Gene Literature Dashboard

Glioblastoma (GBM) is one of the deadliest cancers, and the survival rate has remained low for decades. The aim of the study was the construction of the programmed death-ligand 1 (PD-L1) network, identification of its interactors and over-represented pathways, and analysis of the association between the identified genes and the immunosuppressive microenvironment of GBM. The PD-L1 network was constructed using Cytoscape and Search Tool for the Retrieval of Interacting Genes/Proteins (STRING). Over-representation analysis was performed on WebGestalt using Kyoto Encyclopedia of Genes and Genomes (KEGG), Protein ANalysis THrough Evolutionary Relationships (Panther), and Reactome Pathway Database (Reactome). Gene expression levels were examined in silico using three large datasets (The Cancer Genome Atlas (TCGA), Chinese Glioma Genome Atlas (CGGA), and Rembrandt), as well as with qPCR. The association between PD-L1 gene expression and immune cell infiltration was analyzed using the Tumor Immune Estimation Resource (TIMER 2.0) online tool. Cluster of differentiation 44 (CD44) and tumor necrosis factor superfamily member 14 (TNFSF14) were found to be significantly overexpressed in GBM compared to lower-grade glioma (LGG) and normal brain tissue. Their overexpression was associated with worse overall survival and demonstrated a strong ability to differentiate between GBM and reference brain tissue. Notably, CD44 and TNFSF14 were linked to the mesenchymal subtype of GBM and positively correlated with the presence of regulatory T cells, resting natural killer (NK) cells, and PD-L1 expression. Our findings highlight the overexpression of CD44 and TNFSF14 in GBM and their potential involvement in creating an immunosuppressive microenvironment. Unraveling the PD-L1 interaction network and its associated pathways offers the potential not only to identify novel biomarkers for GBM prognosis but also to pinpoint alternative therapeutic targets that could be more effective in overcoming the immunosuppressive hurdles inherent in GBM treatment.

A protective HIV vaccine will need to induce broadly neutralizing antibodies (bnAbs) in humans, but priming rare bnAb precursor B cells has been challenging. In a double-blinded, placebo-controlled phase 1 human clinical trial, the recombinant, germline-targeting envelope glycoprotein (Env) trimer BG505 SOSIP.v4.1-GT1.1, adjuvanted with AS01_B, induced bnAb precursors of the VRC01-class at a high frequency in the majority of vaccine recipients. These bnAb precursors, which target the CD4 receptor binding site, had undergone somatic hypermutation characteristic of the VRC01-class. A subset of isolated VRC01-class monoclonal antibodies neutralized wild-type pseudoviruses and was structurally extremely similar to bnAb VRC01. These results further support germline-targeting approaches for human HIV vaccine design and demonstrate atomic-level manipulation of B cell responses with rational vaccine design.

ADP-ribosyl-transferases (ARTs) are versatile post-translational regulators. Mammalian ARTs include poly- and mono-ADP-ribosylating enzymes, which transfer ADP-ribose molecules deriving from β-NAD+ to their targets. Mono-ADP-ribosylation (MARylation), which is catalyzed by mono-ARTs such as PARP3, PARP6-PARP12 and PARP14-PARP16, tunes the activity of targets involved in fundamental cell processes and various signaling pathways, ranging from those regulating cell survival and proliferation to those modulating the cellular response to stress and viral infection. Recent advancements of techniques that enable the discovery of MARylation targets across cellular compartments have further expanded our knowledge about the physiological roles of these targets and the potential connection between MARylation and the onset of pathologies. Furthermore, increasing efforts in the development of specific drugs targeting the different MARylating PARP proteins are opening avenues for innovative pharmacological treatments. In this Review, we illustrate the cell cycle progression, intracellular membrane trafficking and cellular stress pathways regulated by mono-ART PARP proteins. We then describe what is known about the roles of MARylating PARP proteins in the context of viral infection and cancer. Finally, we discuss potential future directions towards mapping out the complex network of PARP targets and functions.

