, 3-, and 5-year survival being 0.757, 0.707, and 0.686, respectively. High-risk patients exhibited distinct immune microenvironment suppression, reduced tumor mutation burden, and diminished immunotherapy responsiveness as compared to their low-risk counterparts. Single-cell and spatial transcriptomic analyses identified interactions between cancer cells and stromal cells as potential therapeutic targets.<h4>Conclusions</h4>The integrative approach employed in this study identified prognostic biomarkers and immunotherapy response predictors for patients with LUAD, offering a machine learning-driven framework for risk stratification and precision oncology. The findings underscore the clinical utility of combining genetic epidemiology with advanced analytics to optimize therapeutic strategies.

<h4>Background</h4>Processing bodies (P-bodies) are cytoplasmic granules involved in post-transcriptional gene regulation and play pivotal roles in carcinogenesis. However, the clinical significance of P-body-associated genes in lung adenocarcinoma (LUAD) remains poorly understood. This study aimed to investigate the clinical prognostic significance and potential biological roles of P-body-associated genes in LUAD, and establish a gene-based prognostic model for clinical application.<h4>Methods</h4>Twenty-four core P-body-related genes were curated from a user-friendly database RNAgranuleDB. Using transcriptomic and clinical data from The Cancer Genome Atlas (TCGA) LUAD cohort, univariate Cox regression was performed to screen prognostic genes, followed by least absolute shrinkage and selection operator (LASSO)-Cox regression with ten-fold cross-validation to construct a P-body-related risk score based on five retained genes (<i>MOV10</i>, <i>PCBP1</i>, <i>PCBP2</i>, <i>YBX1</i>, <i>YWHAG</i>). The prognostic value of this score was evaluated in TCGA and validated in five independent Gene Expression Omnibus (GEO) cohorts. Functional implications were explored through gene set variation analysis (GSVA) and reverse-phase protein array (RPPA) analysis. Immune microenvironment characteristics were assessed using single-sample gene set enrichment analysis (ssGSEA).<h4>Results</h4>The P-body-related risk score served as an independent prognostic factor in LUAD, with high score significantly associated with worse overall survival (OS), progression-free interval (PFI), and disease-specific survival (DSS) in the TCGA cohort. These findings were consistently validated across five independent GEO cohorts. Functionally, high scores were associated with enhanced mRNA editing, activation of cell cycle pathways (G2/M checkpoint, E2F targets), PI3K-AKT-mTOR signaling, and DNA repair activity, as well as increased genomic instability markers including tumor mutational burden (TMB), tumor neoantigen burden (TNB), and homologous recombination deficiency (HRD). Regarding the tumor immune microenvironment, low score correlated with an immunologically active phenotype characterized by increased infiltration of CD8⁺ T cells, B cells, and dendritic cells (validated by both ssGSEA and xCell), and upregulation of antigen presentation genes. In contrast, high score was associated with an immunosuppressive phenotype and elevated expression of immune checkpoint molecules including programmed death-ligand 1 (PD-L1) and CD276.<h4>Conclusions</h4>We developed and validated a P-body-related five-gene risk score that independently predicts LUAD prognosis and stratifies immune phenotypes. As these genes have extra-P-body functions, the score serves as a transcriptomic proxy for P-body component enrichment, not a direct measure of P-body activity. Despite this limitation, it offers a clinically useful tool for risk stratification and mechanistic hypotheses.

<h4>Background</h4>Recurrent cervical internal carotid artery vasospasm (RCICAV) is a rare cause of ischemic stroke in young adults. However, its pathophysiology remains largely unknown. RCICAV is a life-threatening condition, sometimes accompanied by coronary spasmodic angina in addition to severe stroke, highlighting the need for further investigation of its etiology and therapeutic approaches.<h4>Aims</h4>This study aimed to identify the genetic basis of Japanese families harboring RCICAV.<h4>Methods</h4>Whole-exome and Sanger sequencing analyses were performed in five Japanese patients from four families (two affected siblings and three unrelated sporadic patients) and seven unaffected members from three families enrolled from October 2016 to May 2025. RCICAV was diagnosed using magnetic resonance angiography, which demonstrated vasospasm of the cervical portion of the internal carotid artery during the event and normalization during remission. <i>In vitro</i> and <i>ex vivo</i> analyses of the detected variants were performed to verify functional deficiencies.<h4>Results</h4>RCICAV was found to be an autosomal recessive disorder caused by biallelic loss-of-function variants in the <i>PTGIS</i> gene, which encodes the prostacyclin synthase. Three splice-site and nonsense variants, c. 1358 + 2T > C, p.W39X, and p. E314X, were identified in five patients from four families in either homozygous or compound heterozygous states. The c.1358 + 2T > C variant caused exon 9 skipping (p.V403Ifs*24), which was confirmed by a minigene assay and reverse transcription polymerase chain reaction using RNA extracted from peripheral blood leukocytes of a homozygous patient. Human carotid artery endothelial cells expressing these truncated variants exhibited markedly reduced <i>PTGIS</i> protein and prostacyclin metabolite production.<h4>Conclusion</h4>The results indicate that prostacyclin synthase deficiency is the core pathology of RCICAV. Given that prostacyclin is a potent vasodilator, impaired biosynthesis is consistent with the disease mechanism and may serve as a promising therapeutic target to prevent life-threatening events, including severe stroke and angina.

The MAPRE3 gene is aberrantly expressed in several cancers. We profiled DNA methylation in tumor tissues from early-stage non-small cell lung cancer (NSCLC) patients and assessed associations with overall survival (OS). Significant CpG probes were validated in The Cancer Genome Atlas (TCGA). The methylation level of cg12821679_MAPRE3 showed significant associations with OS in lung squamous cell carcinoma (LUSC) (HR = 0.32, P = 6.55 × 10^-7), but it was not observed in lung adenocarcinoma (LUAD). In LUSC, MAPRE3 expression was significantly correlated with cg12821679_MAPRE3 (r = 0.17, P = 2.96 × 10^-3) and potential trans-regulated genes were enriched in the Nicotine addiction pathway. Additionally, MAPRE3 expression showed significant associations with OS in both LUAD and LUSC (LUAD: HR_{low vs high} = 2.28, P = 2.40 × 10^-3; LUSC: HR_{low vs high} = 1.61, P = 0.0244). The association between smoking cessation and overall survival was significantly modified by MAPRE3 expression (HR_interaction = 0.69, P = 0.0282). Smoking cessation improved OS only in patients with high MAPRE3 expression (HR = 0.56, P = 2.82 × 10^-3). We conclude MAPRE3 may predict NSCLC prognosis and influence the prognostic benefit of smoking cessation.

BACKGROUND: Ischemia-reperfusion acute kidney injury (IRI-AKI) is a common critical clinical condition characterized by high morbidity and mortality. Renal tubular epithelial cells (TECs) ferroptosis plays a key role in the pathogenesis of IRI-AKI. We observed that olfactomedin 4 (OLFM4) expressed in TECs and was downregulated in the kidney of AKI patients. And the downregulation of OLFM4 was correlated with TECs ferroptosis. Therefore, our present study intended to clarify the role and mechanism of OLFM4 in TEC ferroptosis and IRI-AKI. METHODS: OLFM4-deficient HK-2 cell lines and OLFM4−/− mice were generated. In vitro hypoxia/reoxygenation (H/R) TECs model and IRI-AKI mouse models, were employed. Renal injury and ferroptosis-related markers were evaluated by immunofluorescence, Western blotting, and biochemical assays to evaluate the role of OLFM4 in TECs ferroptosis. Molecular docking, co-immunoprecipitation (Co-IP), and yeast two-hybrid assays were performed to identify the mechanism of OLFM4 on ferroptosis pathway. RESULTS: OLFM4 was markedly downregulated in renal tissues of IRI-AKI mice and HK-2 cells subjected to H/R. OLFM4 deficiency aggravated renal function impairment of the IRI-AKI mice (Cr, BUN), accompanied by elevated ferroptosis markers (Fe²⁺, ROS, and MDA) and reduced GSH, whereas OLFM4 overexpression ameliorated IRI-AKI and suppressed TECs ferroptosis. Additionally, we demonstrated that OLFM4 stabilizes GPX4 by inhibiting its ubiquitination, and GPX4 is a key molecule in the occurrence of ferroptosis. The overexpression of OLFM4 upregulating GPX4 expression, enhancing antioxidant defenses, and attenuating TECs ferroptosis in H/R HK-2 cell model and IRI-AKI mice. CONCLUSIONS: Our present study demonstrates that OLFM4 reduction play an important role in IRI-AKI occurrence. Overexpression could protect IRI-AKI by reducing TEC ferroptosis via GPX4. For the first time, OLFM4 is identified as a ferroptosis suppressor, expanding our understanding of AKI pathogenesis, and highlighting OLFM4 as a potential therapeutic target.

Pompe disease (PD) is a rare genetic disorder, caused by the lysosomal acid alpha-glucosidase enzyme deficiency leading to accumulation of glycogen, primarily in skeletal and respiratory muscles, but also in the heart. In both known PD forms, namely infantile (IOPD) and late-onset (LOPD), the prevalence and nature of myocardial involvement are still to be clarified. Cardiovascular magnetic resonance (CMR) may help to detect unrecognized alterations contributing to patients' management. This study aimed to systematically review PD CMR features. We conducted a systematic search of three electronic databases (PubMed, Scopus, and Web of Science) up to February 2024, without language or time interval restriction. Two reviewers performed the search and selection process, data extraction, and synthesis. We resolved disagreements by consensus and/or involving a third reviewer. The included studies have been classified according to the Oxford Centre for Evidence Based Medicine (CEBM) grading system. Out of 276 articles, 11 papers were finally included: seven about IOPD and four regarding LOPD. According to CEBM, seven were level 5, one was level 4, two were level 3, and one was level 2. Six studies reported qualitative late-gadolinium enhancement tissue characterization, 1 described cardiac infiltration on TIRM sequences, 3 explored changes in ExtraCellular Volume and, among these, 2 evaluated also T1/T2 mapping and perfusion; one study assessed the management after therapy through T1 mapping. Literature data regarding CMR features in PD are limited and based on nonparametric assessment mainly. Further investigations, involving also mapping techniques, are required to better understand PD cardiac involvement.

Neutrophil migration to bacterial infection sites is key for host defense. Host ribosomal protein SA (RPSA) has been recently reported to regulate the anti-infection immunity of immune cells; however, its role in neutrophil migration remains unclear. Here, using myeloid-specific Rpsa-deficient mice, we found that RPSA deletion inhibited neutrophil infiltration and markedly exacerbated Streptococcus suis serotype 2 infection. Adoptive cell transfer and neutrophil depletion assays identified RPSA as vital for the anti-infective function of neutrophils. Mechanistically, RPSA deficiency induced the overexpression of olfactomedin 4 (OLFM4), which in turn inhibited the activation of the RhoA/ROCK1/pMLC2 signaling pathway, reduced MYH9 expression, and caused aberrant MYH9 translocation from the uropod to the cytosol in migrating neutrophils. Ultimately, this disrupted cytoskeletal polarization and uropod extension, thereby abrogating migratory function. Clinically, septic patients' neutrophils exhibited reduced RPSA and elevated OLFM4 expression, a phenotype that correlated with a marked impairment of migratory capacity. Therapeutic targeting of the RPSA-OLFM4 axis restored neutrophil migration and improved disease outcomes in both S. suis 2-infected and septic mice. Thus, our findings demonstrate that RPSA promotes neutrophil migration via downregulating OLFM4 to counter bacterial infection, and establish the RPSA-OLFM4 axis as a critical immune migratory checkpoint in host antibacterial immunity.

Type 2 diabetes mellitus (T2DM) patients with colon cancer have an increased incidence and poor prognosis, which may be closely related to advanced glycation end products (AGEs) and its receptor (RAGE), but the underlying mechanism is still unclear. The aim of this study is to investigate how the AGEs-RAGE axis inhibits ferroptosis in T2DM-associated colon cancer by regulating carcinoembryonic antigen-related cell adhesion molecule 1 (CEACAM1). ELISA and Western blot were used to detect expression levels of CEACAM1 in 155 patients with colon cancer. In vitro experiments examined various parameters in CEACAM1 knockdown/overexpression cells co-cultured with AGEs. The molecular mechanism was validated using multiple detection methods following intervention with FPS-ZM1 and sabatolimab. Serum and tissue analyses revealed that CEACAM1 is up-regulated in colon cancer patients with T2DM. The AGEs-RAGE promotes colon cancer cell proliferation, migration and invasion by upregulating CEACAM1. CEACAM1 overexpression triggers IFN-γ pathway, thereby enhancing SLC7A11/GPX4-mediated ferroptosis resistance. Blockade of RAGE with FPS-ZM1 reverses CEACAM1 driven inhibition of ferroptosis by reducing GPX4. A nude mouse model of type 2 diabetic tumors was established to assess tumor growth. Furthermore, blockade of Tim-3 by sabatolimab disrupts its interaction with CEACAM1, restoring ferroptosis sensitivity and inhibiting subcutaneous tumor growth in these mice. The AGEs-RAGE axis inhibits ferroptosis by driving CEACAM1 overexpression and mediating IFN-γ/SLC7A11/GPX4 signaling pathway, which reveals a new mechanism by which AGEs-RAGE-CEACAM1 promotes tumor survival in diabetic environment. Sabatolimab may provide a therapeutic strategy for colon cancer patients with T2DM.

INTRODUCTION: The incidence of Mycobacterium abscessus complex (MABC) infections is rising, becoming a major pathogen of nontuberculous mycobacteria responsible for pulmonary disease (PD) and extrapulmonary disease (ED). However, studies on the clinical characteristics of MABC-ED remain limited. METHODS: A 7-year retrospective analysis was conducted on MABC-ED cases at Zhongshan Hospital in Shanghai, China. We analyzed predisposing factors, clinical features, metagenomic sequencing (MS) results, drug susceptibility testing (DST), and genomic characteristics of MABC-ED patients, comparing the data with those of PD cases. RESULTS: Among 17 MABC-ED patients, 15 had predisposing risk factors and underlying conditions, with 2 of 3 patients with rheumatic disease showing poor prognosis. The diagnostic performance of metagenomic sequencing for MABC-ED was comparable to that for MABC-PD. However, MABC-ED samples exhibited distinct microbiome features and a more diverse mycobacterial community structure compared to PD. Resistance rates among extrapulmonary MABC isolates were observed as follows: 0% (amikacin), 20% (macrolides), 30% (linezolid), and 40% (cefoxitin). One case showed paradoxical results between erm (41) T28 sequevar and susceptibility phenotype. Genomic analysis revealed no specific dominant circulating clones (DCC) for MABC-ED isolates. CONCLUSION: MABC-ED patients commonly present with risk factors and underlying diseases. Metagenomic sequencing diagnosis of MABC-ED poses challenges, and DST and whole genome sequencing data indicate diversity among MABC-ED isolates. Our study provides detailed data on MABC-ED, contributing to a better understanding of its disease characteristics.

