Gene Literature Dashboard

Cytosolic Abundant Heat-Soluble (CAHS) proteins are thought to be responsible for protection of tardigrades against conditions of extreme environmental stress, in particular desiccation. Hypsibius exemplaris CAHS-8 is intrinsically disordered in solution, undergoing conformational transformation as a function of stress, assembling into fibres that form a hydrogel. Here we present the crystal structure of the fibrils of CAHS-8, comprising a single 101 residue-long helix, forming an atypical 90 amino-acid coiled-coil dimer exhibiting non-canonical periodicities and assembling into fibrils via a second coiled coil interface associating adjacent dimers via the opposing face of the helix. Combination with electron microscopy, atomic force microscopy and disorder modelling provides structural insight into the details of this assembly that is essential for cell survival. Individual fibrils appear to interact in a pairwise manner, possibly via their intrinsically disordered tails, forming straight fibres.

BACKGROUND: Despite advancements in diagnostic techniques and therapeutic strategies, the prognosis for Lung adenocarcinoma (LUAD) patients remains poor. Cell proliferation and cycle dysregulation drive cancer via uncontrolled cell growth. These genes also modulate tumor immune microenvironment (TIME), yet the precise mechanisms in LUAD remain largely unknown. METHODS AND RESULTS: This study aimed to identify key proliferation-cycle genes in LUAD, characterize the TIME associated with proliferation-cycle gene signatures and assess the impact of proliferation-cycle gene signatures on immunotherapy responsiveness. We analyzed The Cancer Genome Atlas (TCGA) LUAD transcriptomic data and identified eight proliferation-cycle-related risk genes (seven up-regulated: FAP, IL2RA, ITGA2, CHORDC1, PIM2, POU3F2, CD180; one down-regulated: FKBP1B). Independent cross-validation using the National Center for Biotechnology Information (NCBI) Gene Expression Omnibus (GEO) dataset confirmed the consistent expression patterns for all eight candidate genes in LUAD tumors. A risk model based on these genes stratified patients into distinct prognostic groups, revealing: (1) Survival disparity: High-risk patients exhibited poorer overall survival (p = 6.2e−05). (2) Immunosuppressive TIME: Elevated risk scores correlated with enhanced immune infiltration (p = 2.9e−12), enriched immunosuppressive populations (Tregs), reduced cytotoxic effectors (CD8+ T cells), and up-regulated immune checkpoint molecules (PDCD1/PD-L1, CTLA4). (3) Scientific implications: Risk signatures exhibited no significant correlation with tumor mutational burden (TMB), yet uncovered novel candidate targets with therapeutic potential, meriting further mechanistic exploration. CONCLUSION: Proliferation-cycle gene signatures are robust biomarkers for LUAD risk stratification, prognosis, and immune landscape prediction. Their mechanistic integration into multi-dimensional oncological models could reveal previously unrecognized layers of antitumor immune regulation.

Protein palmitoylation in the Golgi apparatus is critical for the appropriate sorting of various proteins belonging to secretory and lysosomal systems, and defective palmitoylation can lead to the onset of severe pathologies. HIP14 and HIP14L ankyrin repeat-containing palmitoyl transferases were linked to the pathogenesis of Huntington's disease, however, how perturbation of these Golgi resident enzymes contributes to neurological disorders is yet to be understood. In this study, we investigated the function of Hip14 and Patsas - the Drosophila orthologs of HIP14 and HIP14L, respectively - to uncover their role in secretory and lysosomal membrane trafficking. Using larval salivary gland, a well-established model of the regulated secretory pathway, we found that these PAT enzymes equally contribute to the proper maturation and crinophagic degradation of glue secretory granules by mediating their fusion with the endo-lysosomal compartment. We also revealed that Patsas and Hip14 are both required for lysosomal acidification and biosynthetic transport of various lysosomal hydrolases, and we demonstrated that the rate of secretory granule-lysosome fusion and subsequent acidification positively correlates with the level of Hip14. Furthermore, Hip14 is also essential for proper lysosome morphology and neuronal function in adult brains. Finally, we found that the over-activation of lysosomal biosynthetic transport and lysosomal fusions by the expression of the constitutively active form of Rab2 could compensate for the lysosomal dysfunction caused by the loss of Patsas or Hip14 both in larval salivary glands and neurons. Therefore, we propose that ankyrin repeat palmitoyl transferases act as rate-limiting factors in lysosomal fusions and provide genetic evidence that defective protein palmitoylation and the subsequent lysosomal dysfunction can contribute to the onset of Huntington's disease-like symptoms.

<h4>Background</h4>Tetrahydromagnolol (THM) is a compound isolated from Magnolia officinalis with unique chemical structure and composition. However, its anticancer effect of THM and the underlying molecular mechanisms remain unclear.<h4>Method</h4>Firstly, the anticancer effects of THM on different cancer cell lines in vitro were investigated. Subsequently, the antitumor activity of THM was further evaluated in vivo and in vitro using colorectal and lung cancer models. This assessment involved the effects of THM on cell viability, apoptosis, proliferation, cell cycle progression, and tumor growth inhibition. In addition, the anticancer molecular mechanisms of THM were determined by RNA sequencing, western blot, immunohistochemistry, immunofluorescence, CETSA, and SPR. Meanwhile, the effects of THM on organelles were evaluated by measuring endoplasmic reticulum stress, mitochondrial membrane potential damage, reactive oxygen species (ROS), and calcium ion concentration. Finally, the efficacy of THM in combination with conventional anticancer drugs for colorectal cancer treatment was evaluated in vivo.<h4>Results</h4>The results showed that THM had a significant anticancer activity in colorectal cancer and lung cancer both in vitro and in vivo. THM significantly inhibited cell proliferation and induced PANoptosis-like cell death through GSDME mediated pyroptosis, CASP3 mediated apoptosis, and MLKL mediated necroptosis. In addition, the anticancer potential of THM was also related to elevation of endoplasmic reticulum stress, mitochondrial membrane potential destruction, and increase of ROS and intracellular calcium concentration. Mechanically, we found that THM could directly bind to triplet motif-containing 38 (TRIM38) and induced its upregulation at mRNA and protein levels. Importantly, knockdown of TRIM38 remarkably rescued the anticancer effects of THM and PANoptosis induced by THM treatment, suggesting that TRIM38 played a key role in mediating the antitumor activity of THM. In addition, THM showed significant synergistic therapeutic effects when used in combination with conventional anticancer strategies (Cetuximab, FOLFOX, and FOLFIRI regimens) for colorectal cancer treatment.<h4>Conclusion</h4>Our data suggest that THM exerts its anticancer potential by inducing TRIM38-dependent PANoptosis and it also has synergistic antitumor effects in combination with conventional anticancer strategies. THM will be a promising candidate drug used alone or in combination with other anticancer regimens for cancer treatment.

Resistance to and associated toxic side effects of neoadjuvant chemotherapy remain major obstacles to improving the prognosis of osteosarcoma patients. Consequently, there is an urgent need to discover effective therapeutic agents with lower toxicity. In this study, the patient-derived xenograft (PDX) model was established and single-cell multi-omics sequencing was performed to comprehensively analyze changes in cellular heterogeneity and gene expression patterns of under formononetin treatment. We found that formononetin can significantly inhibit tumor growth in the osteosarcoma PDX model, on which the single-cell sequencing identified MYO1B as a key target mediating the anti-osteosarcoma effects of formononetin. <i>In vitro</i> experiments demonstrated that MYO1B overexpression enhanced the proliferation, invasion, and migration of osteosarcoma cells, while MYO1B silencing exhibited the opposite effects. Further investigation revealed that formononetin treatment markedly downregulated MYO1B expression, effectively suppressing the proliferative, invasive, and migratory phenotypes of osteosarcoma cells. Moreover, single-cell transcriptomic analysis of murine-derived cells showed that formononetin enhanced the cytotoxic activity of NK cells, promoted M1 macrophage polarization and inhibited M2 polarization, and reduced the proportion of senescent neutrophils, thereby alleviating the immunosuppressive state of the tumor microenvironment. Overall, our findings provide a comprehensive single-cell-level elucidation of the molecular mechanisms underlying the anti-osteosarcoma effects of formononetin, primarily involving downregulating the expression of MYO1B and remodeling the tumor immune microenvironment.

Among gynecologic malignancies, ovarian cancer (OC) stands out as a highly aggressive disease with the highest mortality rate and the poorest prognosis. At the beginning stage, it demonstrates high sensitivity to platinum-based chemotherapy. Nevertheless, most patients will encounter recurrence following the initial surgery and chemotherapy. Small extracellular vesicles (sEVs), characterized by a "cup-shaped" morphology and with a diameter of 40 to 160 nm, encompass diverse biologically active substances including nucleic acids (such as DNA, mRNA, microRNA (miRNA), and other non-coding RNAs (ncRNAs)), as well as oncogenic proteins, lipids, and metabolites, which play a crucial role as mediators of intercellular communication. Increasing evidence shows that sEVs promote various cancers' progression (including OC) via transporting molecular cargoes to target cells or organs. It is worth mentioning that existing literature often focuses on sEVs from a single cell type and lacks a comprehensive review of multiple cell sources. In this review, we summarize the biological functions of sEVs derived from different cell types in OC, including regulating cell proliferation, promoting metastasis, mediating drug resistance, inducing angiogenesis, facilitating immune escape, and maintaining stemness. Meanwhile, we focus on exploring the clinical value of sEVs as biomarkers for the diagnosis and prognosis of OC, as well as their application potential in translational medicine fields related to cancer vaccine development, targeted drug delivery, and precision tumor-targeted therapy. Additionally, we analyze the major challenges currently faced in sEV-based OC treatment research and propose potential strategies to overcome these limitations.

Diffuse midline gliomas (DMGs) with H3K27M alterations are rare, aggressive, World Health Organization (WHO) grade 4 tumors arising in midline central nervous system (CNS) structures, characterized by a lysine-to-methionine substitution at histone H3K27, which disrupts epigenetic regulation via global loss of H3K27 trimethylation. The pineal gland is an exceptionally uncommon site of origin. We report the first case of a synaptophysin-positive, H3K27M-altered DMG in the pineal gland of a 7-year-old female, characterized by hypercellularity, moderate atypia, high mitotic activity, and a Ki-67 index of 30%. Immunohistochemistry confirmed positivity for glial markers: glial fibrillary acidic protein (GFAP), oligodendrocyte transcription factor 2 (Olig2), loss of hypotrimethylation of lysine 27 on histone H3 (H3K27me3), and synaptophysin expression, an unusual feature for DMGs. Methylation profiling established the diagnosis. A systematic review identified seven cases of pineal H3K27M-altered DMG (age ranged from 7 to 65 years, with three pediatric and four adult) revealing notable immunohistochemical heterogeneity, limited molecular data (only our case had available methylation profiling) and a synaptophysin expression limited to our pediatric case. Sparse clinical outcome data precluded robust prognostic comparisons. These findings underscore the biological heterogeneity and diagnostic challenges of pineal DMGs and underscore the necessity of comprehensive molecular and immunohistochemical assessments to optimize diagnosis and guide emerging targeted therapies.

Autophagy process is important in the removal of damaged organelles following radiation interaction. Autophagy maintains stability or balance between intracellular environments at the cellular level. High linear energy transfer radiation is a complex and under-researched field of cancer and radiation biology. In-depth investigation in this area is necessary because autophagy has both protective and detrimental effects on cells, and the specific response can vary depending on factors like the type of radiation, the cell type, and the stage of cancer. Ionizing radiation exposure caused significant alterations in the autophagy genes (ATG3, ATG5, ATG7, ATG8, ATG12). Autophagy blocker could prevent cells from autophagic cell death in response to high linear energy transfer radiation. We have outlined how autophagy can reverse complications of serious diseases in tumor cells and tumor microenvironment, which will contribute to a new direction for better treatment of diseases.

ARID1A, a key subunit of the SWI/SNF chromatin remodeling complex, plays a context-dependent function in cancer, acting both as a tumor suppressor and, in certain conditions, as an oncogene. ARID1A, as a tumor suppressor, maintains transcriptional regulation, genomic stability, and cellular differentiation. In breast cancer, ARID1A loss-of-function leads to dysregulation of cell cycle checkpoints and impaired DNA repair and promotes epithelial-to-mesenchymal transition (EMT), jointly accelerating tumor proliferation and increasing therapeutic resistance. Notably, context-dependent ARID1A loss-of-function often concurs with activation of the PI3K/AKT signaling pathway and corresponds with poor prognosis. On the contrary, aberrant ARID1A overexpression can provoke oxidative stress and agitate the cytochrome P450 system, potentially facilitating early tumorigenesis. Consequently, understanding ARID1A's dual and context-dependent role highlights its potential as a biomarker and therapeutic target in precision oncology.

Astrocytes are central to brain homeostasis, supporting neuronal metabolism, synaptic activity, and the blood-brain barrier. With aging, these glial cells undergo molecular and functional changes that weaken support functions and promote neuroinflammation, contributing to neurodegeneration. Yet the systems-level mechanisms by which astrocytes respond to aging-related stressors remain poorly defined in human models. Because aging also heightens risk for cardiovascular disease, cognitive impairment, type 2 diabetes, and systemic inflammation, clarifying shared astrocytic pathways is critical for understanding brain-body crosstalk. Using an in vitro human astrocyte model exposed to sublethal oxidative stress (10 µM H₂O₂) as a proxy for age-related cellular stress, we profiled transcriptomic changes and identified differentially expressed genes across antioxidant defenses, proteostasis, transcriptional regulation, vesicular trafficking, and inflammatory signaling. We then performed network-prioritization analyses on a curated human protein-protein interactome: one seeded with the astrocyte oxidative stress responsive genes and six with phenotype-associated gene sets (Alzheimer's disease, cardiovascular disease, cognitive impairment, type 2 diabetes, oxidative stress, and inflammation). Intersecting the top 5 % scoring genes from each run yielded a 127-gene core shared across all seven, enriched for proteostasis, DNA repair, mitochondrial regulation, and telomere and nuclear envelope maintenance. Structure-guided analyses highlighted vulnerable interfaces, including lamin A/C-lamin B1, α-actinin-filamins, 14-3-3 dimers, and aminoacyl-tRNA synthetase assemblies, where pathogenic variants are predicted to destabilize or aberrantly stabilize protein interactions. Structure-based interface predictions also highlight potential interactions between amyloid precursor protein (APP) and valosin-containing protein (VCP), and between p53 and 14-3-3ζ, potentially linking proteostasis and stress signaling. Together, these analyses identify a conserved astrocyte-centered network signature that may relate neurodegenerative and cardiovascular processes, and prioritize structurally testable candidates for biomarker and intervention hypothesis testing.

<b>Background</b>: The transition from the term non-alcoholic fatty liver disease (NAFLD) to steatotic liver disease (SLD), an umbrella term for several related conditions, offers benefits, particularly in identifying cardiometabolic risk factors more effectively. However, the impact of alcohol consumption on liver disease progression remains significant, leading to the recognition of a new entity: MetALD (metabolic dysfunction-associated steatotic liver disease with moderate alcohol intake). <b>Aim</b>: This study aimed to compare characteristics associated with liver disease progression in diabetic patients diagnosed with metabolic dysfunction-associated steatotic liver disease (MASLD) versus those with MetALD. <b>Materials and Methods</b>: In this prospective study, 286 diabetic patients were followed for 12 months. All patients underwent transient elastography (TE) and ultrasound to assess hepatic steatosis. Participants were classified into MASLD and MetALD groups. The performance of fibrosis-4 index (FIB-4), and NAFLD fibrosis score (NFS) were also evaluated. <b>Results</b>: MASLD was diagnosed in 58.2% (167 patients), of whom 4.9% (7 patients) had TE values suggestive for liver cirrhosis. Among those with MetALD, 17.6% (21 patients) had TE values compatible with advanced fibrosis. MASLD subjects presented a slight decrease in liver fibrosis values from 6.58 ± 2.27 kPa to 6.03 ± 1.57 kPa in the 12 months. On the contrary, MetALD subjects had an increase of liver stiffness measurements (LSM) values from 11.83 ± 6.27 kPa to 12.24 ± 8.66 kPa. <b>Conclusions</b>: in diabetic patients, the coexistence of moderate alcohol intake and cardiometabolic risk factors (MetALD) is associated with more advanced liver fibrosis and impaired long-term glycemic control, compared to MASLD alone.

ROS derived from NADPH oxidase, particularly NOX2, are central to antimicrobial defense, coupling direct pathogen killing with redox signaling that shapes inflammation. This narrative review integrates recent advances on NOX2 structure, assembly, and spatiotemporal control in phagocytes, and outlines how ROS interact with NF-κB, MAPK, and Nrf2 networks to coordinate microbicidal activity and immune modulation. We summarize evidence that both ROS deficiency, as in chronic granulomatous disease, and uncontrolled excess, as in sepsis and severe COVID-19, drive clinically significant pathology, emphasizing the need for precise redox balance. Emerging therapeutic strategies include selective NOX2 inhibitors that limit pathological oxidative bursts, redox-modulating peptides that disrupt upstream activation cues, and Nrf2 activators that enhance endogenous antioxidant capacity, with attention to dosing challenges that preserve host defense while mitigating tissue injury. Key gaps remain in biomarker standardization, real-time in vivo ROS monitoring, and translation from animal models to patients, motivating personalized, combination approaches to redox medicine in infectious diseases.

Nano- and submicron particles (NSPs) with folate for targeting are actively used for the treatment and diagnosis of cancer and inflammatory diseases. Albumin-containing systems have enhanced biocompatibility, circulation time, and colloidal stability, which are important for medical applications. The outstanding binding properties of albumin allow the transport of numerous therapeutic and/or imaging agents. This review summarizes multiple aspects of binding a folate residue (or folic acid) to NSPs and the functioning of folate-albumin-NSPs. Special attention in the review is given to the types of bonds between folic acid and albumin, i.e., covalent and non-covalent, and to the confirmation and quantification of binding by different physicochemical methods. The process of binding, the qualitative and quantitative characteristics of binding and forming product, and its functioning are interconnected with the binding conditions; thus, an analysis of reaction conditions is provided. For the proper functioning of folate-albumin-NSPs, the state of albumin within them is important; thus, considerable focus in the review is placed on the features of structure modification of serum albumin in folate-albumin binding, i.e., the amino acid residues involved in this process and the conformational state of the protein. The stability and the functioning of the protein within folate-albumin-NSPs are discussed. Also, the effectiveness of targeting by folate is viewed as dependent on many characteristics of folate-albumin-NSPs, particularly on the peculiarities of binding between the folic acid residue and albumin. Furthermore, the authors discussed and suggested solutions concerning the shortcomings highlighted in the studies devoted to obtaining folate-modified albumin-containing NSPs.

Whole-blood gene expression analysis is essential for understanding molecular host responses, yet its use typically relies on venous sampling, limiting feasibility for frequent and remote monitoring. In an ancillary study of the COVERAGE France platform trial (NCT04356495), which enrolled at-risk outpatients with mild coronavirus disease 2019 (COVID-19) monitored at home, we compared transcriptomic profiles obtained from paired venous blood (Tempus tubes) and ultralow-volume, self-collected finger-prick capillary samples. We observed moderate to good concordance at the individual gene level and excellent agreement at the gene-set level between the two sampling approaches. High-frequency finger-prick sampling enabled daily resolution of immune dynamics, revealing early interferon responses, sustained neutrophil activation, and evolving erythroid and inflammatory signatures during the initial phase of mild COVID-19. These results demonstrate that finger-prick sampling is a feasible and reliable approach for at-home transcriptomic profiling, offering a powerful tool for longitudinal immune monitoring and advanced clinical research.

High mobility group box 1 is a dynamic nuclear protein that acts as a damage-associated molecular pattern when released from cells and plays key roles in neurodegenerative diseases. This review comprehensively analyzes the related post-translational modifications that affect the dual functions of high mobility group box 1 in neuroinflammation and neuronal survival, including acetylation, phosphorylation, oxidation, S-nitrosylation, lactylation, and ubiquitination. Post-translational modifications play critical regulatory roles in high mobility group box 1 subcellular localization, release processes and the specificity of receptor binding. In Alzheimer's disease, high mobility group box 1 exacerbates the disease through the Toll-like receptor 4/nuclear factor kappa B signaling pathway. Inhibition of high mobility group box 1 acetylation can alleviate neuroinflammation. Parkinson's disease models indicate that the S-nitrosylation of Cys106 is essential for the secretion of high mobility group box 1, which contributes to dopaminergic degeneration through the activation of microglia. In multiple sclerosis, high mobility group box 1 obstructs remyelination by inhibiting the maturation of oligodendrocytes and activating pro-inflammatory pathways. In contrast, high mobility group box 1 can maintain autophagy and DNA repair functions, suggesting its protective role. Therapeutic strategies targeting high mobility group box 1 show potential benefits. Glycyrrhizic acid inhibits disulfide-linked high mobility group box 1, SIRT activators suppress acetylation, and anti-high mobility group box 1 antibodies neutralize extracellular isoforms, thereby improving the results of preclinical studies. However, the diverse functions of high mobility group box 1 and the lack of post-translational modification-specific biomarkers present challenges for clinical translation. Future research should aim to create selective inhibitors that can cross the blood-brain barrier to target harmful forms of high mobility group box 1, and establish post-translational modification-based biomarkers for early detection. This review emphasizes that accurately targeting of high mobility group box 1 post-translational modifications in neurodegenerative diseases could be a new approach that can interrupt neuroinflammatory cascades while maintaining neuroprotective functions.

Neurodegenerative diseases such as amyotrophic lateral sclerosis, Alzheimer's disease, Parkinson's disease, and Huntington's disease are characterized by progressive neuronal loss and chronic neuroinflammation, with current treatments remaining largely symptomatic. This review explores the potential of cell-based immunotherapy as a disease-modifying strategy. Advances in stem cell biology and immune engineering have facilitated the development of therapies using mesenchymal stem cells, chimeric antigen receptor T cells, macrophages, regulatory T cells, modified macrophages, and monoclonal antibodies. These approaches aim to regulate immune mechanisms implicated in neurodegeneration, such as microglial activation, systemic inflammation, and immune checkpoint dysregulation. Notably, macrophage-mediated delivery systems, such as genetically modified cells expressing neurotrophic factors or antioxidant enzymes, have demonstrated neuroprotective effects. Likewise, emerging data support T-cell modulation and monoclonal antibody development as therapeutic targets in amyotrophic lateral sclerosis, Alzheimer's disease, Parkinson's disease, and Huntington's disease. We highlight current preclinical findings, underlying mechanisms, and translational challenges, emphasizing that immunomodulatory cell therapies represent a promising avenue for precision medicine in neurodegenerative diseases.

The devastating neurodegenerative disorder of Alzheimer's disease hallmarks the presence of protein aggregates known as amyloid-β plaques and neurofibrillary tangles, composed of amyloid-β peptides and aberrantly phosphorylated Tau protein, respectively. The accumulation of these inclusions leads to significant alterations in neuronal homeostasis and overall brain function, resulting in a progressive and rapid cognitive decline. Autophagy, the molecular mechanism of cellular waste removal through the lysosomal pathway, accounts for the degradation of both amyloid-β plaques and neurofibrillary tangles in the brain, conferring therefore protection against the pathology. In addition to general autophagy, several lines of evidence have reported the implication of selective autophagy receptors, including sequestosome 1/p62, the neighbor of BRCA1 gene, the nuclear-dot protein 52, and optineurin, in mediating the autophagic clearance of amyloid-β, phosphorylated Tau, or both. Herein, we have highlighted autophagy and selective autophagy as pivotal mechanisms in Alzheimer's disease, underlining selective autophagy receptors as a potential target for treatments in the future.

The permanent functional deficits resulting from the inability of adult mammalian central nervous system neurons to regenerate after injury present a significant clinical challenge. While traditional stem cell transplantation strategies continue to encounter ethical concerns and the risk of immune rejection, this impasse has shifted regenerative medicine research toward targeting endogenous astrocytes. Due to their intrinsic plasticity, widespread distribution throughout the central nervous system, and affinity for neurodevelopmental lineage, astrocytes are a unique target for in situ neuronal regeneration. This review systematically elucidates the core regulatory network governing astrocyte transdifferentiation, identifying 10 key signaling pathways, such as Wnt signaling pathway, that form a cascade regulatory system. Directed overexpression of transcription factors such as NeuroD1, Ascl1, or Neurog2 can directly initiate neuronal phenotypic conversion. Meanwhile, small molecule compounds such as valproic acid combined with CHIR99021 activate endogenous neurogenic programs by inhibiting the bone morphogenetic protein signaling axis. Notably, polypyrimidine tract binding protein 1 ( PTB ) gene silencing significantly enhances transdifferentiation efficiency by suppressing the microRNA 124/re1 silencing transcription factor (miR-124/REST) feedback loop. From a translational perspective, a multidimensional evaluation system based on morphological, molecular marker, and electrophysiological properties has demonstrated considerable therapeutic potential. In stroke models, NeuroD1-mediated transdifferentiation replenished approximately 30% of lost cortical neurons and improved motor coordination, evidenced by enhanced performance in food pellet retrieval, grid walking, and cylinder tests compared with controls. In spinal cord injury studies, SOX2-induced glutamatergic neurons moderately reduced glial scar density by about 25%, permitting regenerating axons to pass through while preserving the supportive structure of scar. In neurodegenerative contexts, PTB inhibition yielded functionally mature dopaminergic neurons and reconstructed nigrostriatal pathways in Parkinson's disease models. In Alzheimer's disease models, adeno-associated virus-delivered NeuroD1 induced whole-brain neural circuit remodeling, generating 500,000 new neurons widely distributed across the cortex and hippocampus, accompanied by improved cognitive performance. Current technical limitations include off-target effects of adeno-associated virus vectors, which cause nonspecific gene expression and require rigorous validation via Cre-loxP lineage tracing. Transdifferentiation efficiency is also highly influenced by regional microenvironments: gray matter astrocytes show higher conversion rates than those in white matter, and oxidative stress increases apoptosis among newly generated neurons. Clinical translation is further constrained by the safety of delivery systems and the aging tissue microenvironment, where transforming growth factor beta 1 is often elevated. Ferroptosis inhibitors have been shown to nearly double the survival rate of transdifferentiated cells, offering a novel strategy to mitigate oxidative damage. Based on current evidence, astrocyte transdifferentiation enables neural functional recovery across multiple disease models through endogenous repair mechanisms. Future advances should focus on optogenetically inducible vectors for spatiotemporal precision, non-viral delivery systems to mitigate vector-related risks, and integration of long-term safety validation in non-human primates with single-cell multi-omics technologies to facilitate the clinical translation of personalized regenerative therapies.

Bile acids emerge as multifunctional signaling molecules with dual hepatic and microbial origins, acting through farnesoid X receptor and Takeda G protein-coupled receptor 5 to influence inflammation and metabolism. Their dysregulation is consistently observed across various neurodegenerative diseases. The microbiota-gut-brain axis is a pivotal conduit for bile acids-driven neuromodulation, while sex-specific bile acid profiles and signaling pathways introduce critical biological heterogeneity. Emerging translational evidence indicates the promise of bile acids as biomarkers and therapeutic targets, yet highlights the critical hurdles that need to be addressed to realize precision interventions. Our core findings are: (1) Bile acids are far more than mere metabolic byproducts. They orchestrate core pathological processes such as neuroinflammation and energy metabolism. Their functions, whether neuroprotective or neurotoxic, are highly context-dependent, varying with cell type and disease-specific pathological backgrounds, thus exhibiting a potent "double-edged sword" effect. (2) The "microbiota-bile acids-brain axis" serves as a crucial bridge linking peripheral metabolic dysregulation to central nervous system pathology. (3) Sexual dimorphism emerges as a fundamental biological variable essential for understanding the heterogeneity in bile acid profiles and disease susceptibility. The primary contribution of this work is the proposal of an integrated "microbiota-bile acids-sex" framework that systematically describes the key scientific challenge of the context-dependent, dual roles of bile acids. Ultimately, this review champions a paradigm shift from a traditional brain-centric view to a systemic, metabolic perspective, establishing the bile acid system as a promising target for future precision therapeutic interventions.

Targeted drug delivery systems represent a promising strategy for enhancing the efficacy and specificity of cancer therapy. In this study, 35 nm folate-targeted gold nanoparticles are presented as nanoparticle-drug conjugates obtained by anchoring on their surface lipoyl terminating doxorubicin prodrug (proDoxo) releasable at the endolysosomal acidic pH to prevent off-site toxic effects. Colloidal stable nanoparticles with a density of proDoxo up to 1000 molecules/particle and 2 kDa mPEG-SH coating were obtained. At pH 5, Doxo was completely released from the nanoparticles in 5 days while only 13% was released over the same period at pH 7.4. The nanoparticle decoration with folic acid as a targeting agent bestowed nanosystems with selective drug delivery to folate receptor (FR)-overexpressing cancer cells and controlled intracellular release. This led to enhanced cancer cell killing by folated nanoparticles compared to their nontargeted counterparts. Moreover, folated nanoparticles were found to distribute more homogeneously inside KB^FR+ cancer cell spheroids than non-targeted nanoparticles, resulting in higher spheroid volume reduction.

Prostate cancer exhibits a strong familial association, and its heritability indicates a significant contribution from germline variants. While genome-wide association studies (GWAS) have identified common germline variants associated with prostate cancer risk, translating these statistical associations into functional mechanisms has remained a long-standing challenge. Consequently, most of our understanding of the genetic basis of prostate cancer stems from extensive studies of somatic mutations, leaving the germline genetic architecture largely unresolved. Because most germline variants lie in the noncoding genome and complex human diseases are predominantly driven by regulatory mutations, we herein asked which prostate cell types mediate the functional effects of germline variants, and thus represent the most genetically vulnerable populations. We generated paired epigenomic and transcriptomic profiles from reference human prostate tissues. Integrating these single-cell data with large-scale GWAS data identified a terminally differentiated luminal epithelial subtype that mediates the strongest germline risk in prostate cancer. We subsequently developed a deep learning model to score ~17 million GWAS variants based on their predicted impact on altering local chromatin accessibility in this vulnerable luminal epithelial subtype, and identified high-confidence candidate loci where high-risk germline variants likely alter promoter accessibility in prostate cancer. The implicated genes were involved in several pathways in tumorigenesis, displayed strong dosage sensitivity, and converged on the androgen receptor (AR)-mediated regulon, a mechanism also observed for somatic mutations. Overall, by unveiling cell types and candidate loci that mediate germline risk, our study defines the cell-type-specific germline architecture in prostate cancer and provides a comprehensive framework for understanding cancer heritability.

BACKGROUND: Polycystic ovary syndrome (PCOS) is the most common endocrine and metabolic disorder among women of reproductive age worldwide. This study aimed to investigate the therapeutic potential and molecular mechanisms of Atractylenolide III (ATL-III), a bioactive compound derived from Atractylodes macrocephala Koidz., in PCOS. METHODS: Follicular fluid and granulosa cells (GCs) were collected from women with PCOS. A PCOS mice model was established using dehydroepiandrosterone (DHEA). The body weight, estrous cycle, glucose tolerance, insulin tolerance, and ovarian morphology were measured to assess ovarian function. The knockdown and overexpression of ferredoxin 1 (FDX1) were conducted in human granulosa-like tumor cells (KGN). Viability and proliferation of GCs were evaluated by CCK-8 and EdU staining. RT-qPCR, Western blotting, and immunohistochemical staining were performed to detect molecule expressions in the PCOS patients, PCOS mice, and KGN cells. RESULTS: FDX1 expression was significantly decreased in both follicular fluid and GCs of PCOS patients. In PCOS mice, ATL-III was first found that improved the estrous cycle, disordered folliculogenesis, glucose tolerance, insulin resistance, and ovarian morphology, and upregulated ovarian FDX1, and proliferating cell nuclear antigen (PCNA) expression. DHEA-induced KGN cells demonstrated FDX1 suppression associated with proliferation inhibition, whereas FDX1 overexpression rescued proliferative capacity. ATL-III restored KGN cell viability and proliferation through FDX1 upregulation. Mechanistically, ATL-III activated PI3K/AKT/mTOR signaling by regulating FDX1 in both PCOS mice and KGN cells. CONCLUSION: Our findings establish ATL-III as a novel regulator of FDX1 that alleviates PCOS via PI3K/AKT/mTOR-mediated GCs proliferation enhancement, positioning it as a promising therapeutic agent for PCOS treatment.

Whole-genome sequencing technologies have advanced, leading to an increase in uncharacterized variants with unknown functions. This study focuses on characterizing cardiomyopathy-associated genes harboring theses rare variants and uncovering their cellular contexts using single-cell transcriptomics. We investigated whole genome sequencing on 245 unrelated Korean patients with either dilated (48.2%) and hypertrophic (47.8%) cardiomyopathy. Rare variants were identified and subjected to burden analysis to detect potentially causative candidates. To understand their biological impact, we profiled single-cell transcriptomes from an independent dataset, highlighting cell populations and interactions relevant to disease pathogenesis. A total of 3584 rare variants were discovered, including 50 pathogenic or likely pathogenic variants in 41 patients. Among the remaining 3534 variants of uncertain significance (VUS), burden analysis revealed 144 gene signatures significantly enriched in pathways related to cardiac muscle tissue development, heart morphogenesis, and endocrine system development (FDR < 0.003). These gene signatures were strongly correlated with cardiomyopathy - associated phenotypes, including HCM (HP:0001639) and DCM (HP:0001644) from the Human Phenotype Ontology (HPO) database. Single-cell transcriptomic analysis of 11,664 heart tissue cells demonstrated that the expression of these gene is influenced by cellular heterogeneity, which may contribute to disease manifestation. Further analysis identified five major cell types in heart tissue, revealing dynamic interactions between cardiomyocytes and endothelial cells. Rare genetic variants, including VUS, are closely linked to cardiac developmental processes and the sarcolemma integrity in cardiomyopathy. Our findings underscore the critical involvement of both cardiomyocytes and non-cardiomyocytes in disease etiology, highlighting the utility of integrating genomic and single-cell data for mechanistic insights.

PIC-SURE is an open-source platform for integrating and analyzing large-scale clinical and genomic data. Part of the NIH NHLBI BioData Catalyst® ecosystem, PIC-SURE enables real-time cohort building and analysis across 1.4 M participants within 273 studies (May 2025). Through a graphical interface and API, researchers can explore and analyze complex datasets in real time. This flexibility supports scalable and reproducible research, lowering the barrier to integrated clinical and genomic data analysis.

BACKGROUND: Infertility is a global health concern, with asthenozoospermia (AZS) being a common male factor. Our previous research demonstrated that the semenogelin I-derived peptide SgI-52 (SgI-52) binds more rapidly and extensively to demembranated sperm and AZS sperm, significantly inhibiting sperm motility and impairing mitochondrial function. However, the specific molecular targets of SgI-52 on sperm and the mechanisms by which it modulates oxidative stress and sperm function remain unclear. This study therefore investigates the interactions of SgI-52 with sperm surface proteins and its impact on oxidative stress. METHODS: Sperm proteins from AZS donors were extracted to identify SgI-52 targets using affinity chromatography and quantitative proteomics. The interaction with PRDX 6 was analyzed using AlphaFold 3 predictions and biolayer interferometry, and their localization on sperm membranes was determined by immunofluorescence and immunoelectron microscopy. Western blot measured SgI-52 in seminal plasma and PRDX 6 on sperm membranes. Functional assays evaluated the effects of SgI-52 on PRDX 6 enzymatic activity, oxidative stress levels, and mitochondrial membrane potential in sperm. RESULTS: SgI-52 bound to PRDX 6 and co-localized on sperm membranes. Both seminal plasma SgI-52 and membrane-associated PRDX 6 were higher in AZS donors than in normal donors. Functional assays showed that SgI-52 inhibited PRDX 6 activity, leading to increased ROS and MDA levels and mitochondrial dysfunction in sperm. CONCLUSION: In vitro study, we observed differential expression of SgI-52 and PRDX6 in normal and AZS samples and revealed their interaction, explaining the preferential binding of SgI-52 to compromised sperm. These findings suggest that SgI-52–derived peptides could serve as molecular probes for sperm quality evaluation and sperm selection in assisted reproductive technologies.

Thermal imaging plays a pivotal role in distinguishing distinct cellular states, assessing dynamic cellular activity in real-time health monitoring, and advancing the design of biofunctional materials for tissue engineering applications. The investigated photoluminescence characteristics of Dy³⁺ ion-doped fluorapatite demonstrate that this material offers stable temperature sensitivity. Our findings support the strategic design of next-generation nanomaterials for regenerative medicine and tissue engineering by elucidating key cellular interactions. Furthermore, our study has begun to address the impact of Dy³⁺ ion-doped nanomaterials on progenitor cell activity, providing valuable insights into their potential applications. A comprehensive description of photoluminescence characterization, including the LIR and S_R parameters, is provided to highlight its high thermal sensing potential. The antimicrobial activity of Dy³⁺ ion-doped fluorapatite is confirmed using the well-diffusion method against six commonly encountered microbial species. In vitro assays on hBMSCs included, among others, the determination of apoptosis profiles and metabolic potential through cytometric assays and molecular analyses (RT-qPCR and Western blot). These findings underscore the potential of Dy³⁺ ion-doped fluorapatite as a biocompatible material for biomedical applications while emphasizing the importance of further understanding its mechanism of action.

BACKGROUND: Peripheral nerve injury (PNI) remains a major clinical challenge, often resulting in incomplete functional recovery despite advances in microsurgical repair. Low-intensity pulsed ultrasound (LIPUS) has recently gained attention as a noninvasive biophysical therapy capable of enhancing nerve regeneration through mechanical stimulation and cellular activation. MAIN BODY: This review synthesizes current preclinical evidence on the effects of LIPUS across in vitro and in vivo models and outlines its underlying mechanobiological mechanisms and translational implications. A systematic search identified thirty-four experimental studies evaluating LIPUS in nerve injury models, including crush, transection, autograft, and conduit repair. Collectively, these studies demonstrate that LIPUS accelerates axonal regeneration, promotes remyelination, enhances Schwann cell proliferation and migration, and increases neurotrophic factor expression. LIPUS also facilitates neural stem cell differentiation and neuronal survival by activating key intracellular signaling pathways. Despite encouraging preclinical results, significant challenges remain in translating LIPUS to clinical practice. Variability in stimulation parameters, injury models, and species hampers reproducibility, and regulatory approval for neural applications is still pending. Future research should focus on standardizing dosimetry, validating mechanistic pathways in human tissues, and conducting multicenter clinical trials to establish safety, efficacy, and practical treatment guidelines. SHORT CONCLUSION: The current evidence supports LIPUS as a safe and adaptable mechanotransductive therapy that can modulate cellular behavior and molecular signaling to enhance peripheral nerve regeneration.

<h4>Background</h4>Mesenchymal stromal cells (MSCs) support tissue repair in osteoarthritis (OA), with migration to damaged tissue being a key strategy in in situ tissue engineering. Their regenerative potential depends on factors such as differentiation, level of senescence, and responsiveness to signaling molecules. However, previous findings on these properties of OA MSCs remain inconclusive. This study integrates multiple aspects and tests feasibility using a well-characterized chemoattractant.<h4>Methods and results</h4>MSCs from non-OA donor (ND) and OA donor were characterized for their trilineage differentiation potential as well as for their senescence level by (immune-) histochemistry, RT-qPCR, microarray analysis, and a bead-based immunoassay for cell culture supernatants. No difference in differentiation and senescence level was observed, the latter being indicated by a similar activity of <i>β</i>-Galactosidase (<i>β</i>-gal), gene expression profiles of cyclin-dependent kinase (CDKN) inhibitor 2A (CDKN2A), CDKN inhibitor 1A (CDKN1A), sirtuin 1 (SIRT1), and matrix metallopeptidase 1 (MMP1), as well as secreted cytokines. Chemokine receptors in OA MSCs were detected using immunohistochemistry and RT-qPCR. Expression of CCR1-CCR7, CCR9, and CXCR1-CXCR6 in OA MSCs was confirmed on gene and protein levels. Both OA and ND MSCs migrated toward 1000 nM CCL25, as evaluated via a Boyden chamber assay. Subsequent genome-wide microarray analysis of OA MSCs after treatment with 1000 nM CCL25 corroborated its influence on migration, proliferation, apoptosis, and differentiation as defined by Gene Ontology terms (GO terms). Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis confirmed this broad impact and emphasized the role of cytokine-cytokine-receptor interaction and metabolic pathways.<h4>Conclusion</h4>Our data indicate that OA MSCs retain their differentiation potential and do not exhibit an increased senescent phenotype. Their chemokine receptor profile is conducive of migration, and both OA and ND MSCs respond to CCL25, highlighting the potential of OA MSCs for directed in situ repair.

<h4>Background</h4>Fine particulate matter (PM2.5) has been significantly linked to the progression of various neurological and neurodegenerative diseases.<h4>Objective</h4>This review aims to elucidate the molecular and pathophysiological effects induced by chronic exposure to PM2.5 in neurological and neurodegenerative diseases, including Alzheimer's, Parkinson's, Huntington's, multiple sclerosis, and epilepsy.<h4>Introduction</h4>PM2.5 penetrates the central nervous system (CNS) via the olfactory nerve or by disrupting the blood-brain barrier (BBB), triggering oxidative stress, neuroinflammation, mitochondrial dysfunction, and epigenetic alterations.<h4>Discussion</h4>In Alzheimer's and Parkinson's diseases, PM2.5 exacerbates the accumulation of β-amyloid, hyperphosphorylated tau, and α-synuclein, while in Huntington's disease, it worsens toxicity mediated by mutant huntingtin. In multiple sclerosis, these particles intensify neuroinflammation and axonal damage, whereas in epilepsy, they promote neuronal hyperexcitability and recurrent seizures. These mechanisms contribute to neuronal damage, symptom progression, and functional decline.<h4>Conclusion</h4>This evidence highlights the urgent need for strict environmental policies to reduce PM2.5 exposure and further research to develop therapeutic strategies that mitigate its effects on neurological diseases, thereby improving the health of vulnerable populations.