Viral protein U (Vpu), an accessory protein of HIV-1, functions by antagonizing or hijacking various host factors to allow the virus to evade host immune surveillance and facilitate viral release. Nevertheless, there is limited understanding regarding the impact of Vpu on the biogenesis of HIV-1 RNAs. In this study, we utilized ascorbate peroxidase 2 (APEX2)-based proximity labeling techniques in combination with mass spectrometry and immunoprecipitation-mass spectrometry (IP-MS) to characterize interactions between HIV-1 Vpu and host proteins. We identified nine cellular targets of Vpu. Among these targets, our research demonstrated the interaction between Vpu and RNA-binding motif protein 10 (RBM10), which results in the degradation of RBM10 through the ubiquitin-proteasome pathway. The expression of RBM10 exerted an inhibitory effect on virus replication by binding to viral RNA and reducing the levels of incompletely spliced HIV-1 transcripts. Additionally, it promoted the transcription of various antiviral genes. The findings elucidate the role of HIV-1 Vpu in RNA replication and identify RBM10 as a novel regulator of HIV-1 transcription.IMPORTANCEA comprehensive analysis utilizing APEX2-MS and IP-MS techniques identified a total of 24 cellular targets of Vpu, three of which have been documented as restriction factors. Vpu-interacting proteins were found to be significantly enriched in pathways related to cell adhesion, RNA transport, and the spliceosome. The identification of RBM10 as a novel regulator of HIV-1 replication and infectivity, and RBM10 regulated transcription of both viral and host RNA transcripts. Vpu interacted with RBM10 and promoted its degradation through the ubiquitin-proteasome pathway.

<h4>Background</h4>Preoperative biliary drainage (PBD) may be performed for jaundiced patients with periampullary tumors. This study aimed to evaluate the impact of PBD on biliary microbiome and perioperative complications in patients undergoing pancreaticoduodenectomy (PD).<h4>Methods</h4>This retrospective study enrolled 323 patients who underwent PD between March 2018 and March 2024. Intraoperative bile specimens were obtained for microbiological analysis of species identification and antimicrobial resistance patterns..<h4>Results</h4>PBD was performed in 191 (59.1%) of the 323 patients. Organ/space surgical site infection (SSI) (51.8% vs 37.9%, <i>p</i> < 0.001) and bacterial colonization of bile (90.6% vs 28.0%, <i>p</i> < 0.001) were significantly more frequent in patients with PBD. PBD was identified as an independent risk factor of organ/space SSI (OR = 1.837, 95% CI: 1.158-2.916, <i>p</i> = 0.010) and associated with increased contamination with polymicrobial mixed flora (47.1% vs 4.5%, <i>p</i> < 0.001), <i>K. pneumoniae</i> (23.6% vs 0.8%, <i>p</i> < 0.001), <i>E. faecalis</i> (14.1% vs 1.5%, <i>p</i> < 0.001), <i>E. faecium</i> (6.8% vs 0.8%, <i>p</i> = 0.009). This shift corresponded to higher resistance to piperacillin-tazobactam (23.1% vs 0.0%, <i>p</i> = 0.038), cefoperazone-sulbactam (25.3% vs 0.0%, <i>p</i> = 0.021), ciprofloxacin (36.1% vs 6.3%, <i>p</i> = 0.006), and levofloxacin (47.4% vs 4.8%, <i>p</i> < 0.001). Patients with positive bile culture had a significantly higher occurrence of organ/space SSI than the negative group (53.3% vs 32.7%, <i>p</i> < 0.001). <i>K. pneumoniae</i> was identified as an independent risk factor for organ/space SSI (OR = 2.636, 95% CI: 1.353-5.137, <i>p</i> = 0.004).<h4>Conclusions</h4>There were fundamental differences in the bile microbiome profile and antibiotic resistance of patients with/without PBD. These findings suggest that adjusting perioperative antibiotic regimens based on biliary culture may be warranted.