BACKGROUND: The efficacy of conventional αβ T cell-based immunotherapies is often limited by tumor immune evasion. γδ T cells can bypass these limitations through MHC-independent tumor recognition, but their function within the tumor microenvironment (TME) remains poorly characterized due to a lack of relevant preclinical models. This study aims to establish a patient-derived tumor organoid-immune cell coculture system to evaluate γδ T-cell reactivity and cytotoxicity in a human-relevant TME (n = 10 PDTO lines; n = 3 independent experiments). METHODS: We developed an innovative coculture system—believed to be the first to integrate patient-derived tumor organoids (PDTOs) and autologous tumor-infiltrating lymphocytes (TILs) or healthy donor-derived allogeneic Vγ9Vδ2 T cell. T-cell activation was quantified by comparing CD137 expression on γδ T cells versus CD4+ and CD8+ T cells via flow cytometry after coculture. The cytotoxicity of Vγ9Vδ2 T cells was evaluated at various effector-to-target (E:T) ratios using a live-cell imaging assay to track BCO infiltration and apoptosis over 24 hours. Secreted effector molecules (IFN-γ, granzyme B, perforin) were measured from coculture supernatants using a cytometric bead array. RESULTS: Baseline analysis revealed that γδ T cells represent the most reactive subset within expanded TILs, with 12.3% expressing CD137 compared to 3.49% of CD8+ T cells (p = 0.0118). Notably, autologous BCO–TIL coculture preferentially enhanced γδ T-cell activation, reaching 24.85% CD137+—a frequency significantly higher than that of CD8 (9.15%, p = 0.0062) and CD4 (9.99%) subsets. The net increase in CD137 positivity for γδ T cells was more than double that of CD8 T cells (12.55% vs. 5.66%, p = 0.0467). Allogeneic Vγ9Vδ2 T cells demonstrated significant, dose-dependent cytotoxicity against BCOs (p < 0.0001). Maximal cell death (RFU fold-change: 5.2 ± 0.5) was achieved at an effector-to-target (E:T) ratio of 10:1 (p < 0.0001). This cytotoxic efficacy was highly correlated with the secretion of lytic effectors, including IFN-γ ( > 1000 pg/mL; p< 0.001), granzyme B ( > 2000 pg/mL), and perforin ( > 350 pg/mL). CONCLUSION: Our findings demonstrate that 3D autologous coculture leads to the preferential stimulation of γδ T cells, establishing them as a primary reactive subset capable of bypassing MHC-restriction bottlenecks. The potent dismantling of tumor architecture by allogeneic Vγ9Vδ2 T cells, supported by high-resolution kinetic and cytokine data, substantiates their development as “off-the-shelf” therapies. Collectively, this platform provides a standardized, high-fidelity engine for the rapid preclinical assessment of next-generation cellular immunotherapies.

<h4>Background</h4>Several reports have indicated an association between insulin resistance and hyperferritinemia, and a recent consensus statement has suggested the term metabolic hyperferritinemia (MHF) for patients with metabolic syndrome and increased serum ferritin. The objective of this study was to examine the association between serum ferritin levels and insulin resistance in an unselected Norwegian population.<h4>Methods</h4>Two thousand people were randomly selected from the public registry in one municipality in Nord-Trøndelag County, Norway. Of the respondents, 1115 completed a screening glucose tolerance test to determine the prevalence of impaired glucose tolerance (IGT)/type 2 diabetes mellitus (T2DM), and these individuals were then selected for further examination. A control group comprising 100 age- and gender-matched individuals with normal glucose tolerance and CRP less than 5 was retrospectively generated from the same cohort.<h4>Results</h4>One hundred and seventy-six people were diagnosed with impaired glucose tolerance or diabetes mellitus type 2 (IGT/T2DM), and 17 (10%) of these patients had elevated ferritin levels according to the MHF definition. In the control group with normal glucose tolerance, 11% also had hyperferritinemia based on the same ferritin cut-off. Multiple regression revealed a significant association between serum ferritin, male sex, 2-hour glucose value after the glucose load and waist circumference among the patients with IGT/T2DM, while there was no association with age or transferrin saturation.<h4>Conclusions</h4>This study demonstrated a prevalence of metabolic hyperferritinemia in one out of ten people with IGT/T2DM; however, a comparable level of serum ferritin was also detected in the matched control group. Only one person in the IGT/T2DM group had ferritin levels higher than 550 mmol/L. Thus, hyperferritinemia is prevalent in this unselected Norwegian cohort. A relationship between s-ferritin and metabolic syndrome parameters was found in patients with IGT/T2DM, but hyperferritinemia is likely not related to iron overload.

Crude glycerin (CG) is an energy-dense ingredient capable of partially or fully replacing corn in high-concentrate diets for ruminants. Its rapid ruminal absorption, favorable fermentative profile, and absence of lactic acid production may support safer adaptation to intensive feeding systems. The aim of this study was to evaluate the effects of replacing corn with CG (300 g/kg DM) on growth performance, feeding behavior, rumen morphometry, and proteomic responses of ruminal papillae in feedlot lambs. Sixty-five Santa Inês × Dorper lambs were assigned to either a control diet or a diet containing CG and were evaluated during pre-adaptation, adaptation, and finishing phases. Replacing corn with CG slightly reduced average daily gain (<i>p</i> = 0.02), without affecting final body weight, dry matter intake, or carcass yield (<i>p</i> > 0.05). Lambs fed CG exhibited lower subcutaneous fat thickness (<i>p</i> = 0.04) and reduced neutral detergent fiber intake during feeding behavior assessments (<i>p</i> < 0.05). Rumen papillae showed higher mitotic index and greater epithelial activity throughout the feedlot period, regardless of treatment. Proteomic analysis revealed upregulation of proteins involved in epithelial integrity (<i>Claudin-1</i>, <i>Occludin</i>) and mitochondrial energy metabolism (<i>ATP synthase β</i>, <i>glycerol kinase</i>) in CG-fed lambs, alongside downregulation of proteins related to oxidative stress and inflammation (<i>HSP70</i>, <i>Annexin A1</i>, <i>SOD1</i>, <i>Peroxiredoxin-6</i>). These findings demonstrate that CG promotes beneficial molecular adaptations in the ruminal epithelium without compromising carcass traits, supporting its use as a safe, functional, and sustainable alternative to corn in lamb finishing systems.

Oxidative stress is one of the few defined causes of male infertility affecting at least one third of patients attending infertility clinics. Human spermatozoa are vulnerable to this form of attack because their stripped-down architecture means that they possess limited antioxidant protection and little capacity for biochemical repair. They also compound their vulnerability by being active generators of reactive oxygen species (ROS) and possessing multiple substrates for oxidative damage. The major sources of ROS in these cells are their mitochondria, an L-amino acid oxidase (IL4I1) and a calcium-dependent NADPH oxidase (NOX5). Spermatozoa tolerate the risks associated with ROS generation because their biology is heavily dependent on redox regulation. ROS are important mediators of sperm capacitation, stimulating the generation of cAMP and prostaglandins, inhibiting protein phosphatases and encouraging removal of cholesterol from the plasma membrane. Furthermore, during fertilization, the ability of ROS to activate metalloproteinases facilitates penetration of the zona pellucida and sperm-oocyte fusion. While ROS are physiologically important for sperm function, the over-production of these metabolites can impair sperm function. Antioxidants have therefore assumed some importance as a possible therapy for the infertile male. However, before this potential can be realized, we need to optimize the composition and dose of reagents used in such formulations and develop improved methods of diagnosing oxidative stress within the patient population.

Lung adenocarcinoma (LUAD) remains a leading cause of cancer-related mortality worldwide, and although targeted therapy and immunotherapy have improved clinical outcomes, their long-term efficacy is frequently limited by therapeutic resistance, underscoring the need to identify additional biologically relevant and potentially actionable targets. In this study, we applied summary-data-based Mendelian randomization (SMR) by integrating lung cancer genome-wide association study (GWAS) summary statistics with cis-expression quantitative trait locus (cis-eQTL) and cis-methylation quantitative trait locus (cis-mQTL) datasets to prioritize candidate genes associated with LUAD risk. The robustness of SMR-significant signals was further evaluated using the heterogeneity in dependent instruments (HEIDI) test and Bayesian colocalization analysis. Independent transcriptomic datasets from the Gene Expression Omnibus (GEO), together with multiple public databases, were used for expression and prognostic validation, while protein-protein interaction (PPI) and functional enrichment analyses were performed to provide mechanistic context. Among 13 genes prioritized by integrative genetic analyses, PSMA4 (Proteasome 20S Subunit Alpha 4) emerged as the leading LUAD-relevant candidate on the basis of convergent multi-omics evidence, colocalization support, reproducible overexpression (OE) across independent cohorts, and adverse prognostic association. Functional experiments demonstrated that PSMA4 knockdown (KD) suppressed LUAD cell proliferation, migration, invasion, and xenograft tumor growth, whereas PSMA4 overexpression produced the opposite effects. Mechanistically, PSMA4 interacted with p53 and facilitated the proteasome-dependent turnover of ubiquitinated p53, as supported by co-immunoprecipitation, immunofluorescence, ubiquitination, proteasome inhibition, and cycloheximide chase assays, thereby attenuating p53 signaling. Collectively, these findings support PSMA4 as a biologically relevant and potentially actionable target in LUAD and provide a mechanistic rationale for further translational investigation.

The problem of lung cancer poses a great threat to the health of people all over the world. This threat is mainly associated with resistance to treatment. There is evidence that shows that ferroptosis fails to take place in lung cancer. There is a high degree of suppression in those that have become resistant to treatment. There seems to be a link between resistance to treatment and suppression of ferroptosis. This led to the development of a strategy to induce ferroptosis. In this review, the mechanisms of ferroptosis in drug-resistant non-small lung cancer cells will be discussed, along with a synopsis of recent studies analyzing combination therapies that involve the pharmacological induction of ferroptosis, thus further validating the notion that the prevention of ferroptosis plays an important role in drug resistance. Based on these principles, we will assess the inadequacies in current ferroptosis-inducing agents regarding their use in the treatment of lung cancer, focusing mainly on the suboptimal targeting of cancer cells and drug delivery efficiency. Conversely, these inadequacies have triggered new approaches in cancer therapy, which include using nanomedicine for improved drug delivery, functional nanomaterials for directly triggering ferroptosis, among others. In addition, we evaluate the validity of nanomaterials for precision medicine in cancer therapy, in addition to utilizing bioactive compounds. Lastly, we discuss the integration with artificial intelligence in single-cell sequence analysis for the development of modeling systems for ferroptosis control.

<h4>Background and aim</h4>The increasing emergence of antimicrobial resistance has intensified the search for safe and effective alternatives to antibiotics in livestock production. Probiotics derived from traditional fermented foods represent a promising strategy due to their adaptability and functional diversity. This study aimed to isolate and characterize a plateau-adapted strain of <i>Enterococcus faecium</i> from traditional yak yogurt and to comprehensively evaluate its probiotic potential through <i>in vitro</i> assays and <i>in vivo</i> validation, with a focus on its applicability in yak health management.<h4>Materials and methods</h4>The strain <i>E. faecium</i> SWUN5732 was isolated and identified using morphological characterization and <i>16S rRNA</i> gene sequencing. Probiotic properties were assessed through acid (pH 2.0 and 3.0) and bile salt (0.3%) tolerance, hemolytic activity, antibiotic susceptibility, antimicrobial activity against common yak pathogens, and adhesion to Caco-2 cells. Safety evaluation included virulence gene screening targeting <i>cylA</i>, <i>esp</i>, <i>asa1</i>, <i>gelE</i>, <i>agg</i>, and <i>efaAfm</i>, as well as acute oral toxicity testing in mice. Functional efficacy was further examined through a 21-day oral administration study in mice to evaluate body weight gain and intestinal secretory immunoglobulin A levels.<h4>Results</h4><i>E. faecium</i> SWUN5732 demonstrated strong gastrointestinal tolerance, with survival rates of 34.43% at pH 2.0, 60.17% at pH 3.0, and 77.07% in 0.3% bile salts. The strain exhibited no hemolytic activity and was susceptible to clinically relevant antibiotics. It showed broad-spectrum antimicrobial activity against <i>Escherichia coli</i>, <i>Salmonella</i> spp., and <i>Staphylococcus aureus</i>, with inhibition zones ranging from 18 to 37 mm. The adhesion rate to Caco-2 cells was approximately 29%, indicating effective colonization potential. Among the tested virulence determinants, only <i>efaAfm</i> was detected, while other genes (<i>cylA</i>, <i>esp</i>, <i>asa1</i>, <i>gelE</i>, <i>agg</i>) were absent. No acute toxicity or pathological alterations were observed in mice. Notably, oral administration significantly enhanced body weight gain and increased intestinal secretory immunoglobulin A levels (p < 0.01), demonstrating immunomodulatory and growth-promoting effects.<h4>Conclusion</h4>The findings indicate that <i>E. faecium</i> SWUN5732 possesses key probiotic attributes, including gastrointestinal resilience, antimicrobial activity, safety, and immunomodulatory capacity. Its origin from high-altitude yak yogurt suggests ecological compatibility with yak production systems. This strain represents a promising candidate as an antibiotic alternative in yak farming; however, further <i>in vivo</i> validation in yaks is required to confirm its practical applicability under field conditions.

The growing challenges of conventional ECM bio-inks for 3D cell culture underscore the development of novel hydrogels that fully recapitulate specialized in vivo cell microenvironments. We developed HA-gel-dex hydrogels by double-crosslinking ECM components (hyaluronic acid and gelatin) with synthetic dextran. Engineered with irreversible amide bonds and dynamic imine crosslinks, the HA-gel-dex enables viscoelastic property modifications to replicate native ECM characteristics, with the highest yield ratio reaching ~1000%. This programmability derive from tunable crosslinking architectures and network dynamics, where controlled HA-gel to dex-CHO ratios and peptide functionalization yield optimized hydrogel viscoelasticity and predictable bioprinting performance. Functionalization with selective cell-ligation groups (RGD, collagen, and laminin peptides) strengthened cell-matrix interaction, promoting increased cell proliferation, differentiation, and micro-organo-sphere formation. Moreover, rheological analyses revealed significantly enhanced stress-relaxing and self-healing hydrogel properties; while showing cytocompatibility, increased cell viability, bioprinting capacity, significant biofilm inhibition properties, and synergism with antimicrobials. Further assays on BALB/c mice showed remarkably improved wound healing, hair follicle regeneration, and nature ECM production abilities, the performance of which are further augmented with the presence and encapsulation of therapeutic mesenchymal stem cells (MSCs). The study shows their future promise as wound dressings and potentially antibiotic treatment scaffolds for infection-associated wounds. Above all, HA-gel-dex hydrogels provide an improved ECM-like, reproducible, and cost-effective alternative to traditional ECM bio-inks for 3D cell culture, 3D bioprinting, and tissue repair studies - advancing synthetic ECM alternatives for translational applications.