<h4>Objectives</h4>Ribosomal protein S6 kinase A2 (RPS6KA2) has been identified as a potential prognostic biomarker in several cancers, including breast cancer, glioblastoma, and prostate cancer. However, its functional significance in ovarian cancer is not well characterized. This study was designed to explore the therapeutic relevance of modulating RPS6KA2 in the context of ovarian cancer, particularly in relation to cisplatin resistance.<h4>Methods</h4>The expression levels of RPS6KA2 and key regulators involved in autophagy and ferroptosis were assessed using quantitative reverse transcription-PCR, immunofluorescence staining, immunohistochemistry, and western blotting. Prognostic associations were conducted using the Kaplan-Meier Plotter database. Autophagy flux assays and visualization of autophagosomes were performed to assess autophagy activity. Ferroptosis-related parameters, including intracellular iron content, glutathione (GSH) levels, reactive oxygen species (ROS) generation, and mitochondrial membrane potential, were measured to determine ferroptotic changes. <i>In vivo</i> experiments were carried out to determine the antitumor efficacy of RPS6KA2 modulation in combination with pathway-specific agents.<h4>Results</h4>Using ovarian cancer cell lines and clinical tissue samples, we demonstrated that RPS6KA2 expression was significantly downregulated in cisplatin-resistant cells and tissues compared to their sensitive counterparts. Low RPS6KA2 expression correlated with unfavorable patient outcomes and enhanced chemoresistance. Mechanistically, RPS6KA2 inhibited autophagy by modulating the phosphatidylinositol 3-kinase-protein kinase B-mammalian target of rapamycin (PI3K-AKT-mTOR) signaling pathway, which in turn increased sensitivity to cisplatin. Additionally, RPS6KA2 facilitated ferroptosis, contributing to its tumor-suppressive function. miR-512-3p was identified as a negative regulator of RPS6KA2, driving cisplatin resistance through suppression of RPS6KA2 expression. <i>In vivo</i> validation confirmed that combining RPS6KA2 targeting with autophagy inhibitors or ferroptosis inducers significantly enhanced cisplatin sensitivity in ovarian cancer models.<h4>Conclusion</h4>These results collectively indicate that targeting the miR-512-3p/RPS6KA2 regulatory axis may offer a novel and effective strategy for overcoming cisplatin resistance in ovarian cancer.

<h4>Background</h4>A significant proportion of patients still cannot benefit from existing targeted therapies and immunotherapies, making the search for new treatment strategies extremely urgent. In this study, we combined integrate public data analysis with experimental validation to identify novel prognostic biomarkers and therapeutic targets for lung adenocarcinoma (LUAD).<h4>Methods</h4>We analyzed RNA and protein databases to assess the expression levels of cytochrome C oxidase 5B (COX5B) in LUAD. Several computational algorithms were employed to investigate the relationship between COX5B and immune infiltration in LUAD. To further elucidate the role of COX5B in LUAD, we utilized multiple experimental approaches, including quantitative reverse transcription PCR assays, western blot, immunohistochemistry, electron microscopy, flow cytometry, and EdU proliferation assays.<h4>Results</h4>We revealed that COX5B was significantly elevated in LUAD and positively correlated with poor prognosis of LUAD patients. Analysis of co-expression network indicated that COX5B may take part in the intracellular adenosine triphosphate (ATP) synthesis through the oxidative phosphorylation pathway. There was a negative correlation between COX5B expression and immune infiltration in LUAD. Furthermore, we validated that COX5B levels were significantly elevated in both LUAD tissues and cell lines. Specifically, immunohistochemistry (IHC) assays revealed a 2.32-fold increase of COX5B in tumor tissues compared to that in adjacent normal tissues (<i>p</i> = 0.0044). Additionally, COX5B knockdown disrupted the redox homeostasis, ultimately suppressed the proliferation of LUAD cells. Subsequent investigations demonstrated that berberine effectively targeted COX5B, diminishing its protein expression and consequently inhibiting cell proliferation and tumor growth in LUAD.<h4>Conclusions</h4>This study established that upregulated COX5B was positive associated with poor patient prognosis in LUAD, elucidating the mechanisms by which berberine targets COX5B to inhibit tumor growth, thereby providing a novel therapeutic target and strategy for the clinical management of LUAD.

Dravet Syndrome (DS) is a severe genetic epileptic encephalopathy caused by mutations in the <i>SCN1A</i> gene that encodes the voltage-gated sodium channel (Na_V1.1) subunit alpha. DS is characterized by intractable seizures, progressive developmental delay, cognitive impairment, and high mortality due to sudden unexpected death in epilepsy (SUDEP). SUDEP is mediated by respiratory dysfunction, but the exact molecular underpinnings are unclear. Though hippocampal metabolic alterations have been reported in DS mice, such changes in brain regions controlling breathing have not been studied. We used <i>Scn1a^A1783V</i>^/<i>WT</i> DS mice to study temporal alterations in the brain metabolome, including analysis of brainstem and forebrain regions. Glycolytic and pentose phosphate pathway intermediates were significantly elevated in the brainstem of DS mice during the period of enhanced susceptibility to mortality (post-natal days P20-30). In older P40-P50 mice, mitochondrial aconitate and the antioxidant glutathione were significantly elevated in the brainstem. Single-nuclei RNA sequencing (snRNA seq) and proteomic analyses revealed alterations in genes associated with neurotransmission, cellular respiration, and protein translation, as well as reorganization of protein kinase-mediated pathways that are specific to the brainstem. These findings suggest that there are widespread metabolic changes in the brainstem of DS mice.

Accumulation of misfolded proteins is implicated in neurodegenerative diseases. One of these is Huntington's disease, which is caused by an expansion of trinucleotide (CAG) repeats in exon 1 of huntingtin gene (<i>HTT</i>). This expansion results in the production of mutant huntingtin exon1 protein (mHttEx1) containing polyglutamine tracks that is prone to cytotoxic aggregation. These mHttEx1 aggregates range from small soluble aggregates to large insoluble inclusion bodies. The mechanisms to clear mHttEx1 aggregates include ubiquitin-dependent proteasomal degradation and autophagy. For the proteasomal degradation of mHttEx1, ubiquitinated protein is first recognized by the Cdc48 complex for extraction and unfolding. For autophagy, mHttEx1 inclusion bodies are engulfed by an autophagosome, which fuses with the vacuole/lysosome and delivers cargo for vacuolar degradation. We name this autophagy IBophagy. In this study, we further show that the ubiquitination of mHttEx1 by the E3 ligase San1, its extraction and unfolding by the Cdc48 complex, and subsequent proteasomal degradation are all essential steps for mHttEx1 IBophagy in budding yeast, revealing a new layer of autophagy regulation and mHttEx1 cytotoxicity.

Lion-head goose is a large-sized breed native to Guangdong Province, China, exhibits remarkable capacity for fatty liver production under overfeeding conditions and is highly valued by local farmers and consumers. However, the molecular mechanisms driving fatty liver development in this breed are still unknown. In this study, we evaluated liver weight differences between normally fed and overfed Lion-head geese and further examined sex-specific differences following overfeeding. Overfeeding significantly increased liver weight more than 340%, and males possess a stronger capacity for lipid deposition under the same feeding regimen compared with females. RNA-Seq analysis identified 1476 differentially expressed genes (DEGs) shared by both sexes, which were mainly enriched in lipid and energy metabolism, oxidative stress, and mitochondrial pathways. In addition, 627 male-specific and 420 female-specific DEGs revealed sex-dependent differences, with males showing stronger transcriptional regulation and females exhibiting enhanced antioxidant and detoxification responses. Weighted gene co-expression network analysis (WGCNA) revealed 320 co-hub genes enriched in lipid and energy metabolism in overfeeding-induced fatty liver, along with 9 co-hub genes related to sex differences. Alternative splicing (AS) analysis detected 131 differentially spliced genes (DSGs). Integration of both approaches identified 7 overlapping genes, <i>HYCC2</i> (Hyccin PI4KA lipid kinase complex subunit 2), <i>AGL</i> (Amylo-Alpha-1,6-Glucosidase and 4-Alpha-Glucanotransferase), <i>CCDC62</i> (Coiled-coil domain containing 62), <i>IGSF5</i> (Immunoglobulin superfamily member 5), <i>MGARP</i> (Mitochondria-localized glutamic acid-rich protein), <i>CD80</i> (Cluster of Differentiation 80), and <i>FPGS</i> (Folylpolyglutamate synthase), as potential key regulators. These findings provide new insights into transcriptional and post-transcriptional regulation of overfeeding-induced fatty liver in geese.

During cerebellar development, granule cell precursors (GCPs) undergo a series of tightly regulated events, including proliferation, migration, and differentiation. Arhgef7, a guanine nucleotide exchange factor for Rac1 and Cdc42, plays a crucial role in these processes. This study investigates the role of Arhgef7 in cerebellar development using conditional knockout (cKO) mice. We demonstrate that Arhgef7 is expressed in GCPs. Loss of <i>Arhgef7</i> in GCPs results in severe cerebellar hypoplasia and foliation defects, particularly affecting lobules VI/VII. <i>Arhgef7</i> cKO mice exhibit reduced postnatal GCP proliferation, disorganized cell layers, delayed differentiation, and impaired tangential and radial migration of GCPs. Our findings highlight the essential role of Arhgef7 in regulating multiple aspects of GCP development, thereby ensuring proper cerebellar morphogenesis.

Lung cancer remains a leading cause of cancer mortality worldwide, highlighting the need for novel therapeutics. Here, we designed and synthesized a series of adamantyl-substituted chalcones and identified CA13 as a lead compound with potent and selective antiproliferative activity against non-small cell lung cancer (NSCLC) cells. CA13 triggered both apoptosis and autophagy in H292 cells. Western blotting and confocal imaging confirmed the activation of complete autophagic flux, while inhibition of autophagy markedly enhanced CA13-induced apoptosis, suggesting a cytoprotective role of autophagy. Mechanistically, CA13 activated JNK phosphorylation in a dose- and time-dependent manner, and pharmacological blockade of JNK significantly attenuated apoptotic signaling. In vivo, CA13 effectively suppressed H292 xenograft tumor growth without apparent systemic toxicity. Collectively, these results demonstrate that CA13 exerts its antitumor effects through JNK-dependent apoptosis accompanied by cytoprotective autophagy, providing a promising structural framework for the development of chalcone-based anticancer agents targeting programmed cell death pathways.

Melanin pigments are ubiquitous biopolymers across diverse life forms and play multifaceted roles in cellular defense and environmental adaptation. The specialized neuromelanin in human brains accumulates mainly within catecholaminergic neurons of the substantia nigra and locus coeruleus, serving as a crucial modulator of brain homeostasis, metal detoxification, and oxidative stress responses. The intricate processes of human melanogenesis, encompassing both cutaneous and neuronal forms, are governed by complex genetic networks. Concurrently, melanin in fungi (synthesized through distinct genetic pathways) confers remarkable resistance to environmental stressors, including ionizing radiation. Recent advancements in omics technologies-including transcriptomics, proteomics, metabolomics, and epigenomics-have profoundly enhanced our understanding of neuromelanin's molecular environment in health, aging, and neurodegenerative conditions such as Parkinson's disease (PD), Alzheimer's disease (AD), and other neurological disorders. This article reviews the genetic underpinnings of human melanogenesis and fungal melanogenesis, explores the convergent and divergent evolutionary pressures driving their functions, and synthesizes the rapidly accumulating omics data to elucidate neuromelanin's critical, and often dual, role in human brain pathology. Moreover, we discuss the intriguing parallels between neuromelanin and fungal melanin, highlighting radioprotection and its potential implications for neuroprotection and astrobiology, with a special emphasis on the need to investigate neuromelanin's potential for radioprotection in light of fungal melanin's remarkable protective properties.

Neurodevelopmental disorders (NDDs) are defined as a group of conditions that result from impaired brain development. Disorders that are commonly classified under NDDs include intellectual disability (ID), autism spectrum disorder (ASD), attention-deficit/hyperactivity disorder (ADHD), communication and learning disorders, developmental delay (DD), brain malformations, cerebral palsy, Down syndrome, schizophrenia, and childhood epilepsies. A significant hinderance in the development of targeted treatments for NDDs are gaps in understanding how underlying genetic changes alter cellular physiology and how these changes may converge or diverge across NDDs with similar symptoms. Here, we focus on the genetic overlap between epilepsy, ASD, and other NDDs to identify common cellular and molecular mechanisms that may inform future treatments for each of these disorders individually or together. We describe several genes-including <i>CDKL5, TSC1/2, SCN1a,</i> and <i>TANC2</i>-that have been associated with epilepsy, ASD, or other NDD phenotypes that play a critical role in regulating one or more stages of brain development or function but differ widely in their disease-causing mechanisms. We also describe genotype-phenotype relationships. Finally, how a gene may cause NDDs through distinct functional pathways, or where different types of pathogenic variants within the same gene can have significantly different phenotypic outcomes is detailed.

Kynurenic acid (KYNA), a small molecule derived from the tryptophan-kynurenine pathway, can readily diffuse across biological membranes and act as an endogenous ligand for receptors such as the aryl hydrocarbon receptor (Ahr). While KYNA dysregulation is implicated in neurodegenerative disorders, the role of the KYNA-Ahr-IL-6 axis in MSC proliferation and differentiation remains poorly defined. We investigated the impact of KYNA on murine bone marrow-derived MSCs (BM-MSCs) at various concentrations (10-200 μM) and time points (8-48 h). The BM-MSC phenotype was assessed via flow cytometry; proliferation, via cell counting; and the gene expression of <i>Ahr</i>, <i>Cyp1a1</i>, <i>Cyp1b1</i>, and <i>Il-6,</i> via quantitative real-time PCR. Multipotency was evaluated through adipogenic, osteogenic, and chondrogenic differentiation assays with histochemical confirmation. KYNA significantly upregulated Ahr mRNA expression. Among the tested concentrations, 100 μM KYNA induced the highest <i>Ahr</i> expression (~19.1 ± 1.5-fold greater than that of the untreated controls, <i>p</i> < 0.005). Notably, 10 and 50 μM KYNA caused moderate induction, whereas compared with 100 μM KYNA, 200 μM did not further increase expression. In addition, KYN treatment increased <i>Cyp1a1</i>, <i>Cyp1b1</i>, and <i>Il-6</i> expression, with increases of ~64.6 ± 4.5-fold, ~43.6 ± 2.3-fold, and ~41.6 ± 1.2-fold, respectively. Compared with no treatment, 100 µM KYNA enhanced BM-MSC proliferation by 1.210 ± 0.02, 1.189 ± 0.03, and 1.242 ± 0.02-fold across passages P3, P4, and P5, respectively (<i>p</i> < 0.05), without altering Sca-1, CD90, or CD45 expression or impairing trilineage differentiation potential. KYNA may activate the AHR-IL-6 signaling axis to promote BM-MSC expansion. This controlled proliferative effect, without loss of phenotypic or functional integrity, highlights the pharmacological potential of KYNA as a small-molecule modulator for stem cell-based therapies.

<h4>Background</h4>Platelets play a central role in hemostatic and inflammatory responses during septic shock, with lipids being essential for their function. However, the specific lipidomic alterations occurring in platelets during septic shock remain poorly understood.<h4>Objectives</h4>This study aimed to characterize platelet lipidomic changes in septic shock and investigate their associations with disease severity.<h4>Methods</h4>In this matched case-control study, platelets were isolated from 49 septic shock patients and 47 nonseptic controls (matched for age, gender, and comorbidities). Lipidomic profiling was performed using untargeted lipidomics to identify significant alterations in the platelet lipidome. Associations among lipid changes, clinical data, and plasma biomarkers of coagulopathy and inflammation were explored.<h4>Results</h4>More than 60% of the annotated platelet lipids were significantly altered in septic shock. Cholesteryl esters, sphingomyelins, lysophosphatidylcholines, and ether-lipids were significantly reduced, while ceramide levels increased. Fatty acyl chain remodeling displayed distinct patterns, with polyunsaturated fatty acids increasing in triacylglycerols and decreasing in phospholipids. Lipid alterations were strongly associated with thrombocytopenia, and lysophosphatidylcholine levels inversely correlated with disease severity, as indicated by the Sequential Organ Failure Assessment score.<h4>Conclusions</h4>Septic shock induces significant disruptions in the platelet lipidome, with the extent of these alterations correlating with sepsis-associated thrombocytopenia severity. The observed changes affect multiple lipid classes, surpassing those reported under physiological conditions or in other diseases. These findings highlight the impact of sepsis-driven dysregulated inflammation and coagulopathy on platelet lipid composition, providing new insights into sepsis pathophysiology.

Membranous nephropathy (MN) is the most common cause of adult nephrotic syndrome. Current treatments rely heavily on immunosuppressants; however, some patients do not achieve the desired therapeutic effect. Therefore, the identification of new drug targets and the development of novel medications are of urgent importance. In this study, we collected protein quantitative trait loci (pQTLs) for 734 circulating plasma proteins (CPPs) from previous research. Using principles of Mendelian genetics, we conducted a Mendelian randomization (MR) analysis using three inferential methods: the Wald ratio, inverse-variance weighted (IVW), and MR-Egger. After assessing heterogeneity, we identified 16 CPPs that were positively correlated with the occurrence of MN and 14 CPPs that were negatively correlated with MN. Enrichment analysis showed that these CPPs are primarily involved in oxidative stress and inflammation pathways, which are closely related to the development and progression of MN. Subsequently, using the Drug Signatures Database (DsigDB), we predicted potential drugs that might interact with these CPPs. Finally, we found that curcumin, a natural compound known for its antioxidant, anti-inflammatory, and immunomodulatory properties, could be a potential therapeutic molecule for the treatment of MN. This study aimed to systematically screen CPPs for causal associations with MN risk using MR, characterize the biological pathways of implicated proteins, and nominate potential therapeutic agents via computational drug repurposing analysis.

Bone tissue engineering is a pillar of regenerative medicine; traditional bone autografts, allografts, and metallic implants face difficulties due to donor site morbidity, risks of infection, and poor integration with the host bone. Nanohydroxyapatite (nHA) has become a popular synthetic tissue-engineered approach, although its brittleness and low fracture toughness limit its applications. This systematic review explores nHA-bioactive glass composites (HAGNs) as promising solutions for bone regeneration, analyzing their biocompatibility, osteoconductivity, and mechanical properties. This systematic review followed the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines. A comprehensive search strategy was employed across multiple databases from January 2000 to February 2024, using MeSH terms related to hydroxyapatite (HA), glass nanoparticles, and bone regeneration. Eligible studies included peer-reviewed articles reporting outcomes relevant to bone healing or tissue engineering using HA glass nanoparticles. The search yielded 57 records, with 15 studies meeting the inclusion criteria after screening and selection. These studies explored various fabrication methods and consolidation techniques for nHA-based scaffolds, highlighting their improved biocompatibility, osteoconductivity, and potential for bone regeneration. Limitations involved long drying periods, the need for specific consolidation conditions, and more studies to confirm long-term performance. Findings underscored the importance of preserving key properties of nHA, such as low crystallinity and nanoscale dimensions, to mimic natural bone tissue effectively. nHA and bioactive glass composites show significant potential in bone tissue engineering. They offer the combination of biocompatibility, osteoconductivity, and mechanical strength necessary for effective bone regeneration. Further research is needed to facilitate the clinical translation of these biomaterials.

Zinner Syndrome (ZS) is a rare congenital anomaly defined by the triad of unilateral renal agenesis, ipsilateral seminal vesicle cyst, and ejaculatory duct obstruction. We report a 55-year-old male presenting with chronic perineal pain and LUTS. Imaging confirmed ZS (right renal agenesis, 3.3 × 3.8 cm seminal vesicle cyst). The patient underwent surgical excision, converting from laparoscopic to open due to dense adhesions. He achieved complete symptom resolution and remained stable at two-year follow-up. ZS should be considered in older males with non-specific LUTS. Surgery is effective, and advanced imaging is crucial for diagnosis.

Stiffening of the extracellular matrix underlying the epithelial cells of the large intestine is associated with aging as well as many diseases. Yet the impact of the stiffened matrix on epithelial physiology remains poorly understood. A 2D and 3D microphysiological model of the large intestine was developed using a collagen scaffold with a physiologic or excessive stiffness (Young's moduli of 2.84 ± 0.85 kPa and 15.9 ± 0.73 kPa) by altering the collagen concentration within the substrate. Diffusion of a 10 and 40 kDa fluorescent dextran was significantly different between the physiologic and stiff scaffold (97.8 vs 79.8<i>µ</i>m²s^-1[10 kDa] and 68.2 vs 56.8<i>µ</i>m²s^-1[40 kDa], respectively). When primary human epithelial cells of the large intestine were grown as a 2D monolayer, cultures on the physiologic scaffold grew to a significantly higher density with more proliferative and fewer differentiated cells than cultures on the stiffened scaffold. Three-dimensional crypt arrays were also fabricated with the physiologic and stiff substrates, populated with cells, and a growth factor gradient applied. The cell density, proliferation, and height-to-width ratio was significantly greater for cells on the physiologic scaffold relative to that of cells on the stiffened scaffolds. Placement of a layer of intestinal fibroblasts below the epithelium on the crypt arrays did not mitigate the impact of the stiffened substrate. Bulk-RNA sequencing revealed 378 genes that were significantly upregulated and 385 genes significantly downregulated in the stiffened vs physiologic scaffolds. This work demonstrates that a molded collagen hydrogel can be used to mimic the biophysical characteristics of a stiffened intestinal stroma, recapitulating physiology observed<i>in vivo</i>. This<i>in vitro</i>model of polarized crypts with a tunable underlying substrate will enable an improved understanding of intestinal epithelial cell morphology, stem cell maintenance and lineage allocation within a stiffened environment.

<h4>Purpose of review</h4>Atherosclerotic cardiovascular disease (ASCVD) remains a leading cause of morbidity and mortality worldwide. Although there is increasing recognition of sex differences in ASCVD epidemiology, pathogenesis, and clinical outcomes, the underlying biological mechanisms are still insufficiently understood. Women often present with distinct disease phenotypes, such as a higher prevalence of fibrous plaques and microvascular dysfunction, compared with the lipid-rich, inflammatory plaques more typical in men. This review examines recent omics research to clarify the molecular basis of these sex-specific patterns and explores their implications for precision cardiovascular medicine.<h4>Recent findings</h4>Advances in genomics, epigenomics, transcriptomics, proteomics, and metabolomics have shown that sex differences in ASCVD arise from complex hormonal, genetic, epigenetic, and molecular interactions. The variety of available omics approaches offers the potential to discover sex-specific regulatory networks and therapeutic targets, thereby addressing persistent knowledge gaps. However, significant challenges remain, including integrating these diverse omics layers, harmonising datasets across platforms, managing substantial computational demands, and navigating ethical constraints related to data sharing. Multiomics technologies provide unprecedented opportunities to dissect sex-specific mechanisms in ASCVD and to refine individualised risk stratification and therapeutic strategies. Overcoming current analytical and infrastructural barriers through collaborative efforts, standardised methodologies, and responsible data governance will be critical to unlocking the full potential of multiomics in precision cardiovascular medicine. This review synthesises recent evidence across omics domains and underscores their potential to improve ASCVD prevention and treatment.

<h4>Importance</h4>This study provides nationally representative data on critical care advanced practice provider (APP) workforce structure and practice, addressing gaps in onboarding, productivity assessment, and administrative roles that influence workforce sustainability.<h4>Objectives</h4>To analyze the composition of the clinical practice and workforce demographics of critical care medicine APPs.<h4>Design, setting, and participants</h4>We utilized a cross-sectional survey, which was distributed via email through the Society of Critical Care Medicine email list, targeting critical care APPs.<h4>Main outcomes and measures</h4>A total of 518 critical care APPs participated in the survey, comprising 63.2% advanced practice registered nurses, 28.8% physician assistants, and 8% clinical nurse specialists. The majority of the sample APPs worked 12-hour shifts covering days, nights, and weekends, with over 50% managing 6-10 patients per day shift. Common billing practices included critical care time (60.6%) and bedside invasive procedures (64.3%). Formal onboarding was reported by 68.6% of respondents, with academic hospitals more likely to offer such programs than community hospitals (72.3% vs. 27.7%; p < 0.001). Administrative APP roles were more prevalent in academic centers (p < 0.001) and were associated with greater access to professional development opportunities (p < 0.001). A significant proportion of respondents (41.5%) reported that their productivity was not formally measured. Notably, 22% of respondents reported an intent to leave the profession, particularly those lacking leadership or professional growth opportunities (p < 0.01).<h4>Conclusions and relevance</h4>This study captures practice trends and demographics of the critical care APP workforce. Day-to-day tasks among all respondents were globally clinically focused, but there were disparities in workload distribution, onboarding, productivity measurement, and administrative roles across types of centers. Additionally, there was significant association between limited professional growth opportunities and intent to leave critical care. These findings underscore the need for healthcare systems to invest in structured support, leadership development, and professional growth to enhance APP retention, engagement, and productivity. Further research is needed to identify efficient workload and staffing models.

<h4>Purpose of review</h4>Cystic fibrosis liver disease (CFLD) is a significant nonpulmonary complication of cystic fibrosis, affecting approximately 5-10% of patients. It encompasses a spectrum of hepatic abnormalities ranging from mild, transient elevations in liver enzymes to advanced CFLD (aCFLD), which is marked by clinically relevant portal hypertension due to cirrhotic or noncirrhotic liver pathology. This review focuses on aCFLD as the clinically meaningful form of the disease and summarizes recent mechanistic insights into its pathogenesis that may inform the development of targeted therapeutic strategies.<h4>Recent findings</h4>CFLD pathogenesis has been traditionally linked to defective bile secretion. Emerging evidence, however, highlights additional contributors, including cholangiocyte immune dysregulation, gut dysbiosis, and intestinal barrier dysfunction, which together promote hepatic inflammation. Furthermore, recent studies underscore the role of vascular alterations independent of cirrhosis, specifically noncirrhotic portal hypertension, as the main clinical feature in aCFLD. These findings support a multifactorial, multihit model of disease in the pathogenesis of CFLD.<h4>Summary</h4>The complex interplay of these factors suggests that effective treatment for aCFLD may require a multifaceted approach. Advances in understanding the gut-liver axis and vascular contributions provide new therapeutic targets. Future research should focus on validating these findings and evaluating the efficacy of cystic fibrosis transmembrane conductance regulator modulators and microbiome-targeted treatments in altering the course of CFLD.

<h4>Background</h4>Racehorses undergo profound physiological changes with training and competition, but current biomarkers inadequately capture the complex molecular dynamics of exercise. This study aimed to identify novel plasma biomarkers of training adaptation and peak load using high-throughput proteomics.<h4>Objectives</h4>We hypothesised that systematic training and racing induce distinct plasma proteomic signatures, enabling the discovery of candidate biomarkers linked to training status, oxidative stress, inflammation and metabolic remodelling.<h4>Study design</h4>In vivo longitudinal study.<h4>Methods</h4>Forty-nine Arabian and Thoroughbred racehorses underwent standardised high-intensity training. Plasma samples were collected at rest, immediately post-exercise and after recovery during three phases: initial training (T1), mid-season conditioning (T2) and race-phase (R). In total, 314 samples were analysed using tandem mass tags based quantitative proteomics and Orbitrap mass spectrometry. Protein abundance changes were assessed with multiple-testing correction (q < 0.05), and pathway enrichment was performed using STRING and ShinyGO.<h4>Results</h4>Proteomic responses differed by phase. T1 showed broad activation of inflammatory (S100A8/A9), antioxidant (superoxide dismutase 1, catalase) and metabolic proteins (glucose-6-phosphate dehydrogenase, phosphoglycerate kinase 1). T2 displayed a more refined profile with remodelling and redox regulators (decorin, thymosin β4, glutathione S-transferase). Racing elicited the strongest response, with over 100 up-regulated proteins linked to energy metabolism, oxidative defense and cytoskeletal adaptation. Several proteins: including S100A8, thymosin β4, prothymosin-α, cofilin-1 and lipocalins, were consistently modulated across phases, highlighting their biomarker potential.<h4>Main limitations</h4>Breed imbalance and incomplete follow-up sampling may affect generalisability. Validation in larger, diverse cohorts with targeted assays is required.<h4>Conclusions</h4>This study identifies a panel of promising plasma proteins as candidate biomarkers of exercise adaptation and overload in racehorses. These findings may support improved monitoring of performance, training load and early detection of overtraining in equine athletes.

We explore strategies to harness ancestral diversity in PsychENCODE Consortium Genotype-Expression (GEx) reference panels (adult and developing brain) and Psychiatric Genomics Consortium GWAS data to improve genetically regulated expression (GReX) models and their use for Transcriptome-wide association study (TWAS) discoveries, uncovering previously unknown aspects of psychiatric functional genomics. We trained multiple GReX models on rigorously constructed GEx panel subsets, generated by downsampling, segregating, and/or mixing samples of Admixed African and European ancestries, and based on disease status. Ancestry-specific GReX genes were enriched in pathways involving mitochondrial functions, organelle structure, and metabolism. These models were integrated with ancestry-specific GWASs to conduct bipolar disorder, major depressive disorder, posttraumatic stress disorder, and schizophrenia TWAS. TWAS signals obtained by applying AA- and EUR-specific GReX models to an ancestry-specific GWAS were largely concordant, and mismatched-TWAS (e.g. AA-GReX applied to EUR-GWAS) revealed biologically meaningful signals missed by matched-TWAS. Shared signals across the four disorders were more prominent in the developing brain, involving genes such as H4C13, ZSCAN12P1, and FLOT1, and pathways related to megakaryocyte and muscle development, and neurotransmitter regulation. Overall, we demonstrate concordance in shared TWAS signals across GReX models and provide insight into GReX-specific detectable genes and pathways.

Aerosols are often hypothesized to invigorate deep convective clouds (DCCs), but observational evidence remains limited and inconclusive. Clarifying this hypothesis is critical for regions vulnerable to thunderstorms and flooding, particularly highly polluted coastal cities. Leveraging a novel causal discovery-inference pipeline and high-resolution observations near Houston, TX, we identify multiple causal pathways among aerosols (mostly organic and sulfate), DCCs, and meteorological factors. However, a direct causal link from aerosols to DCCs is found to be uncommon, occurring in less than 35% of analyzed scenarios, and is characterized by strong conditionality and nonlinearity. When aerosol impacts on DCCs do occur, they can be substantial, enhancing DCC core heights by approximately 1.7 km, with 92% of this effect concentrated in warmer-phase cloud regions. Notably, the presence of sea breezes and the inclusion of all measured aerosol particles each enhance DCCs in over 95% of aerosol-sensitive cases.

Understanding gene-disease associations is important for uncovering pathological mechanisms and identifying potential therapeutic targets. Knowledge graphs can represent and integrate data from multiple biomedical sources, but lack individual-level information on target organ structure and function. Here we develop CardioKG, a knowledge graph that integrates over 200,000 computer vision-derived cardiovascular phenotypes from biomedical images with data extracted from 18 biological databases to model over a million relationships. We used a variational graph auto-encoder to generate node embeddings from the knowledge graph to predict gene-disease associations, assess druggability and identify drug repurposing strategies. The model predicted genetic associations and therapeutic opportunities for leading causes of cardiovascular disease, which were associated with improved survival. Candidate therapies included methotrexate for heart failure and gliptins for atrial fibrillation, and the addition of imaging data enhanced pathway discovery. These capabilities support the use of biomedical imaging to enhance graph-structured models for identifying treatable disease mechanisms.

Neurons of distinct subtypes compartmentalize subtype-specific function in part by differentially localizing and translating specific RNAs, but underlying mechanisms are not understood. Here we investigate messenger RNA localization and stability within subtype-specific growth cones (GCs), leading tips of growing axons, of long-range projection neurons (PNs) of the developing cerebral cortex. Comparison of GC-localized transcriptomes between two subtypes of PNs (interhemispheric-callosal and corticothalamic) across developmental stages identified both distinct and shared subcellular machinery involved in distinct phases of growth, target innervation and synaptogenesis, and enrichment of genes associated with neurodevelopmental and neuropsychiatric disorders. Further, we investigated sequence elements in dynamically GC-localized mRNAs, identifying GC-enriched motifs in 3' untranslated regions. For example, we identified that CPEB4, a translational regulator, regulates axonal branching and that RBMS1 functions dynamically in callosal circuit formation. This work offers generalizable insights for subcellular specialization in other polarized cells, toward elucidating neurodevelopmental and behavioral-cognitive disorders.

Understanding Alzheimer's disease (AD) from its earliest stages is essential for uncovering initial mechanisms of pathology and developing interventions. Here, we use the humanized App^NL-G-F mouse model, which develops early amyloid pathology along a predictable timeline, to characterize epigenetic changes in the brain and blood at early pre-symptomatic, as well as later, stages of disease progression. We identified alterations in chromatin accessibility, gene expression, and DNA methylation before and after amyloidosis, in the absence of advanced age. Despite broadly stable hippocampal cell composition, App^NL-G-F mice exhibit major gene expression differences preceding amyloid plaque deposition, particularly in pathways related to mitochondrial function and protein biosynthesis. In later stages of pathology, immune pathways were upregulated, consistent with established neuroinflammatory processes in AD. Extensive DNA methylation changes were also detected in both blood and hippocampus at early and late pathology stages. Many blood methylation differences at early stages overlapped brain cis-regulatory elements and mapped near differentially expressed hippocampal genes, with enrichment in neuronal development and synaptic pathways, underscoring a potential link between blood methylation and brain physiology and supporting the potential of blood DNA methylation as an early biomarker of amyloidosis. Notably, five genes, including Rbfox1 and Camta1, showed coordinated epigenetic dysregulation in both brain and blood prior to amyloidosis, highlighting them as potential early blood-based biomarkers.

Renal cell carcinoma (RCC), the most common type of kidney cancer, is characterized by a poor prognosis due to its aggressive metastasis and resistance to treatment. Long non-coding RNAs (lncRNAs) have emerged as key regulators of RCC progression, influencing cellular processes such as proliferation, migration, invasion, and apoptosis. LncRNA biogenesis occurs through both canonical and non-canonical pathways, involving RNA Polymerase II-mediated transcription and alternative splicing. LncRNAs like HOTAIR, MALAT1, and GAS5 regulate critical signaling pathways, including the PI3K/AKT/mTOR axis, Wnt/β-catenin, and JAK/STAT, while others, such as RCAT1, DUXAP9, and Lnc-LSG1, modulate protein degradation, impacting tumor growth and metastasis. Additionally, lncRNAs like EGFR-AS1 and MALAT1 enhance the EGFR/AKT and VEGF/Akt pathways, driving RCC cell proliferation and migration. LncRNAs such as HOTAIR and TCL6 also promote epithelial-to-mesenchymal transition, contributing to tumor invasion and therapy resistance. Furthermore, lncRNAs regulate the p53 pathway, with some, like MEG3, acting as tumor suppressors, while others, like SNHG3, suppress p53 activity, accelerating RCC progression. These insights into lncRNA-mediated regulatory mechanisms provide promising therapeutic targets for RCC, suggesting that modulating specific lncRNAs or their associated pathways could offer innovative strategies for treatment and prognosis. This review presents a comprehensive analysis of the biogenesis, functions, and regulatory roles of lncRNAs in RCC, emphasizing their potential as diagnostic biomarkers and therapeutic targets to improve patient outcomes.

<h4>Background</h4>Hepatoblastoma (HB) is the most common pediatric liver malignancy with an increasing incidence. However, the functional roles of 3D chromatin organization, epigenetic regulatory factors, and transcriptional reprogramming in HB pathogenesis remain poorly understood.<h4>Methods</h4>​Integrated multi-omics analyses of HB and matched non-tumor tissues were performed, including Hi-C, H3K27ac CUT&Tag, ATAC-seq, and RNA-seq, to construct high-resolution 3D epigenomic maps and identify genes interacting with HB-specific super-enhancers (SEs). Functional assays of identified targets were conducted in cell lines and animal models. The regulatory mechanisms of SEs and upstream transcription factors (TFs) were investigated using CRISPRi-dCas9, 3C-qPCR, ChIP-qPCR, and luciferase reporter assays.<h4>Results</h4>Comprehensive analysis identified TRIB2 as an HB-specific SE-associated oncogene. Functionally, TRIB2 promoted cell proliferation and accelerated tumor growth both in vitro and in vivo. Patients with high TRIB2 expression exhibited advanced PRETEXT stage and metastasis. Mechanistically, TCF3 directly bound to both the TRIB2-SE and its promoter, promoting TRIB2 overexpression. Moreover, TRIB2 conferred resistance to ferroptosis by disrupting KEAP1-mediated ubiquitination of NRF2, thereby stabilizing NRF2 protein and enhancing antioxidant responses. The TCF3-TRIB2-NRF2 axis showed significant co-expression in HB tissues, effectively distinguished HB from normal liver tissues, and was associated with poorer overall survival.<h4>Conclusions</h4>Our findings reveal that TCF3 and SE mediate TRIB2 overexpression to inhibit ferroptosis via the KEAP1-NRF2 pathway and drive HB pathogenesis, providing potential diagnostic and prognostic markers for HB.

<h4>Background</h4>Depression, anxiety, and cannabis use are growing, interconnected primary care concerns, but remain understudied due few health systems conducting systematic cannabis use screening. This study examines cannabis use and risk of cannabis use disorder (CUD) among primary care patients, comparing outcomes by depression and anxiety diagnoses and psychotropic prescriptions.<h4>Methods</h4>We assessed past three-month cannabis use, reasons for use, and risk of CUD among 170,032 adult primary care patients at a large health system in Los Angeles, CA under a routine screening protocol using the validated, self-administered ASSIST survey. This survey was embedded in the electronic health record (EHR), where data on ICD-10 diagnostic codes for depressive and anxiety disorders, psychotropic prescriptions, and demographics were collected. Logistic regression analysis assessed the association of depression and anxiety diagnoses on risk of CUD.<h4>Results</h4>Median age was 48 years (IQR 35-61), 57.8% were female, and 17.5% reported cannabis use. 24.0% had anxiety diagnoses and 9.0% had depression diagnoses. Cannabis use was higher among patients diagnosed with depression or anxiety than other patients (21.7%-27.4% vs. 15.5%, p < 0.001). Most patients who reported cannabis use and had depression or anxiety diagnoses reported using cannabis to manage emotional symptoms (62.5%-71.3%); notably, 47.3% of these patients had current antidepressant or anxiolytic prescriptions. Moderate-to-high risk of CUD was elevated among patients who reported cannabis use diagnosed with depression (9.8%), anxiety (8.0%), and particularly depression and anxiety (12.9%) (other patients = 4.9%; p < 0.001). After adjusting for age, sex, race and ethnicity, sexuality (LGB + versus heterosexual), employment, and Charlson Comorbidity Index, depression and anxiety diagnoses were associated with elevated risk of CUD [aOR 1.99 (95% CI 1.78-2.23); aOR 1.6 (95% CI 1.51-1.69), respectively], with the highest association among patients with both diagnoses [aOR 2.58 (95% CI 2.34-2.83)].<h4>Conclusions</h4>Clinical depression and anxiety diagnoses were associated with elevated cannabis use prevalence and risk of CUD. Many primary care patients reported using cannabis to manage mental health-related symptoms, even when prescribed psychotropic medications. These findings highlight the need for providers to assess and address cannabis use and potential CUD among primary care patients, especially those diagnosed with depression or anxiety.<h4>Trial registration</h4>Clinical trial number: not applicable.

Necrotizing enterocolitis (NEC) severely affects preterm infants with limited treatments. Although intestinal homeostasis dysfunction is considered a trigger for NEC, the key targets and mechanisms remain unclear. Using lipopolysaccharide-stimulated colonic epithelial cells and hypothermic hypoxia-induced NEC mice (both sexes), we demonstrate that the gut metabolite L-cystine alleviates intestinal inflammation by balancing Th17/Treg responses and inhibiting ferroptosis. Mechanistically, L-cystine directly targets KIF11 to suppress RC3H1 expression, blocking IL-6 transcripts through transcriptional modifications, thereby inhibiting IL-6 secretion and ferroptosis. Conditioned medium from L-cystine-treated cells inactivates IL-6/STAT3 signaling, reducing pro-inflammatory cytokine release and restoring Th17/Treg balance. Notably, microbiota colonization from NEC preterm infants exacerbates intestinal damage, an effect mitigated by L-cystine and IL-6/STAT3 inhibition. Thus, L-cystine attenuates NEC by suppressing ferroptosis in epithelial cells, restoring immune homeostasis, and preserving intestinal barrier integrity. Targeting intestinal metabolites represents a promising prophylactic and therapeutic strategy for NEC, addressing unmet clinical needs in neonatal intestinal injury management.