We introduce a method for the 6,5-hydrofunctionalization of 2-azatrienes with <i>N</i>-phosphinoyl imine electrophiles under CuH catalysis to prepare 1,4-diamines bearing two stereogenic centers with excellent regio-, diastereo-, and enantiocontrol. In comparison to previously disclosed 6,3-hydrofunctionalizations of azatrienes with (Ph-BPE)CuH or (<i>t</i>-Bu-BDPP)CuH, the use of Josiphos SL-J001-1 as the supporting ligand facilitates regiodivergent introduction of the electrophile. We identify the electronic character of each phosphine of the ligand as critical to the observed regioselectivity and the utilization of an <i>N</i>-phosphinoyl group on the imine to be essential. Steric modification of the azatriene's activating group may enhance regioselectivity in challenging cases. Electronic modification of the <i>N</i>-phosphinoyl group can improve chemoselectivity to favor reductive coupling over imine reduction in several instances, a discovery that translates to couplings with other copper catalysts in forming 1,2-diamines. Experiments illustrate the delicate balance in favoring 1,4-diamine formation over all other processes and, along with density functional theory (DFT) calculations, lead us to a mechanistic proposal for this catalytic reaction.

A widely held view is that an increased pulmonary arterial (PA) blood flow at birth is not triggered until the onset of lung aeration, but experimental data indicate that the non-ventilatory events of a reduction in lung liquid volume, complete fetal delivery, and umbilical cord clamping can also increase fetal PA flow. However, the effect of cord clamping strategy on the contribution of these non-ventilatory events to birth-related rises in PA flow is unknown. Accordingly, PA blood flow was measured via transit-time flow probe in anaesthetized, acutely instrumented preterm fetal lambs at baseline, after a ∼35% reduction in lung liquid volume, following complete fetal delivery, and then after (1) delayed cord clamping (DCC) preceded by ventilation lasting ∼100 s (n = 11), or (2) early cord clamping (ECC) followed by either a non-asphyxial (∼35 s, n = 10) or an asphyxial interval (∼100 s, PO2${P_{{{\mathrm{O}}_2}}}$  < 10 mmHg, n = 10) before ventilation. PA flow rose stepwise after reduction of lung liquid volume (P < 0.001) and fetal delivery (P < 0.001), as well as initial ventilation (P < 0.001) and subsequent DCC (P = 0.002). PA flow also rose after ECC (P < 0.001), with flow maintained in the non-asphyxial group, but markedly reduced to near-baseline fetal levels by pulmonary vasoconstriction in the asphyxial group (P = 0.009), before rising with ventilation (P < 0.001). Overall, non-ventilatory events cumulatively accounted for ∼30% of the fetal baseline-to-peak newborn increment in PA flow. These findings suggest that (1) non-ventilatory events substantially contribute to a perinatal rise in PA blood flow with ECC or DCC, and (2) this contribution is negated if an asphyxial level of arterial oxygenation develops after ECC. KEY POINTS: Although a widely held view is that an increased pulmonary blood flow (PBF) at birth is not triggered until onset of lung aeration, reduction of lung liquid volume, complete fetal delivery and umbilical cord clamping also increase fetal PBF. The contribution of these non-ventilatory events to birth-related rises in PBF is unknown, particularly with different cord clamping strategies. Anaesthetized preterm fetal lambs instrumented with central arterial flow probes underwent birth via delayed cord clamping (DCC) preceded by ventilation, or early cord clamping (ECC) followed by either a non-asphyxial or asphyxial interval before ventilation. PBF rose with reduction of lung liquid volume, fetal delivery, ECC and DCC. However, while an increased PBF after ECC was maintained with a non-asphyxial interval, it fell markedly after ECC with an asphyxial interval, before rebounding with ventilation. Cumulatively, non-ventilatory events accounted for ∼ 30% of the perinatal increase in PBF occurring with DCC or ECC birth strategies.