The aberrant aggregation of tau leads to loss of its physiological functions and gain of toxic functions, and plays a crucial role in the pathogenesis of tauopathies including Alzheimer's disease (AD). Targeting tau aggregation is considered a promising strategy for treating tauopathies. The BRICHOS family consists of a variety of proteins containing the BRICHOS domain. Certain endogenous BRICHOS domains may inhibit the pathological aggregation of disease-associated proteins. However, the effects of the BRICHOS domains on tau aggregation remain unknown. Here we revealed that BRICHOS domains from integral membrane protein 2B (ITM2B), tenomodulin (TNMD), and out at first (OAF) bind to tau and inhibit its aggregation in vitro. Intravenous administration of TNMD BRICHOS alleviates tau aggregation, synaptic dysfunction, and memory deficits in Tau P301S transgenic mice. Thus, TNMD BRICHOS may serve as a potential therapeutic approach for the development of treatments for tauopathies.

Arsenic is a dual-natured element, functioning as both a carcinogen and a therapeutic agent in cancer treatment. Arsenic trioxide is a clinically approved drug for acute promyelocytic leukaemia, administered either alone or in combination with all-trans retinoic acid. Additionally, arsenic trioxide and organic arsenic compounds, such as darinaparsin, show promising efficacy against various cancers. Given the multi-targeted nature of metallodrugs, a comprehensive understanding of their molecular mechanisms is critical for the rational design of more potent and less toxic therapeutics. In this review, we first outline the structures and activities of key arsenic-based anticancer agents, encompassing both inorganic and organic forms. We then provide a comprehensive overview of proteome-wide identification of arsenic-binding proteins in cancer cells, utilizing metalloproteomic approaches such as immobilized metal affinity chromatography, continuous-flow electrophoresis coupled with inductively coupled plasma mass spectrometry, biotinylated arsenical pull-down, and fluorescence-based metalloproteomics. Subsequently, we discuss how the identified arsenic proteome facilitates elucidation of the molecular mechanisms of arsenic trioxide and darinaparsin from a systems perspective. We also evaluate studies on the cytotoxicity of arsenic compounds at the single-cell level, highlighting their role in improving our understanding of the mode of action of a drug. Finally, we prospect future research direction in this field. This review aims to stimulate further investigation into the molecular mechanisms of arsenic-based drugs at a systems level, guiding the development of more effective and safer arsenic-based therapeutics for cancer treatment.

<h4>Purpose</h4>To determine the likelihood of obtaining at least one transferable embryo per cycle in patients undergoing IVF with PGT-A + PGT-M for variants in one versus two genes.<h4>Methods</h4>IVF cycles in patients 18-45 years undergoing PGT-A + PGT-M for variants in one or two genes at a single genetics laboratory were analyzed from 11/2019 to 3/2023. Cycles were stratified by SART age category and disease inheritance pattern. Analyses were performed using median and chi-square tests.<h4>Results</h4>A total of 4634 IVF cycles were included: 146 cycles (3.15%) using PGT-A + PGT-M for two genes and 4488 cycles (96.85%) for one gene. After adjusting for the number of embryos biopsied, the likelihood of having at least one usable embryo was significantly lower in patients testing for variants in two genes compared to one in patients aged < 35 (75.0% v 82.5%, p = 0.004) and 35-37 (55.0% v 74.4%, p = 0.025) with no difference in other age groups 38-40 (66.7% v 74.4%, p = 0.586) and 41-42 (42.9% v 51.7%, p = 0.113).<h4>Conclusions</h4>There were significantly lower odds of obtaining a usable embryo when testing for variants in genes in patients aged < 35 and 35-37. No significant difference was seen among patients aged 38-40 or 41-42. These findings provide quantifiable data to counsel patients considering PGT-M testing for variants in two genes.

BACKGROUND: Currently, the American College of Medical Genetics and Genomics (ACMG) carrier screening (CS) guidelines do not recommend routinely screening for conditions with carrier frequencies ≤ 1:200. Advances in DNA sequencing and declining costs, alongside growing awareness, make it increasingly feasible to expand CS to cover a broader spectrum of conditions. We aimed to design a CS panel regardless of carrier frequency to evaluate its utility in less common disease. METHODS: We developed a large CS panel through gene curation, phenotypic severity evaluation, independent of carrier frequency. A cohort of 2010 couples across 24 Chinese cities underwent couple-based simultaneous screening. Gene carrier rate (GCR) was calculated, with subsequent assessment of at-risk couple rate (ACR) across varying GCR thresholds. RESULTS: The results obtained from testing 1736 genes were presented. Among 2010 couples undergoing CS, 106 were identified as having an increased risk of offspring with at least one genetic condition. The initial ACR is 5.3% (106/2010). After excluding couples not carrying high-risk CFTR variants based on the ACMG-recommended 100-variant panel for CFTR, the adjusted ACR was 5.2% (105/2010). Furthermore, after excluding GJB2: c.109 G > A and HFE, the adjusted ACR decreased to 2.2% (45/2,010). Fifty genes with a GCR ≥ 1/200 were identified through individual carrier analysis that were absent from the ACMG Tier 3 list. Setting a GCR ≥ 1/200 threshold would fail to detect 14 high-risk couples associated with 16 rare conditions: two couples carried pathogenic variants linked to two rare conditions each. Notably, 53.3% of identified high-risk couples would remain undetected under ACMG Tier 3-recommended conditions, with the majority of missed conditions representing rare yet severe disorders. The simultaneous couple-screening model maintained manageable ACR while resolving all reproductive concerns in a single phase, thereby reducing genetic counseling demands compared to sequential approaches. This methodology also minimized false-positive interpretations through integrated variant analysis. CONCLUSIONS: This study obtained preliminary Chinese population-specific GCR for both common and rare recessive conditions, providing a foundation for future CS panel optimization. Couples at risk of having a child with rare conditions are often undetectable using conventional CS paradigms that set GCR levels at ≥ 1/200 thresholds, emphasizing the need to expand beyond ACMG Tier 3 standards to address rare severe genetic disorders. For large CS panels, simultaneous couple-based screening is an effective strategy.

Epigenetic modifications, especially DNA methylation, play an increasingly important role in oral cancer. However, their specific contributions to the progression of oral squamous cell carcinoma (OSCC) remain unclear. The present study used the Shiny Methylation Analysis Resource Tool (SMART) database (https://smart.embl.de/smart/change_mode.cgi) to identify methylation‑driven genes associated with OSCC. Among the identified candidates, integrin subunit α4 (ITGA4) exhibited significantly elevated methylation levels in head and neck cancers. A methylation‑specific PCR assay showed that ITGA4 is highly methylated in OSCC cells compared with normal immortalized human normal oral keratinocyte (iNOK) cells. Additionally, the mRNA expression levels of ITGA4 were significantly lower in OSCC cell lines compared with normal iNOK cells. ITGA4 overexpression markedly inhibited the cell proliferation, migratory ability and capacity of colony formation and induced apoptosis in FaDu and YD‑15 cells. In proteomic analysis, ITGA4 suppressed the expression of Sorting Nexin 5 (SNX5), a protein linked to cancer progression. siRNA‑mediated knockdown of SNX5 importantly inhibited cell proliferation, migration, and colony formation in FaDu and YD‑15 cells. Moreover, in a chick chorioallantoic membrane xenograft model, overexpression of ITGA4 or small interfering SNX5 significantly inhibited OSCC tumor growth and angiogenesis <i>in vivo</i>. Collectively, these findings demonstrated that ITGA4 acts as a tumor suppressor in OSCC by downregulating SNX5 and suggested that ITGA4 may serve as a valuable prognostic biomarker and potential therapeutic target for OSCC.

High‑altitude polycythemia (HAP) is classically attributed to erythropoietin (EPO)‑driven erythrocytosis, yet epidemiological and mechanistic evidence increasingly challenges this monocular view. Field data have demonstrated that up to 40% of individuals with a hematocrit level >68% circulate EPO within the sea‑level reference range, whereas multi‑omics studies have revealed sustained HIF activity, mitochondrial oxidative stress, iron dysregulation, gut dysbiosis and epigenetic reprogramming as parallel, EPO‑independent drivers. Hypoxia‑inducible microRNAs, hepcidin suppression, TLR4‑IL‑6 signaling and defective mitophagy converge to lock erythroid precursors into a survival‑plus‑proliferation state even after ambient oxygen levels normalize. The purpose of the present review is to integrate these disparate pathways into a unified molecular framework and to outline a phased, biomarker‑guided therapeutic roadmap for the precise prevention of maladaptive polycythemia at high altitudes.

In this study, an innovative methodology was implemented to combine an isoxazole derivative (ARZ) with hydroxyapatite (HAp), varying several parameters to obtain three distinct composites (ARZ-Hap-1, ARZ-Hap-2 and ARZ-Hap-3). These composites (ARZ-Hap) were synthesized by a dissolution-recrystallization process, thereby facilitating enhanced interaction between ARZ and HAp. This approach promoted homogeneous dispersion of ARZ within the HAp matrix, consequently enhancing the stability and functionality of the resulting materials. The structural characterization of these composites was determined by FT-IR, XRD, BET, TGA, and NMR (¹³C, ³¹P) methods. Subsequently, the composites were subjected to a rigorous evaluation process to ascertain their antibacterial and antifungal activities. Among them, ARZ-HAp-3 exhibited noticeable antimicrobial activity against E. coli, S. aureus, and B. subtilis, demonstrating a measurable inhibitory effect on bacterial growth. In terms of antifungal activity, this composite demonstrated the best results, particularly against C. albicans and F. oxysporum, with minimal MIC and MFC values comparable to those of fluconazole. These results indicate the potential of the ARZ-HAp-3 composite as a promising bioactive material, demonstrating a combination of structural efficiency and targeted biological activity. The release of ARZ from the composites was monitored by UV-Visible spectroscopy, revealing a gradual and controlled profile. The quantity of the substance released is proportional to the initial content of ARZ, with a maximum observed for ARZ-Hap-3 (~900 mg/L). This mechanism, based on slow diffusion, confirms the potential of these materials for sustained release of active ingredients. Furthermore, a molecular docking analysis was conducted to explore potential interactions between ARZ and biological targets associated with the pathogens studied. The results of the study indicated a high binding affinity of ARZ with certain enzymes that are essential for bacterial and fungal survival. This finding corroborates the experimental observations made in the study, thereby providing a scientific rationale for the observed effects.

Pulmonary arterial hypertension (PAH) is a fatal vasculopathy driven by proinflammatory signaling that leads to pulmonary vascular remodeling and ultimately, right ventricular failure. Existing therapies fail to eliminate dysfunctional cells or reverse pathologic remodeling. Cellular senescence, a state of persistent cell-cycle arrest coupled with proinflammatory signaling, has emerged as a compelling, yet complex, contributor to PAH pathophysiology. Human data indicate that senescent cells accumulate within remodeled pulmonary vascular lesions and may promote proliferation and apoptosis resistance in neighboring cells. However, preclinical studies testing senolytic therapies in PAH have yielded conflicting results, raising uncertainty about their translational potential. This review will synthesize the current literature regarding cellular senescence in PAH and propose that heterogeneity in <i>1</i>) senescent cell type, <i>2</i>) senolytic mechanisms and off-target susceptibility, and <i>3</i>) the disease stage at the time of intervention can help explain divergent outcomes. By addressing these three key points, this review will identify practical considerations for advancing "precision senotherapy" in PAH, including dosing strategies favoring intermittent, short-course senolytic regimens, and combination treatment approaches. Together, these concepts provide a framework for designing safer, stage-aware, cell-informed senotherapeutic studies in PAH.

Directional cues are essential for orienting cells during tissue morphogenesis and repair. In peripheral nerve regeneration, Schwann cells (SCs) align longitudinally in the nerve bridge to guide axonal pathfinding, but the mechanisms are not fully understood. We show here that after nerve injury, activated SCs up-regulate the guidance receptor Plexin-B1, enabling membrane plasticity required for SC polarization and longitudinal alignment along the axons. Aligned axon-SC provides positional cues to orient macrophages and extracellular matrix. Loss of Plexin-B1 disrupts SC morphological transformation, contact inhibition of locomotion between SCs, and axon-SC alignment, leading to SC misorientation, excessive inflammation, and delayed axon regeneration and functional recovery. These findings identify Plexin-B1 as a key orchestrator to orient SCs by regulating both SC-SC and axon-SC interactions during nerve repair. Elucidating the mechanisms of spatial guidance in nerve repair after injury has potential implications for therapeutic strategies to enhance neural regeneration.

<h4>Introduction</h4>Adhesion G-Protein Coupled Receptors (aGPCRs) represent the second largest GPCR family and consist of 32 members in humans, of which 19 are orphan receptors. Despite making up a substantial portion of GPCRs, there are no clinically approved therapeutics targeting them or using them in clinical applications. aGPCRs have functional roles in numerous tissues and their dysregulation through mutations or expression is associated with a variety of diseases, including cancer. Therefore, there is strong interest in therapeutic targeting aGPCRs or their controlled pathways.<h4>Areas covered</h4>The authors showcase the currently available approaches (small molecules, antibodies, knockouts/downs, peptides, nanoparticles, and cell-based therapies) that modulate aGPCR function or pathways. Furthermore, the authors discuss aGPCRs with potential for future therapeutic targeting. Of all aGPCRs, 14 are being exploited as targets and 7 have potential. For the remaining 11, a brief description of their function and any known involvement in disease were included.<h4>Expert opinion</h4>The field is currently hampered by lack of knowledge about endogenous ligands and downstream signaling for aGPCRs. Once this knowledge gap is overcome, a clear library of agonists and antagonists can be developed, unleashing successful and widespread aGPCR exploitation and modulation. In the coming years, ligand identification will accelerate, allowing more aGPCR therapeutic advances.

One of the most direct methods for synthesizing internal alkenes is the isomerization of terminal alkenes. Currently, noble metal catalysts are the most commonly used for alkene isomerization. However, the high cost and limited availability of late transition metals have prompted growing interest in replacing these precious metals with more readily available, earth-abundant alternatives. Herein, we report an iron-catalyzed alkene isomerization conducted in absence of solvent. A well-defined mononuclear Fe(II) complex, [HFe(CO)₄SiPh₃] ([Fe-1]), efficiently catalyzes the isomerization of terminal alkenes to internal alkenes under mild conditions. To demonstrate the synthetic utility of this transformation, gram-scale isomerization reactions were successfully performed. Kinetic investigations reveal that the reaction follows approximately first-order kinetics with respect to the iron catalyst, with an initial reaction rate of 7.08 × 10^-3 Mmin^-1. Furthermore, the tandem functionalization of terminal alkenes to aldehyde products was achieved via sequential isomerizing-ozonolysis and isomerizing-hydroformylation reactions. The complementarity of the two catalysts (Fe and Rh) was conclusively demonstrated through mechanistic investigations and control experiments.