Flagellogenesis in mammalian sperm is essential for sperm motility and successful egg fertilization. Multiple morphological abnormalities of the sperm flagella (MMAF) represent a condition characterized by various structural defects in the flagellum. While CFAP57 has been identified as a factor in the pathogenic mechanisms of MMAF, the precise molecular regulation underlying this process remains unclear. Here, we report novel biallelic mutations in the CFAP57 gene identified in two infertile males from two unrelated families. The first patient carried a homozygous nonsense mutation in CFAP57 [NM_001195831.2: c.3250 C > T (p.R1084X)], while the second was compound heterozygous for two missense mutations c.1340T > C (p.V447A) and c.1856G > A (p.R619H). CRISPR-Cas9-generated CFAP57 mutant mice recapitulated human MMAF phenotypes, exhibiting structural flagellar defects and complete infertility. Using immunoprecipitation-mass spectrometry (IP-MS), we identified MYH10, a non-muscle myosin II isoform, as an interaction partner of CFAP57. Immunofluorescence analysis confirmed that both MYH10 and CFAP57 localize to the sperm flagella. We further examined the precise localization of MYH10 at the ultrastructural level using immunoelectron microscopy. Gold particles conjugated to the MYH10 antibody were predominantly detected in the sperm flagella. In sperm with CFAP57 mutations, MYH10 was mislocalized to the mid-piece region while being notably absent from the principal and end pieces. This mislocalization affected the expression of IFT88, a key component of the intraflagellar transport (IFT) system that plays a critical role in mammalian flagellar assembly. Fortunately, ICSI can overcome CFAP57-associated male infertility. Together, our findings establish CFAP57 as an important mediator of sperm flagellogenesis that orchestrates MYH10 and IFT88 positioning and intraflagellar transport dynamics to maintain flagellar integrity, providing molecular insights into MMAF-associated male infertility.

INTRODUCTION: Sarcoidosis is an inflammatory disease driven by immune-mediated mechanisms, characterized by the formation of epithelioid cell granulomas and a wide range of clinical manifestations. Its phenotype is the result of a complex interplay of genetic and environmental factors, the precise roles and interactions of which remain poorly defined. AIM: To identify candidate genes and risk loci associated with sarcoidosis from large population datasets. To estimate the genetic heritability of the phenotype in selected ancestries. POPULATION AND METHODS: Public summary statistics from the FinnGen release 12 (European ancestry), pan UK BioBank Project (UKBB - European and African ancestry), Million Veteran Program (MVP - European and African ancestry), and Japan BioBank (East Asian ancestry) were included for European, African and multi-ancestry meta-analysis through sample size-based analysis. Novel risk loci and single nucleotide polymorphisms (SNPs) significantly associated with the disease were critically reviewed on the basis of the available literature. For each risk locus, SNPs highly correlated with the lead SNP were selected based on Combined Annotation Dependent Depletion (CADD) scores. Genetic heritability (h2) scores were obtained through ancestry-specific linkage-disequilibrium score calculation. RESULT: Overall 9659 cases (7559 European, 1880 African, 220 East Asian) and 1,665,804 controls (1,361,726 European, 126,411 African, 177,667 East Asian) were analysed. Nineteen and two risk loci were identified in European and African ancestry, respectively; h2 scores were 0.25 (European) and 0.19 (African). Candidate non-MHC genes for further explorations through functional studies included IL23R, PUS10, ACOXL, PLCL1, FAM117B, BMPR2, PPARG, ESYT2, ANXA11, CCDC88B, ATXN2, CCL24, RP11−540O11.1, HOMER2, CD19, UBASH3A, RNF215, and others. Interferon gamma signaling, meiotic recombination/condensation of prophase chromosomes, and DNA methylation were the most enriched gene sets in European and multi-ancestry meta-analysis. Multi-ancestry meta-analysis was confronted with FinnGen+UKBB+MVP meta-analysis (released by FinnGen freeze 12) yielding consistent results (18 risk loci identified) CONCLUSION: Nineteen and two risk loci were significantly associated with sarcoidosis for European and African ancestries, respectively. Moderate genetic heritability was observed for both ancestries. A set of significantly associated non-MHC genes and SNPs was obtained to investigate functional validation. Although further studies are warranted, epigenetic alterations may contribute to the risk of developing sarcoidosis

<h4>Purpose</h4>The genetic spectrum and early clinical indicators of familial exudative vitreoretinopathy (FEVR) remain incompletely defined, and few studies have investigated the genetic variants and clinical phenotypes associated with eoHM-FEVR and anisometropia-FEVR patients. The purpose of this study was to screen the pathogenic variations in 11 FEVR families and analyze the refractive status and pathogenic genes in patients with irregular dominantly inherited FEVR.<h4>Methods</h4>The patients with clinical diagnoses of eoHM-FEVR or anisometropia-FEVR were evaluated from October 2019 to August 2022. Comprehensive ophthalmic tests were performed on participants to confirm the phenotype. The genotype was identified using whole-exon sequencing and further verified the results among other family members by Sanger sequencing. Normal protein structures were modeled with AlphaFold, whereas mutant variants were analyzed via PyMOL. Variant pathogenicity followed the American College of Medical Genetics and Genomics (ACMG) guidelines. The protein-protein interaction (PPI) network analysis with STRING and <i>k</i>-means clustering was applied for detecting the interaction of genes in the candidate genes, and the ClusPro Server was used for protein-protein docking.<h4>Results</h4>A total of 11 FEVR families were included in the study, and all the probands were found to have high myopia in both eyes or one eye before the age of 7 years. The pathogenic variants were identified in the genes <i>TSPAN12</i>, <i>LRP5</i>, and <i>FZD4</i> known to be associated with FEVR in six probands. Among 13 eoHM-related genes, <i>FZD4</i> and <i>LRP2</i> encode proteins that can dock together as analyzed by ClusPro software.<h4>Conclusion</h4>This study observed dominant inheritance of an irregular pattern in FEVR families, with asymmetric FEVR presenting as severe anisometropia. The eye with higher myopia often had more advanced FEVR and pronounced fundus changes. PPI network analysis revealed important modules of gene interaction, and the FZD4-LRP2 complex protein was potentially related to high myopia development. For patients with high myopia or with obvious anisometropia in both eyes, more attention should be paid clinically to the comprehensive examination of the peripheral fundus and early genetic testing.

<h4>Objective</h4>Resin composite restorations typically last 6-10 years but often fail due to mechanical fatigue, hydrolytic breakdown, and degradation at the interface. These failures result in frequent replacements, leading to significant clinical and environmental impacts. Extending restoration durability is essential for both patient care and sustainability. This review examines recent advances in self-healing dental biomaterials, emphasising the underlying chemical and physical mechanisms, their integration into resin composites, cements, and adhesives, and their relevance to sustainable restorative practice.<h4>Materials and methods</h4>A narrative review methodology was employed to synthesise the current evidence. Studies on self-healing mechanisms, including extrinsic (capsule- or reservoir-based) and intrinsic (dynamic covalent, supramolecular, or shape-memory polymer) systems, were critically evaluated with emphasis on evidence relevant to dental resin composites, cements, and adhesives. Parallel insights from polymer and material sciences were included where dental-specific research was limited. Supplementary searches were conducted on Google Scholar for additional peer-reviewed articles, books, and preprints.<h4>Results</h4>Experimental resin composites and cements incorporating microcapsule-based self-healing systems exhibited fracture toughness recovery between 65% and 77%, maintaining structural integrity after 6 months of water storage in deionised water at 37°C. Disulfide and Diels-Alder dynamic networks, though mostly investigated in polymer science, show potential for repeatable healing under mild triggers, while supramolecular hydrogen-bonding and bioactive fillers offer adaptive repair and remineralisation at adhesive interfaces. Self-healing strategies align with the four pillars of green dentistry, that is pollution prevention, water conservation, energy efficiency, and waste reduction by potentially halving procedural resource consumption through extended restoration lifespan.<h4>Conclusion</h4>Self-healing biomaterials designed and developed in accordance with sustainability principles have the potential to transform restorative dentistry by facilitating autonomous repair, prolonging restoration lifespan, and minimising the environmental footprint through reduced material usage and clinical waste generation.

<h4>Background</h4>Palmitoylation, a reversible lipid modification, plays a critical role in protein trafficking and signaling and has been implicated in various cancers. However, its function in head and neck squamous cell carcinoma (HNSCC)-a highly aggressive and heterogeneous malignancy-remains largely unexplored. This study aimed to establish a prognostic model based on palmitoylation to improve risk stratification and support clinical decision-making in HNSCC patients.<h4>Methods</h4>The HNSCC training and validation cohorts were obtained from The Cancer Genome Atlas (TCGA) and Gene Expression Omnibus (GEO, GSE41613), respectively. A total of 4,052 palmitoylation-related genes (PRGs) were curated from the Molecular Signatures Database (MsigDB), and 282 differentially expressed PRGs were identified. Univariate Cox regression analysis yielded 49 prognostic genes, of which 16 were further selected by least absolute shrinkage and selection operator (LASSO) regression. Subsequent multivariate Cox analysis led to the construction of an 8-gene PRG-based risk model. Clinical variables, including age, sex, stage, and risk score, were integrated into a nomogram. Kaplan-Meier survival curves, time-dependent receiver operating characteristics (ROC) analyses, immune infiltration profiling, gene set enrichment analysis (GSEA), and drug sensitivity analysis were performed to assess the model's clinical utility.<h4>Results</h4>The model effectively stratified patients into high- and low-risk groups with significantly different overall survival (OS) outcomes. Validation in an independent GEO dataset confirmed its robustness. A prognostic nomogram integrating the PRG-based risk score with clinical variables further enhanced predictive performance, achieving an area under curve (AUC) of 0.709, which was superior to stage (0.578), age (0.568), T stage (0.539), N stage (0.512), and sex (0.469). Immune analysis revealed that the low-risk group had higher ImmuneScores and ESTIMATEScores, along with reduced tumor purity, suggesting stronger immune cell infiltration. GSEA indicated enrichment of cancer-related pathways in the high-risk group. Drug sensitivity analysis based on the Genomics of Drug Sensitivity in Cancer (GDSC) database showed that low-risk patients had lower half-maximal inhibitory concentration (IC50) values for several targeted agents.<h4>Conclusions</h4>Our findings highlight the prognostic significance of PRGs in HNSCC and suggest their potential utility in guiding personalized treatment strategies. Specifically, the PRG-based risk model demonstrated superior predictive performance compared to conventional clinical factors, supporting its value in improving patient stratification. Furthermore, the associations with immune infiltration patterns and drug sensitivity indicate that PRGs may not only serve as prognostic biomarkers but also help identify patients who could benefit from targeted or immunotherapeutic approaches.

<h4>Background</h4>Gastric cancer (GC) remains a major global health challenge with persistently low remission rates, largely because most patients are diagnosed at advanced stages. Although chemotherapy, targeted therapy, and immune checkpoint inhibitors have achieved progress, their benefits remain limited due to the highly immunosuppressive tumor microenvironment (TME), characterized by inadequate T-cell activation and impaired costimulatory signaling. Recent genomic and single-cell studies highlight the need for strategies that can more effectively enhance antitumor immunity. Given these challenges, this study aimed to evaluate whether dual stimulation with interleukin-2 (IL-2) and OX40 ligand (OX40L) could synergistically enhance T-cell activation in GC. Specifically, we analyzed OX40/OX40L expression patterns in GC tissues and tumor-infiltrating lymphocytes (TILs), assessed the immunostimulatory effects of IL-2 and OX40L in PBMCs and TILs, and tested a recombinant adenoviral vector co-expressing IL-2 and OX40L to determine its ability to potentiate T-cell-mediated cytotoxicity against primary GC tumor cells.<h4>Methods</h4>Based on preliminary analyses of The Cancer Genome Atlas (TCGA) database and single-cell RNA sequencing (scRNA-seq) data, we identified the costimulatory molecules IL-2 and OX40L as potential therapeutic targets. We collected tumor tissues and paired peripheral blood mononuclear cells (PBMCs) from 70 untreated GC patients, and used bioinformatics tools to analyze T cell subpopulations and OX40/OX40L expression in the TME. The immunostimulatory effects of IL-2 and OX40L, used alone or in combination, were evaluated in vitro on PBMCs and TILs. A recombinant adenoviral vector co-expressing IL-2 and OX40L was also constructed to assess its pro-apoptotic effect in primary GC tumor cell cultures.<h4>Results</h4>The combination of IL-2 and OX40L significantly enhanced T cell activation in both PBMCs and TILs, and increased the expression of antitumor effector molecules. The IL-2/OX40L adenoviral vector effectively activated TILs, which subsequently induced prominent apoptotic responses in primary GC tumor cells.<h4>Conclusions</h4>Our study demonstrates that a costimulatory immunotherapy strategy combining IL-2 and OX40L significantly enhances antitumor responses in PBMCs and TILs from GC patients, showing strong therapeutic potential as a complementary approach for the treatment of GC.

Telmisartan, an angiotensin II type 1 receptor (AT1R) antagonist, possesses cytotoxic activity towards BRAF-mutated melanoma cell lines. However, its antihypertensive effects limit its use in the population of normotensive patients. To mitigate this shortcoming, a group of eight telmisartan-amino acid conjugates, designed to have reduced or no AT1R affinity with enhanced cellular uptake, were synthesized by the coupling reaction in yields ranging from 34% to 60%. Their cytotoxicity was tested on BRAF V600E-mutated melanoma cell lines (A375 and 518A2), and compounds <b>1</b>, <b>3</b>, and <b>8</b> stood out as the best candidates. These three compounds were also tested on the vemurafenib-resistant (A375R) and normal (HaCaT and MRC-5) cell lines, and compound <b>8</b> showed better cytotoxicity (IC₅₀ = 8.84 ± 1.24 µM) and selectivity (>3.50) when compared to telmisartan (IC₅₀ = 29.23 ± 3.88, selectivity > 2.40). The cellular uptake of compounds <b>1</b> and <b>8</b> was significantly higher than telmisartan, with substantial accumulation in the membrane and nuclear compartments. Unlike telmisartan, compounds <b>1</b>, <b>3</b>, and <b>8</b> did not inhibit angiotensin II-induced Ca²⁺ signaling, which indicates diminished AT1R binding. All three compounds induced cell cycle arrest and disrupted mitochondrial morphology and membrane potential. These findings highlight their potential as non-antihypertensive telmisartan derivatives for melanoma therapy.

<b>Background/Objectives</b>: PHF6 is a chromatin-binding protein located in the nucleus, and it is involved in transcriptional regulation. However, limited research exists on the specific roles and mechanisms of PHF6 across various tumors. <b>Methods</b>: Based on The Cancer Genome Atlas (TCGA) database, we analyzed PHF6 expression in pan-cancer. We first evaluated the relevance between PHF6 and prognosis; then, the relevance between PHF6 and immune cell infiltration in pan-cancer were analyzed. Subsequently, we explored the correlation between PHF6 and cancer heterogeneity, such as tumor mutation burden (TMB) and microsatellite instability (MSI), as well as cancer stemness. Finally, the role of PHF6 was validated in liver cancer and pancreatic cancer cell lines by cell proliferation assays. <b>Results</b>: PHF6 expression was higher in the vast majority of cancers than their normal counterparts. PHF6 was substantially correlated with prognosis and immune cell infiltration in various cancers. Moreover, PHF6 expression showed a strong correlation with cancer heterogeneity and stemness in certain cancer types. Additionally, the depletion of PHF6 inhibited cell proliferation in both liver and pancreatic cancer cells. <b>Conclusions</b>: PHF6 expression was closely associated with the occurrence and development of many types of cancer, and it might promote cancer progression by inhibiting the function of the immune microenvironment, while knockout of PHF6 significantly inhibited the tumor cells proliferation.

The autophagy-lysosome system is a master regulator of cellular homeostasis, integrating quality control, metabolism, and cell fate through the selective degradation of cytoplasmic components. Disruption of either autophagic flux or lysosomal function compromises this degradative pathway and leads to diverse pathological conditions. Emerging evidence identifies the autophagy-lysosome network as a central signaling hub that connects metabolic balance to disease progression, particularly in neurodegenerative disorders and cancer. Although cancer and neurodegenerative diseases exhibit seemingly opposite outcomes-uncontrolled proliferation versus progressive neuronal loss-both share common mechanistic foundations within the autophagy-lysosome axis. Here, we synthesize recent advances on the roles of autophagy and lysosomal mechanisms in neurodegenerative diseases and cancer, especially on how defects in lysosomal acidification, membrane integrity, and autophagosome-lysosome fusion contribute to toxic protein accumulation and organelle damage in Alzheimer's and Parkinson's diseases, while the same machinery is repurposed by tumor cells to sustain anabolic growth, stress tolerance, and therapy resistance. We also highlight emerging lysosome-centered therapeutic approaches, including small molecules that induce lysosomal membrane permeabilization, nanomedicine-based pH correction, and next-generation protein degradation technologies. Finally, we discuss the major challenges and future opportunities for translating these mechanistic insights into clinical interventions.

<h4>Background</h4>Malnutrition, a significant complication, triggers devastating consequences among patients with chronic obstructive pulmonary disease (COPD). Although previous studies have demonstrated the benefits of nutritional support in stable COPD, evidence in hospitalized patients with acute exacerbations remains limited. This study aimed to examine the short-term association between individualized nutritional support and changes in nutritional status and clinical symptoms among inpatients with acute COPD exacerbations.<h4>Methods</h4>Thirty male inpatients diagnosed with acute exacerbations of COPD were enrolled in a 40-day individualized nutritional intervention program. The intervention included tailored dietary counseling based on each patient's energy and protein requirements, alongside hospital-standard medical treatment. Anthropometric measurements, serum albumin levels, and Subjective Global Assessment (SGA) were assessed at the hospital admission (T0), 7 days (T7), and 40 days (T40) of intervention. Clinical symptoms, including fatigue, dyspnea, anorexia, dysphagia, and bloating, were also monitored. Statistical analyses were two-sided, and significance was set at P<0.05.<h4>Results</h4>Thirty COPD patients (mean age 68.4±8.6 years; 66.7% aged >65 years) completed the 40-day individualized nutritional intervention. At baseline, 70% had chronic energy deficiency (CED) and 83.3% were malnourished (SGA-B/C). After intervention, the proportion of well-nourished patients (SGA-A) increased from 16.7% to 80%. Serum albumin rose significantly from 36.4±0.7 to 38.3±0.7 g/L (P=0.005). Mean BMI increased from 17.7±2.2 to 18.1±2.3 kg/m², and CED prevalence declined from 70% to 53.3%. Fatigue, dyspnea, and anorexia markedly improved following intervention.<h4>Conclusions</h4>Individualized nutritional support was associated with notable improvements in nutritional status and clinical symptoms among patients hospitalized with acute exacerbations of COPD. These findings underscore the value of incorporating tailored nutritional care into routine COPD management. However, given the small, male-only sample and short follow-up duration, further large-scale and long-term studies are warranted to verify the durability and broader applicability of these outcomes.

<h4>Background</h4>Parkinson's disease (PD) patients face a higher risk of developing heart failure (HF). The objective of the study was to investigate the hub genes and potential mechanisms linking Parkinson's disease (PD) to heart failure (HF) using multiple integrative bioinformatics tools.<h4>Methods</h4>Integrated bioinformatics analyses were performed. One HF dataset (GSE57338) and three PD datasets (GSE7621, GSE20146, GSE49036) were obtained from the GEO database. Weighted gene co-expression network analysis (WGCNA) was used to identify PD-related genes. Differentially expressed genes (DEGs) between PD and normal samples, as well as between HF and normal samples, were identified. The intersection of DEGs, WGCNA-derived PD-related genes, and genes encoding known secretory proteins was analyzed to find PD-associated secretory proteins. Immune cell infiltration in HF was assessed using CIBERSORT. Protein-protein interaction (PPI) network analysis was conducted to identify hub genes. Key findings were experimentally validated in an MPTP-induced PD mouse model through behavioral tests, ELISA, and immunohistochemistry.<h4>Results</h4>Analysis identified 21 PD-associated secretory proteins. Intersection with HF DEGs revealed 12 common genes, from which 8 functional genes with consistent expression patterns in both conditions were identified. PPI network analysis highlighted three hub genes: <i>RELN, SLIT1</i>, and <i>NTN1</i>. Reactome pathway analysis indicated that NTN1 is involved in cardiac-related processes like muscle contraction and cardiac conduction. Experimental validation in PD model mice confirmed a significant decrease in Netrin-1 levels in the blood, striatum, and heart. Furthermore, a strong negative correlation was found between cardiac Netrin-1 expression and collagen deposition, suggesting its potential role in impacting cardiac function.<h4>Conclusion</h4>These insights highlight the coexistence of PD and HF and suggest new avenues for investigating strategies to prevent HF in PD patients, particularly by exploring the role of Netrin-1 in the heart and its potential for cardioprotection.

Ferroptosis is one of the regulated cell death pathways. Molecular mechanisms underlying ferroptosis involve iron-dependent lipid peroxidation, which results in cell-deleterious membrane damage. Ferroptosis-inducing agents have been identified as attractive candidates for anticancer drug development as they can bypass drug resistance in cancer cells. Among pro-ferroptotic agents are many organometallic complexes, including ferrocenyl compounds. In this review, we demonstrate that suitably designed ferrocene-containing molecules can induce ferroptosis in different cancer cell types both <i>in vitro</i> and <i>in vivo</i>. Their pro-ferroptotic activity is triggered by diverse initiating factors through different mechanisms (<i>e.g.</i> redox activation, thermal and light activation, and GPX4 inhibition combined with ROS overproduction). Moreover, ferrocenyl bioconjugates are often cancer-cell-selective and trigger ferroptosis in combination with other regulated cell-death pathways, such as apoptosis and immunogenic cell death. Dual or multimodal anticancer activity mechanisms are sought after in modern anticancer therapy approaches as they help to overcome the problem of drug resistance. Research on ferrocene-based ferroptosis inducers, however, is still in the early stage. Hence, more time and effort are needed to fully elucidate the potential of ferrocenes as ferroptosis initiators in cancer therapy.

<h4>Background & objective</h4>Emerging viral infections can initiate a global pandemic with high mortality. Understanding the immunopathogenesis of these viruses is critical to developing effective strategies for managing/preventing such outbreaks.<h4>Methods</h4>Transcriptomics analysis was performed in a UAE cohort with respective COVID-19 severities. The findings were correlated with published studies of COVID-19 severity/progression GWAS, and transcriptomics data of patients infected with different respiratory viruses.<h4>Results</h4>The transcriptional profiling distinguished significantly between the infected COVID-19 groups and identified the interferon-induced protein, <i>GBP2,</i> as a common significantly up-regulated gene among the different COVID-19 infection severities (nominal <i>p</i> = 0.0019). Key inflammatory pathways were enriched in the higher-severity groups, including Interleukin-1 family signaling. A remarkable immune signature resulted in a trend of cytokine expression changes between all severities, including <i>CCL19, CCL21, IL-19, IL-20, IL-36RN,</i> and members of the IFNA family. The deconvolution of immune cells showed a trend of an uncontrolled pro-inflammatory state and poor immune function in higher disease severities. A systematic analysis of the transcriptomic and GWAS findings identified common signature genes between COVID-19 infection severities including <i>ALCAM, DKK3, EFNA5, FN1, GABRA5, LPAR1, METTL8, MTHFD1L, SPOCK1, TPM4, VTI1A,</i> and <i>WWC2.</i> Differential regulation in potential genes associated with the Interferon signaling pathway including <i>HERC5, IFFI44L, IFI6, RSAD2</i> and <i>SP100</i> was identified as a common feature in transcriptomes of patients afflicted with different virulent respiratory viruses.<h4>Conclusion</h4>Our findings highlight a direction where changes in immune response and specific biomarker panels could be considered as a strategy for the prediction/prevention of new emerging respiratory virus outbreaks.

Recent advancements in the study of orphan G protein-coupled receptors (oGPCRs) have revealed a large number with high levels of constitutive G protein signaling. Structural studies have suggested a new paradigm in which many constitutively active oGPCRs are auto-activated by their own intrinsic protein motifs, which act as auto-agonists. This includes extracellular loop 2 acting as auto-agonist to promote active-state conformations and G protein signaling. In some cases, the oGPCRs lack inhibitory microswitches that may drive a high level of constitutive activity. In this brief review, we discuss oGPCR constitutive activity, highlighting the auto-activating orphan receptors and overview structural underpinnings of constitutive activity. A discussion into the pharmacological and cell signaling implications of oGPCR constitutive activity is provided. We also propose a new concept in which orphan GPCR constitutive activity sets the baseline tone for cellular signaling and allows for dynamic regulation of cAMP signaling. Taken together, recent mechanistic studies with many oGPCRs indicate high constitutive activity is a common phenomenon that modulates cellular signaling and that can be tuned with pharmacology. SIGNIFICANCE STATEMENT: Recent literature describes a subset of orphan Class A G protein-coupled receptors with high constitutive signaling that auto-activate by their own intrinsic protein motifs. Herein, a new concept is proposed in which oGPCR constitutive activity allows dynamic regulation of cAMP signaling. Recently solved structures and functional studies of constitutively active oGPCRs provide fresh insights into oGPCR signaling with relevance for both health and disease.

Reactive oxygen species (ROS)-induced cell damage contributes to many diseases. However, ROS also contribute to cell signaling and immune defences. As ubiquitous thiol peroxidases, peroxiredoxins (Prdx) play integral roles in balancing ROS functions. High levels of Prdx6 are associated with increased metastasis and resistance to chemotherapy, rendering Prdx6 a therapeutic target for treatment of a broad range of cancers. However, Prdx6 has additional activities, in lipid signaling and selenocysteine metabolism, and it remains unclear how Prdx6's thiol peroxidase activity contributes to disease. Here we have investigated the role/s of Prdx6 in the nematode worm Caenorhabditis elegans. Consistent with a ROS-protective role for PRDX-6, prdx-6 mutant C. elegans exhibit elevated levels of lipid oxidation, more apoptotic corpses in their germline and are more susceptible to the toxicity of diethyl maleate. However, unexpectedly, prdx-6 mutant C. elegans are more resistant to other forms of oxidative stress, long-lived and resistant to infection with two opportunistic human pathogens; the gram-positive bacteria Staphylococcus aureus and the dimorphic yeast Candida albicans. Our data suggest these phenotypes are associated with increased activity of the NHR-49(PPARα/HNF4) transcriptional regulator and intestinal expression of the Flavin monooxygenase, FMO-2. FMO-2 has a conserved, pro-survival function and is up-regulated in response to various stresses, including peroxides and S. aureus infection. Here we reveal that fmo-2 expression is also increased as an NHR-49-dependent protective response to C. albicans. Consistent with increased NHR-49 activity, prdx-6 mutant animals also contain increased levels of mono-unsaturated fatty acids. Accordingly, we propose that elevated expression of fmo-2 and other NHR-49 up-regulated genes contribute to the increased arsenite resistance and innate immunity of prdx-6 mutant animals. These findings further illustrate the complex roles that ROS, PRDX and lipid oxidation can play in oxidative stress resistance, immunity and ageing.

<h4>Context</h4>Regardless of age, metabolic dysfunction-associated steatotic liver disease (MASLD) occurs in 70% of adults with type 2 diabetes (T2D) and increases the risk of metabolic dysfunction-associated steatohepatitis (MASH), fibrosis, and cirrhosis. However, many studies focus on older adults.<h4>Objective</h4>To determine the frequency of screening for MASLD in young adults with T2D attending outpatient clinics.<h4>Methods</h4>A retrospective cohort study on young adults (aged 18-44 years) with T2D who accessed care at a tertiary care center from January 1, 2018, to December 31, 2022. At-risk MASH was diagnosed with vibration-controlled transient elastography, magnetic resonance elastography, or liver biopsy.<h4>Results</h4>Of 6891 participants included, 16% (n = 1100) had a diagnosis of MASLD. Those with MASLD were more likely to have cardiometabolic risk factors (83% vs 72%, <i>P</i> < .001) and cardiovascular disease (22% vs 16%, <i>P</i> < .001). Only 12% of those with MASLD underwent further investigation (1.9% of the total cohort), which was associated with dyslipidemia (odds ratio [OR], 2.4; 95% CI, 1.5-3.8), ALT >40 U/L (OR, 2.1; 95% CI, 1.4-3.0), or the use of 3 or more diabetes medications (OR, 2.1; 95% CI, 1.5-3.1). In those with further workup, 38% had at-risk MASH. The fibrosis-4 index was elevated in 16% of those with MASLD and only 28% of those with confirmed at-risk MASH or worse.<h4>Conclusion</h4>Screening for MASLD in young adults with T2D is frequently missed. There is a lack of accurate noninvasive tools in this population. Increased awareness of screening young adults with T2D for at-risk MASH is urgently needed to prevent progression to cirrhosis.

<h4>Background</h4>Inflammatory bowel disease (IBD) is a chronic condition characterized by immune dysregulation and intestinal inflammation, with anti-tumor necrosis factor α (anti-TNFα) therapies being a key treatment strategy. However, therapeutic responses vary across patients due to genetic factors, necessitating the identification of predictive biomarkers to optimize outcomes. This study employs bioinformatic approaches to investigate the deleterious effects of SNPs in IBD-associated genes, focusing on the structural implications of <i>HFE</i> gene variants and the impact of <i>TNFRSF1B</i> 3' UTR SNPs on miRNA binding site alterations.<h4>Method</h4>The study utilized a systematic literature review to identify genes influencing drug response in IBD, followed by SNP detection. SNPs were filtered based on population frequencies using global databases (gnomAD, ExAC) and Iranome for region-specific data. Structural and functional impacts of missense variants were evaluated using in silico tools and protein domain analysis via InterPro and UniProt. Additionally, molecular dynamics simulation indicated structural changes in HFE gene variants, RNA-seq data analyzed differential gene expression, and miRNASNP-v3 predicted miRNA binding site alterations in 3' UTR regions.<h4>Result</h4>A comprehensive bioinformatics approach identified 41 key IBD drug response genes contributing to cellular pathways, with 69 coding sequence SNPs and 27 3'-UTR SNPs selected for analysis. Coding SNPs, including ABCB1 and <i>HFE</i> variants, were predicted to significantly alter protein structure and stability. RNA-seq analysis revealed <i>TNFRSF1B</i> upregulation in IBD patients, and its 3'-UTR SNPs were shown to affect miRNA binding. Molecular dynamics simulation demonstrated structural instability in the <i>HFE</i> H63D variant, suggesting functional disruption.<h4>Conclusion</h4>This study explores genetic variations influencing anti-TNF therapy responses in IBD patients. Polymorphisms like <i>HFE</i> rs1799945 and <i>TNFRSF1B</i> SNPs are identified as pivotal markers for personalized treatment strategies. Computational findings highlight their clinical potential, emphasizing the need for experimental validation to advance precision medicine and improve therapeutic outcomes and quality of life in IBD management.

Lung development is a highly programmed and energy-dependent dynamic process, with the fetal and neonatal periods representing critical developmental windows. Emerging evidence indicates that mitochondrial quality control (MQC) mechanisms—including mitophagy, mitochondrial dynamics (fusion/fission), and mitochondrial biogenesis—play a central regulatory role in pulmonary development and injury responses during these periods. While, under stressors in the maternal-fetal environment or hyperoxia therapy, transiently upregulate MQC may serve as protective compensatory response, MQC dysregulation can lead to the accumulation of dysfunctional mitochondria, exacerbated oxidative stress, aberrant cellular metabolism, and amplified inflammatory responses. These disruptions contribute to pathological sequelae such as impaired alveolar epithelial differentiation, aberrant pulmonary vascular remodeling, diminished antioxidant capacity, and failed tissue repair. In this review, we summarize recent mechanistic advances in MQC-mediated regulation of lung development across fetal, preterm, and term infant models. Therapeutic strategies targeting MQC have shown promise in preclinical studies, with stem cell therapy and precision-targeted antioxidant delivery emerging as potential approaches to restore mitochondrial homeostasis and promote alveologenesis. We advocate for timely, mitochondria-centered interventions during fetal and neonatal periods to improve pulmonary outcomes and sustain lifelong respiratory health.

Fibrosis is a maladaptive response of tissues or organs to adverse stresses, such as chronic inflammation, infection and mechanical injury. It further promotes parenchymal cell loss, abnormal myofibroblast proliferation and excessive extracellular matrix buildup, eventually triggering scar tissue hyperplasia or organ injury. Although a moderate fibrotic response is beneficial for compensatory tissue repair induced by exogenous or endogenous injury, excessive fibrosis is the basis for the promotion of multiorgan pathologies, such as cardiac hypertrophy, idiopathic pulmonary fibrosis, or renal tubulointerstitial fibrosis. In industrialized countries alone, fibrotic diseases account for ~45% of all‑cause mortality. Consequently, the development of medications that regulate the activation of growth factors, proliferation of fibrotic effector cells and deposition and degradation of the extracellular matrix is essential. Botanical compounds derived from Chinese medicine are generally considered natural tonics. Among these compounds, astragaloside IV (AS‑IV) is a bioactive product isolated from the roots of <i>Astragalus membranaceus</i> Bunge. On the basis of the multitarget therapeutic mechanism of Chinese herbal medicine, AS‑IV may have considerable benefits in improving multiorgan fibrosis and complex fibrotic diseases with multisignal cascades. It can effectively alleviate the fibrosis‑induced dysfunction of major tissues or organs, including the heart, lungs, kidneys and liver, by regulating the signal transduction of reactive oxygen species/caspase‑1/gasdermin D, transforming growth factor‑β/Smads, Wnt/β‑catenin and sirtuin 1‑nuclear factor‑κ B. The present review mainly focused on phytomedicine and highlights the potential of AS‑IV as an antifibrotic medication. It aimed to provide a novel reference for the application of AS‑IV in the nutritional intervention of fibrotic diseases.

<h4>Background</h4>Obesity is a known risk factor for colorectal cancer (CRC), but its impact on prognosis and tumor biology remains unclear. This study aimed to identify molecular biomarkers that reflect obesity-associated tumor characteristics and stratify patient outcomes.<h4>Methods</h4>We conducted a multi-step analysis integrating transcriptomic data, clinical validation, and spatial profiling. Candidate genes were first screened in the TCGA-COADREAD dataset based on expression trends across normal, healthy-weight CRC, and obese CRC samples. Prognostically relevant genes were then validated in an independent cohort using immunohistochemistry (IHC). Finally, spatial transcriptomic analysis using GeoMx DSP was performed to elucidate the tumor microenvironment associated with the top candidate.<h4>Results</h4>Among six shortlisted genes, NNT showed a significant association with overall survival specifically in obese patients and was validated at the protein level by IHC. High NNT expression was independent of TNM stage and associated with improved prognosis. Spatial transcriptomic profiling revealed that NNT-high tumors were enriched for antioxidant, apoptotic, and metabolic programs, while oncogenic and proliferative pathways were suppressed. These patterns suggest that NNT contributes to a redox-balanced and metabolically adaptive tumor state.<h4>Conclusions</h4>Through integrative molecular and spatial analyses, NNT was identified as a potential prognostic biomarker in obesity-associated CRC. This study highlights the importance of combining clinical data with spatial transcriptomics to uncover context-specific tumor biology.

Severe mental disorders have been linked to immune system dysfunction. While a genetic association between mental disorders and autoimmune diseases has been suggested, their genetic relationship remains incompletely understood. Utilizing a complementary set of statistical analyses, we conducted a comprehensive investigation of the genetic architecture between severe mental disorders (major depression (MD), bipolar disorder (BD), and schizophrenia (SCZ)) and seven autoimmune diseases (autoimmune thyroiditis, celiac disease, inflammatory bowel disease (IBD), multiple sclerosis, psoriasis, rheumatoid arthritis, and type 1 diabetes), involving a total of 667,518 cases from 10 genome-wide association studies. While MD was positively genetically correlated with five autoimmune diseases, BD and SCZ were only positively correlated with IBD, suggesting differences in the genetic signal shared with autoimmunity across these mental disorders. A considerable fraction of genetic variants influencing autoimmune diseases (range 17.1-88.4 %) was estimated to overlap with mental disorders; however, this constitutes only a minor part of genetic variants influencing the more polygenic mental disorders. Finally, we identified 172 genetic loci jointly affecting mental disorders and autoimmune diseases, implicating both lipid metabolism and TNF signaling. In conclusion, MD, BD, and SCZ have a small but distinct genetic overlap with autoimmune diseases, which may inform new possible immune targets for treatment in mental illness.

Artificial intelligence (AI) has played an excellent supporting role in novel drug discovery and development. This study introduces a reinforcement learning (RL) model based on the Soft Actor-Critic (SAC) algorithm for AI-driven de novo molecular generation targeting tyrosinase. The model facilitates forward molecular generation design by integrating a chemical reaction template and a molecular building block library, concurrently performing molecular docking and assessing drug-likeness. Through sequential decision-making, signal feedback, and a dynamic learning process, the model generates molecules exhibiting potent target affinity, optimal drug-like properties, and good synthetic feasibility. The AI-generated molecules undergo rigorous manual screening, synthesis, and biological evaluation, culminating in the identification of a prioritized lead compound V. Subsequent structural optimization of compound V reveals a series of compounds with significantly enhanced activity, shifting inhibitory potency from the micromolar to the nanomolar range. The optimized compound, V-24, demonstrates low cytotoxicity and significant anti-melanogenic activity both in cell melanogenesis inhibition and zebrafish anti-pigmentation models. Notably, it effectively reduces melanin content in an ultraviolet light-induced human 3D skin pigmentation model, exhibiting the potential to serve as a promising tyrosinase inhibitor for the treatment of skin pigmentation. More importantly, this "AI de novo Molecular Generation + Expert-Guided Structural Optimization" work demonstrates that integrating an AI algorithm with traditional medicinal chemistry experience is a novel approach and efficiency-redefined strategy for drug discovery.

Iron constitutes an essential micronutrient in living organisms. All iron is absorbed into the body through dietary intake, except for exogenous therapeutic sources. Dietary iron is typically categorized as either heme or nonheme iron. Nonheme iron is essential for regulating iron in the body, as it exists in various forms, including soluble iron, storage iron within ferritin, and iron found in the catalytic centers of a wide range of proteins. Iron homeostasis is carefully managed to ensure that sufficient iron is available for critical biological processes while preventing the harmful effects that can arise from excess iron. The small peptide hormone hepcidin is the main regulator of iron homeostasis. Hepcidin and other iron regulatory molecules are regulated by various signaling pathways, such as IL-6/JAK-STAT, BMP/SMAD, and MAPK. Alterations in regulatory pathways may occur in response to iron accumulation or deficiency. Iron overload in the body can activate JAK/STAT, BMP/SMAD and MAPK pathways, leading to the initiation hepcidin synthesis. Conversely, in iron deficiency, as in hypoxic conditions or EPO-mediated signaling pathways, HAMP synthesis in the nucleus is reduced. Thus, this review provides an update on the possible regulatory pathways that play a role in iron regulation and may be potential therapeutic targets.

<b>Background</b>: Iron deficiency (ID) is prevalent in congestive heart failure (CHF), worsening outcomes. While European guidelines recommend screening using ferritin and transferrin-saturation (TSAT), inconsistent diagnostic criteria, especially regarding functional deficiency (ferritin 100-299 μg/L + TSAT < 20%) and hyperferritinemia, limit prognostic accuracy. This study evaluated iron status definitions, including guideline criteria and a combined Ferritin-TSAT model, for predicting 365-day mortality in hospitalised CHF patients. <b>Methods</b>: This retrospective analysis used MIMIC-IV data from 1839 CHF patients. Iron status within 24 h of admission was categorised using: (1) Guideline ID vs. non-ID; (2) Ferritin categories; (3) TSAT categories; (4) Combined Ferritin-TSAT model (Low: guideline ID; Intermediate: ferritin 100-299 + TSAT ≥ 20%; High: ferritin ≥ 300 μg/L). Adjusted Cox models assessed mortality associations. <b>Results</b>: Guidelines-defined iron deficiency (33.66% prevalence) independently associated with higher 1-year mortality (56.1% vs. 29.4%; adjusted HR 4.36, 95% CI 3.35-5.34). The combined Ferritin-TSAT model showed significant prognostic value, differentiating true iron deficiency (reference) from hyperferritinemia (adjusted HR 0.50 vs. iron deficiency) and intermediate group (adjusted HR 0.36 vs. ID), indicating varying risk relative to the most deficient group. This combined model better distinguished hyperferritinemic and iron-replete subgroups than the binary guideline definition. <b>Conclusions</b>: Iron status, including deficiency and hyperferritinemia, independently predicts 1-year mortality in CHF. While guideline iron deficiency is a strong predictor, a combined Ferritin-TSAT classification offers finer risk stratification by identifying distinct phenotypes (true deficiency, hyperferritinemia, intermediate). Nuanced iron status assessment could improve prognostic evaluation and guide personalised therapies (e.g., IV iron for deficiency, investigation for hyperferritinemia) to enhance CHF outcomes.

Hepatic angiosarcoma (HAS) is an uncommon malignant tumor that is often misdiagnosed because of its nonspecific presentation and overlapping imaging findings with benign vascular tumors. Sinusoidal obstruction syndrome (SOS) is a disorder of hepatic microcirculation, usually associated with myeloablative chemotherapy and hematopoietic stem cell transplantation. The clinical and pathological overlap between SOS and hepatic angiosarcoma may complicate the diagnosis of hepatic angiosarcoma, as illustrated by this case. We report the case of a sixty-year-old man who presented to the emergency department with progressive jaundice, abdominal discomfort, and weight loss exceeding 5% of his baseline. Laboratory workup revealed a predominantly cholestatic pattern, and serologies for viral, autoimmune, and metabolic liver diseases were negative. Also, there were no known risk factors for liver disease or SOS. Repeated imaging studies, including computed tomography (CT) and magnetic resonance imaging (MRI), revealed a large vascular hepatic lesion, suggestive of a benign lesion, and Doppler revealed findings compatible with SOS. Liver biopsy and hemodynamic studies indicated hepatic outflow obstruction. After clinical deterioration and expert multidisciplinary review of imaging and histology, malignant endothelial proliferation consistent with hepatic angiosarcoma was identified. Hepatic angiosarcoma can closely mimic SOS morphologically and hemodynamically. The absence of identifiable risk factors and the progressive evolution of hepatic changes should raise suspicion of malignancy, prompting multidisciplinary reassessment and careful histologic review.