<h4>Background</h4>Lung adenocarcinoma (LUAD) is the most common subtype of non-small cell lung cancer. Although immune checkpoint inhibitors (ICIs) have brought new treatment options for advanced patients, a considerable proportion still shows limited response. Mitochondrial dysfunction plays a crucial role in tumor development and immune evasion, but its regulatory mechanisms in LUAD immune microenvironment remain unclear.<h4>Methods</h4>We integrated 149 mitochondria-related pathways (1,136 coding proteins) to develop and validate the Mitochondrial Pathway Signature (MitoPS) using machine learning approaches across seven independent LUAD cohorts (n=1,231). The system was systematically compared with 129 published LUAD prognostic signatures and validated in seven immunotherapy cohorts (n=451). Multiomics analysis, immunofluorescence staining, and experimental validation were performed to investigate its molecular mechanism.<h4>Results</h4>MitoPS demonstrated consistent predictive performance across validation cohorts, with high scores indicating poor prognosis, outperforming 129 existing prognostic models. In immunotherapy cohorts, MitoPS reliably predicted treatment response and prognosis. Immune microenvironment analysis revealed that low MitoPS scores correlated with higher immune cell infiltration and active immune function. Mechanistic studies identified mitochondria-related gene NDUFB10 as a core gene of MitoPS (r=0.38, p<0.05), where its high expression was significantly associated with immune desert phenotype and worse prognosis. Functional experiments confirmed that NDUFB10 knockdown significantly enhanced ICIs therapy and increased GZMB+CD8+T cell infiltration, indicating NDUFB10's crucial role in regulating tumor immune microenvironment and immunotherapy response.<h4>Conclusion</h4>The MitoPS scoring system reliably predicts prognosis and immunotherapy response in patients with LUAD, providing a novel reference for clinical decision-making. Furthermore, its core gene NDUFB10 regulates tumor immune microenvironment, offering a potential therapeutic target for improving immunotherapy outcomes.

Cryogenic transmission electron microscopy (cryo-TEM) enables the visualization of liquid-phase materials, including nanoparticles and soft/biomaterials, under cryogenic conditions while minimizing radiation damage. Cryo-TEM imaging provides insights into particle size, shape, and dispersion. Beyond such conventional structural information, acquiring elemental composition data allows for a more detailed analysis and evaluation. In this study, we developed a method for elemental mapping of nanoparticles and soft/biomaterials in frozen solvent by integrating electron energy loss spectroscopy (EELS) with energy-filtered (EF) cryo-EM. Cryo-EELS and analysis were performed using the three-window method, following the alignment and correction of stage drift during image acquisition and interexposure drift between different energy windows. This approach, by balancing accurate and reliable signal extraction with electron dose, enabled the generation of elemental maps for nanoparticles as small as 10 nm in frozen solvent. Furthermore, we extended the technique to protein-coated silica nanoparticles and hydroxyapatite (HAp) nanoparticles in vitrified solvent, selecting specific energy loss values to identify multiple constituent elements. As a result, we successfully mapped silica from the nanoparticle cores, carbon from the protein shell of the nanoparticles, and phosphorus and calcium─key light elements in biological systems─within the same imaging area of the HAp particles.

Sphingolipids are essential, complex lipids that are abundant in the cell membranes of eukaryotic cells, particularly concentrated in the myelin and neuronal membranes of the central nervous system (CNS). These lipids are crucial components of the cell membrane, affecting their structure and fluidity, and thus regulating various biological processes, including signal transduction, cell differentiation, apoptosis, and autophagy. The metabolic pathways of sphingolipids are highly complex and conserved, and this metabolic process can produce multiple metabolites. Metabolites such as ceramide (Cer) and sphingosine-1-phosphate (S1P) are vital in CNS signaling, affecting neurodevelopment, myelination, and synaptic plasticity. Thus, disruption of sphingolipid metabolism is closely related to neurological disorders. This article provides the latest studies concerning the known features of sphingolipid and sphingolipid metabolism, highlighting its physiological and pathological roles in the CNS.