Beta-caryophyllene (BCP) is an agonist of cannabinoid receptor 2 (CNR2/CB2), exerting mostly anti-inflammatory effects in rodents, but not explored in cattle. Herein we conducted the first reductionist step to examine the effects of in-vitro supplementation of BCP in bovine peripheral blood mononuclear cells (PBMCs) on the inflammatory response under thermoneutral (TN) or heat stress (HS) conditions. We performed this work in two cohorts: in Study 1, PBMC were supplemented with BCP at 1 µM (n = 8), and in Study 2 PBMC from other cows were supplemented with BCP at 1 mM (n = 7). In each study, PBMC from each cow were assigned to controls (CTL) or BCP-supplemented and incubated in TN (38.5 °C) or HS (41.0 °C) conditions. Then, half of the cells from each cow were treated with lipopolysaccharide (LPS), and media were collected for analysis of cytokines/chemokines and cyclic adenosine monophosphate (cAMP) concentrations as indicator of CB2 activation. In addition, RNA and proteins were extracted from plate-adhered monocyte-enriched-PBMCs (MEPs), for gene-expression and proteomics analyses. In each study, the effects of BCP, LPS, HS, and interactions were analyzed. We found in both studies that LPS and BCP (at 1 µM or at 1 mM) lowered CNR2 gene expression in MEPs. At 1 mM, BCP inhibited gene expression of LPS-induced NFKB1, amplified TNFA, and suppressed IL10, and tended to reduce cAMP in medium, possibly indicating CB2 activation. Post-LPS, BCP (1 mM) increased monocyte-chemoattractant-protein-1 and tended to enhance interleukin-1-alpha, beta, and macrophage-inflammatory-protein-1-alpha in medium; while HS increased macrophage-inflammatory-protein-1-beta. Proteomics of LPS-activated MEPs revealed immune-related proteins and canonical pathways enriched by BCP. Overall, BCP supplementation in vitro modulated immune function in PBMC from dairy cows, with similar effects in TN and HS conditions; thus, BCP could be studied as a potential feed supplement in cattle that may induce moderate immune-modulating effects. Further studies should investigate the in vivo effects of BCP on immune function in cattle.

Epigenome-wide studies of pancreatic islets provide valuable insights into type 2 diabetes (T2D) but lack methylomes from individual cell types. Here we show changes to alpha and beta cell-specific methylomes and transcriptomes from people with or without T2D, using whole-genome bisulfite sequencing and RNA sequencing. We discover 22,544 differentially methylated regions annotated to 7,975 genes in alpha versus beta cells, such as INS, GCG, PDX1 and PCSK1, with ~50% showing differential expression. CRISPR-dCas9-DNMT3A-based epigenetic editing increases INS and TH DNA methylation, while CRISPR-dCas9-TET1-based editing decreases GCG methylation, each altering INS, TH or GCG expression and content in beta cells. Pre-T2D/T2D-associated differentially methylated regions in alpha and beta cells overlap 12-18% of T2D-associated genome-wide association study candidates. Additionally, ONECUT2 is epigenetically upregulated in beta cells from people with pre-T2D/T2D and elevated in male Goto-Kakizaki rat islets. ONECUT2 overexpression in beta cells/islets downregulates gene sets impacting insulin secretion and glucose homeostasis, and reduces mitochondrial activity, ATP/ADP ratio and insulin secretion. We also provide 'alpha-beta-methylome' ( https://alpha-beta-methylome.serve.scilifelab.se/app/alpha-beta-methylome/ ), a resource exploring T2D, age and sex associations on methylation, highlighting cell-specific epigenetic regulation and dysfunctions contributing to T2D.

Metastatic cancer cell dormancy, wherein disseminated cancer cells (DCCs) persist in a quiescent state before reactivating to fuel metastasis, has emerged as a critical determinant of cancer relapse. In this Review, we synthesize recent advances in understanding the microenvironmental drivers of dormancy, including the role of niche-derived signals and extracellular matrix composition in maintaining DCC quiescence, as well as the epigenetic and transcriptional programmes, and chromatin remodelling that enforce and sustain dormancy. We also cover the mechanisms by which dormant DCCs evade immune surveillance, highlighting both innate and adaptive immune interactions, and the strategies tumours use to escape immune-mediated clearance. Although most data come from solid cancers, we also examine the biology of residual cells in haematologic malignancies that share key dormancy and relapse mechanisms with solid tumours. We also discuss how, despite these mechanistic insights, clinical translation remains limited, as available biomarkers or therapies targeting dormancy have yet to be effectively implemented. We conclude that by outlining the challenges and opportunities for leveraging dormancy biology, we may be able to prevent metastatic recurrence and improve patient outcomes.

Diverse risk genes have been identified for neurodevelopmental disorders (NDDs), but how these genes converge on similar biological pathways in neurons, and thus give rise to similar phenotypes, is unclear. Here we apply a pooled CRISPR approach to successfully target 23 NDD loss-of-function genes with roles in chromatin biology and examine convergent effects on gene expression across human induced pluripotent stem cell-derived neural progenitor cells, glutamatergic neurons and GABAergic neurons. Points of convergence vary between these cell types, with the greatest number of convergent genes and strongest convergent networks in mature glutamatergic neurons, where they broadly represent synaptic, epigenetic and, unexpectedly, mitochondrial pathways. The most convergent networks were observed between NDD genes with shared biological annotations, clinical associations and co-expression patterns in human post-mortem brain. Drugs that were predicted to reverse convergent transcriptomic signatures and/or arousal and sensory processing behaviors ameliorated behavioral phenotypes in zebrafish NDD gene mutants. These results suggest that convergent effects of NDD risk genes could provide clinically useful insights.

BACKGROUND: Multi-analyte plasma proteomic panels that can accurately detect initial Alzheimer’s disease (AD) pathology in preclinical populations and simultaneously measure related biological processes relevant for disease risk are critical for advancing early detection and prognosis. METHODS: Using the NULISAseq™ CNS panel, we measured plasma proteomics from 315 clinically unimpaired (CU) older adults across two independent cohorts: the Stanford Aging and Memory Study (SAMS; n = 193) with paired cerebrospinal fluid (CSF) and plasma analyzed with Lumipulse, and the Attention, Memory, and Aging Study at Stanford (AMASS; n = 122) with paired florbetaben (FBB) amyloid PET. We evaluated correspondence of core AD-relevant biomarkers pTau217, pTau181, Aβ42/Aβ40, pTau217/Aβ42, GFAP, and NfL measured using multiplex NULISAseq and single-plex Lumipulse immunoassays. ROC curve analyses compared performance for detecting amyloid-positivity (A+) (a) across platforms in SAMS and (b) across brain-derived (BD) and total-pTau assays in AMASS, leveraging novel NULISAseq immunoassays. Linear models were applied across all NULISAseq CNS proteins to explore proteomic abundance patterns associated with age, sex, APOE-ε4, amyloid burden (CSF Aβ42/Aβ40, amyloid PET), and tau burden (CSF pTau181, PI-2620 tau PET) using an FDR-corrected p-value of < 0.05 to identify significant targets. RESULTS: In SAMS, moderate to high correlations were observed between NULISAseq and Lumipulse plasma biomarkers. NULISAseq pTau217/Aβ42 (AUC: 0.940) and pTau217 (AUC: 0.879) performed as well as corresponding single-plex Lumipulse assays (pTau217/Aβ42, AUC: 0.907; pTau217, AUC: 0.838) for detecting CSF A+ in SAMS. In AMASS, BD-pTau217 (AUC: 0.920) and BD-pTau181 (AUC: 0.920) exhibited the highest performance in discriminating PET A+, providing significant performance gains compared to total-pTau measures (pTau217, AUC: 0.861; pTau181, AUC: 0.763). Exploratory proteomic abundance analyses across NULISA CNS targets revealed pTau isoforms as most differentially expressed with amyloid burden across cohorts, together with upregulation of GFAP and downregulation of Aβ42 in SAMS. Tau burden was associated with upregulation of plasma pTau217, independent of amyloid burden, together with proteins related to astrocyte activation, inflammation, and synaptic integrity. CONCLUSIONS: NULISAseq multiplex immunoassays, including novel BD-pTau assays, accurately detect AD pathology among CU older adults and identify multiple biological pathways related to aging and early biomarker abnormality that may become dysregulated in preclinical AD.

Huntington's disease (HD) is an inherited neurodegenerative disorder caused by an expansion of a CAG trinucleotide repeat in the huntingtin (HTT) gene, which leads to a mutant protein that destroys neurons in the brain. Despite intense effort, there remains no approved disease-modifying therapy for HD. Here, we develop a pan-HTT-targeting CRISPR-Cas9 system that, when delivered to the striatum of R6/2 and YAC128 mice by adeno-associated virus serotype 5 (AAV5), lowered mutant HTT mRNA and protein by 55%-80% via its induction of frameshift-inducing insertion or deletion (indel) mutations in HTT exon 1. Cas9 targeting improved motor coordination and locomotor activity, decreased anxiety-like deficits, reduced clasping and weight loss, limited striatal atrophy, and decreased the formation of intranuclear inclusions immunoreactive for the mutant HTT protein. In Hu21/21 mice, which carry the wild-type human HTT gene in lieu of the mouse ortholog, Cas9 lowered the HTT protein by 44% but induced no measurable behavioral deficits and had no adverse effect on neuronal viability, though its targeting was associated with neuroinflammation. Altogether, our results demonstrate the ability of a newly developed pan-HTT-targeting Cas9 system to affect HD-related phenotypes across models and provide insights into its tolerability.

Pathogenic variants of NOTCH2 that lead to a gain or loss of function are associated with serious clinical consequences. A recently reported novel NOTCH2 4006G > C variant was associated with decreased Notch signaling and skeletal fragility in humans and mouse (Notch2^em1Ecan) lines. In the present study, we analyze the transcriptome of femoral bones from mature control and Notch2^em1Ecan mice following the exclusion of hematological cells. Single cells were analyzed following microfluidic partitioning on a Chromium X instrument using a 3' gene expression library. Uniform manifold approximation and projection (UMAP) for non-linear dimensional reduction defined 15 different cell clusters comprised of macrophages, red blood cells, endothelial, vascular and smooth and striated muscle cells and cells of the osteoblast lineage. Notch2 and Hes1 were the prevalent Notch receptor and target gene in most clusters. Independent clustering analysis of transcriptomes from control and Notch2^em1Ecan femurs revealed a 35-45% decrease in cells forming the osteogenic and a 50-55% decrease in cells comprising the vascular cluster in Notch2^em1Ecan femurs. Expression of Hes1 was decreased in Notch2^em1Ecan cells. Analysis of nascent transcripts (intron/exon sequences) using the scVelo pipeline revealed suppressed velocity in vascular and osteogenic clusters from Notch2^em1Ecan cells implying decreased transcription of genes constituting these clusters. In conclusion, a novel NOTCH2 deleterious variant associated with skeletal fragility alters the osteogenic and vascular transcriptome in femurs from adult mice.

<h4>Background</h4>There is a paucity of data regarding short-term outcomes and treatment responsiveness of myocardial T1 mapping via cardiac magnetic resonance (CMR) in patients with transthyretin amyloid cardiomyopathy (ATTR-CM) undergoing tafamidis therapy.<h4>Methods</h4>We retrospectively studied 60 wild-type ATTR-CM patients who underwent baseline CMR to measure native myocardial T1 value (T1_native) and extracellular volume fraction (ECV), followed by tafamidis treatment. Cardiac biomarkers, including high-sensitivity cardiac troponin T and N-terminal pro-B-type natriuretic peptide (NT-proBNP), were measured at baseline. We followed a one-year composite of worsening heart failure (WHF; hospitalization and/or intensification of diuretic therapy for heart failure). Additionally, 51 patients underwent follow-up CMR and measurements of cardiac biomarkers one-year after the initiation of tafamidis treatment.<h4>Results</h4>Patients with WHF (n = 12) exhibited significantly elevated baseline T1_native and ECV than those without WHF, and their optimal cutoffs in predicting WHF were 1447 ms and 48.7 %, respectively. Multivariate analysis adjusted for Mayo or National Amyloidosis Center stages identified T1_native of ≥ 1447 ms as an independent predictor of WHF, with hazard ratios of 15.2 and 9.3, respectively. A notable proportion of patients exhibited a reduction in T1_native (39 %) and ECV (47 %) after one year of tafamidis treatment. Post-treatment changes in T1_native were correlated positively with changes in NT-proBNP concentration (r = 0.40, <i>p</i> = 0.0036).<h4>Conclusions</h4>In wild-type ATTR-CM patients receiving tafamidis, elevated baseline T1 mapping parameters were associated with one-year WHF. T1 mapping parameters, particularly T1_native, may offer imaging-based evidence of alterations in myocardial characteristics induced by tafamidis.

<h4>Background</h4>Hyperkinetic movement disorders (MDs) in children, including tremor, myoclonus, dystonia, chorea, and tics, often present with overlapping or evolving clinical features, making classification based on observation alone challenging. Neurophysiological techniques such as video-EEG polymyography (P-VEEG) and somatosensory evoked potentials (SEPs) may provide objective markers to improve diagnostic accuracy, yet their application in pediatric populations remains limited.<h4>Objective</h4>To describe the polygraphic characteristics of hyperkinetic MDs in a large pediatric cohort and to assess whether specific EMG patterns can support movement disorder classification beyond clinical inspection.<h4>Methods</h4>Seventy-three patients with pediatric-onset hyperkinetic MDs underwent standardized P-VEEG recordings and SEPs at Besta Neurological Institute between January and October 2024. EMG activity was recorded from antagonist muscle pairs during rest, posture maintenance, and specific tasks according to age. Polygraphic findings were compared with clinical diagnoses. Associations between EMG patterns and clinical variables were analyzed using Chi-square tests and regression analyses.<h4>Results</h4>Mean age at MD onset was 9.4 ± 6.1 years and mean age at recording was 13.5 ± 5.1 years. Polygraphic analysis identified four distinct EMG patterns: (i) regular rhythmic oscillatory pattern (RRO), consistent with tremor, in 40 patients (56%); (ii) repetitive myoclonic pattern (RM) in 11 (15%); (iii) complex mixed repetitive pattern (CR) in 9 (12%); and (iv) complex non-rhythmic pattern (CNR), encompassing chorea, dystonia, tics and other dyskinesias, in 11 (15%). Significant differences among repetitive patterns were observed in rhythmicity (<i>p</i> < 0.001), burst duration (<i>p</i> < 0.001), agonist-antagonist synchrony (<i>p</i> < 0.001), and SEP hyperexcitability (<i>p</i> < 0.03). RM was significantly associated with enhanced amplitude SEPs (<i>p</i> = 0.023), supporting a cortical origin of myoclonus in most cases. A significant association between clinical diagnosis and EMG pattern was found (<i>p</i> = 0.003), yet discrepancies between clinical and polygraphic classification was found in 22 cases (30%), particularly among tremor and myoclonus presentations. RM and CR were prevalent in older patients compared with CNR.<h4>Conclusion</h4>P-VEEG proved to be an easily applicable tool providing valuable objective markers for the characterization of pediatric hyperkinetic MDs, revealing distinct EMG patterns that complement clinical evaluation and improve diagnostic precision. Systematic integration of neurophysiological assessment may enhance classification, diagnostic accuracy and help guide management in childhood movement disorders.