RNaseT2-deficient cystic leukoencephalopathy (CLE) presents with severe psychomotor retardation, cystic brain lesions, white matter alterations, and cerebral atrophy. The Rnaset2^-/- mouse mirrors key features of this disease and represents the first murine model with a distinct neurological phenotype for type I interferonopathies. Rnaset2^-/- mice exhibit activated microglia, perivascular monocyte and CD8 + T cell infiltration, and hippocampal accentuated atrophy. However, the mechanisms linking interferon-driven neuroinflammation to neurodegeneration remain unclear, underscoring the need to clarify which molecular processes contribute to tissue injury in a time-dependent manner. We found a sustained upregulation of interferon-stimulated genes (IRF9, RIG-I) over three to 28 weeks of age in the brains of Rnaset2^-/- mice compared to controls. Expression of the chemokines Ccl2, Ccl5, and Cxcl10 peaked early but declined thereafter. Pyroptosis-related markers (ASC, CASP1, GSDMD) were significantly increased already at three to 6 weeks of age and decreased thereafter, whereas apoptotic markers such as Bax, Bad, Bid, CASP3, CASP8, and PARP were not differentially expressed compared to controls. Finally, Cd3e as well as Tnf peaked later (at 17 weeks of age) and declined at 28 weeks. Interestingly, double IHC confirmed the co-localization of the pyroptosis-related marker ASC with the microglia marker IBA-1. Taken together, these findings support the notion that pyroptosis is an early, disease-associated event restricted to microglia that likely contributes to establishing a proinflammatory milieu prior to T cell infiltration and brain atrophy. Targeting pyroptosis could therefore represent a potential strategy to attenuate neurodegeneration in type I interferon-driven neuroinflammatory disorders.

Mitochondrial aminoacyl-tRNA synthetases (mt-aaRSs) are a group of proteins encoded by nuclear DNA that play a crucial role in mitochondrial protein synthesis. Mitochondrial diseases caused by mt-aaRS variants are phenotypically heterogenous but often present with significant neurological features such as childhood-onset encephalopathy and seizures. As such, these conditions are a diagnostic challenge. We present an approach that systematically quantifies phenotypic similarity of individuals with an mt-aaRS variant to published cases, to aid variant interpretation, in RD-Connect-a large Europe-wide rare disease cohort. Across 98 individuals with a mt-aaRS gene of interest, we prioritised 38 individuals with 63 variants following bioinformatic and manual analyses. We additionally reviewed Exomiser prioritisation using a pre-defined gene list for neurological disorders within the RD-Connect Genome-Phenome Analysis Platform (GPAP). We were able to generate likely diagnoses in 11 individuals and VUS findings in 13 individuals, following careful phenotype similarity analysis using a phenotype-genotype dataset generated from 234 published individuals. Four of these 24 individuals did not have an Exomiser-ranked gene variant in the GPAP. Therefore, this approach, using individual-level curated phenotype-genotype data to support variant interpretation, can highlight potentially significant variants that may not be captured by current pipelines. This workflow can be replicated in other heterogeneous rare diseases to support clinical practice.

As critical precursors to colorectal cancer (CRC), high-risk colorectal adenomas (HR-CRAs) lack effective chemopreventive strategies beyond endoscopic resection. We previously established a standardized protocol for culturing patient-derived HR-CRA organoids (HR-CRA-PDOs), creating a robust platform for targeted drug discovery in colorectal premalignancy. Bioinformatics investigation was conducted to unveil the significant dysregulation of lipid metabolism in HR-CRAs. HR-CRA-PDOs were primarily cultured and exposed to gradient concentrations of atorvastatin, with drug responses evaluated with high-throughput and high-content imaging and ATP-based viability assays. Parallel in vivo validation utilized AOM/DSS-induced mouse model under either normal or high-fat diets. Histological and molecular analyses were conducted to evaluate adenoma dynamics, apoptosis, and lipid metabolism-related gene and protein expressions. Bioinformatics analysis of GEO database (GSE100179 and GSE161277) revealed that HR-CRAs are characterized by dysregulated lipid metabolism, particularly through the upregulation of fatty acid metabolism pathways. In vitro, atorvastatin significantly inhibited HR-CRA-PDO growth in a dose-dependent manner via apoptosis induction and proliferation arrest. Mechanistically, atorvastatin treatment led to significant alterations of gene expression in lipid metabolism pathways including ACOX1, ACOX2, FABP2, NRG1, PPAR-α and SREBF1, concomitant with stemness marker suppression in HR-CRA-PDOs. In vivo, atorvastatin markedly reduced CRA burden in AOM-DSS-induced mouse model, particularly demonstrating enhanced efficacy in high-fat diet contexts. This translational study establishes atorvastatin's dual mechanism in metabolic reprogramming and stemness inhibition, suggesting its potential as a therapeutic strategy for CRA prevention and treatment.

<h4>Background</h4>Chronic obstructive pulmonary disease is a common respiratory disease. The severity of acute exacerbation of chronic obstructive pulmonary disease is related to disease progression and risk of death. However, the existing grading standards mainly depend on indicators, such as respiratory rate, whether to apply assisted respiratory muscles, and changes in consciousness state, and only reflect the subjective judgment. Imaging omics can extract muscle characteristic data for more complex analysis, which helps to provide a more objective and accurate method to assess the severity of disease for clinic.<h4>Objectives</h4>The purpose of this study is to construct a severity prediction model based on the combination of chest CT muscle imaging features and clinical data in hospitalized patients with AECOPD.<h4>Methods</h4>234 hospitalized patients with AECOPD were retrospectively included, divided into 79 grade I, 74 grade II, and 81 grade III. Clinical data and chest CT images were collected. Construction of clinical feature model combined with muscle imaging omics model based on Python machine learning platform.<h4>Results</h4>The number of hospitalizations for acute exacerbation, disease course, risk of acute exacerbation in stable stage, white blood cell count, neutrophil count, creatinine, and N-terminal B-type natriuretic peptide precursor were statistically different among hospitalized patients with AECOPD in the last year (all P < 0.05). The best model to predict the severity of AECOPD by cascade probability combination method is Xgboost model with AUC of 0.890.<h4>Conclusions</h4>The disease grading prediction model of AECOPD inpatients constructed based on clinical data and muscle imaging omics characteristics has good performance, and has great potential in assisting clinicians to more accurately stratify the risk of AECOPD inpatients.

The dynamics of the early steps of protein aggregation remain poorly understood, particularly in the case of α-synuclein (α-syn) aggregation, the hallmark of synucleinopathies. Here, we present a protocol that combines light-inducible protein aggregation (LIPA) with proximity biotinylation using an UltraID construct. We describe the workflow from protein expression to biochemical validation, including the purification of biotinylated proteins prior to liquid chromatography-mass spectrometry (LC-MS) analysis and subsequent validation. This platform provides a powerful strategy to identify proteins interacting with nascent α-syn aggregates. For complete details on the use and execution of this protocol, please refer to Teixeira et al.¹.

<h4>Background and aims</h4>Thalassemia is a hereditary hematological condition which interferes with the production of hemoglobin (Hb). Patients with thalassemia require regular blood transfusion, which makes them highly susceptible to pathogens, and especially viral pathogens. Hepatitis C virus (HCV) is also a big threat in that it is hepatotropic in nature and has the potential of causing hepatic injury. Host genetic factors, such as single nucleotide polymorphisms (SNPs) in the IFN λ3 (IL28B) gene, influence the ability to clear the virus and respond to therapy. In this research, we aimed to determine the frequency of the rs8099917 and rs12979860 polymorphisms of the IL28B gene in HCV-infected thalassemia patients from southern Iran.<h4>Methods</h4>Sixty-five people with β-thalassemia were recruited, including 25 people who are HCV positive and 40 people who are HCV negative. The genotyping of the locus at the rs8099917 and rs12979860 sites was done using the tetra-primer amplification refractory mutation system-polymerase chain reaction (T-ARMS-PCR).<h4>Results</h4>On analyzing the locus at rs8099917, we noted that the TT genotype was common among patients who had HCV, with the proportion of 72% versus 12.5% among HCV-negative individuals. The GG genotype was also rare in the HCV-positive group and not present in the HCV-negative group. On the other hand, TG genotype were found in 20% of the HCV-positive patients with 87.5% of the HCV-negative subjects, presenting a very significant difference (<i>p </i>< 0.001). On the other hand, the distribution of the rs12979860 genotype was not significantly different in the two groups.<h4>Conclusion</h4>Our findings suggest that the rs8099917 SNP of the IL28B gene may serve as a useful baseline predictor of antiviral responses in thalassemia patients infected with HCV.

<h4>Purpose</h4>Peripheral nerve block safety and pharmacology for patients with inherited antithrombin III (ATIII) deficiency remains unclear because of the rarity of the disease. Here, we report a case of unexpectedly prolonged motor and sensory inhibition following single-shot femoral and sciatic nerve blocks in a patient with inherited ATIII deficiency. We hypothesized that venous stasis, microcirculatory impairment, elevated peripheral venous pressure, and increased local tissue pressure might have contributed to prolonged nerve dysfunction and peripheral nerve injury.<h4>Case report</h4>The patient was a 58-year-old man (170 cm, 69 kg) with inherited ATIII deficiency. He was scheduled for 1-hour surgical debridement of recurrent venous stasis ulcers with microcirculatory impairment on his lower left leg. Nerve blocks resulted in motor and sensory deficits in the lower leg that lasted 2 and 8 days, respectively. The prolonged duration of this nerve block might have been related to his inherited ATIII deficiency. This condition can contribute to thrombosis of vessels such as the inferior vena cava, potentially leading to venous microcirculatory impairment in the lower limbs and complex coagulopathy.<h4>Conclusion</h4>While peripheral nerve blocks are widely used for their efficacy and safety, the risk-benefit profile of regional anesthesia may be atypical in patients with coagulation disorders. This case highlights the need for cautious nerve block administration in patients with inherited ATIII deficiency.

Despite advances in bone graft design and surgical techniques, bacterial infection remains a major cause of graft failure, exacerbated by the global rise in antimicrobial resistance. This has intensified the pursuit of antibiotic-free strategies to prevent bacterial colonization. Among these, antibacterial surface nanotopographies have emerged as promising tools, leveraging nanoscale geometries to physically disrupt bacteria upon contact. In this study, we engineered the surface of a calcium phosphate bone graft to confer antimicrobial functionality through a dual approach: the creation of high-aspect-ratio nanotopographies and ionic doping with fluoride. Through controlled hydrolysis of α-tricalcium phosphate by biomimetic and hydrothermal treatments, we generated calcium deficient hydroxyapatite nanoneedle structures whose morphology and biofunctionality were tuned via fluoride incorporation. XRD and Raman spectroscopy confirmed the formation of hydroxy-fluorapatite, with phase composition and surface morphology dependent on fluoride concentration and processing parameters. Fluoride doping significantly altered nanoneedle dimensions and spacing and enhanced bactericidal activity, particularly against <i>P. aeruginosa</i>, and to a lesser extent <i>S. aureus</i>. Notably, fluoride-doping alone showed no antibacterial effects; however, when combined with nanotopography, a synergistic increase in efficacy was observed. Importantly, the antimicrobial surfaces supported the proliferation and osteogenic differentiation of SaOS-2 cells. Co-culture assays modeling pre- and post-implantation infection scenarios demonstrated robust cell adhesion and markedly reduced bacterial colonization. In conclusion, our findings present a multifunctional, synthetic bone graft with both physical and chemical antibacterial properties, offering a promising strategy to mitigate infection risks while supporting osteointegration.

Drug-induced liver injury (DILI) is a leading cause of liver disease globally, with complementary and alternative medicine (CAM) representing a significant contributor due to its widespread use and limited regulation. CAM encompasses diverse practices and products, with regional and ethnic diversity, many of which are used to treat liver disease. Even though a few natural remedies have been utilized for centuries, most lack scientific evidence for their safety and efficacy. This report describes a 42-year-old male who self-treated his fatty liver with CAM, culminating in severe concomitant hepatic and renal dysfunctions. Comprehensive evaluation excluded viral, autoimmune, metabolic liver diseases, and obstructive biliary diseases. The Roussel Uclaf Causality Assessment Method (RUCAM) score indicated a probable causal link between CAM and DILI of hepatocellular pattern. The constituents of the CAM, extracts of <i>Carica papaya</i>, <i>Alstonia boonei</i>, and <i>Tetrapleura tetraptera</i>, have been implicated in hepatotoxicity in animal models. Discontinuing the supplements and supportive care, including N-acetyl cysteine, led to symptomatic and biochemical improvement. This case highlights the importance of clinicians' awareness of CAM-related DILI, emphasizing the need for vigilance and patient education about the potential risks associated with herbal products.

Treatment-induced neuroendocrine prostate cancer (t-NEPC) is a lethal subtype of castration-resistant prostate cancer (CRPC) characterized by unique pathological features and molecular changes, including the loss of androgen receptor (AR) activities and the gain of neuroendocrine gene expression. The incidence of t-NEPC has increased substantially in the last decade, in up to 20 % of CRPC cases, largely due to intensive treatment of advanced prostate cancer (PCa) with AR pathway inhibitors (ARPi). While genomic alterations between CRPC and t-NEPC are largely conserved, their epigenetic programs are markedly distinct. The molecular mechanisms underlying the neuroendocrine transformation (NET) of PCa are rapidly emerging. Here, we first briefly summarize the genetic drivers of t-NEPC and then comprehensively review 2D and 3D chromatin alterations, including changes in DNA methylation, histone modifications, chromatin accessibility, and 3D chromatin organization, during NET of PCa. We then review key molecular regulators, including lineage-specific transcription factors and chromatin modifiers, of such epigenetic programs. Lastly, we discuss evidence that suggests a mixed model of clonal selection and transformation that underlies NEPC progression.

<h4>Background</h4>Autism spectrum disorder (ASD) is associated with immune and inflammatory dysregulation. However, the molecular networks linking peripheral immune signatures to neuroinflammatory processes remain poorly understood. This study aimed to explore inflammation-related molecular pathways in ASD through integrated transcriptomic network analysis and to validate key cytokine genes (IL6, IL10, IL17, NF-κB1) using quantitative real-time polymerase chain reaction (qRT-PCR).<h4>Methods</h4>This was an integrative computational-experimental study. We analyzed 4 gene expression Omnibus (GEO) transcriptomic datasets (GSE18123, GSE111176, GSE87847, GSE6575), constructed protein-protein interaction (PPI) networks, and identified inflammation-related modules. Selected inflammatory genes (IL6, IL10, IL17, NF-ΚB1) were validated by qRT-PCR in peripheral blood samples from ASD (n = 15) and healthy controls (n = 5). Statistical analyses were conducted in R. Data normality was assessed using the Shapiro-Wilk test, and normally distributed variables were compared using t-tests.<h4>Results</h4>Integration of datasets revealed core differentially expressed genes (DEGs) and a connected PPI network (26 nodes, 88 edges), with hub genes such as PUM1, TRRAP, ILF3, INO80, and PTBP1. Functional enrichment indicated cytokine-mediated signaling, leukocyte activation, and neuroinflammation processes. Network analysis highlighted central regulators linking chromatin remodeling, ribonucleic acid (RNA) processing, and immune signaling. qRT-PCR confirmed dysregulation of IL6 (fold change ≈ 12.8, <i>P</i> = 0.049), IL17 (≈ 21.3, <i>P</i> = 0.048), NF-ΚB1 (≈ 42.4, <i>P</i> = 0.039), and IL10 (≈ 0.101, <i>P</i> = 0.038).<h4>Conclusion</h4>The findings suggest an IL-6/IL-17/NF-κB1-centric proinflammatory axis and reduced IL-10-mediated regulation in ASD, implicating peripheral immune activation and transcriptional regulators in neuroinflammatory processes. The identified hub genes and pathways may serve as biomarkers and therapeutic targets for an inflammation-associated ASD subtype. Limitations include small qRT-PCR sample size and lack of protein-level validation; future studies should explore longitudinal and multiomics approaches.

<h4>ABSTRACT</h4> Huntington’s disease (HD) is caused by CAG expansion in HTT , yet how somatic repeat instability and huntingtin aggregation relate to selective cell loss in the human brain remains unclear. We have developed a multimodal spatial transcriptomics approach that enables defining transcriptional programs with subcellular resolution, somatic CAG repeat lengths, and six other pathology marks including huntingtin aggregates in every cell of intact brain sections. Imaging 428,173 cells in HD cortex revealed selective vulnerability: L5–6 NP and L6b deep-layer excitatory neurons undergo >50% loss, closely linked to very large (>380±55) somatic expansions. Intranuclear aggregation was most prevalent at intermediate somatic repeat expansion (220-300 CAGs) and was accompanied by broader transcriptional changes. In contrast, chandelier and somatostatin+ inhibitory interneurons are lost despite only modest repeat expansion or aggregation. These data provide a comprehensive resource and establish a broadly applicable framework for connecting repeat expansion and protein pathology across diverse cell types. <h4>Short Bullet points</h4> Development of a novel, multimodal spatial transcriptomics platform enables definition of RNA transcriptomes with subcellular resolution, somatic repeat expansions, and protein accumulation in cells within tissue sections Generation of two complementary datasets for HD and control cortex: 428,173 cells to quantify comprehensively cell-type vulnerability, and a deeper multimodal dataset in which CAG repeat expansion, huntingtin aggregation, six additional pathological readouts, and expression of 1,128 genes were measured in 185,721 cells, providing a comprehensive resource for the neurodegeneration community. Deep layer excitatory neuron loss (L5–6 NP and L6b) was associated with very large somatic CAG expansions (>380±55), while selective inhibitory neuron loss (chandelier and somatostatin interneurons) occurred with modest CAG repeat expansion or huntingtin intranuclear aggregation Intranuclear aggregation, not somatic repeat expansion, was more predictive of transcriptional changes, including chromatin remodeling and RNA export factors

Reticulophagy, a selective macroautophagy/autophagy process targeting endoplasmic reticulum fragments <i>via</i> receptors, plays a critical role in cellular homeostasis. This study reveals that CCPG1, a reticulophagy receptor, drives bladder cancer (BLCA) tumorigenesis and confers cisplatin resistance. We observed elevated reticulophagy activity in BLCA cells compared to normal counterparts, particularly under conditions of nutrient stress. <i>CCPG1</i> expression was significantly upregulated in BLCA patient samples and correlated with poor prognosis. Functional studies demonstrated that <i>CCPG1</i> knockdown suppressed reticulophagy, leading to decreased cell proliferation and increased apoptosis. Conversely, overexpression of the wild-type <i>CCPG1</i>, but not a MAP1LC3/LC3-binding-deficient variant, rescued reticulophagy and promoted tumor growth. Notably, we found that cisplatin treatment inhibited reticulophagy by downregulating <i>CCPG1</i> expression through the ATM-CHEK2/Chk2 signaling pathway. <i>CCPG1</i> knockdown synergistically enhanced cisplatin cytotoxicity to BLCA cells, while <i>CCPG1</i> overexpression conferred resistance. These findings highlight CCPG1-mediated reticulophagy as a driver of BLCA progression and as a potential prognostic biomarker and therapeutic target.

Foot-and-mouth disease virus (FMDV) can cause a severe infectious disease that primarily affects even-toed ungulates. FMDV is classified into the genus of <i>Aphthovirus</i> in the family <i>Picornaviridae</i>. FMDV's 3C protein is a nonstructural protein and, moreover, is a protease (3C^pro) that adopts a chymotrypsin-like fold and harbors a Cys-His-Asp catalytic triad. The 3C^pro plays crucial roles not only in cleaving the FMDV polyprotein but also in degrading various host proteins. Cleavage of the polyprotein contributes to generating different viral polypeptides. Degradation of host proteins possibly affects cellular signaling pathways, making FMDV impair innate immune responses. Here, we systematically reviewed FMDV 3C^pro concerning its multiple characteristics, including nucleotide and protein sequences, crystal structures, enzymatic activities, anti-3C^pro inhibitors, and more importantly, its functions in cleaving the viral polyprotein and host proteins. This review aims to provide a comprehensive insight into FMDV 3C^pro as a protease functioning in the course of viral propagation.

<h4>Background</h4>Innate lymphoid cells (ILCs) are classically divided into three groups: ILC1, ILC2, and ILC3 to reflect their functional analogy to Th1, Th2, and Th17. There is also an IL-9 single-producing T cell subset, namely the Th9 cell, which plays a dominant role in the onset of allergic diseases compared with traditional Th2 cells. Although a corresponding cell subset of ILCs to different Th cell subsets exists, so far the counterpart of Th9 cells in ILCs has not been reported.<h4>Objective</h4>In this study, we aimed to report the existence of group 9 innate lymphoid cells (ILC9s) and characterize them in allergic rhinitis (AR) and in response to allergen immunotherapy.<h4>Methods</h4>The expressions and characterization of ILC9s were investigated in purified ILCs cultures, PBMCs from patients with AR and responder subcutaneous immunotherapy (SCIT) patients by flow cytometry, scRNA-seq transcriptome, qRT-PCR, siRNA knockdown, and immunofluorescence staining.<h4>Results</h4>IL-9-expressing cells were observed in the nasal mucosa of patients with AR without the co-expression of IL-5 and IL-13. IL-4 and TGF-β induce IL-9 secretion by human ILCs. scRNA-seq of whole ILCs defines an H1R⁺OX-40L^-ILC subset as ILC9 expressing high levels of IL-9 and low levels of the ILC2 transcription factor GATA3. Instead, this new ILC9 subset displays Bach2 as a transcription factor, and IL-9 expression decreases after siRNA inhibition of Bach2. Histamine is an important regulator of ILC9 because ILC9 production increases in response to histamine, and IL-9 levels in ILCs positively correlate with the expression of histamine 1R. An up-regulation of PPARγ was observed in ILCs in response to IL-4 and TGF-β, and ILC9 differentiation was suppressed by the PPARγ antagonist.<h4>Conclusion</h4>ILC9s are highly expressed in the nasal mucosa and PBMCs of patients with AR and were decreased in response to house dust mite-SCIT. Our study sheds light on the newly discovered ILC9 subset and demonstrates a potential target in allergen immunotherapy.

Type II interferon (IFN) immunity is crucial for controlling intramacrophagic infections, driven by the interaction between innate immunity (macrophage-derived IL-12) and adaptive immunity (Th-derived IFN-γ). This study examines the maturation of type II IFN immunity in 55 healthy children (ages 1-18) to enable proper identification of deficiencies as part of the diagnostic evaluation of Mendelian Susceptibility to Mycobacterial Diseases (MSMD). The IL-12/IFN-γ axis was assessed through: (1) cytokine production after mycobacterial stimulation (Luminex and ELISA for IFN-γ, IL-12p70, TNF, CXCL10, IL-1RA, IL-10, IL-1β and IL-6), (2) IFN-γR1/R2 expression on monocytes, and (3) STAT1 phosphorylation/dephosphorylation. T cell maturation (primary IFN-γ source) was evaluated via immunophenotyping (naïve/memory/activated, Th1; Th2; Th17; Th1/17; Tfh) and proliferation assays. Main findings: (1) stable expression/production of key components of the IL-12/IFN-γ axis (IFN-γ, IL-12, TNF, IFN-γR1/2, and STAT1 activity) across ages confirming the stability of innate immune function throughout childhood; (2) increasing responses to IFN-γ with age reflected by increased CXCL10 production, and increase in the IFN-γ counter-acting anti-inflammatory cytokines (IL-10, IL-1RA); and (3) progressive T cell maturation, including Th1, Th17 and Th1/17 subsets, with significant milestones between 6 and 8.6 years, while T cell proliferative capacity remained stable. These observations highlight the stability of IL-12/IFN-γ axis innate components with age, accompanied by enhanced downstream IFN-γ signaling, aligning with the maturation of Th cell compartment. These underscore the limited benefit of age-specific controls in the evaluation of IL-12/IFN-γ axis in MSMD diagnosis, while emphasizing the importance of T cell maturation in the overall type II IFN immunity.

Toxoplasma gondii is a globally prevalent intracellular parasite that infects ~40 million Americans. The murine immune response to Toxoplasma relies on both toll-like receptor (TLR) 11/12 and immunity related GTPase-mediated (IRGs) responses, which humans lack, making it unclear how the human immune response detects and responds to the parasite. We investigated whether human Vγ9Vδ2 T cells, which detect phosphoantigens through the BTN3A1 receptor, shape the early immune response to the parasite. Using primary human peripheral blood mononuclear cells (PBMCs), we show that Vγ9Vδ2 T cells are activated by Toxoplasma-infected cells in a BTN3A1-dependent manner leading to secretion of interferon gamma (IFNγ) and tumor necrosis factor-alpha (TNFα). Additionally, these T cells potentiate IFNγ production by natural killer (NK) cells, via TNFα and interleukin (IL)-12 produced during infection. Active parasite invasion is required to stimulate the IFNγ response, and inhibition of the host mevalonate pathway, which limits the synthesis of the phosphoantigen isopentenyl pyrophosphate (IPP), attenuates the cytokine response, indicating Toxoplasma infection increases host phosphoantigens leading to Vγ9Vδ2 T cell activation. Our findings identify Vγ9Vδ2 T cells as key effectors that potentiate NK cells in the early human immune response to Toxoplasma, bridging innate and adaptive immunity in the absence of TLR11/12 signaling.

Predicting cellular responses to genetic perturbations is critical for advancing our understanding of gene regulation. While single-cell CRISPR perturbation assays such as Perturb-seq provide direct measurements of gene function, the scale of these experiments is limited by cost and feasibility. This motivates the development of computational approaches that can accurately infer responses to unmeasured perturbations from related experimental data. We introduce dbDiffusion, a generative framework that integrates diffusion models with classifier-free guidance derived from perturbation information, operating in latent space through a variational autoencoder. Diffusion models are probabilistic generative models that approximate data distributions by reversing a Markovian diffusion process, progressively denoising Gaussian noise into structured outputs. By exploiting biological similarities in gene expression profiles and relationships among perturbations, dbDiffusion enables the conditional generation of gene expressions for previously unobserved perturbations. In contrast to competing approaches, dbDiffusion does not rely on Large Language Model or foundation models, which have been found to yield unsatisfactory results. Rather, it leverages embeddings derived from measured perturbations to generalize to unseen perturbations, effectively transferring information across related experimental conditions. In benchmarking against state-of-the-art methods on Perturb-seq datasets, dbDiffusion demonstrates superior accuracy in predicting perturbation responses. A methodological innovation of dbDiffusion is the integration of prediction-powered inference, which corrects for biases inherent in generative models and enables statistically rigorous downstream tasks, including the identification of differentially expressed genes. By combining deep generative modeling with principled inference, dbDiffusion establishes a scalable computational framework for predicting and analyzing transcriptomic perturbation responses, significantly extending the utility of Perturb-seq experiments.

Oligodendroglial pathology is a hallmark of multiple system atrophy (MSA), yet it remains unclear whether MRI-detected atrophy reflects underlying biological mechanisms. This study investigated whether regional atrophy aligns with gene expression and neurotransmitter systems. We recruited 65 MSA patients and derived brain atrophy measures from T1-weighted MRIs. Using postmortem data from the Allen Human Brain Atlas, partial least squares (PLS) regression identified gene expression components associated with brain atrophy. Gene enrichment analyses explored biological processes, and annotation mapping identified neurotransmitter systems matching atrophy patterns. Specificity was tested against 57 patients with Parkinson's disease. Atrophy primarily affected the cerebellar white matter, pons, putamen, olive, and substantia nigra. PLS revealed two latent variables explaining 27.5% of the covariance. Atrophic regions overexpressed genes linked to mitochondrial function and oligodendrocytes, showing patterns distinct from Parkinson's disease. These regions also exhibited lower serotonin and GABA levels, and higher acetylcholine and noradrenaline receptor densities. MRI-derived atrophy in MSA is biologically grounded and may inform future therapeutic studies.

Venous thromboembolism is a common fatal disease that includes pulmonary embolism (PE) and deep vein thrombosis (DVT), and many genetic risk factors are associated with its pathogenesis. We describe a patient with compound heterozygous mutations in PROC combined with a heterozygous mutation in THBD, who was diagnosed with DVT. Genetic sequencing identified three missense mutations in the proband: PROC c.565 C > T (p.R189W), PROC c.1218G > A (p.M406I), and THBD c.1456G > T (p.D486Y), this genotype not previously documented in association with thrombotic disease. His father carried heterozygous mutations of PROC p.R189W and THBD p.D486Y, while his mother and maternal grandfather were heterozygous for PROC p.M406I. These mutations were not detected in other family members.

Transfer of antibodies across the placenta provides critical early life protection for neonates against infection. Understanding factors influencing efficient transfer is vital for enhancing neonatal immunity and improving maternal vaccination strategies. Stored maternal and cord serum samples from studies in The Gambia, Guatemala, Mali, The Netherlands, Pakistan, Thailand, UK and Vietnam were processed for measles plaque reduction neutralisation titres (PRNT), and measles, mumps and rubella immunoglobulin G (IgG) (multiplex immunoassay) at a central laboratory (N = 557 pairs). Nutritional indicators (ferritin, soluble transferrin receptor, retinol binding protein) and total IgG, were measured in subsets of maternal serum. Transplacental transfer ratios (TPTRs) were calculated and factors associated with maternal IgG, cord IgG and TPTRs explored in multivariable regression. At delivery, most mothers (range 78 % Guatemala to 100 % Pakistan, Netherlands) and infants (range 86 % Guatemala to 100 % Netherlands, UK) had PRNT above the threshold of clinical protection (0.12 IU/mL). Maternal and cord antibody concentrations across diverse geographical settings were highly correlated. TPTRs were highest in high-income countries; geometric mean range 0.7 (Pakistan) to 2.2 (Netherlands), and were correlated across antigens. However, TPTRs varied widely within low/middle-income countries. In models adjusting for country, higher maternal total IgG was consistently associated with lower TPTR. In multivariable regression, lower iron status, indicated by increasing soluble transferrin receptor concentration, was significantly associated with lower TPTR for measles neutralising antibody. There is marked geographical variation in TPTRs, and following adjustment for this, measurable factors in maternal blood can inform estimates of transplacental transfer efficiency.

Oncogenic KRAS G12D mutations drive pancreatic ductal adenocarcinoma (PDAC) but face therapeutic resistance from pathway reactivation. We synthesized MDP5, a dual BRD4/PI3K inhibitor, to address this issue. When combined with the KRAS G12D inhibitor MRTX1133, MDP5 resensitized resistant cancer cells. This combination synergistically enhanced apoptosis and proliferation inhibition, outperforming the standard-of-care, Gemcitabine (GEM). This dual-inhibition strategy effectively counters resistance mechanisms in KRAS-mutant PDAC, offering a promising therapeutic approach. We formulated MUC4-targeted polymeric nanoparticles co-loading MRTX1133 (8.3%) and MDP5 (7.4%) that exhibited pH-responsive release and uniform morphology. This targeted delivery translated to superior anti-tumor efficacy, as the combination of NPs markedly reduced tumor burden more effectively than single-drug treatments or a polymer-Gemcitabine conjugate. Importantly, this potent therapeutic effect was achieved without inducing detectable liver highlighting the potential, safe, and effective cancer therapy. Mechanistically, dual targeting reduced p-AKT and YAP1, depleted CD44⁺/ALDH⁺ cancer-stem-like cells, and reprogrammed the tumor immune microenvironment by elevating CD8a and CD86 while lowering Ly-6 G. Targeted co-delivery of MRTX1133 and MDP5 simultaneously inhibits the KRAS G12D and PI3K pathways. This dual-action approach aims to overcome therapeutic resistance, offering potentially more durable and precise treatment for patients with KRAS G12D-mutant pancreatic cancer. STATEMENT OF SIGNIFICANCE: Pancreatic ductal adenocarcinoma (PDAC) is commonly driven by KRAS G12D, yet the KRAS G12D inhibitor MRTX1133 faces rapid adaptive resistance and delivery hurdles. We engineered MUC4-targeted polymer nanoparticles that co-deliver MRTX1133 with MDP5, a dual BRD4/PI3K blocker. The particles show uniform size, high loading, and pH-triggered release, increasing intratumoral drug levels versus free drugs. In orthotopic PDAC, the combination outperformed monotherapies and gemcitabine without overt toxicity. Mechanistically, dual inhibition suppresses AKT/YAP signaling, depletes CD44⁺/ALDH⁺ cells, and shifts the tumor immune milieu (↑CD8a, ↑CD86, ↓Ly-6 G). By integrating molecular targeting with multi-pathway blockade, this work addresses two major barriers-resistance and delivery-and outlines a generalizable strategy to improve precision nano therapy for KRAS-mutant pancreatic cancer.

Nanodiamonds (NDs) have emerged as a highly promising nanomaterial due to their intrinsic biocompatibility and remarkable antimicrobial and anti-adhesive properties, which result from their unique surface morphology. NDs serve as an excellent platform for extensive functionalization with diverse chemical groups and complex bioactive molecules, including peptides, photosensitizers, antibiotics and polycations. The antimicrobial potential of NDs has gained considerable attention in recent years across numerous application areas, including drug-delivery platforms, wound dressings, dentistry, surface coatings, biomedical implants, the food industry and water treatment technologies. This article compiles and critically evaluates the current microbiological evidence on ND antimicrobial activity. However, translating these findings into practical guidelines remains challenging due to the wide variability in reported results and the limited diversity of bacterial strains employed. The antimicrobial mechanisms of NDs in the context of Gram positive, Gram negative, and flagellated bacteria are examined, and it is demonstrated that key factors, including particle size, surface charge, and the composition of testing media, profoundly influence experimental outcomes and underlie many apparent contradictions in the field. Moreover, this review summarizes the functionalization strategies available for NDs, their reported biomedical and industrial applications, and current knowledge regarding their cytotoxicity and biocompatibility. Collectively, the article provides an integrated view of the structure-activity relationship governing ND antimicrobial performance.

Our genome-wide association study identified single-nucleotide polymorphisms (SNPs) associated with estimated breeding values (EBVs) for udder traits and longevity in Holstein-Friesian cows. While no SNP was individually associated with multiple EBVs, the functional profiles of the associated genes revealed overlapping biological processes across traits, including cell signaling, transcription regulation, immune response, metabolism, and cellular maintenance. Notably, nearby SNPs BTB-01738708 and ARS-BFGL-NGS-111478 were associated with EBV_longevity and EBV_udder and located near numerous genes, including <i>GPR85</i>, <i>BMT2</i>, <i>IFRD1</i>, and <i>DOCK4,</i> suggesting a potential for shared genetic influence on these traits. Our findings provide insights into the complex genetic architecture of these economically important traits and highlight the need for further research, including fine-mapping and functional genomics, to elucidate the specific variants and their effects.

In higher vertebrates, Tripartite Motif (TRIM) proteins modulate the immune response by coordinating processes related to inflammation such as antiviral restriction, autophagy and inflammasome activation. In fish, TRIM proteins have been reported mainly in cyprinids (e.g., carp-<i>Cyprinus carpio</i> and zebrafish-<i>Danio rerio</i>) and salmonids (i.e., rainbow trout-<i>Oncorhynchus mykiss</i>). However, their molecular mechanisms and functions are still being described in aquatic animals. Thus, our study focused on characterizing novel TRIM proteins involved in the innate immunity of gill cells from rainbow trout (RTgill-W1 and primary cultures) stimulated with lipopolysaccharide (LPS) or polyinosinic-polycytidylic acid (poly I:C). Furthermore, an <i>in vivo</i> experiment with rainbow trout was performed to detect TRIM proteins after the challenge with <i>Flavobacterium psychrophilum</i> (a major bacterial pathogen affecting Chilean salmonid industry). <i>In vitro</i> results showed that OmTRIM25 triggered an LPS-induced expression of pro-inflammatory cytokines such as TNF-α2 and IL-1β. Moreover, in the fish experiment, OmTRIM25 and finTRIM2 were up-regulated in the gills two days post-infection (dpi), whereas IL-1β and TNF-α2 had a higher gene expression at four and six dpi, respectively. To investigate the immunological role of OmTRIM25, a gene silencing strategy using RNA interference (RNAi) was used, confirming the immunomodulatory function of OmTRIM25.

<h4>Background</h4>Pancreatic cancer represents a significant global health burden. Although dysregulated lipid metabolism and its associated inflammation drive tumorigenesis, their molecular interplay remains incompletely understood. This bioinformatics study investigates lipid metabolism-related genes (LMRGs) for prognostic prediction and treatment guidance in pancreatic cancer.<h4>Methods</h4>LMRGs were obtained from the Gene Set Enrichment Analysis (GSEA) database, while messenger ribonucleic acid (mRNA) expression profiles and clinical information were downloaded from The Cancer Genome Atlas (TCGA), Genotype-Tissue Expression (GTEx), and International Cancer Genome Consortium (ICGC) databases. Cox regression analysis and the least absolute shrinkage and selection operator (LASSO) regression analysis were employed to screen prognosis-related genes, followed by the construction of a risk prediction model. Patients were stratified into high- and low-risk groups for prognosis and immune infiltration comparison. Potential therapeutic drugs for pancreatic cancer were predicted using the DSigDB database based on the identified LMRGs.<h4>Results</h4>We successfully established and validated a prognostic prediction model for pancreatic cancer patients based on six LMRGs (<i>AGT</i>, <i>AHR</i>, <i>PLA2G6</i>, <i>PTGS2</i>, <i>TNFRSF21</i>, and <i>VDR</i>). The 1-, 3-, and 5-year area under the receiver operating characteristic (ROC) curve values were 0.623, 0.698, and 0.720, respectively. Immune infiltration analysis showed that after prognostic risk stratification using the six-gene signature, the high-risk group had higher proportions of M0 macrophages and neutrophils. Furthermore, the expression of eight immune checkpoint-related genes was significantly increased in the high-risk group. DSigDB database analysis revealed four possible therapeutic drugs for pancreatic cancer: prolinedithiocarbamate, isoliquiritigenin, aspirin, and resveratrol.<h4>Conclusions</h4>The risk score based on the six LMRGs provides prognostic insights for pancreatic cancer. High-risk pancreatic cancer populations are potentially associated with an immunosuppressive microenvironment. Candidate drugs screened based on LMRGs offer new possibilities for personalized treatment of pancreatic cancer.

Combined heart-liver transplantation (CHLT) is a rare and highly complex procedure reserved for patients with end-stage failure of both organs. We report the first successful CHLT performed in Chile, involving a 55-year-old man with severe dilated cardiomyopathy and chronic liver disease who underwent sequential bicaval heart transplantation followed by piggyback liver transplantation. The postoperative course was complicated by acute neurological deterioration due to an acute subdural hematoma with tentorial extension requiring decompressive craniectomy, after which the patient recovered fully. More than one year after transplantation, he remains in excellent health, physically active, and fully reintegrated into work, with excellent function of both grafts. This case highlights the feasibility of CHLT in Chile and underscores the importance of multidisciplinary evaluation and coordinated perioperative management in optimizing outcomes for patients with advanced multi-organ failure.

In this study, we characterized the holin-like protein ORF70 from the cyanophage MaMV-DC, offering valuable insights into its role in phage-mediated host cell lysis. ORF70 shares key features with class III holins, such as a hydrophobic transmembrane domain and membrane-associated localization, which are crucial for its bacteriolytic activity. Subcellular localization studies suggested its association with the membrane, supporting its classification as a holin-like protein. Overexpression of ORF70 in <i>E. coli</i> resulted in significant growth inhibition, increased β-galactosidase leakage, and visual confirmation of cell death through live/dead staining. Additionally, ORF70's sensitivity to the energy toxin 2,4-dinitrophenol (DNP) further indicated its holin-like activity by promoting membrane depolarization. Transmission electron microscopy and Gram staining revealed characteristic morphological changes in <i>E. coli</i> cells, including membrane disruption, consistent with damage caused by holins. These results suggest that ORF70 acts as a holin-like protein that disrupts the host membrane, leading to bacterial cell death. Our study provides evidence supporting the holin-like activity of ORF70 from cyanophage MaMV-DC. This research significantly enhances our understanding of phage-host interactions and opens new avenues for developing phage-based therapies, offering promising alternatives to traditional antibiotics amidst the growing challenge of antibiotic resistance.