CRISPR-Cas-based genome imaging opened a new era of genome visualization in living cells. While genomic loci with repetitive sequences, such as centromeres and telomeres, can be reliably imaged, applying the technique to nonrepetitive genomic loci has remained challenging. Recent advancements in the design of CRISPR RNAs and Cas proteins, the development of novel fluorophores and the combination of CRISPR-Cas with other molecular machinery amplified target-specific signals and suppressed background signals, revolutionizing this unique genome imaging technique and enabling the tracking of genomic loci with a small number of CRISPR-Cas complexes, down to a single complex. Here we review the latest advancements in CRISPR-Cas-based genome imaging techniques and their application to imaging nonrepetitive genomic loci. The challenges that these techniques are currently facing are the cellular toxicity and genomic instability induced by the expression of CRISPR-Cas and its interference with DNA metabolism, which impacts DNA replication and genome maintenance. Recently reported adverse effects of CRISPR-Cas-based genome labeling are discussed here, along with perspectives on how to overcome the problem.

<h4>Background</h4>PPCS deficiency disorder (PPCS DD) is an ultra-rare, autosomal recessive form of dilated cardiomyopathy (DCM) caused by pathogenic variants in PPCS, which encodes the enzyme catalyzing the second step in the coenzyme A (CoA) biosynthesis pathway. To date, only six patients worldwide have been identified.<h4>Methods</h4>Whole-exome sequencing was performed to identify pathogenic PPCS variants in affected individuals. Protein stability was assessed by Western blotting. CoA levels were quantified using a microplate-based assay in patient-derived fibroblasts, cardiac progenitor cells, and cardiomyocytes. Functional evaluation of cardiac cells and engineered heart patches was conducted to investigate contractile performance and arrhythmogenicity. Pantethine was tested as a potential therapeutic agent both in vitro and through long-term clinical follow-up in patients.<h4>Results</h4>Causative PPCS variants are identified in six individuals with DCM and variable associated features, including neuromuscular and neurological symptoms. Identified variants lead to reduced PPCS protein stability and decreased cellular CoA levels. Cardiac cells exhibit impaired contractility and arrhythmias, which are partially rescued by pantethine treatment. Clinically, patients receiving pantethine show sustained improvement over time.<h4>Conclusions</h4>Our study expands the genetic and clinical spectrum of PPCS deficiency disorder, identifying six new cases with diverse phenotypes. Functional investigations reveal reduced CoA levels and dysfunction in patient-derived cardiac cells. Pantethine treatment shows promise in partially rescuing DCM phenotypes, both in vitro and in patients. However, complete reversal may require early intervention. These findings underscore the importance of timely diagnosis and treatment in PPCS DD. Future research should focus on optimizing pantethine supplementation and exploring additional therapies to enhance CoA levels and cardiac function in affected individuals.

<h4>Introduction</h4>People with metabolic-dysfunction associated steatotic liver disease (MASLD) had higher risk of extrahepatic multimorbidity, and fibrosis is the strongest prognostic factor for mortality in MASLD. This study aimed to investigate how fibrosis was associated with multimorbidity and their relationships with all-cause mortality.<h4>Methods</h4>We utilized data from the UK Biobank. MASLD was identified via a Fatty Liver Index ≥ 60 and the presence of cardiometabolic risk factors. The fibrosis-4 (FIB4) score was used to measure liver fibrosis. Multimorbidity was defined as having two or more long-term conditions (LTCs) from a prespecified list of 47 LTCs. Logistic regressions estimated the cross-sectional association between FIB4 scores and multimorbidity prevalence, while Cox models assessed the prospective association between FIB4 scores, multimorbidity and mortality.<h4>Results</h4>We included 127,470 participants with MASLD (41.7% female, age 57.4 years, 21.3% with multimorbidity, 2.2% with high FIB4 scores). 14,471 deaths were recorded during 13-year follow-up. Compared to low FIB4 scores, high FIB4 scores were associated with 41% higher prevalence of multimorbidity (OR 1.41 (95%CI 1.30-1.54)), and 94% higher all-cause mortality (HR 1.94 (95%CI 1.77-2.13)), while adjusting for multimorbidity reduced the association by 10%, primarily driven by contributions from cardiovascular diseases, extrahepatic cancers, and chronic kidney disease.<h4>Conclusion</h4>FIB4 scores were positively associated with higher multimorbidity and mortality in MASLD patients. Adjustment for multimorbidity reduced 10% the relationship between fibrosis and mortality, with cardiovascular diseases, cancers and chronic kidney disease contributing notably to this reduction. These findings underscore the importance of managing both fibrosis and multimorbidity in MASLD patients.