Runx2 is a key regulator of osteoblast differentiation and chondrocyte maturation. However, the distinct functions of the two functional isoforms remain to be clarified. Transcription of two isoforms, Runx2-I and Runx2-II, starts from the proximal and distal promoters upstream of exons 2 and 1, respectively. To investigate the functions of the two isoforms, we generated a novel mouse model (<i>Runx2</i>-br^mut/mut), in which intron 1 splicing for <i>Runx2</i>-II was disrupted by mutating the branch site essential for splicing. <i>Runx2</i>-II was severely reduced but <i>Runx2</i>-I was increased in <i>Runx2</i>-br^mut/mut mice as compared with those in <i>Runx2</i>-br^wt/wt mice. Although <i>Runx2</i>-II was about three times higher than <i>Runx2</i>-I in E15.5 limbs and newborn calvaria of <i>Runx2</i>-br^wt/wt mice, <i>Runx2</i>-II was extremely lower than <i>Runx2</i>-I in <i>Runx2</i>-br^mut/mut mice. Endochondral ossification was retarded in <i>Runx2</i>-br^mut/mut mice, but the delay was milder than in <i>Runx2</i> ^+/- mice, and the primary spongiosa formation was impaired due to the reduced osteoblasts. The development of calvaria in the newborn was retarded similar to <i>Runx2</i> ^+/- mice, which showed cleidocranial dysplasia, but it was much less affected than in <i>Runx2</i> ^+/- mice at 8 weeks of age, and the suture mesenchymal cell proliferation increased. Furthermore, clavicle development was less affected than that in <i>Runx2</i> ^+/- mice throughout their lives. The trabecular and cortical bones in the femurs of <i>Runx2</i>-br^mut/mut mice were lower than those of <i>Runx2</i>-br^wt/wt mice owing to the reduced bone formation, and the strength of the bones was also weaker. Osteoblast differentiation was impaired in <i>Runx2</i>-br^mut/mut mice. Overexpression of <i>Runx2</i>-II failed to affect endogenous <i>Runx2</i> expression <i>in vitro</i>, and <i>Runx2</i> knockdown by siRNA failed to affect the proximal promoter activity. These findings indicated that both isoforms contribute to endochondral ossification, <i>Runx2</i>-I can compensate for <i>Runx2</i>-II in endochondral ossification, Runx2-II plays important roles in osteoblast differentiation, and Runx2-I plays important roles in the development of calvaria and clavicles, at least in part, by enhancing suture mesenchymal cell proliferation. Our findings also showed that a minimal amount of Runx2-II is necessary for the efficient function of Runx2-I, indicating a basal requirement of Runx2-II in bone development, but that <i>Runx2</i> expression is not autoregulated by Runx2.

Epstein-Barr virus (EBV) is associated with a wide spectrum of lymphoid and epithelial malignancies. This review provides a comprehensive pan-tumor perspective on the convergent oncogenic mechanisms, immune microenvironment characteristics, and shared therapeutic vulnerabilities across EBV-related cancers. We systematically summarize EBV infection and latency patterns, highlighting key shared signaling networks (e.g., NF-κB, PI3K/AKT) alongside nuanced immune evasion strategies, such as HLA modulation and steric immune blockade. By bridging these molecular insights with clinical translation, we evaluate emerging therapeutic paradigms, including immune checkpoint inhibitors, EBV-specific T-cell therapies, neutralizing antibodies, and mRNA vaccines. Ultimately, this synthesis underscores the potential of exploiting shared viral vulnerabilities to develop pan-tumor targeted therapies, offering a strategic roadmap for future clinical development.

Hematologic disorders, including malignant and autoimmune conditions, present persistent clinical challenges characterized by relapse, treatment resistance, and profound immune dysregulation. While conventional immunotherapies have advanced, their efficacy is frequently limited by HLA downregulation and effector T cell exhaustion. In this context, γδ T cells offer a promising therapeutic alternative. Recognizing antigens independently of MHC restriction, γδ T cells possess intrinsic tissue-homing capabilities and exhibit dual cytotoxic and immunoregulatory functions. These properties make them highly suitable candidates for allogeneic, "off-the-shelf" cellular therapies where αβ T cells face alloreactive limitations. This review systematically synthesizes the immunobiology of γδ T cells, exploring the functional heterogeneity of specific subsets and their regulation within the tumor microenvironment (TME). We critically evaluate recent preclinical and clinical evidence supporting adoptive transfer, CAR-γδ T strategies, and combination regimens across acute leukemias, lymphomas, multiple myeloma, and immune cytopenias. Furthermore, we address critical translational barriers-including <i>in vivo</i> persistence, subset exhaustion, and manufacturing variability-and discuss rational engineering strategies, metabolic preconditioning, and epigenetic modulation as solutions. Ultimately, advancing γδ T cell therapies requires overcoming these hurdles to transition them effectively from the bench to mainstream clinical practice.

With limited therapeutic progress despite aggressive multimodal treatment, glioblastoma (GBM) remains one of the deadliest primary brain tumors. Emerging evidence suggests that GSCs are key drivers of tumor initiation, intratumoral heterogeneity, therapeutic resistance, and recurrence. GSCs retain self-renewal capacity, multilineage differentiation potential, and remarkable plasticity, enabling them to adapt to diverse microenvironmental cues. These properties are upheld by dysregulated developmental and oncogenic signaling pathways, including Notch, Wnt/β-catenin, Hedgehog, PI3K/AKT/mTOR, and STAT3, as well as epigenetic and metabolic reprogramming. In addition, dedicated niches such as hypoxic and perivascular microenvironments critically support GSC maintenance and immune evasion. In this review, we summarize the current understanding of the molecular pathways governing GSC biology, examine their interactions with the tumor microenvironment, and discuss emerging therapeutic strategies targeting GSCs, including pathway inhibition, differentiation therapy, immunotherapy, and nanomedicine-based drug delivery. We highlight key challenges and future directions for translating GSC-targeted approaches into effective clinical interventions for GBM.

Peripheral nerves provide a direct connection between the brain and the tumor microenvironment. This connection allows the nervous system to influence processes associated with the development, progression, and metastasis of different tumor types. Therefore, tumor innervation by peripheral nerve fibers is currently emerging as a characteristic that contributes to multiple hallmarks of cancer. Several experimental studies have shown that cancer progression involves actively inducing the ingrowth of autonomic and sensory nerve fibers into tumor tissue. In this process, known as neoaxonogenesis, cancer and other cells in the tumor microenvironment play an important role by synthesizing and releasing neurotrophic factors (e.g., nerve growth factor, brain-derived neurotrophic factor, glial cell line-derived neurotrophic factor), axonal guidance molecules (netrins, semaphorins, ephrins, slits), exosomes (containing microRNA and axonal guidance molecules), and other molecules present in the tumor microenvironment (e.g., granulocyte colony-stimulating factor, leukemia inhibitory factor), which modulate the ingrowth of nerve fibers into the tumor. This results in an increased nerve supply to tumor tissue, which is primarily linked to its growth. However, there are also studies demonstrating the protective effects of increased nerve fiber density against processes associated with cancer progression in certain types of cancer. The findings from these studies contribute to the complexity of neuro-cancer interactions, which is probably based on the type of cancer and the physiological specializations of the nerve fibers in a given organ. Despite contrasting findings, the stimulatory effects of nerve fibers on cancer growth are supported by several studies that described reducing the negative impact of nerve fibers on tumors and thus inhibiting cancer progression. The most significant approaches to reducing neural effects appear to be denervation, the administration of neurotransmitter receptor antagonists, the administration of local anesthetics, and the administration of antibodies against neurotrophic factors. Other significant approaches include methods that improve quality of life, such as psychotherapy and heart rate variability biofeedback. Despite their therapeutic potential, there are several limitations to using approaches that manipulate cancer innervation in clinical practice. These limitations include impaired normal tissue function and nervous system function, as well as the problematic direct application of the therapeutic agent to the tumor site, dosage-dependent, cancer type-dependent, cancer stage-dependent, duration-dependent, and timing-dependent effects. Procedures that modify neoaxonogenesis and nerve fiber signaling appear to be a promising new therapeutic approach in oncology. However, more research is needed to better understand their effects on cancer progression. In the future, the assessment of the presence and density of nerve fibers in tumors, as well as the evaluation of approaches aimed at reducing their negative impact, could be part of personalized anticancer therapy. As part of this therapy, a fresh tumor sample would be collected from the patient to generate patient-derived organoid models to test and consider the possibility of using supportive therapy and to predict its efficacy. Based on these results, it would be possible to evaluate the applicability of nerve-fiber-targeted therapy for a given patient. This review article summarizes and describes the current knowledge concerning the significance of nerve fibers in cancer progression, with a particular emphasis on neoaxonogenesis in tumors and the various factors that influence this process.

Delayed cerebral ischemia (DCI) is a major complication of aneurysmal subarachnoid hemorrhage (aSAH), strongly associated with neurological deterioration and poor outcomes. Its pathophysiology remains incompletely understood and involves multiple interacting processes. Increasing evidence highlights the role of redox imbalance triggered by hemoglobin breakdown and the subsequent generation of reactive species, leading to vascular dysfunction, impaired nitric oxide signaling, and inflammatory activation This review aims to summarize current knowledge on redox-related mechanisms involved in DCI and to explore the potential role of the peroxiredoxin (PRDX) family in this setting. A narrative review of experimental and preclinical studies was performed, focusing on molecular pathways associated with vascular regulation, cellular injury, and antioxidant defense. Particular attention was given to the distribution and biological functions of PRDX isoforms within the central nervous system. This work addresses a topic not previously systematically discussed, the potential involvement of PRDX proteins in aSAH-related complications. By integrating available data, it provides a conceptual framework linking PRDX to mechanisms relevant for DCI. The manuscript serves as a starting point for future research, particularly translational and clinical studies in humans, which are necessary to verify the relevance of these findings and to better understand their potential clinical implications.

Cartilage is a strong, yet flexible, type of connective tissue found in various parts of the body. Cartilage that covers the ends of bones in joints, referred to as articular cartilage, is frequently damaged during injuries and osteoarthritis (OA). Lack of blood vessels and nerves in the cartilage limits its self-renewal capacity, rendering cartilage regeneration a critical challenge. Three-dimensional (3D) bioprinting provides a promising approach for cartilage regeneration by depositing bioink layer by layer to create a 3D structure of more complex tissues such as cartilage, with clinically relevant sizes and mimicking the native shape and natural microenvironment. The present review discusses the potential of bioprinting materials to direct the fate of induced pluripotent stem cells (iPSCs) towards chondrocytes, while simultaneously enhancing lubrication. This will be achieved by incorporating iPSCs, natural polymers, synthetic polymers and lubricants such as lubricin, growth factors and signaling molecules that are involved in chondrogenesis. By optimizing bioink formulation and other parameters in bioprinters, the production of tissues in a precisely controlled manner is ensured, and limitations in current treatments for degenerative diseases such as OA can be avoided. Researchers have developed various complex and functional tissues and organs using bioprinting with successful animal trials, and are in the process of developing patient-specific iPSCs to generate personalized cartilage grafts having high and prolonged regenerative ability with further enhanced lubrication. The present review aimed to provide hope for patients suffering from cartilage degeneration by demonstrating the potential of 3D bioprinting for effective cartilage regeneration.

The use of minor cannabinoids has been advanced, in part, by the idea of providing relief from pain and inflammation without the unwanted psychogenic effects associated with delta-9-tetrahydro-cannabinol (Δ⁹THC). With a focus on peripheral nociception, 4 common minor cannabinoids: cannabidiol (CBD), cannabigerol (CBG), cannabinol (CBN), and cannabichromene (CBC) were studied in primary cultures of mouse dorsal root ganglion (DRG) neurons. We queried whether calcium responses induced by the 4 cannabinoids differed in potency of activation, neuronal size preference, and dose-response relationships. Additionally, we determined the dependence of CBD and CBN on key channel receptors that are known to mediate pain and/or antinociception. Individually, CBD, CBG, and CBC directed greater response magnitudes when compared with CBN. All 4 minor cannabinoids activated overlapping size populations of sensory neurons. CBD and CBG activated the widest range of DRG neuron sizes (smaller-larger), overlapping with smaller capsaicin-sensitive neurons. In contrast, CBN and CBC activated predominantly larger sensory neurons. CBD diverged from other minor cannabinoids in directing a linear dose-response profile, whereas CBG and CBC directed sigmoidal profiles and CBN activated DRG neurons with an inverted U-shaped dose-response relationship. CBD-induced activation of DRG neurons was dependent on coexpression of the nociceptive channel transient receptor potential cation channel subfamily Vanilloid member 1 (TRPV1) plus cannabinoid receptor 1 (CB₁R), whereas CBN-induced activation was independent of TRPV1 and CB₁R. Overall, we observed that minor cannabinoids CBD, CBG, CBN, and CBC directed unique activation properties across a diverse population of sensory neurons. Such differences underlie the hypothesis that a combination of minor cannabinoids can direct complementary antinociceptive activity. SIGNIFICANCE STATEMENT: Minor cannabinoids CBD, CBG, CBN, and CBC differ in their dose-dependent properties of sensory neuronal activation. CBD-induced activation of small to large sensory neurons was dependent on coexpression of the nociceptive channel TRPV1 plus CB₁R and directed a linear dose-response profile. In contrast, CBN activated predominantly larger sensory neurons in an inverted U-shaped dose-response profile and was independent of TRPV1 and CB₁R. A combination of minor cannabinoids is hypothesized to direct complementary antinociceptive activity.

Modified peptide derivatives (<b>D1-D15</b>) containing hydrophobic (phenylalanine and tryptophan) and positively charged (histidine) amino acid residues, were designed and synthesized. The cytotoxicity of the compounds was evaluated in healthy (CCD1079Sk) cell lines, revealing no significant cytotoxic effects and indicating favorable safety profiles. Molecular modeling studies were performed for all compounds and indicated that compounds <b>D13</b>, <b>D14</b>, and <b>D15</b> form favorable binding interactions with the Nsp3 macrodomain 1 (Mac1) active site. The affinity of these compounds is expected to be moderate to high, although lower than that of ADP-ribose. Subsequent bioactivity assays, performed using a cell-based SARS-CoV-2 replicon system, revealed that compound <b>D15</b> displays the most potent inhibition of viral replication (IC₅₀ = 22.2 μM (95% CI: 15.4-35.7 μM)), while compound <b>D14</b> shows the second most potent inhibitory activity (IC₅₀ = 61.2 μM (95% CI: 39.2-143.9 μM)). Considering these results, it has been shown that compounds with a histidine side chain exhibit higher biological activity, suggesting a structure-activity relationship driven by positively charged residues.