Acne vulgaris is a prevalent chronic inflammatory skin disorder affecting over 85% of adolescents. Emerging evidence indicates that <i>Cutibacterium acnes</i> phylotype IA₁ contributes to acne initiation and progression, yet its precise mechanisms in epidermal keratinocytes remain unclear. This study investigated <i>C. acnes</i> IA1's effects on keratinocyte behavior using an in vitro HaCaT cell model. Cells were co-cultured with live <i>C. acnes</i> IA₁ (CICC 10864) for 24 h. Transcriptomic profiling identified 769 differentially expressed genes (DEGs; adjusted <i>p</i> < 0.05, |log2FC| > 1), including 392 upregulated and 377 downregulated. The protein-protein interaction network analysis via Cytoscape revealed key hub genes (HNRNPA2B1, HNRNPM, RBM39). Enrichment analyses (GO, KEGG, Reactome, DO) highlighted significant involvement of the C-type lectin receptor (CLR) signaling pathway. Validation experiments showed cellular morphological changes, altered structure, and markedly elevated interleukin-6 (IL-6; <i>p</i> < 0.01), underscoring its role in inflammation. These findings suggest <i>C. acnes</i> IA₁ drives acne pathogenesis by regulating hub genes that influence sebaceous gland inflammation, immune activity, and keratinocyte proliferation, positioning them as potential biomarkers for microbiome-targeted therapies. Limitations include the in vitro model's lack of in vivo skin microenvironment complexity and use of only one representative IA₁ strain.

CaMoO₄:Eu³⁺-functionalized hydroxyapatite-silica (HAp-SiO₂-CaMoO₄:Eu³⁺) core-shell nanocomposites were synthesized for the first time and evaluated as a multifunctional drug delivery and imaging platform. HAp-SiO₂ nanocomposites were prepared via a hydrothermal route and subsequently functionalized with a luminescent CaMoO₄:Eu³⁺ shell using the Pechini sol-gel method. Structural analyses confirmed the successful coexistence of HAp, SiO₂, and CaMoO₄:Eu³⁺ phases, while electron microscopy revealed spherical core-shell morphologies. Dynamic light scattering measurements showed average hydrodynamic particle sizes of approximately 1085 nm for HAp-SiO₂ and 1427 nm for HAp-SiO₂-CaMoO₄:Eu³⁺ nanocomposites, indicating particle clustering in aqueous media, which is consistent with the low surface charge of the particles. The Eu³⁺-doped shell exhibited a strong red emission centered at 615 nm, demonstrating suitability for luminescence-based imaging. Zoledronic acid (ZA) was efficiently loaded onto the nanocomposites under supercritical CO₂ conditions, providing high loading efficiency and sustained release behavior. In vitro release studies in phosphate-buffered saline (pH 7.4, 37 °C) followed the Korsmeyer-Peppas kinetic model (n = 0.83), indicating a non-Fickian diffusion mechanism. Cytotoxicity assays on Saos-2 osteosarcoma cells demonstrated that ZA-loaded nanocomposites exhibited enhanced antiproliferative activity, with an IC₅₀ value of 56.33 μM after 48 h. These results highlight the potential of HAp-SiO₂-CaMoO₄:Eu³⁺ nanocomposites as an integrated theranostic system for targeted bone cancer therapy. Overall, the results demonstrate that the proposed nanocomposite design successfully translates the intended theranostic concept into experimentally validated structural, optical, and biological performance.

Ewing sarcoma is an aggressive bone tumor of adolescence characterized by a hallmark EWSR1::ETS fusion oncogene. The resulting chimeric oncoprotein drives tumorigenesis by reshaping transcriptional and epigenetic landscapes. However, how it is transcriptionally regulated and whether additional master transcription factors (MTFs) form a core regulatory circuit (CRC) in Ewing sarcoma remain unclear. Using an extensive panel of Ewing sarcoma cell lines and primary tumors, we mapped super-enhancers and identified enrichment of GGAA microsatellites, confirming their specificity to Ewing sarcoma as compared to other pediatric cancers and normal tissues. Integrating transcriptomic, epigenetic, 3D chromatin conformation, and dependency data, we predicted a set of MTFs potentially forming a CRC. However, functional validation demonstrated that these MTFs neither establish auto-regulatory loops nor confer robust proliferative dependencies typical of CRCs in other pediatric tumors. Instead, EWSR1::FLI1 emerged as an "hegemonic" oncoprotein, regulating expression of these MTFs without reciprocal regulation. Knockdown of EWSR1::FLI1 strongly shifted H3K27ac profiles toward mesenchymal states, whereas silencing individual or combined MTFs did not alter cell growth or EWSR1::FLI1 expression. These findings highlight the absence of a classical CRC in Ewing sarcoma and emphasize EWSR1::FLI1 as the dominant oncoprotein and a major vulnerability in this disease.

Clinical studies have shown a paradox of the usage of platelet-rich plasma (PRP) on treating fatty infiltration (FI) in injured muscles. However, the underlying reason is still unclear, partially owing to unknown effective components and confounders. Here we found that exosomes derived from PRP (thereafter named PRP-exos) most efficiently prevented FI in injured muscles by inhibiting the adipogenesis of fibro-adipogenic progenitors (FAPs). Importantly, we found aging largely impaired the therapeutic effects of PRP-exos. Mechanistically, miRNA cargoes in PRP-exos mediated the effects of PRP-exos on adipogenesis of FAPs as well as FI in injured muscles, of which, hsa-let-7f-5p and hsa-miR-16-5p were the two most important components. TGFBR3 was identified as a new cotarget gene of these two miRNAs and a new regulator to control the adipogenesis of FAPs. The FI in muscles can be significantly reduced after conditional knockout of TGFBR3 in FAPs. In addition, we further investigated that TGFBR3 regulated the activation of ERK-PPARγ pathway through directly inducing the degradation of KRT10, and thus impacted the adipogenesis of FAPs. Interestingly, PRP-exos or these two miRNAs can preserve the viability and promote the proregenerative supporting capacity of FAPs by targeting TGFBR3 to facilitate muscle regeneration. Collectively, our findings identified the effective components in PRP to inhibit FI and support muscle regeneration. Furthermore, the negative influence of aging on clinical applications of PRP cannot be neglected.

Chronic cadmium exposure is linked to esophageal squamous cell carcinoma (ESCC) progression, treatment resistance, and poor prognosis, but its molecular mechanisms remain unclear. Based on whole-transcriptome sequencing (RNA-seq) on chronic cadmium-treated (CCT) and untreated human EC109 ESCC cells, we performed lncRNA-mRNA co-expression and protein-protein interaction (PPI) network analyses to screen the hub genes related to cadmium exposure. Then, Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses depicted that these genes were mainly enriched in nucleocytoplasmic transport, ferroptosis, Huntington disease, and cellular senescence. Based on the lncRNA-miRNA-mRNA potential interaction, three lncRNAs (AC107068.1, TTN-AS1, MAPKAPK5-AS1) and ten mRNAs (IPO5, NUDCD1, OSTM1, CCNB1, FANCD2, TFRC, POLR2B, HTT, NUP43, NBN) were selected for competitive endogenous RNA (ceRNA) network construction. Connectivity map (CMap) analysis for the target mRNAs showed that JAK3-inhibitor-I was the most promising therapeutic drug for cadmium-induced ESCC progression. The prognostic value and abnormal expression of the candidate mRNAs were then validated in TCGA cohorts, cadmium-treated cancer cells and 41 ESCC specimens. Moreover, migration and invasion assays were performed to assess the effects of the identified gene on cell malignant phenotypes. As a result, IPO5 was identified as the pivotal gene and MAPKAPK5-AS1-hsa-miR-379-5p-IPO5 was deemed as a potential ceRNA regulatory mechanism for cadmium carcinogenesis in ESCC. Using TIMER and EPIC algorithms, IPO5 was correlated with increased infiltration of CD4+ T cells and macrophages, while negatively associated with CD8+ T cells and NK cells in ESCC tissues. Collectively, our study provides valuable information in understanding the molecular mechanisms involved in cadmium-induced ESCC progression and treatment resistance. Furthermore, we predict potential agents for cadmium carcinogenicity prevention and treatment.

BACKGROUND: Trigeminal neuralgia (TN) is a severe neuropathic pain disorder characterized by sudden, electric shock-like facial pain. It frequently co-occurs with depression and these conditions often exacerbate each other. However, their shared pathogenic mechanisms remain poorly understood METHODS: In this study, neuroimaging data were integrated with whole-genome sequencing data to identify common genetic risk factors. Initially, single nucleotide polymorphisms (SNPs) associated with both conditions were identified through whole-genome analysis. Gene-tissue associations were then elucidated using the S-PrediXcan and summary data-based Mendelian Randomization (SMR) methods. These findings were further validated through neuroimaging analysis and animal experiments. RESULTS: Whole-genome analysis identified SNPs shared by TN and depression. The neurocan (NCAN) gene was significantly associated with both disorders and showed a cerebellum-specific expression pattern. Neuroimaging revealed significant differences in brain function and structure between TN and depression patients, primarily localized in the temporal lobe, frontal lobe, and cerebellum. Animal experiments corroborated these findings, showing that elevated cerebellar NCAN expression is closely linked to the onset and progression of both TN and depression. CONCLUSIONS: The study identifies the NCAN gene and cerebellar dysfunction as shared potential pathogenic factors between TN and depression. These findings provide new insights into the common mechanisms underlying these comorbid conditions and suggest potential implications for developing targeted therapeutic strategies.

<h4>Objective</h4>This study aimed to investigate differences in protein expression, protein function, and epigenetic modifications related to histone phosphorylation in peripheral blood mononuclear cells among patients with chronic fatigue syndrome (CFS) presenting two common Traditional Chinese Medicine (TCM) syndrome types compared with healthy controls.<h4>Methods</h4>Three groups of participants were selected from the Medical Examination Center of the Affiliated Hospital of Shandong University of Traditional Chinese Medicine between January 2024 and December 2024. The first group consisted of 10 CFS patients with the Qi and Blood Deficiency syndrome pattern; the second group included 10 CFS patients with the Liver Depression and Spleen Deficiency syndrome pattern; and the third group comprised 10 healthy controls. Phosphopeptide enrichment analysis was employed to detect and identify differentially expressed proteins, followed by Gene Ontology (GO) enrichment analysis.<h4>Results</h4>Differences were observed between both TCM syndrome types of CFS (Qi and Blood Deficiency, Liver Depression and Spleen Deficiency) and the healthy control group in various biological processes and cellular components, including intracellular signal transduction, actin filament bundling, and cortical cytoskeleton organization.<h4>Conclusion</h4>Multiple processes, structures, and functions-such as intracellular signaling, presynaptic modulation of chemical synaptic transmission, actin filament bundling, cortical cytoskeleton organization, SH3 domain binding, and profilin binding-are associated with the occurrence and development of CFS.

<h4>Background & aims</h4>In response to injury, pancreatic acinar cells undergo acinar-to-ductal metaplasia (ADM), marked by loss of acinar identity and acquisition of ductal features. Although ADM can resolve to support tissue repair, it may also persist and serve as a precursor to pancreatic cancer. Whether diverse pancreatic stressors drive a shared or context-specific ADM program remains unclear. We sought to comprehensively define metaplastic responses to clinically relevant exocrine pancreas diseases known to increase cancer risk.<h4>Methods</h4>We profiled ADM and the surrounding microenvironment across mouse models of exocrine disease-including acute, recurrent, and chronic pancreatitis, as well as in the setting of oncogenic Kras-capturing over 300,000 single cells. To enable high-quality transcriptomic profiling in enzyme-rich tissue, we leveraged FixNCut, a method that preserves RNA integrity in the exocrine pancreas. Findings were validated in human pancreas tissue using CosMx spatial transcriptomics.<h4>Results</h4>We identify a conserved acinar response across disease contexts that gives rise to previously unrecognized distinct metaplastic states, including a "gateway" ADM population that precedes more advanced metaplastic states marked by complete loss of acinar identity. In pancreatic intraepithelial neoplasia (PanIN) precancerous lesions, we detect classical-like and basal-like states, suggesting that pancreatic cancer subtypes are specified much earlier than previously appreciated. In Kras-mutant tissue, we identify a second wave of inflammation and the emergence of an immunosuppressive niche, coinciding with PanIN formation.<h4>Conclusions</h4>Our findings define a conserved program of acinar plasticity across exocrine pancreas diseases. We further link unresolved ADM to immune remodeling during precursor lesion formation and observe the emergence of pancreatic cancer subtypes in early PanIN lesions.

<i>Eurotium cristatum</i>-Fermented White Tea (FWT) significantly alters white tea (WT) composition, increasing caffeine while decreasing polyphenols and amino acids. FWT effectively ameliorated dextran sulfate sodium (DSS)-induced murine colitis symptoms (reducing weight loss, colon shortening). Mechanistically, FWT suppressed TLR4/Myd88/NF-κB signaling and pro-inflammatory cytokines (TNF-α, IL-6, IL-1β) while upregulating tight junction proteins (ZO-1, occludin, claudin-1), MUC2, and E-cadherin. Single-cell/spatial transcriptomics revealed that FWT treatments augment enterocyte, goblet cell, and stem cell populations, optimize goblet function, restructure stem cell differentiation, and induce epithelial REG3B (antimicrobial) and LYPD8 (motility inhibitor), plus immunomodulator GM42418 lncRNA across cell types, repairing the barrier. FWT intervention was also associated with an increase in beneficial bacteria (<i>Akkermansia</i>, <i>Lactobacillus</i>, <i>Bifidobacterium</i>), restoration of microbiota balance, and elevated levels of short-chain fatty acids (SCFAs) and was associated with alterations in caffeine-related metabolite profiles. Collectively, these multi-scale changes correlate with the alleviation of UC, suggesting an integrated mechanism involving mucosal barrier repair, immune-stromal modulation, microbiota-metabolism regulation, and cellular reprogramming.

Prostate cancer is the second most common cancer in men worldwide, highlighting the urgent need for effective and targeted chemotherapeutic approaches. This study reports the development and optimization of 5-fluorouracil (5-FU)-loaded poly-(lactic-<i>co</i>-glycolic acid)-polyethylene glycol-folic acid (PLGA-PEG-FOL) nanoparticles designed for folate receptor-mediated targeted therapy. The PLGA-PEG-FOL conjugate was synthesized via a stepwise carbodiimide coupling reaction and confirmed by FT-IR analysis. Nanoparticles were formulated via a modified emulsification-solvent evaporation method and optimized through a Box-Behnken design. The optimized formulation demonstrated a particle size of 178.47  ±  3.26 nm, a narrow polydispersity index (0.119  ±  0.008), a zeta potential of -23.4  ±  0.35 mV, a high entrapment efficiency (78.93  ±  1.05%), and sustained release of 5-FU for up to 72 h. In vitro cytotoxicity assays in PC-3 prostate cancer cells revealed a 1.6-fold reduction in the IC₅₀ value compared with that of free 5-FU, indicating enhanced therapeutic potency. In vivo efficacy was evaluated in testosterone-induced prostate cancer in male Wistar rats. Compared with the control, treatment with 5-FU-loaded PLGA-PEG-FOL nanoparticles significantly reduced the prostate index and produced a 2.2-fold decrease in serum PSA levels and a 1.9-fold decrease in serum testosterone levels. Histopathological examination confirmed the attenuation of hyperplastic and dysplastic lesions in the nanoparticle-treated group. These findings suggest that PLGA-PEG-FOL nanoparticles are a promising targeted delivery platform for enhancing the therapeutic efficacy of 5-FU in prostate cancer treatment.

The purpose of this in vitro study was to evaluate the remineralization effect of five preventive treatments on bovine enamel after artificial caries challenge. Sixty sound bovine incisors were randomly distributed into six experimental groups (<i>n</i> = 10). Each group received the following daily preventive treatments for two weeks during pH cycling to assess remineralization efficacy: Group 1-no treatment (control), Group 2-CPP-ACPF treatment, Group 3-fluoride-containing bioactive glass treatment (BioMin™F), Group 4-SnF₂ treatment, Group 5-toothpaste containing fluoride and hydroxyapatite (HA), and Group 6-toothpaste containing HA. Surface hardness changes were evaluated using a nanoindentation tester. Scanning electron microscopy was used to observe changes in surface morphology, and the mineral composition of enamel in each group was analyzed using energy-dispersive X-ray spectroscopy (EDS). Groups 2, 3 and 6 exhibited the highest surface hardness values after pH cycling, with no statistically significant differences among them (<i>p</i> > 0.05), whereas groups 4 and 5 presented significantly lower values (<i>p</i> < 0.05). Although all treatments demonstrated significant remineralization potential-resulting in an 18.7-35.2% increase in surface hardness-none of them fully restored the hardness loss caused by demineralization. SEM observations revealed precipitations mainly in groups 2-4 after the treatments. EDS showed a similar elemental composition on enamel across the groups with no differences compared to the control. Additionally, line scans of the Ca and P content from the surface to the deeper layers resulted higher values in the tested groups compared to the control corresponding to the surface hardness values. The tested remineralizing treatments may be useful to limit the demineralizing effect during caries formation.

Cell-based drug delivery has emerged as a powerful strategy to improve therapeutic targeting while reducing systemic toxicity. This approach is particularly valuable for anticancer agents, which are often limited by severe side effects arising from off-target activity and non-specific distribution. By using cells as carriers, drugs can evade immune clearance, achieve prolonged circulation, and improve pharmacokinetic profiles, ultimately enhancing therapeutic efficacy. This review surveys the current landscape of cell-mediated drug delivery in oncology, emphasizing both fundamental principles and practical applications. We discuss the design and preparation of cellular carriers, examine the unique characteristics of commonly used cell types, and highlight recent technological innovations that are expanding their theranostic potential, focusing on strategies for delivery to challenging anatomical sites, with a dedicated focus on the brain. By consolidating recent advances and insights, this review aims to provide a comprehensive perspective on the promise and future directions of cell-based drug delivery for cancer therapy.

ATAD2 is a widely conserved, homohexameric ATPase that, in humans, features a bromodomain tier. It plays a role in chromatin remodelling in embryonic stem and germ cells but is frequently upregulated in many cancers, making its bromodomain an attractive drug target. While ATAD2-like proteins generally modulate nucleosome density to control genome compartmentalization and gene expression across eukaryotes, their precise molecular functions vary. We investigated LmxBDF7, the ATAD2 ortholog in the important human pathogen Leishmania mexicana , the causative agent of cutaneous leishmaniasis. Unlike its human counterpart, the LmxBDF7 bromodomain is predicted to be occluded and non-canonical. BDF7 null mutants were unable to develop into infectious amastigote forms and could not infect macrophages, demonstrating it is essential for lifecycle progression. Chromatin Immunoprecipitation sequencing (ChIP-seq) suggested low affinity for chromatin, aligning with its atypical bromodomain. Instead, Proximity-Biotinylation (XL-BioID) suggested a role in ribosome maturation within the nucleolus. RNA-sequencing (RNA-seq) of the Δ bdf7 mutant revealed widespread disruption of gene expression during growth and differentiation. Crucially, key genes required for amastigote survival, such as ribosomal protein genes and glutamine synthetase, were downregulated. This downregulation was spatially biased, preferentially affecting genes on Chromosome 23. Our combined data suggest that while BDF7 is essential, its functions have diverged from other ATAD2-like factors found in opisthokonts. <h4>Author Summary</h4> ATAD2 is a protein that helps cells balance the number of nucleosomes bound to DNA in the nucleus of a cell. Occurring at important sites or times this provides a helper function that ensures other protein complexes can operate on chromatin effectively. In some cancers ATAD2 is disrupted and therefore is being explored as target for new medicines. Orthologues of ATAD2 have been characterised in mammals and several species of yeast. We have sought to identify if an ATAD2-like protein can be found in the important human pathogen, Leishmania mexicana – which is evolutionary distant from humans and yeast. Indeed, we were able to find an ATAD2-like protein called BDF7. Interestingly, BDF7 has a bromodomain which is predicted to be non-functional in terms of being able to bind histones. We were able to make strains of Leishmania mexicana that lacked BDF7 which were viable, but unable to complete the differentiation step required for infecting macrophages. Intriguingly we found BDF7 had a poor association with chromatin and inhabited a protein neighbourhood defined by factors which facilitate ribosome biogenesis. Lastly, RNA-seq analysis of the cells revealed that those lacking BDF7 were potentially depleted for glutamine synthetase which would prevent them developing into fully functional amastigotes.

<h4>Background</h4> The Allan-Herndon-Dudley syndrome (AHDS) is a rare, X-linked human genetic disorder caused by mutations in the monocarboxylate transporter 8 (MCT8), essential for thyroid hormone (TH) transport across the blood-brain-barrier. The resulting central TH deprivation disrupts brain maturation and function, leading to intellectual disability and movement disorders. Cortical development, highly dependent on TH, is particularly affected and contributes significantly to AHDS pathologies. <h4>Methods</h4> To elucidate disrupted cortical processes, we conducted single nucleus RNA sequencing in a mouse model engineered to mimic the central TH deficiency characteristic of human AHDS. This murine AHDS model features the concomitant deletion of both MCT8 and OATP1C1, a T4 transporter largely absent in human brain capillaries that in mice plays a role in TH transport. The phenotype of dKO mice bears striking resemblance to the pathologies observed in human AHDS patients. <h4>Results</h4> Single nuclei were isolated from the cortex and attached striatum of 21-day old WT and MCT8/OATP1C1 dKO mice and sequenced using the 10x Genomics workflow. Cell proportion analyses on the resulting 48 clusters suggested elevated numbers of GABAergic striatal D1 and D2 neurons in the dKO mice. Diminished levels of mature oligodendrocytes coincided with a bifurcation within the oligodendrocyte lineage trajectory, leading to distinct subpopulations of WT and dKO oligodendrocytes. Differentially expressed gene (DEG) patterns align poorly with Slc16a2 and Slco1c1 mRNA levels in the respective clusters, but closely with prior published cortical bulk RNAseq data of mice with systemic hypothyroidism or MCT8/OATP1C1 deficiency. These parallels confirm the reliability of our data and provide new insights by pinpointing TH-responsive DEGs to specific cellular clusters. Moreover, inferred cell-cell communication using NeuronChat suggested a disbalance in GABAergic versus glutamatergic signaling. We further uncovered perturbed primary cilia formation in several GABAergic and glutamatergic clusters of the dKO cortex. <h4>Discussion</h4> Molecular signatures and perturbations uncovered by our snRNAseq study reveal new molecular characteristics of the AHDS. The imbalance in GABAergic versus glutamatergic cell-cell-communication, perturbed primary cilia formation, and bifurcation of the oligodendrocyte lineage align with pathologies observed in AHDS patients and highlight the role of TH signaling in maintaining neuronal network homeostasis in the cortex.

Immune exclusion is associated with adverse prognosis in tumors, and this phenomenon may be facilitated by a physical barrier constituted by the extracellular matrix (ECM). Hypoxia and acidity promote immune exclusion, however the detailed mechanism remains unclear. This study demonstrate that G-protein-coupled-receptor-4 (GPR4), as a proton sensor, induces immune exclusion in colon cancer by promoting collagen fiber alignment and deposition. Specifically, GPR4 facilitates collagen alignment via LOXL2 regulation and enhances collagen I by modulating TGF-β, both mediated by the JAK2/STAT3 pathway. We further validate our findings in a male animal model, observing that elevated GPR4 expression in colon cancer results in an immune-excluded microenvironment. Inhibition of the JAK2/STAT3 pathway and LOXL2 function effectively reverse immune exclusion and enhance immunotherapy efficacy. Collectively, our findings elucidate a mechanism of immune exclusion and propose a potential target for improving the therapeutic efficacy of immunotherapy through the remodeling of the tumor ECM.

Many pediatric cancer survivors experience chemotherapy-induced cognitive impairment (CICI), which negatively impacts their quality of life. However, while some patients develop CICI, others do not, suggesting that genetic variants may contribute to the development of CICI. The Apolipoprotein E (ApoE) E4 allele has been identified as a risk variant for CICI among pediatric cancer survivors. However, the mechanisms by which ApoE4 contributes to the development of CICI remain unknown. Using a commonly used chemotherapeutic agent known to induce CICI, doxorubicin, we treated five-week-old rats homozygous for either the human ApoE3 or ApoE4 allele with doxorubicin (2 mg/kg/week for 4 weeks) or saline. Behavioral assessments revealed that ApoE4 rats were more susceptible to doxorubicin-induced impairments in visual and spatial memory compared to ApoE3 rats. Pathophysiological analyses showed a significant reduction in hippocampal neurogenesis of ApoE4 doxorubicin-treated rats relative to the other groups. Serum levels of GFAP were significantly increased in ApoE4 doxorubicin-treated rats. These findings suggest that the ApoE genotype influences vulnerability to CICI and highlight a potential mechanistic link through impaired neurogenesis, laying the groundwork for genotype-specific therapeutic strategies.

This study developed a simple and label-free fluorescent probe for deferiprone quantification in exhaled breath condensate (EBC) samples. For that, UiO-66, synthesized in a one-step process with the solvothermal method, along with ferric ions, provided a selective system for deferiprone detection. Deferiprone formed a complex with ferric ions, and the resulting complex quenched the fluorescence intensity of UiO-66 through an inner filter effect process. The probe demonstrated good sensitivity, with a detection limit of 0.005 mg.L^- 1 and a linear range of 0.016-1.0 mg.L^- 1, and performed reliably in the analysis of deferiprone in EBC samples of the patients.

<h4>Abstract</h4>Sickle cell disease (SCD) is an inherited red blood cell disorder that affects an estimated 70,000 to 100,000 people in the United States, most of whom are of African descent. SCD causes acute and chronic pain and widespread end-organ damage, resulting in acute and chronic complications, high morbidity, and early death. Advances in research and treatment have improved patient outcomes for people living with SCD. Over the past 10 years, several new medications and curative therapies have been approved by the U.S. Food and Drug Administration for people with SCD, and more medications are in the pipeline. This review includes historical perspectives, current treatment options, and future directions for this population. With the ongoing rapid changes in SCD care, nurses are well positioned to play a role in patient education, research, advocacy, and the implementation of these emerging therapies to advance the care of those affected by SCD.

Macrophages are well known for their widespread distribution, diverse roles, and involvement in multiple pathophysiological contexts, thereby constructing an immunological front line. Meanwhile, constant efforts over the past few decades have unveiled diverse reprogramming patterns of lipid metabolism as crucial, response- and context-specific drivers of macrophage functions and fate. Here, we take a bird's-eye view of major fields across the research landscape of lipid-regulated macrophages; review the latest advances in understanding how alterations in several lipid subclasses, especially their fatty acyl composition and oxidative status, direct macrophage-mediated responses and pathology outcomes; and summarize representative insights that have deciphered the lipidome composition of macrophages or profiled specific lipid species under different scenarios. We hope that this review provides readers with a handy grip to learn and explore the field of lipid-regulated immunobiology, exemplified by but not limited to macrophages.

The progesterone receptor (PR) expression is associated with disease-free survival in breast cancer. Yet, PR is known to elicit the activation of pro-tumor or anti-tumor signalling pathways. To gain a comprehensive understanding of intrinsic activities of PR, we conducted a global profiling of PR-regulated proteins using Tandem Mass Tag (TMT) proteomics in MCF-7 cells with elevated PR levels. The analysis identified reliably and reproducibly 4,915 PR-regulated proteins and 678 phosphorylated peptides in response to progestin R5020. Consistent with its growth inhibitory activity, PR broadly reduced levels of proteins for cell division, including CDKs, cyclins, DNA replication factors, and proteins involved in chromatin condensation, spindle assembly and chromosome segregation. PR also induced previously reported upregulation of pro-growth proteins such as EGFR, IRS2, and CCND1, but the upregulations are functionally futile due partly to inhibitory phosphorylation. Importantly, PR regulated 200 mitochondrial proteins including proapoptotic factors BNIP3, NIX, AIF/AIFM1, AIFM2, ENDOG, HtrA2/Omi, and Smac/DIABLO, culminating in mitochondria-mediated apoptosis independent of caspases. In conclusion, this proteomics study achieved to date the most comprehensive understanding of PR-regulated molecular networks that are strongly anti-proliferative and proapoptotic with pivotal involvement of mitochondria. PR agonists warrant evaluation for the treatment of breast cancer with high PR expression.

Diabetic cardiomyopathy (DCM) requires a systematic approach for effective management. Focusing on key signaling pathways in myocardial fibrosis could offer novel therapeutic approaches for DCM. The objective of this research is to investigate new biomarker and the underlying mechanisms by which baicalin intervenes in the progression of DCM. Swiss Target Prediction databases and weighted correlation network analysis (WGCNA) were employed to collect and analyze targets of baicalin and DCM. Intersection targets were obtained through venn online website, network visualization was conducted through STRING database and Cytoscape 3.7.2 software. Molecular docking validation was completed via AutoDock Vina software. Western blotting, RT-qPCR and histology were carried out to validate the direct targets of baicalin and explore the molecular mechanism during DCM progression. In mice induced to DCM, we observed that disruptor of telomeric silencing 1-like (DOT1L) was upregulated. Baicalin (BC) reduced myocardial fibrosis in mice as indicated by the reduced expression levels of fibrosis markers COL-1 and TGF-β. The inhibitory effect of BC was reversed by overexpressed DOT1L and further enhanced by silenced DOT1L. Our results indicate that cardiomyocyte DOT1L exerts a pathogenic role in DCM and BC specifically targets DOT1L/COL-1 signaling to reduce myocardial fibrosis in DCM.

BACKGROUND: Mirror therapy (MT) and augmented reality (AR) are gaining popularity in stroke rehabilitation. MT uses mirror visual feedback to promote bilateral brain coupling and increase primary motor cortex excitability. AR offers an interactive context of practice for promoting motor and cognitive recovery. MT and AR may complement each other for hybrid interventions in stroke rehabilitation. This study investigated the benefits of MT-primed AR (MT + AR) versus AR group, relative to conventional therapy (CT) for individuals with stroke. METHOD: The study randomly assigned 45 stroke survivors to the MT + AR group, the AR, or the CT group, and 44 of them completed the experiment and were included in the analysis. Each treatment session was 90 min, 3 times a week, for 6 weeks. All assessments were administered before, immediately after treatment, and at 3 months. Primary outcome measures were the Fugl-Meyer Assessment-Upper Extremity (FMA-UE) and the Berg Balance Scale (BBS). Secondary outcome measures were the revised Nottingham Sensory Assessment (rNSA), Chedoke Arm and Hand Activity Inventory (CAHAI), Motor Activity Log (MAL), and Stroke Impact Scale Version 3.0 (SIS). Adverse events were monitored before and after each session. RESULTS: After 6 weeks of treatment, the three groups demonstrated significant improvements in the FMA-UE, BBS, CAHAI, MAL, and SIS. In the between-group comparisons, MT + AR and AR groups demonstrated significant advantages in the BBS, proprioception scale of rNSA and SIS, compared with the CT group. Only the MT + AR group, not the AR group, showed significantly better improvements in the FMA-UE and tactile scale of rNSA than the CT group. The MT + AR and AR alone showed differential benefits in the FMA-UE, tactile scale of rNSA, and SIS; the MT + AR rendered significantly better benefits. There were no significant differences among the three groups in the stereognosis scale of rNSA and MAL. No adverse effects were observed. CONCLUSION: MT + AR and AR both effectively enhanced sensorimotor functions, balance and postural control, task performance, and life quality in patients with stroke with moderate-to-severe motor impairments. The results showed that MT + AR and AR were more beneficial for improving stroke survivors’ balance, functional mobility, proprioception recovery, and quality of life than the CT group. Furthermore, the MT + AR revealed better outcomes in the upper limb motor function and tactile sensory recovery. Between the MT + AR and AR comparisons, the MT + AR was more beneficial for improving upper limb motor function, tactile sensory recovery, and quality of life. Trial registration NCT05993091.

<h4>Background</h4>Preeclampsia is a hypertensive disorder of pregnancy with major maternal and fetal consequences. While the molecular basis of early-onset preeclampsia is well studied, the mechanisms underlying late-onset disease-and how they differ by fetal sex-remain poorly understood. Placental transcriptomic profiling at term can reveal persistent molecular alterations reflecting cumulative disease processes.<h4>Methods</h4>We conducted a cross-sectional observational analysis of placental gene expression using RNA sequencing in a subset of 58 term placentas (21 male-bearing and 37 female-bearing pregnancies) drawn from two large prospective birth cohorts. Pregnancies were classified based on a clinical diagnosis of late-onset preeclampsia (diagnosed ≥ 20 weeks' gestation according to ISSHP criteria) or as uncomplicated pregnancies. We then assessed for differential gene expression. Cell type proportions were estimated using CIBERSORTx from a placenta-specific reference single-cell dataset. Weighted gene co-expression network analysis identified modules of co-expressed genes associated with late-onset preeclampsia and fetal sex.<h4>Results</h4>Differential gene expression analysis identified 150 genes with altered expression in male-bearing placentas from pregnancies with late-onset preeclampsia compared to those from uncomplicated pregnancies. No differentially expressed genes were identified in female-bearing placentas. Cell type deconvolution revealed increased abundance of CD14 + monocytes and CD8 + activated T cells (log odds of 1.42 and 1.44 respectively) and reduced fetal GZMK natural killer cells (log odds of 0.60) in male-bearing placentas from affected pregnancies. In female-bearing placentas, late-onset preeclampsia was associated with increased fetal nucleated red blood cells and maternal plasma cells (log odds of 1.33 and 1.40 respectively). Male-specific co-expression analysis identified gene modules enriched for biological processes including RNA processing, immune regulation, and metabolism.<h4>Conclusions</h4>Placental transcription and cellular responses to late-onset preeclampsia differ by fetal sex. Evidence of altered immune cell composition and gene co-expression in male-bearing placentas suggests a sex-specific vulnerability. These findings highlight the importance of considering fetal sex in molecular investigation and clinical management of preeclampsia. Preeclampsia is a common pregnancy complication marked by high blood pressure, but how it affects the placenta, especially in later pregnancy and depending on the baby's sex, is not well understood. In this study, we analysed placental tissue from pregnancies with and without late-onset preeclampsia using RNA sequencing. By separating the data based on whether the neonate was male or female, we found striking differences in gene expression. Only placentas from male-bearing pregnancies showed significant changes in gene expression linked to preeclampsia. These changes involved genes related to immune response, metabolism and vascular function. We also used computational tools to estimate what types of cells were present in each placental sample. In male-bearing pregnancies affected by late-onset preeclampsia, there was a notable increase in certain immune cells, suggesting an altered immune response and increased inflammation. In contrast, female-bearing pregnancies affected by late-onset preeclampsia showed an increase in cell composition for two blood related cell types, but no significant gene expression differences. By grouping genes that worked together into networks, we identified several groups, especially in placentas from male-bearing pregnancies, that were strongly associated with biological processes known to be disrupted in preeclampsia, such as blood vessel formation, extracellular matrix remodelling, and hormone regulation. These findings emphasise the importance of considering fetal sex in pregnancy research and could help guide future sex-specific diagnostic or treatment strategies.

<h4>Background</h4>Neddylation modifications in immune and tumor cells are linked to poor tumor prognosis. This study identifies prognostic genes associated with neddylation-related genes (NRGs) in colorectal cancer (CRC) using single-cell and spatial transcriptome (ST) sequencing, aiming to advance CRC treatment strategies.<h4>Methods</h4>Datasets included TCGA-CRC (training/internal validation, 7:3 split), GSE28722 (external validation), GSE132257 (scRNA-seq), and GSE226997 (ST). Single-cell analysis annotated seven cell types, with epithelial cells identified as key. Differentially expressed genes (DEGs) from key cells [DEGs(sc)] and bulk analysis of TCGA-CRC [DEGs(bulk)] were intersected with 247 NRGs to yield candidate genes. Regression analyses screened prognostic genes for risk model construction, validated internally and externally. Pseudotime trajectory and ST mapping visualized gene expression, while molecular networks and drug predictions were generated.<h4>Results</h4>In scRNA-seq dataset, seven cell types were annotated, and epithelial cells were the key cells. A sum of 32 candidate genes were obtained by intersecting 5,131 DEGs(sc)(key cells), 9,089 DEGs(bulk), and 247 NRGs to produce PSMD12, PSMB2, and FBXL5 as prognostic genes. Both prognostic risk model and nomogram model were predictive of CRC. At the ST samples, PSMD12 was expressed at a low level in all sections, whereas PSMB2 and FBXL5 were expressed at a slightly higher level in the sections. In addition, a lncRNA-miRNA-mRNA network and a drug-prognostic gene network were created, getting some potential drugs like bortezomib.<h4>Conclusion</h4>A novel three-gene prognostic model for CRC was developed and validated, offering therapeutic insights through molecular networks and drug predictions.

The detection and identification of intracranial tumours is limited by the lack of accurate biomarkers and requires invasive biopsy procedures. We investigated a machine perception liquid biopsy approach to detect and identify intracranial tumours from peripheral blood and to discover biomarkers responsible for the predictions. Quantum well defect-modified single-walled carbon nanotubes stabilized with single-stranded DNA, interrogating 739 plasma samples from brain tumour patients, were used to train and validate machine-learning models to detect intracranial tumours with 98% accuracy and identify tumour type. The protein corona of the top model-contributing nanosensor was interrogated using quantitative proteomics, resulting in the identification of tumour ecosystem-secreted factors, both previously reported and newly discovered, originating from intracranial tumour cells, the tumour microenvironment and the innate immune system of patients with glioblastoma and meningioma. Newly discovered factors elicited linear nanosensor responses and were elevated in one or both tumour types, matching the original protein corona enrichment. This investigation reveals that a perception-based detection of disease in blood can identify biomarkers responsible for the signal and also amplify cancer detection signals by detecting factors beyond tumour cells, thereby recruiting the entire tumour ecosystem for cancer diagnosis.

<h4>Background</h4>Gastrointestinal acute radiation syndrome (GI-ARS) is characterized by disruption of the intestinal barrier function, leading to bacterial translocation and sepsis. Intestinal stem cells are highly radiosensitive and dramatically reduced after radiation injury. Clusterin (Clu)-positive revival stem cells contribute to the restoration of intestinal stem cells. Ghrelin, a gastric peptide hormone, has been shown to improve intestinal integrity in models of inflammatory enteropathy. In this study, we investigated the effects of ghrelin on intestinal stem cell recovery and its potential to mitigate radiation-induced intestinal injury.<h4>Methods</h4>Mice were subjected to 12 Gy partial body irradiation (PBI). Ghrelin at the doses of 2 to 6 nmol per mouse was administered daily for 4 consecutive days, starting at 24 h post-PBI, and survival was monitored for 30 days. To assess intestinal histology, cell proliferation, and intestinal stem cell markers, mice were treated with 6 nmol of ghrelin on days 1, 2, and 3 post-PBI, and on day 4 jejunal samples were collected for qPCR, immunofluorescence, and microcolony assays. Intestinal permeability was assessed in vivo by the leakage of gavage-fed 4-kDa FITC-dextran into the circulation.<h4>Results</h4>Ghrelin administration significantly improved 30-day survival rate following 12-Gy PBI in a dose-dependent manner. Treatment with ghrelin restored villus length and enhanced intestinal barrier integrity. Ghrelin also significantly increased the expression of proliferation markers in the jejunum. Microcolony assays revealed that ghrelin reversed the decrease in BrdU-positive cells following PBI. The mRNA and protein expression of intestinal stem cell markers was decreased after PBI but was restored by ghrelin treatment. Finally, ghrelin significantly increased the population of Clu⁺ population following irradiation.<h4>Conclusions</h4>These findings indicate that ghrelin mitigates radiation-induced intestinal injury by promoting the expansion of Clu⁺ revival stem cells and the recovery of intestinal stem cells. This study highlights the therapeutic potential and identifies the mechanism of action of ghrelin as a medical countermeasure against GI-ARS.

Hundreds of computational methods for predicting ligand binding pockets exist, but the problem of finding druggable pockets throughout the human proteome persists. Different strategies for pocket-finding excel in different use cases. Ensemble models that leverage multiple different pocket-finding strategies can best capture diverse pockets at scale. Despite this, no publicly available human-proteome-wide datasets of pocket predictions from multiple pocket-finding methods exist. We present the Human Omnibus of Targetable Pockets (HOTPocket), a dataset of over 2.4 million predicted pockets over the entire human proteome that utilizes both experimentally-determined and computationally-predicted protein structures. We assembled this dataset by running seven diverse, established pocket-finding methods over all PDB and AlphaFold2 structures of the canonical human proteome. We created a novel pocket scoring method, hotpocketNN, which we used to filter candidate pockets and assemble the final proteome-wide dataset. Our hotpocketNN method is able to recover known ligand binding pockets, including those which are dissimilar from any pocket seen in its training set. The hotpocketNN method outperforms all constituent methods, including P2Rank and Fpocket, when assessing the precision with DCA criterion on the Astex Diverse Set and PoseBusters dataset. Additionally, hotpocketNN was able to identify recently-discovered druggable pockets on KRAS and the mu opioid receptor. We make both the HOTPocket dataset and the hotpocketNN method freely available.

Despite rapid advances in mapping genetic drivers and gene expression changes in hematopoietic stem cells (HSCs), few studies exist at the protein level. We perform a deep, multi-omics characterization (epigenome, transcriptome, and proteome) of HSCs in a mouse model carrying a loss-of-function mutation in Tet2, a driver of increased self-renewal in blood cancers. Using state-of-the-art, multiplexed, low-input mass spectrometry (MS)-based proteomics, we profile TET2-deficient (Tet2^-/-) HSCs, revealing previously unrecognized molecular processes that define the pre-leukemic HSC molecular landscape. Specifically, we obtain more accurate stratification of wild-type and Tet2^-/- HSCs than transcriptomic approaches and identify extracellular matrix (ECM) molecules as being dysregulated upon TET2 loss. HSC expansion assays using ECM-functionalized hydrogels confirm a selective effect on the expansion of Tet2-mutant HSCs. Taken together, our study represents a comprehensive molecular characterization of Tet2-mutant HSCs and identifies a previously unanticipated role of ECM molecules in regulating self-renewal of disease-driving HSCs.