<h4>Background and purpose</h4>Microfluidic technology provides enhanced standardization for formulating nanoencapsulated drugs compared to traditional bulk methodologies. Here, microfluidic nano-niosomal deferoxamine (Nn-DFO) was compared to micro-niosomal (Mn-DFO) and liposomal (L-DFO) formulations synthesized by conventional thin-film hydration.<h4>Experimental approach</h4>The formulations were subjected to physicochemical analyses. <i>In vivo</i> functional efficacy and safety were analyzed in mouse models of iron overload. Urinary and fecal iron excretion were also assessed.<h4>Findings/results</h4>Mean particle sizes were 180.89±74.34 nm for L-DFO, 87.06±2.67 nm for Nn-DFO, and 7.41 ± 0.19 μm for Mn-DFO. Encapsulation efficiencies of L-DFO, Nn-DFO, and Mn-DFO formulations were 64.54%, 57.1%, and 70.72%, respectively. The 4-h drug release rates for L-DFO, Nn-DFO, and Mn-DFO formulations were 20.95%, 31.34%, and 26.30%, respectively, compared to 59.46% release from free DFO (F-DFO) within 4 h. Animals treated with F-DFO, L-DFO, Nn-DFO, and Mn-DFO showed significant reductions in the iron content of the liver (45.03%, 49.36%, 41.63%, and 23.52%, respectively). Urinary iron excretion on day 1^st and fecal iron excretion on day 3^rd after drug administration were the highest in the groups treated with F-DFO and Nn-DF O compared to other groups. Fecal iron excretion on day 7^th was the highest in mice receiving Nn-DFO compared to different formulations.<h4>Conclusion and implications</h4>Nn-DFO synthesized by the microfluidic approach showed iron chelation efficacy and immediate iron excretion profile comparable to F-DFO, while being superior to Mn-DFO and L-DFO regarding delayed iron excretion.

Extracellular vesicles (EVs) serve as critical regulators of intercellular communication by carrying diverse molecular cargos which reflect cellular states associated with environmental stressors. We investigated transcriptomic profiles of EVs derived from human skeletal muscle tissue chips flown abroad the SpaceX CRS-25 mission to characterize age- and microgravity-induced adaptation. EVs were isolated using a novel magnetic capture-release method (ExCy) from conditioned media of muscle chips cultured in microgravity or ground control conditions. Next-generation sequencing and differential gene expression analysis were performed on EV RNA. In this exploratory study, we identified age- and spaceflight-specific transcriptomic signatures. Young donor-derived EVs in microgravity appear to exhibit upregulation of oxidative stress and signaling pathways, while older donor-derived EVs showed markers of mitochondrial quality control, ER stress, and proteostasis failure. Muscle-derived EVs capture environment- and age-dependent molecular signatures in space, supporting their use as non-invasive biomarkers for physiological stress and tissue adaptation under microgravity.

Structurally complex plasmonic nanoarchitectures represent an emerging class of nanomaterials with properties that extend beyond those of conventional spherical nanoparticles. Their distinctive structural motifs generate dense near field electromagnetic hot spots, expand interfacial surface area, and create biophysical environments at the nano-bio interface that can actively engage cellular signaling networks relevant to neural regeneration and aging. Despite growing interest in these platforms, a systematic, omics-guided synthesis that links nanoparticle structural features to transcriptomic programs and regenerative outcomes has been lacking. In this review, we summarize recent advances in high complexity plasmonic nanoparticle engineering and integrate published omics-based evidence of their cellular effects, organizing the discussion. Across these studies, transcriptomic analyses of nanoparticle treated neural systems consistently highlight three convergent biological themes: mitigation of oxidative stress and activation of antioxidant pathways, suppression of neuroinflammatory signaling, and induction of neuronal developmental and plasticity programs. Collectively, the omics-guided findings synthesized here suggest that structural complexity in plasmonic nanoarchitectures is not merely a synthetic achievement but a tunable determinant of cellular state, with important implications for the rational design of regenerative nanomedicines targeting neurodegenerative diseases and age-related neuronal decline.

<h4>Background/objectives</h4>Hepatic diseases frequently present with ocular manifestations that aid diagnosis, provide prognostic data, and guide therapy. Despite the clear utility of the liver-eye axis, the literature lacks reviews that categorize these manifestations by etiology. This review evaluates current evidence to identify ocular findings that serve as clinical tools for diagnosis, prognosis, and therapeutic monitoring of hepatic pathologies.<h4>Methods</h4>A narrative review was conducted using PubMed and Google Scholar to identify English-language articles addressing ocular manifestations associated with liver disease. The primary search encompassed publications from 2000 to 2025, with inclusion of select foundational works published prior to 2000 when they represented seminal studies establishing diagnostic criteria, pathophysiological mechanisms, or natural history data not superseded by subsequent research. Search terms included combinations of liver, hepatic, hepatitis, cirrhosis, cholestasis, eye, ocular, retina, cornea, sclera, conjunctiva, ophthalmic manifestations, and specific disease names. All study designs were eligible. Society guidelines, systematic reviews, and studies from high-impact journals were prioritized. The final selection comprised 59 references representing the most authoritative sources across the spectrum of hepatic conditions.<h4>Results</h4>A spectrum of ocular findings linked to distinct hepatic conditions was identified. Manifestations with established clinicopathologic associations were categorized into congenital and acquired etiologies. Congenital liver pathologies included metabolic disorders (Wilson disease, galactosemia, lysosomal storage disorders) and syndromic/genetic causes (Alagille syndrome, hereditary hemochromatosis). Acquired liver diseases encompassed infectious (hepatitis B/C), drug-induced and iatrogenic (interferon, immune checkpoint inhibitors), nutritional (vitamin A deficiency), neoplastic (metastatic hepatocellular carcinoma), and cirrhotic causes.<h4>Conclusions</h4>Specific ocular signs raise clinical suspicion for underlying liver disease and warrant targeted hepatic evaluation. Recognizing these associations facilitates earlier diagnosis and improves outcomes. Systematic screening for these signs is supported in at-risk populations, and prospective validation studies should establish their sensitivity and specificity.

BisphenolA (BPA) is a common endocrine disruptor that impairs male fertility through oxidative stress and alterations in membrane lipids. This study evaluated the protective effects of <i>Myrtus communis</i> L. essential oil (EOMC) on BPA-induced sperm toxicity in Wistar rats in vitro. BPA significantly decreased sperm motility and viability. It also increased lipid peroxidation, depleted thiols, and reduced the activity of antioxidant enzymes (SOD, CAT-like and GPx-like). Concomitant treatment with low and intermediate doses of EOMC (0.5-1 µL/mL) restored sperm function, reduced oxidative stress, and preserved membrane phospholipids. However, the highest dose (5 µL/mL) further impaired sperm function and disrupted membrane phospholipids. BPA also altered amino acid profiles and accumulated intracellularly, effects partially reversed by EOMC, which redistributed free BPA into the culture medium. Bioavailability analysis revealed selective absorption of α-pinene, while d-limonene and 1,8-cineole were undetectable. Molecular modeling indicated strong binding of BPA to antioxidant enzymes, potentially disrupting their structure and activity. Overall, these results show that EOMC protects sperm from BPA-induced damage in a dose-dependent manner through antioxidant, membrane-stabilizing, and redistribution mechanisms. This highlights its potential application in phytotherapy for male reproductive health.

<b>Background/Objectives</b>: Early detection through minimally invasive approaches is critical for timely patient stratification and optimal therapeutic decision-making in colorectal cancer (CRC). Liquid biopsy, based on the analysis of tumor-derived components in blood and other body fluids, has emerged as a promising strategy to overcome current limitations in CRC diagnosis and follow-up. This review evaluates the current landscape of liquid biopsy clinical trials in CRC, focusing on predictive biomarker detection, prognostic assessment, and disease monitoring. <b>Methods</b>: ClinicalTrials.gov was searched using the terms "colorectal cancer" and "liquid biopsy" yielding 153 registered trials. After manual screening, 44 trials were excluded for not using liquid biopsy for CRC management, leaving 109 trials for analysis. Of these, 25 were completed, and 13 had publicly available results related to liquid biopsy. <b>Results</b>: The included trials were conducted across 27 countries on four continents. Overall, 119 biomolecules assessments and 167 different endpoints were reported across 109 clinical trials. Because individual trials could evaluate multiple biomolecules and endpoints, counts exceed the total number of trials. Cell-free DNA (cfDNA) was evaluated in 92/109 trials (84%) and accounting for 77% of all biomolecule assessments. Circulatingtumor cells (CTCs) were analyzed in 9/109 trials (8%, representing 8% of all the biomolecules analyzed), and microRNAs (miRNAs) in 8/109 (7%, representing 7% of all the biomolecules analyzed). Treatment sensitivity was the most common endpoint (57/109, 52% of the clinical trials; representing 34% of all the 167 different endpoints analyzed), followed by disease progression (28/109, 26%; representing 17% of all the different endpoints analyzed) and diagnostic applications (21/109, 19%; representing 12% of all the different endpoints analyzed). Among the 25 completed studies, 10/25 (40%) were interventional and 15/25 (60%) observational, spanning 14 countries. The majority of completed trials (21/25, 84%) used cfDNA. Interventional studies were predominantly phase II (5/10), with fewer phase III trials (2/10), primarily evaluating treatment response, particularly in relation to EGFR inhibitors and RAS/BRAF mutation status. Four observational studies (4/15) investigated emerging biomarkers, including long noncoding RNAs and miRNAs. <b>Conclusions</b>: Current clinical trials highlight cfDNA as the dominant and most clinically advanced liquid biopsy biomarker in CRC, primarily used for treatment guidance and disease monitoring. In contrast, CTCs and RNA-based biomarkers remain underrepresented. The limited number of randomized late-phase trials, heterogeneity in study design, and technical challenges associated with emerging biomarkers underscore the need for standardized methodologies and robust validation before routine clinical implementation.

Serotoninomics, a nascent emerging discipline within the field of omics, provides a transdisciplinary framework for understanding reproductive toxicology via serotonergic signalling. This research investigates the neuroendocrine effects of permethrin, a commonly used pyrethroid insecticide often considered to pose a low risk to humans, and positions it as a model compound for evaluating reproductive susceptibility beyond conventional endocrine endpoints. It is hypothesized that serotonin, traditionally examined in neuropsychiatric contexts, plays an essential role in gonadal function, hormonal regulation, and emotional resilience. Although permethrins are generally regarded as safe, acute exposure may subtly interfere with serotonergic pathways, potentially resulting in molecular, biochemical, behavioural, and reproductive alterations. These effects could extend beyond immediate exposure, including during gestation, considering permethrins' ability to cross the placental barrier and influence foetal development. By synthesizing evidence across molecular, organismal, and environmental domains, we advocate for a serotonergic approach to facilitate a more comprehensive assessment of risk and resilience. We emphasize the importance of fostering a transdisciplinary dialogue to redefine reproductive health through the perspectives of serotonergic vulnerability and systemic resilience.

Tendon disease is a highly prevalent musculoskeletal disorder characterized by extracellular matrix (ECM) disorganization and fibroblast activation, contributing to fibrotic tissue healing and impaired function. While extracellular vesicles (EVs) have emerged as key mediators of intercellular communication and drivers of fibrosis in various tissues, their role in tendon pathology remains poorly understood. In this study, we developed a physiologically relevant 3D in vitro model that recapitulates biophysical features of healthy and fibrotic tendon microenvironments to investigate EV-mediated contributions to tendon remodeling. Primary tendon-derived cells on diseased scaffolds had increased proliferation, higher collagen III and fibronectin protein expression, and inferior cellular alignment. Proteomic profiling of EVs revealed temporally regulated, microenvironment-dependent cargo reflective of disease progression in vitro. Diseased EVs were enriched in cytoskeletal, ECM-remodeling, and inflammatory proteins, including vimentin (VIM), Annexin A2 (ANXA2), MMP2, and INHBA, suggesting an EV-mediated role in promoting matrix remodeling, fibroblast activation, and chronic inflammation. Notably, the temporal analysis demonstrated the late-stage emergence of stress-responsive and myofibroblast-associated proteins such as ENO1 and DES, underscoring the model's ability to capture the progressive nature of tendon pathology. In contrast, EVs from the healthy mimetic model demonstrated cargo associated with metabolic homeostasis and lipid transport, including APOA2 and CKM. Collectively, these findings highlight the utility of our tendon model as a dynamic platform for studying tendon pathology and establishing EVs as both sensitive indicators of microenvironmental state and potential mediators of fibrotic progression. This work provides a foundation for future studies exploring the diagnostic and therapeutic potential of EVs in tendinopathy.

<h4>Introduction</h4>Hidradenitis suppurativa (HS) is a chronic inflammatory skin disease associated with obesity and a high burden of psychiatric comorbidity. Glucagon-like peptide-1 receptor agonists (GLP-1RAs) and metformin are commonly prescribed for metabolic disease and have anti-inflammatory effects, prompting interest in their use among patients with HS. However, recent reports have raised concerns regarding potential psychiatric adverse effects of GLP-1RAs. We evaluated psychiatric outcomes following GLP-1RA versus metformin initiation among adults with HS.<h4>Methods</h4>We conducted a retrospective cohort study within the NIH All of Us Research Program (Controlled Tier v8) using a new-user, active-comparator design. Adults with HS were identified using validated SNOMED concepts. Incident users of GLP-1RAs were compared with incident users of metformin after a 365-day washout. The primary outcome was the first psychiatric diagnosis within 12 months of treatment initiation. Stabilized inverse probability of treatment weighting (IPTW) was used to control for confounding. Effect estimates included 12-month cumulative risks, risk differences (RD), risk ratios (RR), and IPTW-weighted Cox proportional hazards models.<h4>Results</h4>The weighted analytic cohort included 77 GLP-1RA users and 233 metformin users. At 12 months, psychiatric diagnoses occurred in 61.3% of GLP-1RA users versus 51.1% of metformin users (RD + 10.2 percentage points; RR 1.20; 95% CI: 0.95-1.52). The hazard ratio for time to first psychiatric diagnosis was 1.26 (95% CI: 0.90-1.78). Divergence between groups occurred primarily within the first 90 days after initiation.<h4>Conclusion</h4>Among adults with HS, GLP-1RA initiation was associated with a modest, nonsignificant increase in psychiatric diagnoses compared with metformin. These findings are hypothesis-generating and warrant further investigation.

Earlier menopause is a risk factor for several age-related diseases, including dementia. The biological pathways linking menopause timing to later-life brain aging are not understood. Leveraging large-scale plasma proteomics in postmenopausal women from the UK Biobank ( N =15,012), earlier menopause was associated with upregulation of pro-inflammatory and extracellular matrix degradation pathways, plus accelerated aging across proteomic clocks of organ and cellular aging, including brain and oligodendrocyte aging. Elevated GDF15, a canonical aging marker, was the top protein correlate of earlier menopause. We observed robust replication of menopause timing proteomic shifts in the Women’s Health Initiative Long Life Study ( N =1,210). In UKB, proteins associated with earlier menopause, including GDF15, exhibited concordant associations with incident dementia risk and brain atrophy, cerebral small vessel disease burden, and white matter microstructural integrity. Collectively, our findings identify proteomic signatures linking ovarian aging to brain aging, providing a framework to inform interventions to reduce dementia risk.

BackgroundIdentifying genetic variants conferring resilience to Alzheimer's disease and related dementia (ADRD) may hold promise for developing therapeutics.ObjectiveTo determine genetic associations with being dementia-free at age 85 (DF85).MethodsWe examined genetic associations, using whole genome sequencing data, with DF85 in three Trans-Omics for Precision Medicine cohorts and the Alzheimer's Disease Sequencing Project Phenotype Harmonization Consortium. We tested common variants individually and aggregation of rare (MAF ≤ 1%) coding and non-coding variants in DF85 participants (n = 3657) against individuals who were not DF85 (n = 20,010). We verified associations using a stricter control set who developed dementia before age 85 (n = 5552).ResultsWe observed an association at <i>APOE</i> (rs429358, MAF = 0.21, odds ratio [OR] = 0.49, 95% confidence interval [CI] = 0.46-0.53, p = 1.0 × 10^-92) as well as for two common variants (rs16892237-A near <i>MAL2</i>, MAF = 0.08, OR = 1.34, 95% CI = 1.21-1.48, p = 1.1 × 10^-8 and rs8004018-G near <i>GCH1</i>, MAF = 0.16, OR = 1.24, 95% CI = 1.15-1.34, p = 1.7 × 10^-9) and an aggregate of rare loss of function and disruptive missense variants in <i>FBXW10</i> on chr 17 (p = 1.4 × 10^-7) associated with DF85.ConclusionsThrough a genome-wide assessment of a resilience-focused outcome, we identified common and rare genetic variants contributing to DF85 status. Genes associated with DF85 may delay onset of ADRD and provide translational impact.

β-thalassemia is a common inherited hemoglobin disorder caused by reduced or absent β-globin production, leading to ineffective erythropoiesis, chronic anemia, and, in severe cases, lifelong transfusion dependence. Although allogeneic hematopoietic stem cell transplantation can be curative, its use is limited by donor availability and transplant-related complications. In recent years, gene therapy has emerged as a promising alternative and has rapidly changed the treatment landscape for β-thalassemia. In this review, we summarize both established and emerging gene-based strategies, including lentiviral gene addition to restore HBB expression and gene editing approaches aimed at reactivating fetal hemoglobin. We discuss key targets such as the erythroid-specific BCL11A enhancer, repressor-binding sites in the HBG promoters, and other regulatory elements involved in globin switching. We also highlight the growing potential of newer technologies such as base editing and prime editing, which may offer greater precision and reduce the risks associated with double-strand DNA breaks. Finally, we address the major challenges that still need to be resolved, including safety, durability, technical complexity, and access to treatment. Overall, gene therapy is moving β-thalassemia closer to a broadly applicable curative approach.

Donnai-Barrow syndrome (DBS) arises from loss-of-function (LoF) variants in the endocytic receptor low-density lipoprotein receptor-related protein 2 (LRP2; or megalin) and is characterized by low-molecular weight proteinuria and developmental abnormalities. Urinary proteomics of 9 patients with DBS revealed that the urinary proteome of a DBS patient with the missense variant LRP2 p.C1400R was indistinguishable from that of patients with splice site, nonsense, or frameshift mutations. A CRISPR mouse model of the variant was generated to determine the mechanism of LoF and proteinuria. The mutant LRP2 was expressed and observed to dimerize and localize to the proximal tubule apical membrane. However, both fluid-phase and receptor-mediated endocytosis was impaired in the context of a general perturbation of endocytic flux. Immunofluorescence revealed aberrant endocytic recycling with mislocalized RAB11+ and TFR1+ compartments and enlarged lysosomes. Structural modeling showed that the LRP2 assembly likely tolerates the cysteine-to-arginine substitution at the cell surface, but at endosomal pH the variant introduced steric clashes that may disrupt intramolecular interfaces and disturb receptor recycling. These findings point to the importance of LRP2 recycling for global endocytic flux and offer a blueprint for leveraging patient-specific alleles to dissect proximal tubule function.

<h4>Background</h4>Genetic mechanisms that predispose people to type 2 diabetes (T2D) and cardiovascular disease (CVD) remain poorly understood, partly because of a lack of sufficient data on non-European ethnic groups. Extending these evaluations to diverse cohorts is essential for gaining insights into the molecular pathways involved in disease development among human populations. In this study, we aimed to evaluate the genetic connection between the human lipidome and cardiometabolic disorders. We conducted a metabolite genome-wide association study (mGWAS) in a Punjabi population from India, along with multi-layer replication studies using the UK Biobank and other independent European and non-European cohorts.<h4>Methods and findings</h4>We performed mGWAS using 516 lipid metabolites in 3,000 Punjabi Sikh individuals, and validation was performed in 1.13M Europeans and 15K individuals from Asian Indian ancestry using independent cohorts of the UK Biobank, GeneRISK, DIAMANT, PROMIS, and other studies. We identified 609 SNP-metabolite associations representing 236 SNP-metabolite pairs that attained genome-wide significance (p </= 5 × 10-8). Of the 36 SNP-lipid metabolite signals that survived multiple testing correction (p </= 1.92 × 10-10), 33 associations were not reported before, and 3 associations were confirmed to be ancestry-specific. Using colocalization analysis, polygenic risk scores, and Mendelian randomization approaches, we identified a causal association of LPC O-16:0 with T2D, represented by a lead variant in CD45, a key regulator of T- and B-cell antigen receptor signaling, and is already used as a therapeutic target. Another possible causal relationship of PC 38:4 (C) in protecting against coronary artery disease risk in Asian Indians, attributed to a variant in the untranslated region in the FADS1/2 genes, may be specific to ancestry and/or could not be confirmed in Europeans because of extensive pleiotropy in this region. The main limitation of this study was the absence of an independent validation cohort of Asian Indians from India.<h4>Conclusions</h4>The mGWAS of Asian Indians offers new insights into the diverse molecular origins of cardiometabolic diseases and suggests potential pathways for innovative treatments. Our findings highlight the need for additional research on human lipidomics to better understand the downstream effects of the genome and its impact on cardiometabolic health.

Down syndrome is a genetic condition that causes intellectual disability and is characterized by early-onset delays in motor, cognitive, and language development. The molecular mechanisms underlying these neurodevelopmental impairments remain poorly understood. We used single-nucleus multiomic sequencing to simultaneously profile gene expression and chromatin accessibility in the Down syndrome prefrontal cortex during early postnatal development, a critical period for synaptogenesis, neural maturation, and developmental neuroimmune interactions. Our findings reveal widespread dysregulation of chromatin accessibility and gene expression, with deficits spanning metabolic and synaptic pathways, oligodendrocyte lineage progression, and a pronounced neuroinflammatory signature. We present a molecular atlas of Down syndrome neuropathology at a critical stage of brain development, highlighting convergent neurodevelopmental and neurodegenerative pathways and informing potential targeted therapies for Down syndrome-associated neuroinflammation.

Neither rodent models nor in vitro studies of human cells adequately describe the molecular ontogeny of human glial progenitor cells (hGPCs). Here, we use scRNA-seq together with scATAC-Seq and CUT&TAG assessment of chromatin accessibility to track the in vitro genesis and in vivo differentiation of hGPCs from pluripotent stem cells (PSCs). In vitro, the hGPC pool comprises 4 transcriptionally distinct subpopulations, each associated with a distinct pattern of chromatin accessibility and histone modification of stage-dependent genes. After the neonatal transplant of these cells into myelin-deficient shiverer mice (MBP^shi/shi), they differentiate further as astrocytes and oligodendrocytes. A combination of gene co-expression, motif enrichment, cell-trajectory, cell-cell interaction, and spatial transcriptomic analyses reveals that the host environment potentiates the context-dependent differentiation of the hGPCs, via their activation of distinct gene regulatory networks. Together, these data describe the process and pathways by which human PSC-derived GPCs are generated in vitro and diversify in vivo to mature as astrocytes and oligodendrocytes.

Head and neck squamous cell carcinoma (HNSCC) shows marked heterogeneity in clinical behavior, treatment response, and immune contexture. Existing molecular and immune classification systems are often derived from small, homogeneous cohorts and lack integration of multimodal data, limiting clinical relevance. We performed an integrated multi-omics analysis of transcriptomic and genomic data (copy number alterations and single-nucleotide variants) from 1149 tumors across 1102 HNSCC patients spanning multiple treatment settings. Using Similarity Network Fusion (SNF) on immune-related gene profiles, we identified immune subtype clusters (ISCs) and characterized them through mutational, transcriptional, immune cell enrichment, hypoxia, and established molecular subtype analyses. Clinical associations, including progression-free survival, were evaluated in first-line and post-metastatic settings. Four ISCs were defined: ISC1, immune-cold and EMT-enriched; ISC2, immune-activated; ISC3, mixed immune-regulatory with stromal enrichment; and ISC4, immunosuppressed. These subtypes showed distinct clinical outcomes and differential responses to checkpoint inhibitors, chemotherapy, and combination therapies. Analysis of 44 patients with matched pre- and post-treatment tumors revealed dynamic immune-state transitions, with checkpoint inhibition promoting immune activation and chemotherapy enriching immunosuppressive signaling. This study establishes a clinically relevant immune subtyping framework for HNSCC that captures dynamic tumor-immune states linked to treatment response and survival, supporting its utility in guiding immune-based therapies.

Leveraging existing genomic data to opportunistically screen for secondary findings (SFs) can identify individuals at increased genetic risk who may be missed by criteria-based testing. While some guidelines support returning actionable SFs with professional support, there is a gap in consistent practice regarding the return process. This study reports the outcomes of opportunistic genomic screening in an Australian biobank. Whole genome sequencing data from 1057 healthy participants in the Tasmanian Ophthalmic Biobank (TOB), all of white European ancestry, underwent opportunistic screening for pathogenic (P) or likely pathogenic (LP) variants affecting genes in the ACMG SF v3.0 list. Variants of interest were manually curated, and only P/LP variants were returned. Actionable SFs (P/LP variants) were identified in 3.6% (38/1057) of participants. The most common genes were HFE (haemochromatosis), LDLR (Familial Hypercholesterolemia), and TP53 (Li-Fraumeni syndrome). Of the 38 participants with a variant, 27 received their result, with two-thirds being newly informed. Ten participants were referred to clinical genetics for diagnostic confirmation, while seven declined to proceed. Opportunistic screening identified a clinically significant incidence of actionable SFs in a healthy biobank cohort. There was high participant interest in receiving results, although subsequent uptake of clinical referral remains a challenge.

Activation of oncogenes by hijacking immunoglobulin gene loci (IG) enhancers via chromosomal translocation is a common pathogenetic mechanism in B-cell malignancies, affecting 5-10% of chronic lymphocytic leukemia (CLL). The oncogenic partners in many of these cases remain unidentified. Therefore, we conducted a comprehensive analysis of 144 CLL samples with IGH-translocation excluding IGH::BCL2, IGH::CCND1, IGH::BCL3 and IGH::MYC. By combining fluorescence in situ hybridization (FISH) with whole-genome, targeted sequencing, and RNA expression profiling, we identified 25 IG-translocation partners; 12 were previously unreported. Of 142 cases, 107 (75%) displayed an unmutated IGHV. Genetic profiling showed a heterogenous distribution of chromosomal aberrations and recurrently mutated genes across the groups. Of 41 informative cases, 32 (78%) exhibited breakpoints driven by aberrant class-switch recombination (CSR), with prominent involvement of IGHM (9/41) and IGHG3 (9/41). Three cases with unmutated IGHV carried a juxtaposition of the IGH locus 5' to the intact NKX2.6 gene in chromosome 8p21.2 due to illegitimate VDJ recombination, associated with significant ectopic upregulation of NKX2.6 transcriptional expression (FDR < 0.001, logFC: 15). Similarly, METRNL, located at the telomere of chromosome 17q25, was identified as a translocation partner gene in four cases. Our findings expand the spectrum of the oncogenic translocation partners targeting IGH in CLL.

BACKGROUND: The evolution of human genome has been inextricably linked to germ cell infectivity and retrotransposition activity of endogenous retroviruses (ERVs). In result, 8% of the modern human DNA is occupied by human ERVs (HERVs), which although non-infectious and replication incompetent, are transcriptionally active in several cases (such as viral infections and cancer), and are able to produce functional mRNAs, non-coding RNAs and proteins, which on one hand are utilized for cell processes and on the other hand have been reported to be implicated in various pathologies. RESULTS: We used Nanopore Direct RNA Sequencing (DRS) technology to study the transcriptome of the transcriptionally active HERV family, HERV-K HML-2 (HK2) in a teratocarcinoma cell line. We developed a unique pipeline of HK2 DRS data analysis, which along with DRS itself, enabled us to investigate the true HK2 transcriptional profile on a single-molecule basis, unveiling alternative and non-canonical splicing patterns, length variants distribution and polyadenine tail length estimation. CONCLUSIONS: We uncovered a plethora of non-annotated HK2 transcriptional isoforms and read-through transcripts that utilized HK2 splicing sites, promoters and polyadenylation signals to assemble copies of the surrounding genes. This finding supports the observation that HERVs are involved in regulation of cellular transcription.

BACKGROUND: Krabbe disease (KD) is caused by mutation of the galactosylceramidase (GALC) gene, leading to deficient sphingolipid metabolism, which is essential for functional myelination. The twitcher (Galctwi/twi) mouse, a KD model with a premature termination codon (PTC) caused by a single-nucleotide G-to-A substitution at the 355th codon of the Galc gene, is a model candidate for treatment with adenine base editors (ABEs). ABEs have emerged exclusively among genome editing systems as viable therapeutic candidates to correct mutant genes. METHODS: To confirm base editing efficiency of ABEs, mouse embryonic fibroblasts (MEFs) or mutant GALC HEK293T cells treated with three ABE variants (ABEmax, ABE8eWQ, ABE8e) were assessed using targeted deep sequencing. Each split-ABE8e vector was packaged into a dual-vector adeno-associated virus serotype 9 (AAV9) system and delivered to twitcher mice via intracerebroventricular injection on postnatal day 1. Thereafter, motor functions and survival rate were evaluated by rotarod test, clasping test and lifespan analysis. Various methods, including next-generation sequencing (NGS), qRT-PCR, enzyme activity assay, and flow cytometry, were used to measure the base correction rate of the target gene and verified restoration of GALC enzyme activity in the brain of ABE8e-treated mice. Additionally, myelin recovery was evaluated in the brain using histological analysis, magnetic resonance imaging (MRI), diffusion tensor imaging (DTI), and transmission electron microscopy (TEM). RESULTS: The ABE8e-treated MEFs and mutant GALC HEK293T cells showed the most effective editing among the ABE variants tested. Three weeks after dual-AAV9 injection, the PTC was corrected in approximately 0.5% of genomic DNA and 5% of mRNA in twitcher mice. ABE8e treatment restored GALC enzymatic activity to approximately 5% of wild-type (WT) levels, while reducing the accumulation of psychosine—a major neurotoxic metabolite—by approximately 47% relative to WT. Moreover, histological analysis, TEM and, DTI and T2-weighted MRI showed preserved myelination and axonal integrity, along with amelioration of myelin deficits in the corpus callosum of ABE-treated twitcher mice. Five weeks after ABE8e administration, body weight recovered to approximately 64% of WT levels, accompanied by an extension of lifespan. In addition, clasping scores and rotarod performance improved to approximately 23% and 64% of WT levels, respectively. CONCLUSIONS: These data demonstrate a reliable application of base editing technology using ABEs as a potential treatment option for KD, progressing the development of therapeutics treating various genetic diseases.