<h4>Introduction</h4>Long-term brain health profiles following exposure to repetitive head impacts and/or concussions in contact sports are a public health focus and the subject of a national debate. The true prevalence rates of mild cognitive impairment (MCI) or neurobehavioural dysregulation are unknown in the nearly 20 000 current/living former professional football players. Here, we describe the procedures and methodology of the prevalence study of cognitive function in former professional football players from the Brain Health Initiative at the University of Pittsburgh. The objective is to define the prevalence of normal cognitive function versus neurodegeneration in former professional football players through clinical, neuroimaging and biomarker assessments.<h4>Methods and analysis</h4>Participants include former professional football players aged 29-59 years at study onset who played a minimum of three professional football games in three professional seasons and non-exposed controls. Participants are recruited by two mechanisms, a random and non-random sample. The full study protocol includes a 3-4-day, multidomain assessment (eg, neurological, neurocognitive, psychiatric, sleep, vestibular, orthopaedic and cardiovascular) for neurodegenerative disease and overall health and function, including MRI, positron emission tomography scans, analysis of blood plasma and cerebrospinal fluid, neurocognitive assessments, applanation tonometry, overnight sleep study and informant interview. A multidisciplinary clinical panel conducts a blinded diagnostic consensus conference to adjudicate the presence of MCI and/or traumatic encephalopathy syndrome, which serve as the study's primary and secondary outcomes, respectively. Point prevalence of these for both the exposed and unexposed cohorts will be calculated as the primary statistical analysis.<h4>Ethics and dissemination</h4>The University of Pittsburgh Institutional Review Board approved the study prior to recruiting human subjects (protocol numbers STUDY19010008: sIRB - Brain Health Initiative (Part 1) and STUDY19030211: sIRB - Brain Health Initiative (Part 2)). The results will be disseminated in peer-reviewed journals and as presentations at national and international scientific conferences.

Amino acid conjugates are progressively becoming popular as a potent tactic to enhance the pharmacological efficacy of drugs, especially in the areas of cancer and antimicrobial therapy. By taking advantage of the intrinsic biological attributes of amino acids, their conjugates facilitate drug stability, selective accumulation, and enhanced therapeutic efficacies. In particular, the structural analogy of amino acids to physiological substrates enables these conjugates to use solute carrier transporters, commonly overexpressed in tumour cells, which allow for targeted and effective drug delivery. This review considers how amino acid properties like chirality, hydrophobicity and steric bulk can be modulated to maximize drug conjugates. We emphasize important design aspects, such as selection of linkers and coupling reagents, and how these have an impact on drug release and biodistribution. Specific focus is given to d-amino acid, which increases proteolytic stability and bioactivity for both anticancer and antimicrobial uses, and to l-amino acid, which is responsible for receptor recognition, metabolic compatibility and amino acid decorated nanoparticle formulation. The existing drawbacks of antibody-drug conjugates (ADCs) and peptide-drug conjugates (PDCs) are immunogenicity, enzymatic degradation and poor tissue penetration. Amino acid conjugates provide a strong rationale with higher chemical versatility and potential for better pharmacokinetics and less toxicity. By harnessing the insights from chemistry, transporter biology and therapeutic design, this review presents a strategy for the creation of next-generation amino acid conjugates that bridge molecular accuracy to clinical utility.

Methamphetamine addiction remains a major public health concern in the United States that has paralleled the opioid epidemic. Psychostimulant use disorders have a heritable genetic component that remains unexplained. Methamphetamine targets membrane and vesicular transporters to increase synaptic dopamine, norepinephrine, and serotonin. We previously identified Hnrnph1 (heterogeneous nuclear ribonucleoprotein H1) as a quantitative trait gene underlying methamphetamine behavioral sensitivity. Hnrnph1 encodes the RNA-binding protein hnRNP H1 that is ubiquitously expressed in neurons throughout the adult brain. Gene-edited mice with a heterozygous frameshift deletion in Hnrnph1's first coding exon showed reduced methamphetamine-induced dopamine release and behaviors. To potentially inform the mechanism linking hnRNP H with methamphetamine behavior, we surveyed the mRNA targetome of hnRNP H via cross-linking immunoprecipitation coupled with RNA-sequencing in striatum at baseline and at 30 min post-methamphetamine in wild-type female and male C57BL/6 J mice. Methamphetamine induced changes in RNA-binding targets of hnRNP H in mice, including 3'UTRs of enriched mRNAs involved in synaptic plasticity. Targetome, transcriptome, and spliceome analyses triangulated on Cacna2d2 which showed methamphetamine-induced changes in hnRNP H binding, gene expression and splicing. Pre-treatment with pregabalin, an inhibitor of CACNA2D2 and CACNA2D1 voltage-gated calcium channel subunits, attenuated methamphetamine-induced locomotor activity, suggesting CACNA2D2 could contribute to methamphetamine locomotor stimulant sensitivity. Our study identifies a dynamic hnRNP H RNA targetome that can respond rapidly to methamphetamine and could potentially contribute to synaptic plasticity and behavior. Given our discovery-based findings, future studies will require directly validating the link between methamphetamine-induced changes in hnRNP H binding, gene regulation, synaptic plasticity, and behavior.

Skeletal remains from mummies of the Egyptian Collection of the National Museum of Brazil, damaged by fire in 2018, were analysed by vibrational spectroscopy (infra-red, Raman and inelastic neutron scattering). Four different mummies were probed, including one from the Roman period which was one among eight worldwide. The present data delivered the burning conditions to which the mummies were subjected and allowed us to determine the extent of the heat-induced effects and the way they were impacted by the mummification process. Some of the mummies were found to have been subjected to different temperatures, depending on the location of the bone fragment in the skeleton. Several contaminants were identified; both compounds used during mummification (including salts and pigments) and construction materials from the building collapse. The results enabled us to characterise the mummified skeletal remains recovered after the fire and are expected to help establish the most suitable preservation methods.

<b>Background/Objectives:</b> Atrial fibrillation (AF), characteristic of chaotic atrial electrical activity along with ineffective atrial systole, remains the most frequent sustained cardiac dysrhythmia, with an overall lifetime risk for AF being approximately 15% to 40% in the global population. AF is associated with substantially enhanced risks for multiple adverse clinical outcomes, including thromboembolic cerebral stroke, dementia, chronic kidney disease, myocardial infarction, cardiac failure, and even premature cardiac demise. Although remarkable advances have been achieved toward unravelling the complex hereditary etiopathogenesis underpinning AF, it has become increasingly clear that inherited determinants predisposing to AF in a vast majority of individuals are still uncertain. <b>Methods:</b> A Chinese pedigree with idiopathic AF and another group of 236 cases suffering idiopathic AF along with 312 unrelated healthy volunteers were prospectively recruited. Exome-wide sequencing and Sanger sequencing assays were implemented in research participants. The functional effects of the discovered variations in the <i>SOX5</i> gene were explored through dual-luciferase reporter analysis. <b>Results:</b> Two novel <i>SOX5</i> mutants, NM_006940.6: c.355C>T; p.(Gln119*) and NM_006940.6: c.640G>T; p.(Glu214*), were identified in the AF pedigree and one of the 236 unrelated patients affected with AF, respectively. These two heterozygous truncating SOX5 variations were absent from the 624 control chromosomes. Quantitative luciferase reporter assays unraveled that both Gln119*- and Glu214*-mutant SOX5 lost the ability to transactivate <i>GJA1</i>. Additionally, the two variations abolished the synergistic transactivation of <i>SCN5A</i> by SOX5 and SHOX2. <b>Conclusions:</b> The current findings indicate <i>SOX5</i> as a novel gene contributing to AF, which adds more insight to the molecular pathogenesis of AF, and provides a potential target for personalized precision medicine.

Rare genetic diseases (RGDs) affect individuals, families, and healthcare systems worldwide. Population-scale genomic data remain largely restricted to Western cohorts with an estimated 10,000 RGDs. South Asian populations remain underrepresented in molecular, clinical, and genomic databases. This study presents the first preliminary molecular genetic characterization of RGDs in the Punjabi population of Pakistan. Data were collected from the provincial RGD registry at the Punjab Thalassemia and Other Genetic Disorders Prevention and Research Institute (PTGDPRI), Lahore. Families diagnosed using next-generation sequencing (NGS) between 2021 and 2023 were enrolled. Structured questionnaires captured clinical, demographic, and socioeconomic information, and statistical and genetic analyses were performed to assess allele frequencies, and disease distribution. The registry included 167 families with 72 distinct RGDs, with a mean burden of 0.81 ± 0.24 affected children per family. Niemann-Pick disease (NP), progressive familial intrahepatic cholestasis (PFIC), and mucopolysaccharidosis (MPS) were the most common diseases. Consanguinity was observed in 89% of families, 77% of which involved first-cousin marriages, and was significantly associated with RGD incidence. Most families belonged to low-income groups despite high literacy rates, underscoring inequity in healthcare. The primary and secondary variants included 131 variants, including copy number variants (CNVs) and single nucleotide variants (SNVs), annotated as pathogenic, likely pathogenic, or variants of unknown significance (VUS) across 109 genes, including 24 South Asian-enriched variants. This study provides the first genomic and epidemiological overview of RGDs in the Punjabi population. The findings reveal how genetic, socioeconomic, and cultural factors converge to amplify the RGD burden and highlight the need for affordable molecular diagnostics, inclusive genomic databases, and regional genomic surveillance initiatives in South Asia.

Non-invasive glucose monitoring remains a significant challenge in diabetes management, with existing approaches often limited by poor accuracy, high cost, or patient discomfort. Microwave-based biosensors offer a promising label-free alternative by exploiting the dielectric contrast between glucose and water. This paper presents a compact, dual-band concentric square-shaped split-ring resonator (SRR-type) biosensor fabricated on a low-cost FR-4 substrate for aqueous glucose detection. The sensor leverages electric field confinement in inter-ring gaps to transduce glucose-induced permittivity changes into measurable shifts in resonance frequency and reflection coefficient. Experimental results demonstrate a linear, monotonic response across the clinical range up to 250 mg/dL, with a frequency-domain sensitivity of 1.964 MHz/(mg/dL) and amplitude-domain sensitivity of 0.0332 dB/(mg/dL), achieving high coefficients of determination (R² = 0.9956 and 0.9927, respectively). The design achieves a normalized size of 0.137 λ_g², combining high sensitivity and compact size within a scalable platform. Operating in the UWB-adjacent band (2.76-3.25 GHz), the proposed biosensor provides a practical, reproducible, and PCB-compatible solution for next-generation label-free glucose monitoring.

In avian species, the chorioallantoic membrane (CAM) is a vital, highly vascularized extraembryonic structure that supports embryonic respiration, calcium transport, and innate immune defense. In this study, we applied LC/MS/MS-based proteomics to CAM tissue harvested at embryonic days (ED) 6, 8, 10, and 12 to characterize its protein profile during the expression of different CAM functionalities during embryonic development and gain insight into possible sex-based distinctions. A total of 2688 proteins were identified, with 2347, 2265, 2351, and 1267 proteins detected at ED 6, 8, 10, and 12, respectively. Notably, 1191 common proteins were identified across all stages, while 124, 47, 86, and 2 proteins were uniquely expressed at ED 6, 8, 10, and 12, respectively. Functional annotation revealed correlations with abundant CAM protein constituents (as per their emPAI); for example: calcium mobilization - v-type proton ATPase subunit E1 (ATP6V1E1) and G1 (ATP6V1G1); intracellular transport-calcium-binding protein 39 (CAB39); vascular system and gaseous exchange - annexin A2 (ANXA2); lymphatics-actin, gamma 1 (ACTG1); blood elements-hemoglobin subunit alpha-1 (HBA1); immune defense-cathelicidin-1 (CATH1), cathelicidin-2 (CATH2); and protection against luminal toxic contents-thioredoxin (TXN). Notably, a sex-specific analysis identified 614, 320, 314, and 212 proteins that were uniquely expressed in female embryos, and 212, 273, 144, and 56 proteins only in male embryos at ED 6, 8, 10, and 12, respectively. The identification of sex-linked proteins during early CAM development may provide insight into their functional roles and highlight the CAM's potential as a target for the development of <i>in-ovo</i> sex identification technology.

<b>Background/Objectives:</b> Hydroxyapatite (HAp)-based implants and HAp-titanium (HApTi) composites are widely used in orthopedic and dental applications, but their long-term success is limited by peri-implant bone loss. Local delivery of osteoactive molecules from implant surfaces may enhance osseointegration and reduce periprosthetic osteolysis. This study combined in silico modeling and experimental assays to compare calcium fructoborate (CaFb), sodium alendronate, and calcium alendronate as functionalization agents for HAp and HApTi implants. <b>Methods:</b> Molecular docking (AutoDock 4.2.6) and 100 ns molecular dynamics (MD) simulations (AMBER14 force field, SPC water model) were performed to characterize ligand-substrate interactions and to calculate binding free energies (ΔG_binding) and root mean square deviation (RMSD) values for ligand-HAp/HApTi complexes. HAp and HApTi discs obtained by powder metallurgy were subsequently functionalized by surface adsorption with CaFb or alendronate salts. The amount of adsorbed ligand was determined gravimetrically, and in vitro release profiles were quantified by HPTLC-MS for CaFb and by HPLC after FMOC derivatization for alendronates. <b>Results:</b> CaFb-HAp and CaFb-HApTi complexes showed the lowest binding free energies (-1.31 and -1.63 kcal/mol, respectively), indicating spontaneous and stable interactions. For HAp-based complexes, the mean ligand RMSD values over 100 ns were 0.27 ± 0.17 nm for sodium alendronate, 0.72 ± 0.28 nm for calcium alendronate (range 0.35-1.10 nm), and 0.21 ± 0.19 nm for CaFb (range 0.15-0.40 nm). For HApTi-based complexes, the corresponding RMSD values were 0.30 ± 0.15 nm for sodium alendronate, 0.72 ± 0.38 nm for calcium alendronate and 0.26 ± 0.14 nm for CaFb. These distributions indicate that CaFb and sodium alendronate maintain relatively stable binding poses, whereas calcium alendronate shows larger conformational fluctuations, consistent with its less favorable binding energies. Experimentally, CaFb exhibited the greatest chemisorbed amount and percentage on both HAp and HApTi, followed by sodium and calcium alendronate. HApTi supported higher loadings than HAp for all ligands. Release studies demonstrated a pronounced burst and rapid plateau for both alendronate salts, whereas CaFb displayed a slower initial release followed by a prolonged, quasi-linear liberation over 14 days. <b>Conclusions:</b> The convergence between in silico and adsorption-release data highlights CaFb as the most promising candidate among the tested ligands for long-term functionalization of HAp and HApTi surfaces. Its stronger and more stable binding, higher loading capacity and more sustained release profile suggest that CaFb-coated HApTi implants may provide a favorable basis for future in vitro and in vivo studies aimed at improving osseointegration and mitigating periprosthetic osteolysis, although direct evidence for osteolysis prevention was not obtained in the present work.

A 10-week growth experiment was conducted to evaluate the physiological effects of dietary phosphorus supplementation on red swamp crayfish (<i>Procambarus clarkii</i>) feeding diets with high <i>Clostridium autoethanogenum</i> protein (CAP) levels. Six isonitrogenous and isolipid diets were formulated: The FM diet contained 10% fishmeal, which is equivalent to a dietary phosphorus level of 1.41%, and the CAP, CAPSP1, CAPSP2, and CAPSP3 diets substituted all fishmeal with CAP and supplemented with 0, 2.5%, 3%, and 3.5% Ca(H₂PO₄)₂, respectively (corresponding to dietary phosphorus levels of 0.66%, 1.27%, 1.40%, and 1.52%). A total of 600 crayfish with an initial mean weight of (5.01 ± 0.02) g were selected and randomly assigned to 15 cages for feeding and sampled at the end of the experiment. Results indicate that high-dose CAP replacing fishmeal caused abnormal hepatopancreatic tissue structure in crayfish, exacerbating lipid deposition and oxidative stress. Compared with the CAP group, the specific growth rate (SGR) of crayfish in the CAPSP2 and CAPSP3 groups significantly increased (<i>p</i> < 0.05). The activities of antioxidant enzymes and lipid-degrading enzymes in the hepatopancreas, along with the relative expression of related genes, were significantly enhanced (<i>p</i> < 0.05). Metabolomic analysis demonstrated significant differences in major differential metabolites and metabolic pathways between the CAP group crayfish and the CAPSP2 group (<i>p</i> < 0.05). CAPSP2 group crayfish exhibited a higher content of phosphatidylcholine (PC) and lysophosphatidylcholine (LPC), with significant enrichment in glycerophospholipid metabolism and fatty acid metabolism pathways (<i>p</i> < 0.05). Overall, supplementing dietary phosphorus levels to 1.40-1.52% effectively mitigated growth retardation, oxidative damage, and lipid metabolism disorders induced by high-proportion CAP replacement of fishmeal.

<b>Objectives</b>: The repurposing of existing drugs as anticancer agents has attracted attention in cancer drug discovery. This study aimed to examine the anticancer efficacy of rosiglitazone (RSG) against cholangiocarcinoma (CCA) and its underlying mechanisms. <b>Methods</b>: The effect of RSG on the viability of KKU-100 CCA cells was examined. The possible molecular targets were identified using proteomic analysis and verified by a series of cell-based assays. Furthermore, the expression of PPARγ protein in CCA tissues was also assessed. <b>Results</b>: RSG exhibited a cytotoxic effect against KKU-100 cells. Proteomic analysis demonstrated a significant different expression protein pattern of the 100 μM RSG-treated group compared to the control group. Significant alteration of several proteins was found, including the up-regulation of calcium-binding, cytoskeletal, and metabolic proteins, concomitant with the down-regulation of antioxidant enzymes. Detailed analyses revealed that RSG induced apoptosis in CCA cells, accompanied by increased caspase 3/7 activities, reactive oxygen species (ROS) generation, and disruption of mitochondrial function. RSG altered the expressions of annexin A1 and antioxidant enzymes, according to Western blot analysis. GW9662, a PPARγ antagonist, did not affect the viability and apoptosis of KKU-100 cells caused by RSG. Immunohistochemistry analysis revealed that PPARγ expression in CCA patients was associated with sex, but not with other common clinicopathological parameters. Its expression did not correlate with patients' overall survival time. <b>Conclusions</b>: RSG induced apoptotic cell death in CCA cells, which was accompanied by increased ROS levels and impaired antioxidant defense. Its apoptosis-inducing effect is independent of PPARγ activation. These findings underscore the therapeutic potential of RSG for CCA treatment.

Intestinal stem cells (ISCs) play critical roles in the self-renewal and regeneration of the intestinal epithelium under physiological conditions and after injury, respectively. However, the underlying mechanisms are not fully understood. In this study, we investigate the role of the G protein-coupled receptor formyl peptide receptor 2 (FPR2) in intestinal epithelium homeostasis and regeneration. In mice, knocking out <i>Fpr2</i> in either intestinal epithelial cells (IECs) or ISCs significantly reduces villus height and crypt depth by impairing ISC and transit-amplifying (TA) cell proliferation and differentiation, primarily TA cell differentiation. Mechanistic studies using intestinal organoid culture and bulk and single-cell RNA sequencing revealed that activation of FPR2 promotes proliferation and differentiation of ISCs and TA cells by activating the wingless/integrated (Wnt), Notch, and Hippo signaling pathways via protein kinase C (PKC)-extracellular signal-regulated kinase (ERK). Under physiological conditions, the Wnt and Notch signaling pathways mediate the regulation of ISC proliferation and differentiation by FPR2. <i>Fpr2</i> deficiency in mouse IECs exacerbates X-ray- and 5-fluorouracil-induced villus and crypt injury, and delays intestinal epithelium regeneration by reducing ISC and TA cell proliferation. Administering an FPR2 agonist to mice significantly increases survival rates and accelerates intestinal epithelium regeneration after irradiation. Taken together, these results demonstrate that intestinal epithelial FPR2 plays a key role in intestinal epithelium homeostasis and regeneration by promoting ISC and TA cell proliferation and differentiation. FPR2 is a potential therapeutic target against chemotherapy- and radiotherapy-induced intestinal injury.

Climate-driven changes are reshaping the ecology of <i>Vibrio vulnificus</i> in European waters. Here, we present a retrospective genomic and phenotypic analysis of pre-2018 isolates belonging to lineage 4 (L4), a phylogenetic group historically confined to the Mediterranean Sea and now detected in northern Europe. Using a lineage-specific multiplex PCR combined with whole-genome sequencing, we identified 49 clinical and environmental L4 isolates from German coastal waters. Comparative genomics revealed extensive genetic plasticity in L4, indicative of frequent recombination and horizontal gene transfer, including three MARTX toxin architectures, fourteen distinct capsular genotypes, two type VI secretion systems, and multiple prophages. Notably, nearly half of the L4 isolates encoded a previously undescribed MARTX variant (type H), apparently derived from recombination within a type C toxin and containing a novel calmodulin-dependent NADase (CdN) domain with potential functional implications for virulence. One strain also harboured the plasmid-borne genes <i>ftbp</i> and <i>fpcrp</i>, which confer resistance to fish innate immunity and the ability to cause sepsis, thereby extending the distribution of the <i>piscis</i> pathovar to all five <i>V. vulnificus</i> lineages. Functional assays showed that most L4 strains withstood the bactericidal activity of iron-overloaded human serum, consistent with a capacity to cause sepsis in susceptible individuals. Collectively, these findings redefine <i>V. vulnificus</i> as a multi-host climate-responsive marine pathogen and establish L4 as a newly adapted European lineage whose northward expansion exemplifies how genomic diversification and ocean warming jointly drive the evolution of high-risk marine pathogens within a One Health framework.

Macrophages are pivotal regulators of immunity, and due to their ability to differentiate into either pro-inflammatory (M1) or anti-inflammatory (M2) phenotypes with remarkable plasticity, they can be used as strategies for treating diseases ranging from cancer to chronic inflammatory conditions. However, despite extensive research, the mechanisms driving macrophage polarization and the role of extracellular vesicles (EVs) in regulating these processes remain not fully understood. In particular, the specific signaling pathways and clinical applications of EV-mediated macrophage reprogramming are still under active investigation. Many recent studies have shown that EVs alter the immune environment by reprogramming macrophages to transition between M1 and M2 polarized states, thereby exhibiting therapeutic activity in a variety of diseases, including cancer. In addition, EV engineering aimed at improving macrophage reprogramming capabilities has been shown to enhance therapeutic efficacy, providing unprecedented opportunities to overcome previously untreatable diseases. This review addresses ongoing challenges by examining the latest findings on EV-mediated macrophage reprogramming and highlighting gaps in our understanding of the signaling mechanisms and clinical applications. We explore the signaling pathways and therapeutic potential of EVs in macrophage phenotyping, analyze evidence from disease models, and discuss how EV engineering strategies, including cargo loading and surface modification, expand their clinical use. Additionally, we consider critical factors for clinical translation, such as standardized production, immunogenicity, and safety. Overall, this review emphasizes the potential of EV-mediated macrophage reprogramming as a promising therapeutic strategy for immune-related diseases, while also addressing challenges and future directions for clinical application.

Cell-to-cell communication via Delta-Notch signaling is widely used in various tissues and organs to regulate development and patterning; however, the mechanisms regulating Notch signaling for precise cell fate decisions remain poorly understood. Similar to mammals, the intestinal stem cells in the adult <i>Drosophila</i> midgut generate both absorptive and secretory cell progeny, guided by differential levels of Notch activation. Here we performed a forward genetic screen in <i>Drosophila</i> and identified glucosylceramide synthase (GlcT), a rate-limiting enzyme for glycosphingolipid (GSL) production, whose mutation causes the development of secretory cell tumors. Genetic analysis of the GSL synthesis pathway, combined with metabolite rescue experiments, revealed that the tumor formation is linked to a deficiency in Mactosylceramide/Lactosylceramide. This deficiency impaired the endocytic recycling of the Delta, subsequently reducing Notch signaling activation. Conditional knockout of <i>Ugcg</i>, the mammalian ortholog of <i>GlcT</i>, in mouse small intestine caused an excessive differentiation of goblet cells, phenotypes similar to these caused by Notch inhibition. Our study suggests an evolutionarily conserved role for a specific GSL metabolite in modulating Notch signaling during stem cell fate decisions and provides a molecular connection between ceramide metabolism and Notch signaling in regulating tissue homeostasis and tumor formation.

The metabolic reprogramming of immunosuppressive tumor microenvironment (ITME) greatly influences the anti-tumor immunity. Bioinformatic analysis demonstrates that indoleamine 2,3-dioxygenase 1 and 3-hydroxy-3-methylglutaryl coenzyme A reductase (key enzymes of kynurenine (Kyn) and cholesterol metabolism, respectively), are overexpressed in human colon adenocarcinoma tissues. Herein, biomimetic and pH/ROS dual-responsive nanoparticles (PTSK@CRM) loaded with simvastatin and kynureninase (KYNase) are prepared to regulate Kyn and cholesterol metabolism, thereby enhancing the immunotherapeutic efficacy of PD-1 antibody (αPD-1). The monodisperse spherical PTSK@CRM is stable at pH 7.4, while it can release simvastatin and KYNase under low pH and high H₂O₂ concentration. PTSK@CRM achieves excellently homologous tumor targeting to CT26 cells and induces cell apoptosis more effectively than PTSK. Moreover, PTSK@CRM significantly reduces the contents of Kyn and cholesterol and decreases the activation of the Kyn-AhR pathway in tumor metabolism. In vivo experiments show that PTSK@CRM possesses a favorable tumor-targeting ability to effectively suppress tumor growth and increase the infiltration of immune cells, including CD8⁺ T cells, CD4⁺ T cells, M1-like macrophages, and mature dendritic cells. Further, PTSK@CRM reduces the infiltration of immunosuppressive cells, thereby reversing ITME to improve the therapeutic efficacy of αPD-1. Overall, this immune-metabolic therapeutic strategy provides a potential route for remodeling ITME to enhance tumor immunotherapy.

In mice, postnatal endothelial cell (EC)-specific knockout of the genes coding for transforming growth factor-beta receptor (TGFBR)1 and/or TGFBR2 eliminates TGF-beta signaling in vascular ECs and leads to distinctive central nervous system (CNS) vascular phenotypes. Knockout mice exhibit (1) reduced intraretinal vascularization, (2) choroidal neovascularization with occasional anastomoses connecting choroidal and intraretinal vasculatures, (3) infiltration of diverse immune cells into the retina, including macrophages, T-cells, B-cells, NK cells, and dendritic cells, (4) a close physical association between immune cells and retinal vasculature, (5) a pro-inflammatory transcriptional state in CNS ECs, with increased ICAM1 immunoreactivity, and (6) increased smooth muscle actin immunostaining in CNS pericytes. Comparisons of the retinal phenotype with two other genetic models of retinal hypovascularization - loss of Norrin/Fzd4 signaling and loss of vascular endothelial growth factor (VEGF) signaling - show that the immune cell infiltrate is greatest with loss of TGF-beta signaling, more modest with loss of Norrin/Fzd4 signaling, and undetectable with loss of VEGF signaling. The phenotypes caused by loss of TGF-beta signaling in ECs recapitulate some of the cardinal features of retinal and neurologic diseases associated with vascular inflammation. These observations suggest that therapies that promote TGF-beta-dependent anti-inflammatory responses in ECs could represent a promising strategy for disease modulation.

BackgroundRecent evidence suggests that accumulation of mutant exon 1 protein (HTT1a) may be critical to HD pathogenesis, but the relation of this to differential regional and cellular vulnerability in HD is unknown.ObjectiveWe assessed the contribution of the accumulation of the mutant huntingtin HTT1a to the regional and cellular variation in HD brain pathology by determining if more vulnerable regions and neuron types were relatively enriched.MethodsWe performed immunolabeling using the novel monoclonal antibodies 11G2 and 1B12 against the C-terminal proline 90 (P90) neoepitope of huntingtin HTT1a, which detect accumulation of monomeric, oligomeric and aggregated mutant HTT1a, on forebrain of Q175 and R6/2 mice and human HD cases.ResultsDiffuse nuclear and aggregate immunolabeling increased in abundance in Q175 with age, with striatal projection neurons showing immunolabeling earlier than cortical neurons, and only neuropil immunolabeling prominent in pallidal regions. Nonetheless, some regions less affected in HD, such as hippocampus, were rich in mutant HTT1a as well. In humans, striatal immunolabeling was sparser than in mouse, and mainly in the neuropil, but sparser in striatal target areas. In human HD cortex, the P90 antibodies detected predominantly neuropil aggregates, which appeared to, in part, localize to dendrites. Immunostaining in mouse and human could be blocked with HTT1a target peptide, demonstrating antibody specificity.ConclusionsOur results indicate that mutant HTT1a burden appears to partly account for overall differential forebrain regional vulnerability in HD, but additional factors may contribute to vulnerability differences among forebrain regions and between specific neuron types.

Neuronal function requires fine-tuned and coordinated activity of several ion channels and transporters. One member of this ensemble is the KCC2 potassium-chloride cotransporter. Because KCC2 expression is required for GABA-dependent inhibitory synaptic transmission, mutations in the gene encoding KCC2 (<i>SLC12A5</i>) have been linked to several diseases that also arise from defects in GABA signaling, including epilepsy, schizophrenia, and autism spectrum disorders. Although characterization of the corresponding mutant proteins is ongoing, KCC2 mutants may reside at the cell surface but lack function, they may remain trapped intracellularly and are thus unable to function at the cell surface, or they may be readily degraded. In this article, we summarize these data and emphasize the importance of protein degradation and protease activity during KCC2 quality control, i.e. the pathway that ensures only properly folded and mature KCC2 can traffic to and function at the cell surface. We also highlight how proteolysis regulates the amount of active KCC2 at the cell surface, i.e. KCC2 quantity control. Finally, because previously unidentified KCC2 mutants are continuously being discovered, we discuss the use of predictive pathogenicity algorithms to provide researchers with information on potential disease outcomes.

Gasdermins (GSDM) are pore-forming proteins that mediate pyroptosis, an inflammatory form of programmed cell death characterized by membrane permeabilization and the release of intracellular contents. Beyond their roles in host defense and immunity, recent studies have revealed critical contributions of GSDMs, particularly GSDMD and GSDME, to the pathogenesis of neurodegenerative disorders. Their functional scope has now expanded beyond executing cell death to roles in tissue regeneration and food tolerance. The recent discovery that intact, full-length GSDMs can form pores is prompting a reevaluation of long-standing models of gasdermin activation. How post-transcriptional modifications (PTMs) regulate this unconventional activity, and under what physiological or pathological contexts these alternative mechanisms are engaged, remains an open question. Moreover, the development of neutralizing biologics that specifically target GSDM pores opens new avenues for therapeutic intervention. In light of these emerging insights, this review will provide a comprehensive and up-to-date overview of recent breakthroughs in GSDM research. We highlight advances in the structural basis of GSDM activation and pore assembly. We also discuss how these mechanisms are involved in the pathogenesis of neurodegenerative diseases and therapeutic strategies based on the emerging small-molecule inhibitors and neutralizing biologics.

SARS-CoV-2 infection has raised significant concerns regarding its impact on assisted reproductive technology. We found that oocyte retrieval during acute SARS-CoV-2 infection significantly reduced the rates of good-quality blastocyst formation, but the underlying molecular mechanisms remain poorly understood. To address this, we investigated the effects of maternal acute SARS-CoV-2 infection on preimplantation embryo development and the early offspring hematopoietic system. Using single-cell RNA sequencing (scRNA-seq), we identified developmental delays in morphologically normal blastocysts from infected mothers, characterized by prolonged expression of zygotic genome activation-related genes, downregulation of mTORC1 signaling, and altered energy metabolism, including suppressed oxidative phosphorylation (OXPHOS) and enhanced glycolysis. We further revealed that maternal acute infection induced abnormal methylation/demethylation patterns in preimplantation embryos. To assess the potential long-term impact on offspring, we conducted integrated multi-tissue analyses, including bulk RNA-seq and genome-wide DNA methylation profiling of placental tissues, along with scRNA-seq of umbilical cord blood (UCB) cells from neonates delivered by SARS-CoV-2-infected mothers. Neonates exhibited elevated levels of inflammatory cytokines and an increased abundance of monocytes, indicating an activated myelopoiesis response. In addition, hematopoietic stem and progenitor cells (HSPCs) from UCB showed reduced OXPHOS activity and a skewed differentiation bias toward the myeloid lineage, potentially impacting long-term immune function. Collectively, these findings reveal that maternal acute SARS-CoV-2 infection impairs preimplantation embryo development and leaves a lasting imprint on offspring hematopoietic health through dysregulated energy metabolism, epigenetic modifications, and altered immune responses.

Ubiquitination, a critical posttranslational modification (PTM), involves the enzymatic covalent attachment of ubiquitin to target proteins. This process is fundamental for maintaining cellular homeostasis and regulating key biological functions. The ubiquitination pathway, orchestrated by ubiquitin and its associated enzymes, offers remarkable versatility, acting as a cellular sentinel to ensure precise spatiotemporal control of essential molecular processes. Importantly, the components and mechanisms of ubiquitination can be finely tuned in various ways. Dysregulation of this system can disrupt normal biological processes and contribute to the development of various serious human diseases. These findings underscore the importance of investigating ubiquitination to understand disease mechanisms and develop effective treatment strategies. In this review, we summarize the historical developments and key milestones in ubiquitination research, with a focus on its roles in both health and disease. We explore the components and mechanisms involved, the relevant signaling pathways and their crosstalk, and the multilayered regulatory functions of ubiquitination under physiological and pathological conditions. The pathological contexts discussed include cancer, neurodegenerative disorders, cardiovascular diseases, inflammatory conditions, autoinflammatory disorders and developmental disorders. Enhancing our understanding of ubiquitination could provide novel insights into disease pathogenesis and identify new therapeutic targets. We also highlight emerging strategies for cancer treatment, such as proteolysis-targeting chimeras (PROTACs) and molecular glues. Furthermore, we review therapeutic targets and recent progress in clinical research, including ongoing clinical trials and FDA-approved drugs, aimed at leveraging the ubiquitination pathway for disease treatment.

Dearomatization offers a powerful route towards three-dimensional spirocyclic architectures, which are pivotal in bioactive molecules and pharmaceuticals, particularly for enantioselective construction of congested quaternary spirocenters. However, biocatalytic dearomatization towards these attractive spirocyclic frameworks under mild and green conditions remain underexplored. Here, we report a photoenzymatic platform synergizing visible-light activation with engineered nicotinamide-dependent ketoreductase (KRED) catalysis to achieve highly enantioselective dearomative spirocyclization of N-hydroxyphthalimide esters. Semi-rational engineering of KRED P2-D12 enables the synthesis of diverse chiral spirocyclohexadienones, also including the challenging ortho-cyclohexadienones, with exceptional functional group tolerance and stereocontrol (up to >99% ee). The utility of this methodology is underscored significantly by the enantioselective total synthesis of the cytotoxic natural product (+)-denobilone A (93% ee). Overall, this work establishes a chemobiocatalytic platform for the synthesis of complex and chiral spiroarchitectures with high enantioselectivity, expanding the biocatalytic toolbox in sustainable synthetic chemistry.

BACKGROUND: Several long non-coding RNAs (lncRNAs) have been identified as oncogenic factors in hepatocellular carcinoma (HCC). This study aims to investigate the biological function and prognostic value of lncRNA RHPN1-AS1 in HCC. METHODS: HCC data were drawn from TCGA database. Coexpression, enrichment, and immune infiltration analyses were performed to identify the roles and mechanisms of RHPN1-AS1 in HCC. Kaplan‒Meier analysis was used to determine overall survival (OS) and progression-free survival (PFS). Univariate and multivariate Cox regression analyses were employed to discover independent prognostic indicators. A prognostic nomogram was established and validated. RESULTS: The expression of RHPN1-AS1 was significantly upregulated in HCC samples, and was associated with poor OS and PFS. Univariate Cox regression analysis demonstrated that RHPN1-AS1, AJCC stage, T stage, and M stage were correlated with OS. In multivariate Cox regression analysis, RHPN1-AS was significantly linked to HCC prognosis. The predictive model based on RHPN1-AS1 and clinical factors reached good accuracy. In addition, RHPN1-AS1 was positively co-expressed with TONSL, CHRAC1, JRK, RECQL4, STK3, and CYHR1. Functional enrichment analysis of RHPN1-AS1 co-expressed genes were indicated significant involvement in the cell cycle, DNA replication, and double-strand break repair. CONCLUSIONS: LncRNA RHPN1-AS1 was identified as a potential independent prognostic indicator for HCC patients. A prognostic nomogram integrating RHPN1-AS1 expression and T and M stage was constructed to predict the survival of HCC patients. Furthermore, our findings suggested the involvement of the cell cycle in the action of RHPN1-AS1, providing new insights into the molecular mechanism underlying HCC progression.

<h4>Background</h4>Migraine is a complex neurological disorder with poorly understood molecular mechanisms. Despite advances in genetic and omics research, the shared mechanisms between central and peripheral nervous systems in migraine pathogenesis remain unclear.<h4>Methods</h4>We employed a multi-omics approach, integrating human trigeminal ganglion (TG) single-nucleus RNA sequencing (snRNA-seq) data and expression quantitative trait loci (eQTL) data from eight major cortical cell types. Mendelian randomization (MR) analysis was used to prioritize susceptibility genes, followed by functional enrichment, molecular network mapping, and computational drug screening. Key findings were experimentally validated in the primary sensory cortex hindlimb area brain region and TG, given their established roles in pain processing.<h4>Results</h4>We identified 586 migraine-associated genes in TG and 1,108 in the cortex, with 109 overlapping genes. These overlapping genes converge on pathways including autophagy and neuroinflammation, suggesting shared mechanisms of central and peripheral nervous systems. Five hub genes - HSP90AB1, EGFR, ERBB3, MET and ATG7 - were implicated in both TG and cortical tissues. Experimental validation identified five hub genes strongly linked to migraine, with ATG7 emerging as a key candidate. Immunofluorescence co-localization revealed ATG7's prominent expression in both cortical astrocytes and neurons, suggesting its dual role in glial and neuronal pathways underlying migraine pathophysiology. Western blot analysis revealed that in the S1HL brain region of migraine model mice, the protein level of LC3-II showed an increasing trend, while the expressions of both LC3-I and p62 exhibited decreasing trends compared to the control group. Furthermore, both the LC3-II/LC3-I ratio and the LC3-II/p62 ratio were significantly elevated in the model group, suggesting that the upregulation of ATG7 promotes the activation of autophagic flux in the migraine model, with the autophagic flux remaining unobstructed.<h4>Conclusions</h4>Our study provides novel insights into migraine's central and peripheral mechanisms, highlighting cell-type-specific genetic contributions and potential therapeutic targets. The integrative framework combining snRNA-seq, eQTL, GWAS, and MR enhances the understanding of migraine biology and accelerates drug discovery, offering a pathway toward more effective treatments.

Lean non-alcoholic fatty liver disease (NAFLD) presents a unique clinical phenotype increasingly recognized within the evolving nomenclature of metabolic dysfunction–associated fatty liver disease (MAFLD) and metabolic dysfunction–associated steatotic liver disease (MASLD). This study evaluated the diagnostic applicability of both frameworks in lean Egyptian patients and explored their capacity to encompass metabolically healthy lean phenotypes. Ninety ultrasonography confirmed lean NAFLD patients (BMI < 25 kg/m²) were reassessed using MAFLD and MASLD criteria. Clinical, metabolic, and biochemical parameters, alongside non-invasive fibrosis indices and transient elastography, were compared across classifications. Among all patients, 87.8% met MASLD and 77.8% fulfilled MAFLD criteria, showing substantial overlap yet minor classification differences. MASLD identified a slightly higher proportion of lean cases, whereas MAFLD more stringently captured those with overt metabolic dysfunction. Clinical, biochemical, and fibrosis parameters were comparable among groups (p > 0.05). However, 12.2% of patients remained unclassified under either definition, representing a metabolically healthy lean with steatotic liver disease (MHL-SLD) subgroup. In lean steatotic liver disease, MASLD and MAFLD show substantial diagnostic overlaps and identify similar patient profiles with no significant metabolic or fibrosis differences. MASLD includes slightly more patients, yet without clear clinical distinction. Refinement of criteria may be needed to better capture metabolically healthy lean individuals with liver steatosis (MHL-SLD).

<h4>Objective</h4>Prematurity is the leading cause of under-five mortality worldwide. The first hours after birth (Golden Hours, GH), are critical for preterm infants, as early interventions impact morbidity and mortality. While GH-protocols aim to optimize stabilization and NICU admission, their components and outcomes vary. This study systematically reviews GH-protocols to identify common elements, implementation differences, and clinical outcomes, informing best practices and highlighting knowledge gaps.<h4>Study design</h4>JBI and PRISMA-ScR guidelines were followed to map GH-protocol content and outcomes for preterm infants. Three-step search strategy included database searches and reference screening. Two reviewers extracted data on study characteristics, GH-protocol components, and outcomes. Study quality was assessed using the QI-MQCS checklist.<h4>Results</h4>Of 2,656 records, 34 studies were included, mostly from the USA (n=17). Infants had gestational ages from 24.4-30.2 weeks, and birth weights from 670-1,418 grams. GH-protocol content varied, with respiratory support (31/34 studies), cardiovascular support (26/34), thermoregulation (24/34), and team-based interventions (24/34) being most common. Improved protocol adherence was reported, yielding decreased mechanical ventilation, and quicker GH completion. Clinical outcomes showed improvements in mortality, bronchopulmonary dysplasia, intraventricular hemorrhage, necrotizing enterocolitis, and sepsis. Additionally, 7/9 studies observed significant increase in delayed cord clamping.<h4>Conclusion</h4>GH-protocols for preterm infants demonstrate beneficial effects on morbidity and mortality, yet exhibit variability. Differences in populations, hospital resources, and methodologies hinder cross-study comparisons. Standardized guidelines and outcome measures are needed to refine GH-protocols and identify effective interventions. Future research should evaluate short- and long-term effects of GH-protocols across diverse healthcare settings to enhance neonatal outcomes.