BACKGROUND: Ovarian cancer (OC) is often characterized by poor prognosis due to paclitaxel (TAX) resistance, with ferredoxin 1 (FDX1) emerging as a key mediator of copper metabolism. The present study aimed to elucidate the role of FDX1 in TAX resistance and to evaluate the efficacy of pH/reactive oxygen species (ROS)-responsive nanomicelles (PPD/PDP@si-FDX1) in reversing this resistance. METHODS: TAX-resistant A2780 and SKOV3 cells were established by gradient exposure, and FDX1 expression was assessed using Western blotting. FDX1 was either overexpressed or silenced, and resistance was evaluated using CCK-8, clonogenic, scratch, and Transwell assays. Autophagic activity was examined through Western blotting, immunofluorescence, and transmission electron microscopy. Mechanistic validation involved the ULK1 activator BL-918 and the copper chelator TTM. PPD/PDP@si-FDX1 nanomicelles were prepared via self-assembly, with structural and responsive characteristics analyzed by transmission electron microscopy (TEM), dynamic light scattering (DLS), and drug release profiling. Tumor penetration and in vivo antitumor efficacy were examined using multicellular spheroids and subcutaneous xenograft models. RESULTS: FDX1 overexpression elevated intracellular copper, activated the ULK1/ATG13 autophagic axis, and enhanced TAX resistance; silencing FDX1 reversed these effects. Copper chelators or ULK1 inhibition phenocopied FDX1 silencing. PPD/PDP@si-FDX1 demonstrated pH/ROS-responsive tumor accumulation and enhanced si-FDX1 delivery. In vivo, it significantly suppressed tumor growth and restored TAX sensitivity, outperforming free si-FDX1. CONCLUSION: FDX1 drives TAX resistance via copper-dependent ULK1/ATG13 activation; PPD/PDP@si-FDX1 nanomicelles effectively reverse resistance, offering a promising strategy for OC therapy.

Depot-specific fat deposition influences carcass yield and meat quality in ducks. To explore regulatory mechanisms, we profiled adipogenic progenitor cells (APCs) from pectoral muscle (PM), subcutaneous (SAT), and visceral adipose tissue (VAT) using single-cell RNA sequencing. Nine major cell types were identified, with APCs being most abundant in VAT and least abundant in PM, consistent with histological findings. Transcriptomic comparisons revealed depot-specific microenvironments: PM was enriched with myogenic-related pathways, SAT with immune and inflammatory responses, and VAT with pro-adipogenic signaling. Next, APCs were regrouped into 5 subclusters. Cluster 0 (PI16^high PDGFRB^high) enriched in SAT/VAT showed activated signaling of BMP and glycolysis, indicating strong adipogenic potential. Cluster 1 (PDGFRA^high F3^high) enriched in PM was associated with Wnt signaling, which maintains APCs in a progenitor state. Cell cycle, RNA velocity, and pseudotime analyses indicated that APCs of SAT/VAT were more advanced in differentiation, while APCs of PM retained stronger proliferative capacity. Cell-cell communication analysis further revealed more active paracrine regulation within SAT/VAT, while APCs from PM showed limited signaling interactions. Key depot-specific regulators were identified, including NEGR1 (PM), CD36 (SAT), and RSPO2 (VAT), which may contribute to regulations of the adipogenic process. This study provides the first single-cell atlas of duck APCs across depots, revealing molecular heterogeneity, lineage dynamics, and niche-specific regulation. These findings advance understanding of depot-specific adipogenesis and offer targets to enhance IMF while limiting excessive SAT and VAT, thereby improving both meat quality and production efficiency.

Parkinson's disease (PD) is a complex neurodegenerative disorder characterized by both motor and non-motor symptoms, primarily attributed to dopaminergic dysfunction in the substantia nigra pars compacta. However, growing evidence indicates that serotonergic and noradrenergic alterations also contribute significantly to PD pathophysiology and progression. This growing understanding has driven the development of advanced neuroimaging techniques aimed at visualizing not only dopaminergic deficits but also serotonergic and noradrenergic alterations, providing deeper insights into PD pathophysiology and progression. Positron emission tomography (PET) and single-photon emission computed tomography (SPECT) have been instrumental in visualizing dopaminergic deficits, particularly dopamine transporter loss, aiding in diagnosis and disease progression tracking. While PET and SPECT remain crucial in assessing dopaminergic dysfunction, novel MRI techniques, including neuromelanin-sensitive MRI, iron-sensitive MRI, diffusion-weighted imaging, and pharmacological MRI, have emerged as promising non-invasive alternatives for evaluating monoaminergic dysfunction in PD. In this narrative review, we have discussed the recent neuroimaging advancements in assessing monoaminergic dysfunction in PD, highlighting how these advances not only improve our understanding of the distinct contributions of dopaminergic, noradrenergic, and serotonergic systems to motor and non-motor symptoms, but also offer promising biomarkers for disease progression and treatment response.

Since the 1980s, 70 kDa heat shock proteins (HSP70s) have been recognized as central regulators of proteostasis, with diverse roles in cellular physiology and pathology. Recent research has significantly expanded our understanding of these molecular chaperones, revealing functions that extend beyond their classical roles in proteostasis. In this review, we integrate these emerging insights with foundational knowledge by outlining the biology of HSP70s, with particular emphasis on recent discoveries, such as new data on the substrate specificity and molecular dynamics of HSP70-client interactions. In addition, increasing evidence highlights their noncanonical anti-inflammatory properties, as well as other nonimmune functions, including the promotion of adipose tissue browning and the enhancement of angiogenesis through extracellular HSP70 activity. Finally, although HSP70s have long been known to regulate mRNA degradation in a transcript-specific manner, new findings demonstrate their ability to bind double-stranded RNA, further broadening their functional repertoire.

Recent studies highlight the potential of nanoparticles to enhance in vivo drug efficacy. Polylactide-<i>co</i>-glycolide acid (PLGA) is widely used for nanoparticle fabrication due to its biocompatibility, and PEGylation further improves its circulation time by reducing rapid reticuloendothelial system (RES) clearance. Folate-modified nanoparticles can additionally enable active targeting, as many cancer cells overexpress folate receptors. In this study, dual-drug-loaded and folate-targeted PLGA-polyethylene glycol (PEG) nanoparticles were developed, and their enhancing effects on cellular uptake, cytotoxicity, and apoptotic response were evaluated. The nanoparticles were prepared via the nanoprecipitation method. They exhibited a particle size of 276 ± 6 nm, a polydispersity index (PDI) of 0.5 ± 0.02, and a ζ potential of -19 ± 2 mV. The drug-loading capacity was 8.8% for ε-viniferine (EV) and 2.6% for vincristine sulfate (VS). The use of EV-VS-loaded PLGA-PEG-folate nanoparticles enhanced cytotoxicity and apoptosis compared to the free drug, reducing HepG2 cell viability from 94.3 to 53% at 10 μM EV + 1.64 μM VS and increasing early apoptotic cells from 15.3 to 31%, indicating their potential as a targeted drug delivery system for cancer cells.

Osteoporosis (OP) is primarily characterized by reduced bone mass, microarchitectural deterioration, and an increased susceptibility to fragility fractures. A central pathological feature of OP is the progressive impairment of osteogenic differentiation and bone matrix production in osteoblasts. The endoplasmic reticulum (ER), a pivotal organelle responsible for secretory protein folding, lipid/sterol biosynthesis, and intracellular Ca²⁺ storage, is subjected to a substantial secretory burden during osteogenic differentiation and functions as a critical regulatory hub integrating metabolic stress, inflammatory signaling, and mineralization-associated calcium signaling. Emerging evidence indicates that disruption of ER homeostasis regulates osteogenic differentiation through the three canonical branches of the unfolded protein response (UPR), including PERK-eIF2α-ATF4, IRE1α-XBP1, and ATF6 signaling pathways. In parallel, selective ER autophagy (ER-phagy) dynamically regulates ER quality control during osteogenic differentiation through removal of damaged ER domains and misfolded substrates such as procollagen. In addition, ER Ca²⁺ stores and STIM/ORAI-mediated store-operated Ca²⁺ entry (SOCE) cooperatively maintain calcium homeostasis during osteogenesis and regulate spatiotemporal expression of osteogenic transcription factors, including Runx2 and Sp7, through Ca²⁺ oscillatory signaling. ER membrane lipid composition further modulate osteogenic fate by influencing membrane contact site dynamics and cellular metabolic adaptation. In this review, we systematically summarize the crosstalk among ER stress, ER-phagy, Ca²⁺ homeostasis, and lipid metabolism during osteogenic differentiation from the perspective of ER structure-function coupling. We further discuss potential therapeutic strategies targeting ER stress regulation, including chemical chaperones and UPR/autophagy modulators, to provide new insights for targeted therapeutic approaches for osteoporosis.

<h4>Introduction</h4>This study conducted a meta-analysis across three large European cohorts (UKBB, FinnGen, and REPAIR), including 12,660 rheumatoid arthritis (RA) cases, 2,446 radiographic axial spondyloarthritis (r-axSpA) cases, and over 530,000 shared controls.<h4>Methods</h4>Ten independent SNPs in <i>CARMIL1</i>, <i>GRM4</i>, <i>ITPR3</i>, <i>PRSS16</i>, <i>ZNF322</i>, <i>HTT</i>, <i>IKZF1</i>, <i>MANEA</i>, and <i>MGAM2</i> were analyzed, and functional characterization was performed through cytokine and protein assessments as well as eQTL analyses.<h4>Results</h4>Ten independent SNPs were significantly associated with both RA and r-axSpA. Risk alleles included <i>HTT</i> _rs363075A, <i>IKZF1</i> _rs12718261A, <i>MANEA</i> _rs72920280T, and <i>MGAM2</i> _rs73158426G, while <i>CARMIL1</i> _rs72831267C, <i>GRM4</i> _rs2495964G, <i>ITPR3</i> _rs77601296A, <i>ITPR3</i> _rs9469540T, <i>PRSS16</i> _rs72843633T, and <i>ZNF322</i> _rs6901425G had protective effects. Functional analysis showed that <i>GRM4</i> _rs2495964G was linked to decreased CCL25 levels (p = 0.00030), and <i>ITPR3</i> _rs9469540T to reduced IL10 production after LPS stimulation (p = 1.3×10^-4). The ZNF322rs6901425G allele was associated with reduced TNFB and increased TGM2 levels (p = 9.60×10^-4 and p = 3.00×^10-4), both involved in immune signaling and tissue remodeling. Disease-specific associations were found in <i>BTN2A1</i>, <i>BTN3A2</i>, and <i>H2BC11</i>. The <i>BTN2A1</i> _rs1977199A allele was protective in RA (OR = 0.93) but increased r-axSpA risk (OR = 1.23), and was associated with reduced IL22 (p = 0.00016) and elevated HO-1 in obese individuals (p = 6.73×10^-6). In contrast, <i>BTN3A2</i> _rs9393716G and <i>H2BC11</i> _rs66462181C increased RA risk but were protective in r-axSpA, linked to decreased HO-1 and IL6 (p = 2.43×10^-5, 3.287times;10^-4, 1.18×10^-4). These SNPs also acted as eQTLs for immune-related genes such as <i>BTN3A2</i>, <i>HMGN4</i>, and <i>TRIM38</i>.<h4>Discussion</h4>Our findings highlight novel shared and disease-specific variants and key immunoregulatory mediators-IL10, IL22, IL6, CCL25, and HO-1-offering insights for disease stratification and therapeutic targeting.

Tuberculosis (TB) remains a major global cause of morbidity and mortality. Current tools for monitoring treatment response rely on sputum-based microscopy and culture, which are often insensitive, time-consuming, and impractical in extrapulmonary or pediatric TB and in individuals unable to produce sputum. Metabolomics has emerged as a promising approach for identifying host-derived biomarkers that reflect treatment-associated immunometabolic changes; however, the available evidence remains heterogeneous and has not been comprehensively synthesized. We conducted a comprehensive literature review of human studies evaluating metabolomic biomarkers in relation to TB treatment response or outcomes. PubMed, Scopus, and EMBASE were searched for human studies evaluating targeted or untargeted metabolomics (NMR, LC-MS, GC-MS, CE-MS) in relation to treatment response or outcomes. Two reviewers independently screened studies, extracted data, and assessed risk of bias using QUIPS and PROBAST. Findings were synthesized using a structured framework organized across treatment stages and outcomes. Of 218 records identified, 139 titles and abstracts were screened and 42 full texts assessed; 15 studies met the inclusion criteria. Recurrent treatment-associated signals involved amino acid metabolism, particularly the tryptophan-kynurenine pathway, as well as vitamin and cofactor metabolites (pyridoxate, nicotinamide, trigonelline). Plasma studies frequently reported lipid remodeling and bile acid perturbations, whereas urine studies highlighted polyamine metabolism (e.g., N¹,N¹²-diacetylspermine) and fatty acid β-oxidation markers. Common limitations included inadequate adjustment for confounders and, in prediction models, small sample sizes and limited external validation. Metabolomics reveals reproducible but heterogeneous immunometabolic changes during TB therapy. Key pathways include tryptophan-kynurenine metabolism, vitamin and cofactor metabolism, lipid remodeling, and urine polyamine pathways. Standardization and prospective multicenter validation are needed for clinical translation.

<i>Background</i>: Chromosomal rearrangements involving lysine methyltransferase 2A (KMT2A) define a genetically distinct subset of acute myeloid leukemia (AML) in 10% of cases in adult patients; the frequency of <i>KMT2A-r</i> is higher in pediatric AML. Translocations involving the <i>KMT2A</i> locus at chromosome 11q23 result in the formation of a chimeric oncogene partner, where the N-terminal part of <i>KMT2A</i> is fused to a variety of translocation partners. The leukemogenic activity of <i>KMT2A</i>-fusion partners is related to their capacity to hyperactivate the expression of <i>HOX-A</i> and <i>MEIS1</i> target genes, which stimulate the proliferation of hematopoietic stem cells. The oncogenic activity of <i>KMT2A</i> fusion proteins requires the binding with Menin, and this interaction can be targeted pharmacologically by small molecules acting as potent and selective Menin inhibitors. <i>Methods</i>: A search of the literature showed a marked development of experimental studies exploring the molecular pathogenesis of AML with <i>KMT2A-r</i> and of clinical studies evaluating new induction intensive treatments and the development of a targeted therapy based on Menin inhibitors. <i>Results and Conclusions</i>: In the present review article, we summarize our current understanding of the biology of <i>KMT2A-r</i> in AML development and the recent consistent progress made in the treatment of <i>KMT2A-r</i> AML through new chemotherapy regimens and targeted therapy using Menin inhibitors. However, the prognosis of older <i>KMT2A-r</i> AML patients remains poor and could be improved by drug combination studies including Menin inhibitors. Many encouraging observations derived from ongoing clinical trials with Menin inhibitors need to be confirmed through randomized clinical trials.