BACKGROUNDS: Significant heterogeneity was observed in the epithelial-derived endometrial cancer (EC), which contributes to different prognostic outcomes. While the mechanisms involved in this heterogeneity in the tumor microenvironment (TME) of early-stage EC remain poorly defined. METHODS: Single-cell RNA sequencing, spatial transcriptomics, and multiplex immunofluorescence (mIF) were applied to characterize cellular subpopulations in early-stage EC. EC cell proliferation was assessed by CCK-8 and flow cytometry (FC). Furthermore, the epithelial dynamics and macrophage interactions were also investigated in vitro by molecular docking and GST pull-down mass spectrometry. Phagocytic activity was also evaluated by FC and immunofluorescence. Moreover, the upstream transcription factors were identified using CUT&Tag. For in vivo study, an organoid–macrophage co-culture model was established to mimic the EC microenvironment, and altered organoid proliferation and macrophage phagocytosis were confirmed. CD47 was evaluated as a potential prognostic marker by proteomic analysis and immunohistochemistry. RESULTS: We identified a previously unrecognized epithelial subpopulation (CD47⁺CDK1⁺) that was strongly linked to cell-cycle signaling and malignant proliferation. This subpopulation interacts closely with macrophages through the CD47–Galectin 9 (LGALS9) receptor-ligand pair. Functionally, CD47 inhibition suppressed EC cell proliferation and induced apoptosis, while CD47 overexpression enhanced the proliferative capacity of EC patient-derived organoids (PDOs). EC cells and PDOs engaged macrophages through the CD47–HCK (Hemopoietic Cell Kinase) axis, driving the secretion of immunosuppressive molecules LGALS9, IL-10, and TGF-β1 and forming an immunosuppressive TME. In turn, macrophage-derived LGALS9 reinforced EC cell and PDO proliferation via CD47, establishing a positive feedback loop involving CD47–HCK–LGALS9. Furthermore, estrogen-related receptor gamma (ERRγ) was identified as an upstream transcriptional regulator of CD47, and its expression was suppressed by progesterone. CONCLUSIONS: Regulated by ERRγ, an optimistic therapeutic target, the CD47-HCK-LGALS9 axis modulated cellular proliferation-immunosuppression coupling in early-stage EC.

Hypoxia plays an important role in multiple biological processes, including inflammation, and modulates the T helper 17 (Th17) and regulatory T-cell (Treg) imbalance that often contributes to inflammatory diseases. Although the transcription factor (TF) hypoxia-inducible factor 1-alpha (HIF-1α) drives gene expression changes that promote Th17 differentiation, whether hypoxia modulates chromatin openness in Th17 and Treg cells has not been well characterized. Here, we applied assay for transposase-accessible chromatin using sequencing (ATAC-seq) and RNA sequencing (RNA-seq) to investigate how hypoxia alters the chromatin accessibility of Th17 and Treg cells, and how this correlates with the transcriptomic consequences that impact Th17/Treg balance. Our integrated analysis led to the identification of several factors that could contribute to shifting Treg differentiation toward a Th17 phenotype. Hypoxia induced extensive gene expression changes in Treg cells, compared with Th17, including a stronger upregulation of Hif1a, and increased expression of signal transducer and activator of transcription 3 (Stat3) mRNA and protein. This crosstalk between hypoxia and STAT3 in Treg cells suggests a previously unknown potential regulatory mechanism influencing Treg cell differentiation. Furthermore, we highlight TFs protein C-ets-1 (ETS1), interferon regulatory factor 1 (IRF1), runt-related transcription factor 2 (RUNX2) and cyclic AMP-dependent transcription factor ATF-3 (ATF3), which could be relevant modulators of T helper (Th) cell differentiation in hypoxic environments, such as inflamed tissue. Our study shows that hypoxia contributes to regulating the Th17 and Treg balance by increasing accessibility of key loci that are involved in Th cell differentiation and favor a Th17 phenotype. Our results provide valuable insight into how hypoxia alters chromatin accessibility and gene expression of TFs that, in addition to HIF-1α, could modulate Treg differentiation and shift the Th17/Treg balance in hypoxic environments.

X-linked dystonia parkinsonism (XDP) is a progressive adult-onset neurogenerative disorder caused by the insertion of a SINE-VNTR-Alu (SVA) retrotransposon in TAF1. One element of the SVA is a tandem polymorphic CCCTCT repeat tract whose length inversely correlates with the age of disease onset. Previous observations that the repeat exhibits length-dependent somatic expansion and that XDP onset is modified by variation in DNA repair gene MSH3 indicated that somatic repeat expansion is an important disease driver. Here, we sought to uncover genetic modifiers of CCCTCT instability in XDP individuals and to provide a mechanistic link between somatic instability and disease. We determined quantitative metrics of both repeat expansion and repeat contraction in blood. Using genetic association analyses of exome sequencing data and directed sequencing of a variant MSH3 repeat, we found that MSH3 modifies repeat expansion and contraction in blood as well as age at onset. MSH3 alleles associated with earlier disease onset were associated with more expansion and less contraction. Conversely, alleles associated with later disease onset were associated with less expansion and more contraction. Notably, MSH3 repeat alleles were also similarly associated with expansion and contraction in brain tissues. Our findings provide key evidence that the role of MSH3 in CCCTCT repeat dynamics underlies its impact on clinical disease and indicate that therapeutic strategies to lower or inhibit MSH3 are predicted to both slow CCCTCT expansion and promote CCCTCT contraction, impacting the disease course prior to clinical onset.

The emergence of next-generation sequencing techniques has led to the genetic characterization of numerous congenital erythroid disorders, emphasizing crucial pathways in both normal and pathological erythropoiesis. In this study, whole exome sequencing of a single patient with atypical congenital pure red cell aplasia revealed a mutation in the <i>CDAN1</i> gene, typically associated with congenital dyserythropoietic anemia type 1 (CDAI), together with a previously unreported mutation in the <i>MMS22L</i> gene. Combined <i>mms22l</i> and <i>cdan1</i> haploinsufficiency results in severe anemia in a zebrafish model. In human erythroid progenitors, loss of <i>MMS22L</i> leads to proliferation and differentiation arrest associated with activation of the p53 pathway and global epigenetic alterations, showing that MMS22L plays an indispensable role in erythropoiesis. Furthermore, MMS22L and CDAN1 are involved in the same protein complex whose nuclear import is mediated by the importin 4 (IPO4) protein, and MMS22L nuclear import is impaired in CDAI patients due to a defective interaction between CDAN1 and IPO4. Overall, through the genetic description of a single case characterized by digenic inheritance, we identified MMS22L as a novel key factor in erythropoiesis and brought new insights into normal erythropoiesis regulation and CDAI pathophysiology.

Sepsis is a life-threatening syndrome caused by dysregulated host response that culminates in organ dysfunction. Recognizing that many prior reviews treated neutrophils as largely homogeneous or discussed their functions in isolation, we present a framework centered on emergency granulopoiesis that unifies groundbreaking single-cell multi-omics discoveries, bone marrow niche remodeling, and bedside biomarkers such as the immature-to-total neutrophil ratio and sepsis endotypes to explain the shift from defense to dysfunction. As a hallmark adaptation, emergency granulopoiesis rapidly replenishes neutrophils but paradoxically yields immature neutrophils with impaired chemotaxis, excessive neutrophil extracellular trap formation (NETosis)/reactive oxygen species, and immunosuppressive activity that aggravate tissue damage, organ failure, and late immunoparalysis. We synthesize subset-level diversity and map this diversity to transcriptional shifts coupled with epigenetic and metabolic reprogramming as well as stromal inflammation. Clinically, elevated immature-to-total neutrophil ratios and these immature subsets align with disease severity and mortality, supporting biomarker-guided stratification. Stage-specific therapeutic strategies are evaluated along the continuum of emergency granulopoiesis, calibrating NETosis, and modulating metabolic reprogramming and epigenetics to restore neutrophil homeostasis. Furthermore, we delineate testable priorities for future research such as standardizing immature neutrophil phenotyping across cohorts and defining harmonized immature-to-total neutrophil ratio thresholds. By explicitly centering emergency granulopoiesis and integrating single-cell biology with bedside indicators, this review clarifies why a protective program becomes maladaptive in sepsis and outlines practical avenues for targeted interventions and trial design.

Schizophrenia (SCZ) is a highly heritable psychiatric disorder, yet the mechanisms linking genetic risk to pathogenesis remain unclear. This study employs context-specific expression quantitative trait loci (eQTL) analysis using the BrainSeq Phase 1 dataset to dissect schizophrenia-associated regulatory dynamics. We identified widespread loss and gain of regulatory associations in schizophrenia group versus controls, alongside consistent eQTLs. A notable target gene switching phenomenon emerged, where specific SNPs regulated distinct genes across disease states, indicative of genetic pleiotropy mediated by competition for shared regulatory elements. Pleiotropic SNPs exhibited stronger schizophrenia associations, localized farther from target genes, and were enriched in repressive chromatin domains marked by H3K27me3. Transcription factor binding site analysis implicated EZH2, a polycomb repressive complex component, in mediating these regulatory shifts. Integration of schizophrenia-specific eQTLs with GWAS data via Mendelian Randomization prioritized risk genes like ANKRD45, which showed disease-context regulation and links to behavioral deficits. Overexpression of ANKRD45 inhibited neuronal differentiation, whereas knockdown promoted it. This study presents context-specific eQTL dynamics as a crucial factor in the genetic landscape of schizophrenia, enhancing our understanding of non-coding risk variants and their role in disease susceptibility, and emphasizing the importance of utilizing context-specific eQTL data in elucidating the mechanisms of mental illness.

Exosomes, nanosized extracellular vesicles that play key roles in intercellular communication, have great potential in diagnostic and therapeutic applications. However, isolating exosomes from complex biological matrices such as human serum remains challenging due to limitations in scalability, cost, and purity of current methods. In this study, we developed and optimized a polyethylene glycol (PEG) precipitation method as a cost-effective alternative to the commercial ExoQuick kit for efficient exosome enrichment from pretreated human serum. The efficiency and reproducibility of PEG precipitation were systematically evaluated and compared with ExoQuick using various analytical techniques, including size exclusion chromatography (SEC), scanning electron microscopy (SEM), nanoparticle tracking analysis (NTA), Western blotting, and nano-liquid chromatography-tandem mass spectrometry (nano-LC-MS/MS). SEC confirmed comparable exosome recovery between PEG and ExoQuick, while SEM and NTA analyses verified intact morphology and consistent size distribution of exosomes. Western blotting confirmed the presence of characteristic exosomal markers (CD63 and CD9) and a reduction of contaminating proteins such as albumin and apolipoprotein A1. Moreover, proteomic analysis by nano-LC-MS/MS identified diverse exosomal proteins, confirming the isolation of intact exosomes. Repeated PEG precipitation up to four cycles further improved exosome purity without compromising yield. These results demonstrate that PEG precipitation is a scalable, accessible, and cost-effective method that provides comparable exosome enrichment efficiency to ExoQuick. The optimized protocol offers a practical solution for analytical laboratories to enable cost-effective exosome-based studies in diagnostics and therapeutics.<h4>Supplementary information</h4>The online version contains supplementary material available at 10.1186/s40543-025-00521-0.

<h4>Background</h4>Frailty and falls significantly impact the health and quality of life of older adults with type 2 diabetes (T2D), increasing morbidity and dependency. However, their association in Vietnamese populations remains understudied. This study aimed to examine the association between frailty and fall risk and identify additional fall-related factors among older Vietnamese outpatients with T2D.<h4>Methods</h4>A cross-sectional study was conducted among 740 outpatients with T2D aged ≥60 years at a geriatric hospital in Hanoi, Vietnam, from March 2024 to December 2024. Fall risk was assessed using the 21-item Fall Risk Index (21-FRI), and frailty was determined by the Clinical Frailty Scale (CFS). Four multivariable logistic regression models evaluated frailty's association with falls, adjusting for demographics, diabetes-related factors, and geriatric syndromes.<h4>Results</h4>Among 740 older participants with T2D, 17.8% were frail, and 16.8% reported at least one fall in the past year. Frailty was strongly associated with a higher fall rate (p<0.001). In the fully adjusted logistic regression model, frailty emerged as a powerful and independent predictor of falls, increasing the odds by over four-fold (AOR 4.47, 95% CI 2.00-5.29). Other significant risk factors included advanced age, poor glycemic control, diabetic complications, and co-existing geriatric syndromes (depression, urinary incontinence, cognitive impairment, and sarcopenia).<h4>Conclusion</h4>Routine frailty screening should be integrated into T2D care to identify high-risk older adults and implement tailored fall prevention strategies.

<h4>Introduction</h4>Obesity affects a large percentage of the population, and it is associated with many comorbidities. These pathologies often coincide with the presence of a low-grade chronic inflammatory state in individuals with obesity that can be measured using circulating inflammatory proteins. The objective of this study was to use a noninvasive approach to determine obesity-induced inflammation by measuring the urinary proteome of individuals with and without obesity.<h4>Methods</h4>Morning urine samples were taken from normal-weight controls (n = 30) with a mean body mass index (BMI) of 22.8 kg/m² and people with obesity (n = 58) with a mean BMI of 32.9 kg/m². Further data regarding high-density lipoprotein, fat distribution, blood pressure, and waist circumference were available from the individuals with obesity. Using these samples, normalized protein expression data were obtained using the Olink Explore 384 inflammation panel. These data were analyzed using R to elucidate the differences in protein expression between the individuals with and without obesity.<h4>Results</h4>Of the 384 inflammation proteins, 48 proteins had a <i>P</i> < .05 with false discovery rate correction between the persons with obesity and the persons without obesity. Network analysis revealed 5 different clusters of proteins with several clusters associated (<i>P</i> < .05) with circulating concentrations of high-density lipoprotein, waist circumference, and fat distribution in individuals with obesity.<h4>Conclusion</h4>This paper shows that urine may represent a novel noninvasive approach to measure the state of inflammation in individuals with obesity using Olink targeted proteomics.

The reasons for the aggressive clinical phenotype of signet ring cell carcinoma (SRCC) have not been fully elucidated. Previous studies suggest similarities in the genotype of colorectal and gastric SRCC and a clear distinction from non-SRCC. The immune microenvironments of gastric and colorectal SRCC have not been comprehensively examined. We isolated RNA from formalin-fixed, paraffin-embedded (FFPE) sections of 34 tumor specimens, 10 colorectal SRCC, 24 gastric SRCC, 4 non-SRCC colorectal (CCC), and 3 gastric adenocarcinoma (GCC) samples. The PanCancer Immune Profiling Panel was used to evaluate the expression of 770 immune-related genes. We compared the expression profiles of colorectal and gastric SRCC and non-SRCC adenocarcinoma. We found that the immune-related gene expression profiles (GEPs) of colorectal SRCC (CR-SRCC) and gastric SRCC (G-SRCC) were distinct from the non-SRCC. A total of 127 genes were upregulated and 32 downregulated in CR-SRCC compared to CCC. Only two genes (CCL27 and LAIR2 reached statistical significance (<i>p</i>-adj < 0.05)) among the differentially expressed genes in G-SRCC compared to GCC. None of the clinically relevant immune checkpoints were significantly differentially expressed in SRCC vs. non-SRCC. Overall, we noted a relative abundance of CD8+ cells in CR-SRCC and G-SRCC and relative overexpression of genes involved in innate immune response including the complement pathway. Finally, we identified IL13RA2 as a potential biomarker and therapeutic target candidate for CR-SRCC. The immune microenvironments of CR-SRCC and G-SRCC are distinct from non-SRCC. Broadly, both CR-SRCC and G-SRCC are characterized by a complex immune microenvironment that features cytotoxic cells and innate immune activity that may facilitate immune evasion.

The intestinal epithelium maintains tissue homeostasis through a dynamic balance of stem cell proliferation and differentiation. This process is spatially regulated along the crypt-villus axis, with intestinal stem cells in the crypt regions proliferating and progenitor cells differentiating as they migrate toward the villus tips. Because the lumen of the gut contains very low levels of oxygen (i.e., hypoxia), an oxygen gradient is established within the crypt-villus axis, placing the crypt regions under normoxic conditions while the villus tips reside under hypoxic conditions. Hence, intestinal epithelial cells encounter distinct oxygen microenvironments throughout their life span as they migrate along the crypt-villus structures during their proliferation and differentiation process. To investigate how oxygen availability influences intestinal stem cell proliferation and differentiation, we cultured patient-derived human ileum organoids (i.e., enteroids) under normoxic (20% oxygen) or hypoxic (1% oxygen) conditions. Under hypoxia, enteroid growth was reduced, and expression of several stem cell markers, such as <i>OLFM4</i> and <i>LGR5,</i> was decreased. Bulk and single-cell RNA sequencing revealed that hypoxia suppressed Wnt signaling pathways and reduced stem cell activity. Importantly, pharmacological stabilization of HIF-1α under normoxic conditions recapitulated the hypoxia-induced loss of stemness, demonstrating that HIF-1α is a key mediator of oxygen-dependent stem cell regulation in enteroids. These findings establish that physiological hypoxia in the intestinal epithelium directly regulates stem cell fate through HIF-1α stabilization, providing mechanistic insight into how oxygen availability along the crypt-villus structures controls intestinal homeostasis.

Mandibular prognathism (Class III malocclusion) is a craniofacial anomaly characterized by an anteriorly positioned mandible, a concave facial profile and impaired mastication, and appears unusually frequently in Dolang sheep (<i>Ovis aries</i>). We combined clinical phenotyping and three-dimensional (3D) genome profiling to investigate this trait in a Dolang sheep flock. We examined 959 animals using standardized criteria, estimated a local prevalence of 10.3%, and assembled a 200 affected/200 unaffected case-control cohort for genomic analyses. As an exploratory pilot study of 3D genome architecture, we generated in situ Hi-C datasets from mandibular bone of two affected and two control sheep. At 40 kb resolution, global topologically associating domain (TAD) organization and boundary strength were broadly conserved between groups, but sliding-window analyses identified a small number of 1 Mb hotspots where affected animals showed increased TAD-boundary density and strengthened insulation. These UNDER-enriched windows lay near genes with plausible roles in craniofacial development, including <i>ROBO2, COL27A1, VRK2</i> and a cytokine cluster (<i>IL22/IL26/IFNG</i> with <i>MDM1</i>). Together, our data indicate that mandibular prognathism in Dolang sheep is associated with localized remodeling of chromatin insulation at a restricted set of gene-proximal loci and highlight candidate regions and mechanisms for integration with whole-genome sequencing, association and transcriptomic data.

Endometriosis has a significant impact on the social, psychological, psychosomatic, and physical aspects of women's lives. There is increasing evidence that endometriosis has to be seen as a systemic and complex disorder with a multifactorial etiology, accompanied by numerous other pathologies, such as mental disorders and even cancer. Herein, we analyzed Disability-Adjusted Life Years (DALYs) and Years Lived with Disability (YLDs) generated from the Global Burden of Disease Study (GBD 2021), which are key metrics used to measure the worldwide impact of diseases. Besides, differential gene expression data generated from the Turku Endomet Database were calculated. Briefly, log2-transformed gene expression counts were investigated using linear modeling with the function expression ~ condition to generate log2 fold changes and <i>p</i>-values for each gene. This enabled a precise comparative analysis of mRNA expression levels between control endometrium and various endometriosis-affected tissues, including ovarian endometrioma, peritoneal lesions, and deep endometriosis. Expression patterns of specific genes related to pain and malignant turnover within endometriosis samples and controls have been analyzed. The identification of upregulated genes like <i>FOS</i>, <i>DES</i>, <i>SIRT1</i>, <i>SBDS</i>, <i>SRF</i>, <i>SPN</i>, <i>P2RX1</i>, <i>TEAD3</i>, and <i>SLITRK3</i>, alongside downregulated genes such as <i>KIF22</i>, <i>KIF25</i>, <i>GAS2L2</i>, and <i>HINT3</i>, highlights a broad transcriptional reprogramming within endometriotic tissues. The clustering analysis, which reveals pain-related genes (<i>SRP14</i>/<i>BMF</i>, <i>GDAP1</i>, <i>MLLT10</i>, <i>BSN</i>, and <i>NGF</i>), further solidifies the genetic basis for the chronic and often debilitating pain experienced by patients with endometriosis. In 2021, women with endometriosis experienced the highest rates of total YLDs at 19.98%, with anxiety contributing 17.21% and major depression 8.12%, equating to mean YLDs of 15-24 years. In conclusion, our findings reinforce the need for adopting a holistic, psychosomatic approach to managing endometriosis. The identified genetic markers related to pain provide a biological basis for the profound physical suffering. At the same time, the robust DALYs and YLDs data quantify the devastating impact on mental health, particularly highlighting the significant burden of depression and anxiety.

Osteocalcin (OC) is a small protein (49 aa) produced by the osteoblast and the most abundant noncollagenous protein in bone. Studies using the osteocalcin-depleted knock-out mouse (OC-/-) reported that osteocalcin affects bone mineral properties and bone strength. Other reports indicated osteocalcin was a hormone regulating glucose metabolism by increasing insulin secretion and sensitivity. These results were controversial. Several years ago, a couple of groups generated osteocalcin knock-out mice using different methods and failed to observe an effect of osteocalcin on bone mineral properties, bone strength or glucose metabolism. One review claimed that the previously reported results from OC-/- mouse studies were inconsistent. Within the last 2 years, additional OC-/- mouse studies have been reported, and they confirm the role of osteocalcin in glucose metabolism, bone mineral properties and strength. Some of the new data reported clears up any controversy. Published studies on the role of osteocalcin in bone and glucose metabolism using OC-/- mice will be reviewed, and results will be compared. It is shown that most of the data on the effect osteocalcin on bone properties and strength is consistent and corroborated when comparing valid data from similar techniques and similar regions of the bone. Data corroborating the role of osteocalcin in glucose metabolism will be presented, and reasons for conflicting results will be discussed. Diabetics and the elderly have reduced osteocalcin levels and are prone to bone fractures, while diabetics are glucose-intolerant. Osteocalcin may be of therapeutic use to these populations.

<h4>Background</h4>Left atrial appendage aneurysm (LAAA) is a rare cardiac abnormality associated with thromboembolic events and arrhythmias. This systematic review aimed to provide a comprehensive evaluation of literature reports on the demographics, clinical presentation, electrocardiographic and imaging findings, treatment, and outcomes of patients with LAAA.<h4>Methods</h4>A literature search was conducted using the PubMed, MEDLINE, and Scopus databases through September 2025. Only case reports and series explicitly describing LAAA were included. Extracted data included age, sex, clinical symptoms, electrocardiogram (ECG) characteristics, imaging findings, associated cardiac abnormalities, treatment modalities, and outcomes.<h4>Results</h4>A total of 216 cases were included. The mean age at diagnosis was 30.41 ± 22.39 years, with a slight predominance of males (50.5%). Symptoms included palpitations (32.4%), dyspnoea (17.2%), and thromboembolic events (7.8%). Atrial fibrillation and flutter were the most commonly detected arrhythmias. Echocardiography was the most frequently used initial diagnostic tool, with computed tomography (CT) and magnetic resonance imaging (MRI) providing additional anatomical details. Chest X-rays often yielded non-specific findings. The mean aneurysm diameter was 6.87 ± 2.64 cm. Surgical treatment, mainly aneurysm resection, was the most commonly used approach (72.7%), while conservative and device-based therapies were applied selectively. Concomitant cardiac anomalies were present in 13.7% of cases and influenced case management. The mortality rate was 4.6%, although significant morbidity was observed. Multivariate logistic regression analysis revealed that atrial fibrillation/flutter was the sole variable significantly linked with clot formation/embolism (<i>p</i> < 0.05).<h4>Conclusion</h4>LAAA is a rare, although clinically significant, entity with variable presentation and management challenges. However, early recognition and individualized treatment are essential. Further research is needed to define standardized diagnostic criteria and treatment guidelines.

No abstract available.

Psoriasis is a chronic immune-mediated disorder characterized by immune dysregulation and abnormal keratinocyte proliferation. Dysbiosis of the skin microbiota is a key contributor to its pathogenesis, and the expansion of pathogenic type III innate lymphoid cells (ILC3s) plays a central role in disease progression. <i>Cutibacterium acnes</i> (<i>C. acnes</i>) has shown therapeutic potential in alleviating psoriatic symptoms. Here, we engineered a fusion membrane by combining the bacterial cell wall of <i>C. acnes</i> with M2-type macrophage (M2) membranes, and encapsulated Cu/Zn-based metal-organic frameworks (Cu/Zn-MOFs) with antimicrobial and antioxidant properties to form the nanoparticle CM@MOF. In an imiquimod (IMQ)-induced mouse model of psoriasis, CM@MOF significantly reduced <i>Staphylococcus aureus</i> colonization, restored microbial homeostasis, enhanced skin barrier integrity, and suppressed oxidative stress and proinflammatory cytokine expression. Furthermore, CM@MOF effectively reduced the accumulation of pathogenic ILC3s in psoriatic lesions and reestablished local ILC homeostasis. These findings highlight CM@MOF as a promising therapeutic strategy for the treatment of psoriasis.

Late blastocyst development before implantation is a unique feature of ungulates, during which the epiblast proliferates and maintains pluripotency while extra-embryonic tissues expand dramatically, elongating to several tens of centimeters. The mechanisms coordinating these processes are not well understood. We performed single-cell omics profiling of porcine blastocysts from the hatched stage (E7) through early (E9) and late ovoid stages (E11). From 15,370 cells, we identified distinct embryonic and extra-embryonic populations with characteristic chromatin accessibility profiles. We reconstructed gene regulatory networks using enhancer-based eRegulons and validated them through motif occupancy analysis. Extra-embryonic tissues showed strong shifts in gene regulatory module activity at the onset of elongation, reflecting major transitions in morphogenesis and differentiation and the activation of pathways linked to cell morphology, proliferation, metabolism, trafficking, and biomolecule transport. In contrast, epiblast cells retained a stable transcriptional and regulatory identity from day 7 to day 11, immediately preceding the onset of gastrulation.

Ferroptosis is a distinct necrotic form of regulated cell death caused by a breakdown in membrane redox homeostasis. Accumulating evidence highlights a central role for ferroptosis in both acute and chronic kidney diseases, with proximal tubule cells being the primary target. It is tightly controlled by an intricate network of metabolic pathways for iron, lipid, and redox homeostasis, all of which are highly affected by kidney diseases. Moreover, recent studies have demonstrated that several human kidney disease genes modulate cellular susceptibility to ferroptosis by altering these metabolic pathways, underscoring ferroptosis as a potential therapeutic target to improve patient outcomes. Mechanistic studies have defined the cysteine-glutathione-glutathione peroxidase 4 (GPX4) axis as the central defense against ferroptosis. GPX4 detoxifies membrane phospholipid hydroperoxides, thus preventing iron-dependent lipid peroxidation chain reactions and damage to the plasma membrane. When GPX4 is overwhelmed, toxic lipid peroxides accumulate and disrupt membrane integrity-a process known as ferroptotic stress-ultimately leading to plasma membrane rupture and cell death. In this review, we provide a conceptual framework for understanding how ferroptotic stress contributes to kidney disease progression and how it can be therapeutically targeted. We highlight recent evidence that ferroptotic stress not only triggers cell death but also significantly affects the surviving proximal tubule cells. We discuss sex-specific differences in ferroptosis and explore the implications of female resilience to ferroptosis for identifying new therapeutic strategies. By integrating mechanistic insights into ferroptotic stress with new experimental observations, this review underscores ferroptosis as both a pathogenic driver and a promising therapeutic target in kidney disease.

<b>Background/Objectives</b>: Many studies in the literature are increasingly focusing on how genes influence the development of individual behaviors and personality traits through genome sequencing. Most research indicates that complex behaviors and their characteristics are influenced by multiple genes, highlighting the crucial role of genetic studies in this field. Behavioral genetics, as a scientific discipline, investigates how genetic factors shape individuals' behaviors and personality traits. The concepts of violence and aggression, observable in various contexts, have been extensively studied, with a particular focus on the underlying causes of these behaviors. In sports, where physical strength plays a significant role, regulations designed to prevent violent behaviors and aggressive attitudes contribute to the establishment of appropriate behavior patterns and discipline. <b>Methods</b>: This study aims to identify correlations between polymorphisms found in athletes and their responses to questionnaires, focusing on candidate genes known to influence personality and behavior traits, such as <i>catechol-O-methyltransferase (COMT)</i>, <i>serotonin transporter (5-HTT)</i>, <i>monoamine oxidase (MAO-A)</i>, and <i>serotonin 1A transporter (5-HT1A)</i>. A total of twenty licensed athletes participated in the study. Participants completed three standardized instruments: the Sportsmanship Behavior Scale (27 items), the Sports Emotion Scale (22 items), and the Anger-Control Scale (34 items). Following the acquisition of informed consent, buccal swab samples were collected for single nucleotide polymorphism (SNP) analysis targeting the <i>COMT, MAO-A, 5- HT1A, and 5-HTT genes</i>. Subsequent to sample collection and questionnaire administration, statistical analyses were conducted to evaluate the relationships among behavioral measures and genetic variants. <b>Results</b>: Overall, the findings point to gene-specific patterns in <i>5-HTT, MAO-A</i>, and <i>COMT</i>, while no clear pattern emerged for <i>5-HT1A</i>. <b>Conclusions</b>: Ultimately, this study provides an early exploration of aggression-related genetic patterns within the context of forensic sciences, highlighting preliminary trends and potential associations that may inform the design of future research.

Goose astrovirus 2 (GAstV-2) infection leads to visceral gout and swollen kidneys in goslings, causing a 5-50% mortality rate and significant economic losses for goose flocks. While most studies on the virus's pathological damage have focused on the kidneys, few reports have examined the effects of this fecal-oral pathogen on the digestive system. This study investigated GAstV-2 localization, cellular targets, and its impact on intestinal structure and homeostasis in orally infected goslings. Twenty 1-day-old goslings were randomly assigned to the infected and control groups. Clinical signs, organ lesions, viral distribution, histopathology, and alterations in intestinal cell populations, cytokine expression, and signaling pathways were assessed at 7 days post-infection. GAstV-2 was detected in the duodenum, jejunum, ileum, cecum, and rectum, with the highest viral load in duodenal crypt cells. Infection induced crypt cell necrosis, reduced villus height, decreased villus-to-crypt ratio, and lowered numbers of goblet cells and Lgr5+ intestinal stem cells. In contrast, Paneth cell abundance, Bmi1+ stem cells, and tight junction-related gene expression increased. Inhibition of stem cell differentiation into goblet cells was observed, mediated by modulation of the Notch signaling pathway. Proinflammatory cytokines, including <i>IL-1β</i>, <i>IL-6</i>, <i>IL-8</i>, <i>IL-22</i>, and <i>TNF-α</i>, were markedly upregulated, indicating a strong inflammatory response. These results demonstrate that GAstV-2 preferentially targets duodenal crypt cells, disrupts epithelial renewal, and impairs mucosal barrier function, while triggering compensatory regenerative and immune mechanisms. This study provides new insights into the intestinal pathogenesis of GAstV-2 and identifies potential targets for interventions to mitigate intestinal injury and economic losses in gosling production.

Computational pathology (CPath) leverages histopathology images to enhance diagnostic precision and reproducibility in clinical pathology. However, publicly available datasets for CPath that are annotated with extensive histological tissue type (HTT) taxonomies at a granular level remain scarce due to the significant expertise and high annotation costs required. Existing datasets, such as the Atlas of Digital Pathology (ADP), address this by offering diverse HTT annotations generalized to multiple organs, but limit the capability for in-depth studies on specific organ diseases. Building upon this foundation, we introduce ADPv2, a novel dataset focused on gastrointestinal histopathology. Our dataset comprises 20,004 image patches derived from healthy colon biopsy slides, annotated according to a hierarchical taxonomy of 32 distinct HTTs of 3 levels. Furthermore, we train a multilabel representation learning model following a two-stage training procedure on our ADPv2 dataset. By leveraging the VMamba model architecture, we achieve a mean average precision of 0.88 in multilabel colon HTT classification.. Finally, we show that our dataset is capable of an organ-specific in-depth study for potential biomarker discovery by analyzing the model's prediction behavior on tissues affected by different colon diseases, which reveals statistical patterns that confirm the two pathological pathways of colon cancer development. Our dataset is publicly available here: Part 1, Part 2, and Part 3.

Amino acid-based nanostructures represent an emerging class of biomolecular building blocks for next-generation biomaterials. Unlike native amino acids that lack structural complexity and mechanical integrity, their nanostructured forms, such as nanoparticles, nanofibers, nanotubes, hydrogels, peptides, and protein-based self-assemblies, offer a multifunctional scaffold design that actively participates in autonomous tissue repair. This perspective delves into the polar and nonpolar amino acid nanostructures and their roles in autonomous tissue repair through dynamic interactions with the cellular microenvironment, therapeutic delivery, and stimuli-responsiveness. The potential of advanced amino acid nanostructures, such as hydrogels and protein assemblies, is also discussed, as hydrogels provide hydrated, bioactive networks that mimic extracellular matrix functions, while protein nanostructures bring structural precision and inherent bioactivity to regenerative systems. Importantly, in our perspective, we highlight that amino acid nanostructures represent a dual closed-loop framework comprising a functional loop where these materials demonstrate adaptive response by sensing microenvironmental cues as well as a life cycle loop wherein green fabrication methods and biodegradation support sustainability. We believe that this dual functionality positions amino acid nanostructures as state-of-the-art regenerative platforms that integrate biological intelligence with sustainability, hence bridging the gap between material design and clinically translatable therapeutics.

Critical-sized bone defect repair remains a major challenge in orthopaedics and tissue engineering. Polyetheretherketone (PEEK) has attracted wide attention due to its excellent mechanical compatibility and radiological transparency; however, its inherent bioinertness and insufficient antibacterial properties restrict its clinical utility. In recent years, the incorporation of graphene oxide (GO) has markedly improved the biological performance of PEEK. GO can increase surface hydrophilicity and roughness, enhance protein/ion adsorption, and promote osteoblast adhesion and differentiation, while simultaneously strengthening antibacterial and immunomodulatory effects without compromising, and in some cases even enhancing, mechanical performance. In vitro studies demonstrate that GO-PEEK stimulates osteogenic gene expression and mineralized nodule formation, while in vivo animal models confirm superior osseointegration and new bone formation compared with controls. Synergistic modifications, such as combination with hydroxyapatite, metallic ions, or antimicrobial peptides, further amplify both osteogenic and antibacterial outcomes. Nevertheless, clinical translation of GO-PEEK remains hampered by challenges including long-term stability, potential particulate-related risks, the dynamic balance between antibacterial and osteogenic functions, and issues of manufacturing scalability, consistency, and sterilization compatibility. Future research should focus on establishing a "structure-property-safety" design paradigm, developing temporally programmed multifunctional strategies, and advancing 3D-printed personalized fabrication, with low-load applications such as alveolar or cranial bone repair as potential pioneer indications. Overall, GO-PEEK composites exhibit significant promise in contexts such as post-tumour bone reconstruction, dental implantation, and spinal or joint implants, and are expected to achieve successful clinical translation under evidence-based validation and standardised manufacturing pathways. <b><i>The Translational Potential of this Article</i></b> : The findings of this review highlight the potential of graphene oxide-modified PEEK (GO-PEEK) composites as next-generation orthopaedic biomaterials. By integrating enhanced osteogenic activity, antibacterial efficacy, and immunomodulatory capacity into a mechanically compatible and radiolucent polymer, GO-PEEK offers a multifunctional platform for bone repair. Importantly, its promising performance in vitro and in vivo provides a foundation for translation into clinical contexts such as dental implants, spinal fusion cages, and tumour-related bone defect reconstruction. Addressing challenges in long-term stability, sterilization compatibility, and large-scale manufacturing will be critical to establish a clear regulatory and translational pathway from laboratory research to clinical practice.

<h4>Background & aims</h4>Transitioning from yeast to hyphal morphology enables Candida albicans (C. albicans) to secrete candidalysin, invade the intestinal mucosa and translocate to the blood stream. Patients with alcohol-associated hepatitis show increased intestinal abundance of C. albicans, and the candidalysin-encoding gene is associated with reduced survival. Paneth cell-derived peptide YY (PC-PYY) inhibits hyphal growth of C. albicans. In this study, we evaluated the potential of different C. albicans strains isolated from patients with alcohol-associated hepatitis to cause systemic infections and explored the therapeutic potential of PC-PYY in ethanol-induced liver disease in mice.<h4>Methods</h4>C. albicans strains isolated from fecal samples of patients with alcohol-associated hepatitis (n = 105) were co-cultured with intestinal epithelial Caco-2 cells to assess in vitro cytotoxicity. Caco-2 cells and primary mouse hepatocytes were incubated with C. albicans in the presence or absence of PC-PYY. Mice were subjected to a chronic plus binge ethanol-feeding model.<h4>Results</h4>C. albicans strains isolated from stool of patients with alcohol-associated hepatitis induced significant cytotoxicity in Caco-2 cells, and high cytotoxicity was associated with worse 30-day survival (log-rank p = 0.032). This cytotoxicity was primarily mediated by the hyphal form and largely driven by candidalysin. PC-PYY significantly reduced C. albicans-induced cytotoxicity in Caco-2 cells (Wilcoxon rank-sum test, p = 0.015) and in primary mouse hepatocytes (p = 0.03) compared with a scrambled peptide control, by inhibiting hyphal morphogenesis. The peptide YY-to-chromogranin A ratio in intestinal crypts was significantly increased in ethanol-fed mice compared with both isocaloric (p = 0.005) and antifungal-treated controls (p = 0.009), indicating that fungal overgrowth stimulates PC-PYY release. In ethanol-fed mice, PC-PYY administration attenuated liver injury (p = 0.032) and steatosis (p = 0.0498) and reduced fecal hyphae formation (p = 0.0159).<h4>Conclusion</h4>PYY inhibits filamentous growth of C. albicans in vitro and alleviates ethanol-induced liver disease in mice, highlighting its potential as a therapy for patients with alcohol-associated liver disease.<h4>Impact and implications</h4>Candida albicans (C. albicans) and particularly its toxin candidalysin are associated with poor outcomes in patients with alcohol-associated hepatitis but the extent to which the cytotoxicity of individual C. albicans strains influences patient survival, and the role of Paneth cell-derived PYY (PC-PYY) in this context remains elusive. This study identifies a link between the cytotoxic effect of patient-derived C. albicans strains and survival in patients with alcohol-associated hepatitis and demonstrates that PC-PYY plays a protective role in ethanol-induced liver disease by limiting candidalysin-producing hyphae. Our work provides insight into why some patients with alcohol-associated liver disease have worse outcomes and highlights the potential of PC-PYY as a therapy for patients with alcohol-associated liver disease.

Background and Purpose: Aminoglycoside antibiotics such as gentamicin cause nephrotoxicity in 2-55% of patients, particularly the elderly and those with chronic kidney disease. Although essential for treating severe infections, strategies to mitigate this toxicity are lacking. Although antibiotic combinations are routinely used to enhance antimicrobial efficacy, their potential to reduce gentamicin-induced nephrotoxicity has not been previously investigated. Experimental Approach: The effects of N6-methyladenosine (m 6 A) RNA methylation on lysosomal function, endosome-to-Golgi signalling, and necroptosis were examined in vitro and in vivo models of gentamicin toxicity. The ability of combined gentamicin and minocycline therapy to mitigate kidney injury was also assessed. Key Results: Our results support an integrative model of gentamicin-induced renal toxicity linking ribotoxic stress to lysosomal disruption via Fe 2+ and phospholipid accumulation, oxidative stress, stress-vesicle formation, and altered RNA methylation. Within this model, gentamicin activated a previously unrecognised toxic pathway marked by increased cytoplasmic fat mass and obesity-associated protein (FTO), decreased methyltransferase-like 3 (METTL3), and impaired m 6 A methylation, diminishing YTH N6-methyladenosine RNA binding protein 1 (YTHDF1)-dependent translation of peroxiredoxin 6 (PRDX6). PRDX6 loss elevated 4-hydroxy-2-nonenal, driving receptor-interacting protein kinase 3 (RIPK3)-dependent necroptosis and transcription factor EB (TFEB)-mediated lysosomal dysfunction with autophagosome accumulation. Co-treatment with minocycline restored m 6 A-mediated PRDX6 translation and reduced renal injury. Conclusion and Implications: We observed an unexpected spatial coupling of m 6 A-modified RNAs with stress granules and acidic lysosomes in gentamicin-induced renal injury. Importantly, co-treatment with minocycline reduced this toxicity by modulating m 6 A RNA methylation, suggesting a promising route to improve antimicrobial therapy in patients.

In targeted protein degradation (TPD) the cell’s quality control machinery is co-opted to degrade proteins of interest. Currently TPD is limited by the availability of small-molecule binders for targets and the many potential human E3 ubiquitin ligases. Consequently, other approaches are needed to tackle new targets and to exploit degradation pathways fully. The natural binding epitopes for E3s and other degradation machineries are often short linear peptide motifs, which offer routes to new biologics-based degraders. Here, targeting the anti-apoptotic BCL-x L and initiation of apoptosis, we show that several protein-degradation pathways—ubiquitination, direct proteasome, autophagy-lysosome recruitment—can be leveraged by integrating target-binding and degradation motifs into single-polypeptide constructs. This exploits versatile and varied de novo coiled-coil scaffolds as adaptable molecular glues by presenting motifs multivalently and in diverse arrangements. The constructs are small (≈80 residues) and provide a plug-and-play platform to map and optimise target–degradation space using rational design. We call this Co iled C o ils for induced Prox imity (CoCoProx) and propose its use for targeted degradation and other induced-proximity reactions.

Increased understanding of the functional complexity of the genome has led to growing recognition of the role of non-sequence-based regulatory variation in disorders of the human central nervous system. Most genomic analyses of the brain are limited by the use of bulk tissue, which comprises a heterogeneous mix of different neural cell types with distinct epigenetic profiles, thereby limiting the ability to attribute regulatory changes to specific cell populations. Given the limited availability of human post-mortem tissue resources and the importance of integrating multi-omic data from the same samples, there is a critical need for methods that enable parallel, cell-type–resolved genomic profiling. We present optimised protocols using fluorescence-activated nuclei sorting (FANS) to isolate nuclei from different human and mouse brain cell types for downstream multi-omic analysis. Our approach enables the robust purification of neuronal, oligodendrocyte, microglial and other glial-origin nuclei from both adult and fetal brain tissue. We demonstrate that FANS-isolated nuclei are compatible with a wide range of genomic assays, including profiling of DNA modifications, histone modifications, chromatin accessibility, and gene expression. This protocol maximises the utility of limited post-mortem tissue resources and provides a unified workflow for comprehensive, cell-type–specific interrogation of molecular mechanisms involved in the brain.

The Hippo-YAP/TEAD pathway plays a central role in melanoma progression by regulating tumor cell proliferation, survival, and migration. Using a NanoLuc Binary Technology (NanoBiT) protein-protein interaction assay, we screened honokiol-based small molecules and identified several analogues that disrupt the YAP-TEAD interaction. HK03 was the most effective analogue, leading to a pronounced reduction in Cyr61 levels and diminished Erk and Akt phosphorylation in B16-F10 melanoma cells. HK03 also blocked epithelial-mesenchymal transition (EMT) and impaired melanoma cell migration in wound-healing assays. <i>In vivo</i>, HK03 treatment markedly reduced metastatic burden in a B16-F10 lung metastasis model. These findings suggest that honokiol derivatives, particularly HK03, represent potential lead compounds for targeting the YAP-TEAD axis in melanoma therapy.

Mitochondrial metabolism plays a central role in organismal physiology and aging. In Caenorhabditis elegans, FAHD-1 (oxaloacetate decarboxylase) and PYC-1 (pyruvate carboxylase) catalyze opposing reactions that influence oxaloacetate homeostasis within the tricarboxylic acid cycle. To dissect their functional interplay, we analyzed single- and double-knockout strains generated by CRISPR/Cas9 alongside the classical allele. Fahd-1 mutants exhibit impaired mitochondrial respiration, reduced motility, and early egg-laying onset, whereas pyc-1 mutants display increased locomotion and enhanced metabolic flexibility. Paradoxically, although each single mutantion extended lifespan, combining them restored wild-type lifespan and partially normalized respiratory function, suggesting a compensatory interaction. These findings establish FAHD-1 and PYC-1 as antagonistic mitochondrial enzymes whose balance governs locomotion, reproduction, and lifespan in C. elegans, providing a conceptual framework for conserved links between mitochondrial metabolism and aging.

The gut microbiota plays a pivotal role in maintaining gastrointestinal (GI) homeostasis by influencing epithelial integrity, immunity, and metabolism. Recent studies have uncovered that gut microbiota can directly or indirectly modulate the behavior and function of adult stem cells across the GI tract, which are essential for tissue regeneration and disease prevention. Moreover, key microbial metabolites including short-chain fatty acids (SCFAs), tryptophan-derived indoles, succinate, secondary bile acids, and retinoic acid exert diverse effects on stem cell quiescence, proliferation, and differentiation. This review provides current knowledge on the interaction between gut microbiota and host stem cells in the stomach, intestine, and colon.

Necrotizing enterocolitis (NEC) is a serious condition in premature infants, driven by dysbiosis-induced inflammation and barrier damage. This study screened probiotics for efficient 2'-fucosyllactose (2'-FL) utilization and intestinal repair potential. <i>Lactiplantibacillus plantarum</i> JY039 (<i>L. plantarum</i> JY039, JY039) was selected from 25 strains (<i>p</i> < 0.05) based on 2'-FL metabolism, intestinal adhesion, and <i>Lgr5</i> induction in TNF-α-damaged organoids. JY039 showed versatile carbon metabolism and sensitivity to 11 antibiotics, indicating its clinical safety. In TNF-α-injured organoids, it elevated intestinal stem cell (ISC) markers (Olfm4 and Ascl2, 4-fold) and proliferation markers (c-Myc, Ki67, and cyclins, 3-fold), with EdU intensity increasing 4-fold versus NEC models. In NEC rats, JY039 (10⁹ CFU/mL) reduced pathology scores and permeability, restored tight junctions, modulated cytokines (TNF-α, IL-6, myeloperoxidase decreased; and IL-10 increased, <i>p</i> < 0.05), activated ISC regeneration, and enriched <i>Lactococcus</i> and <i>Akkermansia</i>. JY039 effectively maintained epithelial regeneration and improved the microbial imbalance, representing a safe NEC candidate. It also offers guidance for producing a functional infant formula.

Bipolar Disorder (BD) is a highly complex and heterogeneous disorder. As such, accurate diagnosis is often delayed, and effective treatment options are limited. The Integrated Network, a program of Breakthrough Discoveries for thriving with Bipolar Disorder (BD²), was designed as a platform for longitudinal deep phenotyping of a diverse group of people with bipolar disorder to define disease trajectories and to gain insight into the biological drivers of the illness. Biosamples, including whole blood, serum, plasma, and peripheral blood mononuclear cells (PBMCs), will be collected longitudinally and will also be made available. The Integrated Network is designed to be the largest and most comprehensive prospective longitudinal study conducted in bipolar disorder, allowing for the development of precision-based treatment strategies that optimize quality of life for people living with bipolar disorder.

Recent epidemiological evidence showed that mutations to the HFE-63 allele of this hemochromatosis-associated iron-assimilation protein improve chances of avoiding Alzheimer's disease (AD). This is unexpected since increased brain ferroptosis in gray-matter increases the risk for vascular dementia and AD. However, diffusion tensor imaging from a key Alzheimer's Disease Neuroimaging Initiative biomarker study showed that the hemochromatosis H63D allele protected white matter tracts and improved cognitive performance in individuals when <i>APOE4</i> accelerates AD. H63D-carrying individuals exhibit elevated serum ferritin levels. We suggest coordinate increased levels of H-ferritin in iron-rich oligodendrocytes in H63D carriers generates sufficient neuroprotection to enhance myelin sheath integrity in white matter axons.

Huntington's disease (HD) is a severe neurodegenerative disorder that causes motor impairment and ultimately death. Yet, safe and effective disease-modifying treatments of HD are scarce, and delivery to the HD brain is challenging. Here, we present a small, ∼11 nm polyethylene glycol-coated gold nanoparticle (NP) for brain delivery and treating HD in R6/2 mice. Upon intravenous injection, this NP crosses brain barriers, preferentially accumulates in the brain of R6/2 mice than healthy littermates without pronounced liver or kidney sequestration, diffuses to the cortex and striatum, and enters neurons and microglia. Devoid of known chemical or biological drugs, this NP improves motor deficit as effectively as tetrabenazine (standard therapy for symptomatic relief) and prolongs survival, without long-term brain retention or systemic toxicity. This NP activates the oxidative phosphorylation pathway, blocks p38α mitogen-activated protein kinase phosphorylation and pyruvate dehydrogenase kinase 1 activity, and inhibits pyroptosis. Our data offer molecular insights into gold nanomedicines against neurodegenerative diseases.

Tertiary lymphoid structures (TLSs) have been associated with favorable clinical outcome and improved responses to immune checkpoint inhibitors in soft-tissue sarcomas (STSs). However, due to their rarity and high heterogeneity, information regarding the mechanisms involved in TLS formation and contribution to antitumor immunity in STS are extremely elusive. To address this gap, we integrated immunohistochemistry and transcriptomic analyses from 31 treatment-naïve STS specimens from an independent cohort of patients with primary or locally recurrent disease. Further validation was conducted using external bulk, single-cell and spatial transcriptomics data from 5 publicly available datasets. We found TLSs to be highly heterogeneous in terms of amount, localization, maturation status and cellular composition, and corroborated previous findings showing that their presence, along with B cells in the STS microenvironment, are good indicators of favorable prognosis. Transcriptomic analysis showed that high expression of germinal center (GC) B cell-related genes was associated with TLS presence and with an upregulation of signatures specific for T helper 17 (Th17) cells in STS and other cancer types. Conversely, genes signatures discriminating for immunosuppressive M2-like macrophages were enriched in tumors with low expression of GC B cell-related genes. Immunohistochemistry showed distinct spatial patterns for Th17-like cells and M2-like macrophages within TLS areas, with IL17A⁺ cells predominantly localized within intratumoral mature TLSs, and CD163⁺ macrophages mainly observed in immature TLSs. Integrating these findings, we identified tumors with high expression of GC B cell- and Th17 cell-related signatures together with low fractions of M2-like macrophages, to have superior survival outcomes. Herein, our findings point out to two cellular players (Th17-like cells and M2-like macrophages) with potentially opposite roles in STS-associated TLS formation and maturation, thus providing the basis for future research efforts aiming at the development of therapeutic immunological interventions to enhance TLS-mediated antitumor immunity in STS.

Although the effects of noise-induced hearing loss (NIHL) on cognitive functions have been widely investigated, the cognitive effects of noise-induced hidden hearing loss (NIHHL), particularly its impact on memory, remain poorly understood. The Dnah11 gene, which encodes a dynein motor protein involved in synaptic development, may play a role in NIHHL-related cognitive impairment. We aimed to investigate whether NIHHL induces memory impairment and explore the role of Dnah11 expression in this process. Behavioral experiments identified the peak of memory impairment at 1 month following noise exposure. To elucidate molecular changes, hippocampal gene expression was analyzed using transcriptomic sequencing, reverse transcription quantitative polymerase chain reaction (RT-qPCR), and immunofluorescence. RNA sequencing revealed significant Dnah11 upregulation, with immunofluorescence confirming DNAH11 overexpression in hyperactivated CaMKIIα-positive excitatory neurons. Stereotaxic injection of recombinant adeno-associated virus (rAAV) vectors to knock down hippocampal Dnah11 expression improved memory performance in NIHHL mice without improving hearing loss. This cognitive improvement was accompanied by partial restoration of synaptic plasticity-related proteins, including SYN and PSD95. These findings indicate that Dnah11 upregulation in hippocampal excitatory neurons contributes to NIHHL-induced memory impairment, and targeting Dnah11 may offer a therapeutic strategy for memory impairment associated with hidden hearing loss.

<h4>Purpose</h4>High programmed death-ligand 1 (PD-L1) expression is associated with unfavorable clinical outcomes in epidermal growth factor receptor (EGFR)-mutated lung adenocarcinomas (LUAD) patients treated with tyrosine kinase inhibitors (TKIs) or anti-PD-1/PD-L1 therapy, yet the underlying mechanisms are less explored.<h4>Methods</h4>Bulk RNA sequencing (RNA-seq) datasets were analyzed to investigate intertumoral transcriptional variations linked to PD-L1 expression. Immunohistochemistry (IHC) was utilized to quantify PD-L1 expression on tumor cells. Digital spatial profiling (DSP) was performed on 23 EGFR-mutated LUAD tissue samples to characterize transcriptomic differences in tumor cell (TC), immune cell (IM), and macrophage (MA) compartments between PD-L1 high and low groups. Furthermore, a publicly available DSP dataset was analyzed and IHC was conducted for validation.<h4>Results</h4>Analysis of RNA-seq datasets identified differentially expressed genes, signaling pathways, and immune profiles associated with PD-L1 expression. Compared to low PD-L1 tumors, high PD-L1 tumors exhibited increased infiltration of T regulatory cells (Tregs) and enhanced immunosuppressive signatures. DSP analysis revealed compartment-specific molecular disparities: TC segment in high PD-L1 tumors showed upregulated signatures of cell proliferation, invasion, and metastasis. IM segment displayed increased infiltration of immunosuppressive cells, including Tregs and myeloid-derived suppressor cells and upregulated expression of inhibitory immunomodulators CD276, HAVCR2, and LGALS9C.<h4>Conclusion</h4>Combining bulk and spatial RNA-seq, this study characterized the molecular and immunological hallmarks of EGFR-mutated LUAD in the context of PD-L1 expression, providing new insights into the development of tailored therapeutic strategies for EGFR-mutated LUAD with high PD-L1 expression.

The role of plasma and serum proteomics in characterizing human disease, identifying biomarkers, and advancing diagnostic technologies is rapidly increasing. However, there is an ongoing need to improve proteomic workflows in terms of accuracy, reproducibility, and cost-effectiveness, and to achieve cross-platform transferability. Based on large serum and plasma proteome studies, we generate the Charité Open Peptide Standard for Plasma Proteomics (OSPP), an open, versatile peptide internal standard for targeted and untargeted mass spectrometry-based proteomic studies. The OSPP includes 211 concentration-matched stable-isotope-labeled peptides selected for consistent quantification across a large number of plasma and serum proteome studies, and synthetic accessibility. We show they are consistently quantified across serum and EDTA, citrate, and heparin plasma using multiple LC-MS platforms. Despite being selected for technical parameters, the OSPP peptides represent proteins that function in a wide range of biological processes, are used in routine clinical tests, or are targets of FDA-approved drugs, making OSPP able to serve as an expandable clinical marker panel. We demonstrate the utility of OSPP in a COVID-19 inpatient cohort study for improving analytical performances, for cross-platform alignment of proteomic data, disease stratification, and biomarker discovery.

Chronic stress is a risk factor for neuropsychiatric disorders, making the ability to adapt to repeated stress a crucial determinant of mental health. It remains unclear whether repeated stress exposure leads to habituation or new adaptive responses. Here, we investigated how the transcriptomic response to acute restraint stress changes - in the mouse ventral hippocampus - with repeated exposure. Using bulk RNA-seq across multiple timepoints, we observed widespread transcriptional habituation: stress-induced gene expression was blunted without the emergence of new response profiles or changes in baseline gene expression. Temporally resolved single-nucleus multi-omic profiling (RNA and chromatin accessibility) confirmed that the transcriptional habituation occurs across cell types, revealing cell type-specific patterns. We identify gene clusters whose expression habituates with repeated exposure, which appear primarily related to two distinct mechanisms: an early blunting of cAMP-associated genes chiefly linked to a reduced number of activated cells, and a shortened transcriptional response of corticosterone-associated genes, independent of the number of activated cells. Our study provides a comprehensive resource, accessible as an interactive app, that characterizes the dynamic transcriptomic response to stress in the mouse hippocampus and how it habituates with repeated exposure to stress.

<h4>Introduction</h4>Antithrombin (AT) deficiency, often caused by mutations in the SERPINC1 gene, is a well-established risk factor for venous thromboembolism (VTE), which can lead to serious complications such as deep vein thrombosis (DVT), pulmonary embolism (PE), and cerebral venous sinus thrombosis (CVST). Syndecan-4 (SDC4), an endothelial cell membrane protein, enhances AT's anticoagulant function by facilitating its interaction with the vascular endothelium, thereby supporting hemostatic balance.<h4>Methods</h4>Our team enrolled a patient diagnosed with CVST, and subsequent genetic testing identified the SERPINC1 p.Phe155del variant. This study sought to elucidate the interaction between AT mutations and SDC4 in VTE pathogenesis using clinical case analyses, molecular docking, co-immunoprecipitation (Co-IP), and immunofluorescence assays.<h4>Results</h4>The patient was diagnosed with Type I hereditary AT deficiency, with a five-generation family pedigree constructed. Molecular docking simulations showed the SERPINC1 p.Phe155del variant weakened the protein's interaction with SDC4. Cell-level co-immunoprecipitation and immunofluorescence results were consistent with molecular docking findings, confirming that the SERPINC1 p.Phe155del mutant binds less strongly to SDC4 than the wild-type.<h4>Conclusion</h4>This reduced binding may underlie hereditary AT deficiency and increased venous thrombosis risk. The findings from this research deepen understanding of the connection between hereditary hypercoagulable states and VTE -with CVST as one possible clinical manifestation-offering new insights to inform clinical diagnosis and treatment.

<h4>Background</h4>Infants born preterm are at high risk of anemia, red blood cell transfusions, and iron deficiency, all of which may negatively influence long-term neurodevelopment. To ameliorate these complications of prematurity, we developed a Phase II trial to determine whether treatment with an erythropoietic-stimulating agent, darbepoetin (Darbe), plus a slow-release intravenous (IV) iron preparation (ferumoxytol (FMX) or low-molecular-weight iron dextran (LMW-ID)) might decrease transfusions while maintaining iron sufficiency.<h4>Methods</h4>This single-center study is a parallel design, prospective, randomized controlled Phase II trial of 120 infants born 24-0/7 to 31-6/7 weeks of gestation cared for in the University of Washington Neonatal Intensive Care Unit. After informed consent, infants less than 72 h of age are randomized to one of five treatment groups: (1) Oral iron (standard care), n = 40, or weekly Darbe 10 µg/kg/dose IV or SQ plus; (2) FMX - 10 mg/kg/dose IV, n = 20; (3) FMX - 20 mg/kg/dose IV, n = 20; (4) LMW-ID - 10 mg/kg/dose IV, n = 20; or (5) LMW-ID - 20 mg/kg/dose IV, n = 20. Infants will be followed to 2-year corrected age with sequential developmental testing. Our primary outcome is ferritin level at 34-36 weeks postmenstrual age. Secondary outcomes include other hematologic assessments, drug safety, evaluation of the gut microbiome, and neurodevelopment to 2 years corrected age.<h4>Discussion</h4>This trial will determine whether darbepoetin plus a slow-release IV iron preparation is safe, which iron preparation and dose best maintain iron sufficiency and decrease or eliminate transfusions, whether IV iron will result in a more diverse, less pathogenic microbiome when compared to oral iron supplementation, and, finally, whether these treatments affect neurodevelopment to 2 years corrected age.<h4>Trial registration</h4>National Clinical Trial (NCT) NCT05340465. Registered on March 1, 2022.

T cell immunoglobulin and mucin-domain containing-3 (TIM-3) is an emerging immune checkpoint receptor. Blocking immune checkpoint signals is a promising strategy for cancer immunotherapy. While TIM-3 blockade is currently under clinical investigation, its context-dependent role remains poorly understood. This study investigates the molecular consequences of TIM-3 inhibition using an experimental murine breast tumor model. Contrary to therapeutic expectations, administration of an anti-TIM-3 monoclonal antibody led to accelerated tumor growth and a significant increase in liver metastases. Flow cytometry revealed a paradoxical increase in tumor-infiltrating CD8⁺ T cells, accompanied by a reduction in CD3⁺ and Foxp3⁺ T cells. Cytokine profiling showed elevated levels of IFN-γ, TNF-α, and IL-17 in the serum, with increased IL-10 and IL-1β in tumors and altered cytokine expression in the spleen. Proteomic analysis identified 1371 dysregulated proteins, and gene set enrichment analysis revealed upregulation of PI3K/Akt-mTORC signaling, which promotes CDK4-mediated proliferation and tumor stemness via B2M and CD44. Gene ontology analysis indicated suppression of autophagy and apoptosis pathways, including downregulation of the proinflammatory protein complex calprotectin (S100A8/A9). Notably, TIM-3 blockades enhanced epithelial-to-mesenchymal transition (EMT) and c-MYC signaling, potentially driven by Foxp3 downregulation. Proteomics data are available via ProteomeXchange with identifier PXD065028. This finding challenges the prevailing view that immune checkpoint blockade uniformly suppresses tumor growth. Instead, this study demonstrates that TIM-3 inhibition may paradoxically exacerbate tumor progression and metastasis.

The regulation of midline crossing of axons is of fundamental importance for the proper development of nervous system connectivity in bilaterian animals. A number of conserved axon guidance signaling pathways coordinate to attract or repel axons at the nervous system midline to ensure the proper regulation of midline crossing. The attractive Netrin-Frazzled/DCC (Net-Fra) signaling pathway is widely conserved among bilaterians, but it is not clear whether the mechanisms by which Net and Fra promote midline crossing are also conserved. In Drosophila, Fra can promote midline crossing via Netrin-dependent and Netrin-independent mechanisms, by acting as a canonical midline attractive receptor and also through a non-canonical pathway to inhibit midline repulsion via transcriptional regulation. To examine the conservation of Fra-dependent axon guidance mechanisms among insects, in this paper we compare the midline attractive roles of the Frazzled receptor in the fruit fly (Drosophila melanogaster) and flour beetle (Tribolium castaneum) using CRISPR/Cas9-mediated gene editing. We replace the Drosophila fra gene with sequences encoding Drosophila Fra (DmFra) or Tribolium Fra (TcFra) and examine midline crossing of axons in the ventral nerve cord of embryos carrying these modified alleles. We show that Tribolium Fra can fully substitute for Drosophila Fra to promote midline crossing of axons in the embryonic nervous system, suggesting that the mechanisms by which Frazzled regulates midline axon guidance are evolutionarily conserved within insects.

A rare prothrombin variant (c.1787G > A, p.Arg596Gln), also known as the prothrombin Belgrade variant, has been associated with an increased predisposition to thrombosis through resistance to antithrombin. This variant has been previously reported in individuals from Serbia, Japan, China, and India. In this case report, we described the first reported case of heterozygosity for the prothrombin Belgrade variant in a Mexican-American family. Affected individuals had negative results on standard hypercoagulable studies; however, they exhibited a history of early-onset and recurrent venous thromboembolism (VTE). Although rare, the prothrombin Belgrade variant-and other prothrombin variants associated with antithrombin resistance-may be underrecognized in patients with recurrent thrombotic events, particularly among individuals from ethnic backgrounds not previously associated with this variant. These findings support the consideration of comprehensive genetic thrombophilia testing, including full sequencing of the prothrombin gene, in patients with negative standard hypercoagulable studies but a strong personal and/or family history of VTE.

Tolerogenic dendritic cells (tolDC) are currently in clinical trials for the treatment of autoimmune diseases such as rheumatoid arthritis and multiple sclerosis. The methods for producing therapeutic tolDC vary widely, with little being known about the commonalities and differences of distinct cell products in terms of their regulatory actions on CD4⁺ T cells. We compared human monocyte-derived tolDC generated with vitamin D3 alone or in combination with dexamethasone. We found marked differences in the surface expression of HLA-DR and immune regulatory molecules, but also found commonalities, e.g. a strongly reduced capacity to produce interleukin-12 and a concomitant decreased ability to induce interferon-γ secretion by allogeneic CD4⁺ T cells. To gain a deeper understanding of how these tolDC types exert their regulatory effects, we co-cultured them with CD4⁺ T cells from rheumatoid arthritis patients or healthy controls and analysed the gene expression profile and function of the responding T cells. We found that tolDC generated with vitamin D3 alone, but not in combination with dexamethasone, induced potent cytotoxic activity in the responding CD4⁺ T cells as demonstrated by an enhanced cytotoxic gene signature, increased levels of intracellular granzyme B, and superior cytotoxic activity towards myeloid and B cells. These data identify cytotoxicity as an atypical CD4⁺ T helper cell effector function induced by some but not all tolDC types, with implications for their individual clinical applications.

Huntington's disease has traditionally been considered a motor disorder, but it is currently classified as a multisystem neurodegenerative disease that involves brain regions, such as the amygdala, and causes depression. The aim of the present study was to analyse the distribution of huntingtin in the human amygdaloid basolateral complex, considering its nuclei, sex, triplet repeats and Vonsattel score, as well as to characterize the cellular relationships between huntingtin and associated copathologies. The present study included 23 human brain samples from patients (males and females) with and without Huntington's disease, Parkinson's disease and Alzheimer's disease. An unbiased stereology approach was used to quantify huntingtin deposits. Multiple immunofluorescence experiments were conducted to analyse the relationship between huntingtin and glial populations. Immunohistochemistry against pathological markers of other neurodegenerative diseases was also carried out. Quantification data did not reveal differences among different nuclei (basomedial, basolateral or lateral) in the basolateral complex or according to sex. Huntingtin deposits did not correlate with cytosine-adenine-guanine (CAG) repeats. However, these deposits were positively correlated with pathological Vonsattel grades. Additional aggregates of other pathological proteinopathies were also observed. This correlation between the human basolateral amygdaloid complex and the Vonsattel stage provides a new perspective for neuropathological diagnosis and helps in understanding nonmotor symptoms such as depression.

This study examined the effects of long-term <i>Saccharomyces cerevisiae</i> supplementation on feed intake, milk production, milk composition, and selected reproductive indicators in lactating dairy cows. Twenty-four multiparous Holstein-Friesian crossbred cows were blocked by parity and randomly allocated to three treatments: a control group without supplementation (CON; n = 7), live yeast supplementation for 60 days (YS-60; n = 10), and live yeast supplementation for 90 days (YS-90; n = 7). Dry matter intake and body weight gain were significantly higher in cows receiving live yeast, with the greatest responses observed in the YS-90 group (<i>p</i> < 0.05). Milk yield and energy-corrected milk were increased by supplementation, particularly in YS-90 cows (<i>p</i> < 0.01), along with higher milk fat and lactose concentrations. Somatic cell count was consistently lower in YS-90 cows throughout the 14-week experimental period. Body condition score differed among treatments (<i>p</i> < 0.01), with higher values observed in yeast-supplemented cows. Feed efficiency did not differ among treatments. Reproductive parameters, including estrus detection and pregnancy rate, were not significantly affected by live yeast supplementation, although plasma progesterone concentration was higher in supplemented cows (<i>p</i> < 0.05). Given the limited number of animals per treatment, reproductive outcomes should be interpreted cautiously. Overall, extended live yeast supplementation improved production performance and udder health, while its effects on reproductive performance warrant further investigation.

Proteolysis-targeting chimeras (PROTACs) have emerged as a revolutionary therapeutic modality that enables degradation of therapeutically relevant proteins through the protein disposal machinery, the ubiquitin-proteasome system (UPS). Unlike traditional small-molecule inhibitors, PROTACs harness bifunctional molecules to induce targeted protein degradation, offering advantages such as increased specificity, catalytic activity, and the potential to address previously undruggable targets. Since their conception 20 years ago, PROTACs have made significant strides in target protein degradation (TPD), and today, PROTACs are on the verge of their first clinical approval. This review presents a detailed overview of PROTAC targets, clinical development progress, and the design and detailed synthesis of degrader molecules that have advanced to clinical trials.

<h4>Background</h4>The ABO blood group locus is a well-established genetic determinant of venous thromboembolism (VTE) risk in both individuals of European and East Asian ancestry. Recent studies have identified key <i>ABO</i> haplotypes tagged by four common SNPs-rs2519093 (A1), rs1053878 (A2), rs8176743 (B), and rs8176719/rs41302905 (O1/O2)-that influence both incident and recurrent VTE risk in Europeans. However, marked differences in ABO allele frequencies and haplotype structures across ancestries may render European findings inapplicable to East Asians, highlighting a critical gap in understanding the genetic basis of VTE in this population.<h4>Methods</h4>We conducted a haplotype-based association study using ABO-tagging SNPs (including rs512770 that distinguishes between O1.1 and O1.2) in 1576 VTE cases from China Pulmonary Thromboembolism Registry Study (CURES) and 17,535 ancestry-matched controls, adjusted for age, sex, and genetic principal components to evaluate the effects of <i>ABO</i> haplotypes on VTE risk and recurrence.<h4>Findings</h4>Our analyses revealed key population-specific differences: in East Asians, the rs1053878-A allele is consistently co-inherited with the rs2519093-T allele, precluding its use as a specific marker for the A2 blood group, unlike in Europeans. Furthermore, all non-O1 haplotypes were homogeneously associated with a ∼1.4-fold increased risk of VTE (<i>p</i> = 5.2 × 10^-20) and a ∼1.7-fold increased risk of recurrence (<i>p</i> = 0.023), compared to the O1.1 group. Notably, the O1.2 blood group was also associated with a 1.7-fold increased risk of recurrence (<i>p</i> = 0.039).<h4>Interpretation</h4>These findings highlight fundamental differences in <i>ABO</i> haplotype structure and disease associations between East Asians and Europeans. Our study provides a population-specific SNP panel-rs8176719, rs2519093, rs1053878, rs8176743, and rs512770-for accurate genetic risk assessment of VTE in East Asians, underscoring the importance of ancestry-tailored approaches to thrombotic disease prediction.<h4>Funding</h4>This study is funded by the Chinese Academy of Medical Science Innovation Fund for Medical Sciences (No. 2024-I2M-TS-035, No. 2021-I2M-1-061), National Key Research and Development Program of China (No. 2024YFE0101900, No. 2023YFC2507200), National Natural Science Foundation of China (No. 82470046, No. 82241029) and Noncommunicable Chronic Diseases-National Science and Technology Major Project (No. 2024ZD0528700).

<b>Background and Clinical Significance:</b> Histiocytic sarcoma (HS) is a rare and aggressive form of malignant histiocytosis, often associated with poor prognosis. The diagnosis and management of HS are challenging due to the complexity of its pathogenesis, molecular profile, and the unclear cellular origin of histiocytic neoplasms, compounded by the limited literature on treatment strategies. <b>Case Presentation:</b> We report the case of a young patient with HS localized to the lymph nodes, spleen, and liver, who also presented with hemophagocytic lymphohistiocytosis (HLH) documented on bone marrow biopsy. Initial treatment with CHOEP-21 and ICE-21 chemotherapy resulted in only a partial metabolic response, as evidenced by a Fluorodeoxyglucose-Positron Emission Tomography (FDG-PET)/CT scan. Given the aggressive nature of the disease and the presence of HLH, an allogeneic hematopoietic stem cell transplantation (HSCT) from a matched unrelated donor was performed as consolidation therapy, leading to a progressive complete response without significant toxicity. A suspected relapse at 18 months post-transplant was excluded following a mediastinal lymph node biopsy, which revealed a benign intravascular papillary endothelial hyperplasia (IPEH). Over five years post-diagnosis and more than four years after transplantation, the patient remains in complete remission with full functional recovery. <b>Conclusions:</b> This case highlights the diagnostic and molecular challenges of HS and demonstrates the curative potential of early allogeneic HSCT, even when only partial remission is initially achieved.

Systemic hypertension arises from the interplay of numerous common and rare genetic variants spanning vascular, renal, endocrine, metabolic, and immune pathways. Modern genomic approaches triangulate evidence from candidate gene studies, biobank-scale genome-wide association studies (GWAS), and whole-exome or whole-genome sequencing, enabling stronger mechanistic inference. In this narrative synthesis, we focused on recent human studies emphasizing candidate gene analyses, GWAS, and sequencing efforts in hypertension, extracting data on study design, populations, key variants, and implicated biological pathways. Across methodologies, genetic evidence consistently supported central roles for endothelial nitric-oxide biology (NOS3) and oxidative or tonic regulation of arteriolar resistance (PRKG1, CYBA, and CYP4A11), alongside contributions from lipid-handling genes (ApoB and PCSK9) and mitochondrial or smooth-muscle regulators (HSG and MFN2). GWAS conducted across diverse ancestries repeatedly mapped blood pressure variation to vascular calcium dynamics (ATP2B1 and CACN* loci), renal tubular transport mechanisms (UMOD and SLC4A7), renin-angiotensin-aldosterone system-related steroidogenesis (CYP17A1 and CYP11B2), and immune remodeling pathways (SH2B3), with several loci demonstrating sex- or ancestry-specific modulation and enrichment in resistant-hypertension cohorts, particularly within calcium-handling and steroidogenic pathways. Sequencing studies further identified rare, functional, and ancestry-specific variants, including large blood pressure-lowering alleles and signals enriched in Middle Eastern populations, that refine biological mechanisms and support population-tailored risk stratification. Overall, convergent evidence across genetic approaches highlights four translationally actionable systems, such as vascular calcium handling, renal salt and bicarbonate transport, adrenal steroidogenesis, and immune or inflammatory tone, supporting the development of ancestry-aware polygenic risk tools, genetic sub-phenotyping (including resistant hypertension), and mechanism-aligned therapeutics as key steps toward precision hypertension care.

Celiac disease (CeD) is a chronic, immune-mediated enteropathy triggered by dietary gluten in genetically susceptible individuals, with environmental and epigenetic factors also contributing to its pathogenesis. Once considered a rare pediatric malabsorptive disorder, CeD is now recognized as a systemic condition that can manifest with both gastrointestinal and extraintestinal symptoms across the lifespan. Although strict adherence to a gluten-free diet (GFD) remains the cornerstone of treatment, up to 30-40% of patients experience persistent symptoms and/or ongoing mucosal injury despite reported compliance. This therapeutic gap, combined with advances in molecular understanding of disease mechanisms, has driven the development of novel strategies targeting key pathogenic pathways. Intraluminal interventions include gluten-degrading enzymes and gluten-sequestering agents, while other approaches target tissue transglutaminase 2, induce antigen-specific immune tolerance, or modulate cytokine-driven inflammation, with particular emphasis on interleukin-15 (IL-15) signaling. Additional strategies aim to inhibit lymphocyte trafficking to the intestinal mucosa and enhance intestinal barrier function through zonulin modulation. Adjunctive therapies under investigation include nutraceuticals, microbiota-targeted interventions, and vaccine-based approaches. More recently, advanced experimental and computational platforms, such as human intestinal organoids, organ-on-chip systems, and machine learning-driven analytics, are being leveraged in efforts to accelerate translational research and support the rational design of precision medicine approaches. This narrative review synthesizes current evidence for therapies beyond the GFD, examines challenges in clinical implementation, and discusses how technological innovations may reshape the future therapeutic landscape of CeD.

Transdermal drug delivery relies heavily on the skin permeability of therapeutic agents. In order to develop a peptide-based delivery strategy for promoting transdermal absorption, the key physicochemical factors influencing skin permeability are first identified through cell-penetrating peptides (CPPs) screening and computational simulation. Penetratin exhibits the most outstanding permeability and safety among CPPs from various origins, and positive surface patch area emerges as the key property correlated with skin permeability of the peptides. Based on these findings, a precise model to predict skin permeability of the peptides is established, leading to the computational redesign of penetratin's amino acid sequence. The transdermal delivery efficiency of optimized penetratin derivative (589WP) is significantly improved <i>in vitro</i> compared with wild-type penetratin and visualized through <i>in vivo</i> imaging. Furthermore, the anti-metabolic drug floxuridine (FUdR) is covalently conjugated with 589WP <i>via</i> ester linkage, leading to accelerated FUdR release due to esterase degradation. Subsequently, this conjugate is formulated into an anhydrous gel, which significantly inhibits melanoma growth with topical application, outperforming a higher dose of free FUdR without observed skin irritancy or toxicity. The peptide prediction and design approaches established herein hold great potential for advancing transdermal drug delivery.

<h4>ABSTRACT</h4> Targeted protein degradation (TPD) is a therapeutic strategy to remove disease-causing proteins by routing them to the ubiquitin-proteasome, autophagy, or lysosme machineries. For instance, proteolysis-targeting chimeras (PROTACs) are synthetic hetero-bifunctional small molecules that simultaneously bind the target and an E3 ubiquitin ligase to drive ubiquitination and degradation by the proteasome. Despite considerable success, designing such molecules is challenging and the number of currently addressable ubiquitin E3 ligases is limited. Here we demonstrate hetero-bifunctional de novo designed proteins as alternatives for TPD to access more targets and ligases. First, we develop a stable and highly adaptable helix-turn-helix scaffold for presenting different binding sites. Next, we use computational protein design to incorporate and embellish hot-spot-binding sites to target BCL-x L , plus short linear motifs (SLiMs) for KLHL20 ligase recruitment. The resulting mono- and bi-functionalised proteins bind the targets in vitro , and the latter degrade BCL-x L in cells leading to apoptosis.

Huntington’s disease (HD) is caused by expansion of a polyglutamine tract in the huntingtin (Htt) protein, leading to aggregation of the exon 1 fragment (HttEx1) into amyloid fibrils. HttEx1 forms one of the lowest-complexity amyloid cores known, its fibril core consists of a single amino acid, glutamine. With emerging therapies improving patients’ prospects by silencing expression of HTT, tools to monitor HttEx1 aggregation become essential for timely intervention and next-generation therapeutics. Here, we show that the peptide FibrilPaint1 selectively binds HttEx1Q44 fibrils without interacting with monomeric protein, allowing to measure and trace HttEx1 amyloid fibrils. Using the FibrilRuler assay, we tracked fibril formation from early species to larger clustered assemblies. The non-fluorescent variant, FibrilPaint20, was used to recruit the E3 ubiquitin ligase CHIP to HttEx1 fibrils, enabling site-specific ubiquitin tagging. However, unlike Tau fibrils, ubiquitinated HttEx1 fibrils resisted proteasomal degradation. This reveals a fundamental difference in how amyloids with extremely low–complexity cores respond to cellular clearance machinery. Together, our findings establish the FibrilPaint peptide family as a toolset for the detection and molecular targeting of amyloids, providing new opportunities to study protein aggregation and act as building blocks for future diagnostic and therapeutic strategies in neurodegenerative diseases. <h4>Highlights</h4> FibrilPaint1 selectively binds HttEx1Q44 amyloid fibrils and allows monitoring of fibril growth using the hydrodynamic radius (FibrilRuler). FibrilPaint20 recruits the E3 ligase CHIP to Htt fibrils, enabling ubiquitination. Despite successful ubiquitination, Htt fibrils resist proteasomal degradation in vitro, highlighting structural barriers. FibrilPaint provides a scaffold for functional targeting of amyloids with diagnostic and therapeutic potential. <h4>Graphical abstract</h4> Graphical abstract <h4>The FibrilRuler Test: FibrilPaint enables measurement of Huntingtin fibril size during aggregation</h4> After a short lag-phase following removal of the protective MBP tag by Factor Xa, fibrillation proceeds rapidly. Subsequent fibril clustering further accelerates growth, leading to exponential increases in aggregate size.

Horizontal gene transfer (HGT) via generalised transduction is a major driver of antimicrobial resistance (AMR) in Staphylococcus aureus , yet the genetic barriers regulating phage-mediated transfer remain poorly defined. Using a co-culture gene transfer rate assay (COGTRA) that quantifies phage-dependent resistance gene exchange under competitive, antibiotic-free conditions, we screened 1,920 mutants from the Nebraska Transposon Mutant Library for elevated transfer. We identified 32 validated high-transfer mutants that act as HGT barriers, 66% of which clustered within two chromosomal regions that we term S. aureus Transfer Islands (SauTI1 and SauTI2). SauTI loci restricted generalised transduction bidirectionally, and SauTI1 genes were widespread across S. aureus genomes yet showed lineage-specific variation. SauTI1 encodes an SMC2–Cbf1 condensin–nuclease module whose disruption markedly increases generalised transduction and plasmid transfer, resembling Wadjet-like restriction systems. Together, these findings identify clustered defence loci that restrict HGT in S. aureus and identify SauTIs as key regulators of AMR evolution.

<h4>ABSTRACT</h4> 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 AAV5, lowered mutant HTT mRNA and protein by 55-80% via its induction of frameshift-inducing 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 for a newly developed pan-HTT-targeting Cas9 system to affect HD-related phenotypes across models and provides insights into its tolerability.

<h4>Background</h4>Intestinal epithelial stem cells (SCs) and their transit-amplifying (TA) progeny are critical for mucosal repair and regeneration. However, their behaviour under chronic inflammatory conditions, such as those observed in Inflammatory Bowel Disease (IBD), remains incompletely understood.<h4>Methods</h4>We investigated the impact of chronic inflammation on intestinal stem/progenitor cells by integrating bulk RNA sequencing from the largest IBD biopsy cohort to date with single-cell transcriptomic analysis and experimental assays using patient-derived intestinal organoids.<h4>Results</h4>Active inflammation was associated with a reduction in canonical LGR5⁺ intestinal stem cells and a concurrent expansion of OLFM4⁺ populations, consistent with an inflammation-induced epithelial repair program. Notably, SC/TA cells from both inflamed and non-inflamed IBD tissues exhibited persistent transcriptional changes that were distinct from those in healthy controls. Single-cell analysis identified transcriptionally heterogeneous SC/TA subpopulations, including a previously uncharacterized inflammation-associated cluster enriched in immune signalling pathways. Pseudotime trajectory analysis demonstrated a shift in differentiation toward deep crypt secretory (Paneth-like) cell lineages under inflammatory conditions.<h4>Conclusions</h4>Chronic intestinal inflammation reshapes the epithelial stem and progenitor cell compartment, promoting altered differentiation and the emergence of immune-responsive epithelial states. These findings highlight the plasticity of the human intestinal epithelium in IBD and point to new avenues for therapeutic strategies aimed at maintaining epithelial integrity during chronic inflammation.