Neuroendocrine prostate cancer (NEPC), an aggressive and highly malignant subtype of castration-resistant prostate cancer (CRPC), arises through drug resistance mechanisms involving genomic alterations, epigenetic remodeling, tumor microenvironment (TME) reprogramming, and lineage plasticity. A hallmark of NEPC is its independence from the androgen receptor (AR) pathway, evidenced by diminished or absent AR expression-a key barrier to effective clinical management. Therefore, a comprehensive understanding of NEPC pathogenesis and progression is essential, as it facilitates the identification of potential therapeutic targets and the development of more effective therapies. In this review, we summarize the typical characteristics of NEPC and describe its clinical diagnosis, relevant imaging modalities, current treatment strategies, and associated therapeutic difficulties. We also highlight the core events driving NEPC formation. Furthermore, we discuss potential therapeutic targets for the disease and review pharmacological agents that have demonstrated efficacy against NEPC, aiming to offer innovative perspectives and potential research directions for future NEPC treatment.

Astrocytes play a vital role in maintaining homeostasis and function in the central nervous system, including their involvement in reparative processes. Here, we examined how a common mutation in the homeostatic iron regulatory gene, H67D HFE, impacts the antioxidant mechanism of astrocytes and migration under normal and reparative conditions. Previous data from our group suggested that this mutation may modify disease progression through an antioxidant mechanism involving nuclear factor erythroid 2-related factor 2 (Nrf2), glutathione peroxidase 4 (GPX4), and ferritin. In this study, we used primary murine astrocytes with either the H67D or wild-type HFE genotype to determine whether astrocytes contribute to the antioxidant protective mechanism previously observed. We analyzed their antioxidant profile and migration both at baseline and after a scratch wound injury. We found that H67D HFE astrocytes expressed the HFE protein and exhibited an enhanced antioxidant profile, marked by increased glutathione and GPX4 at baseline, and a reduced migration length into three-dimensional granular hydrogel scaffolds. However, following scratch wound injury, these astrocytes exhibited a shift in migratory behavior, leading to faster wound infiltration. Moreover, their antioxidant response became even more pronounced after injury, with increased expression of Nrf2, GPX4, and H-ferritin (FTH1). These results suggest that the mechanism underlying HFE mutation neuroprotection in disease processes involves an antioxidant profile in astrocytes, which is increased upon insult to activate the astrocytic reparative mechanism.

Lung adenocarcinomas (LUAD) are a pressing global health problem with enduring lethality and rapidly shifting epidemiology. Proteogenomic studies integrating proteomics and post-translational modifications with genomics can identify clinical strata and oncogenic mechanisms, but have been underpowered to examine effects of ethnicity, smoking and environmental exposures, or sex on this heterogeneous disease. This comprehensive proteogenomic analysis of LUAD tumors and matched normal adjacent tissues from 406 patients across diverse geographic and demographic backgrounds explores the impact of understudied driver mutations, prognostic role of chromosomal instability, patterns of immune signaling, differential and sex-specific effects of endogenous mutagens and environmental carcinogens, and pathobiology of early-stage tumors with "late-like" characteristics. Candidate protein biomarkers are proposed for unstable tumors with highly fragmented genomes and for carcinogen exposures, and a LUAD subtype-specific atlas of therapeutic vulnerabilities is presented. These observations and the associated data resource advance the objective of precision management strategies for this devastating disease.

Immunotherapy has shown limited success in recurrent ovarian cancer (OC), with prognostic insights largely derived from treatment-naive tumors. We analyzed 697 tumor samples (566 primary and 131 recurrent) from 595 OC patients across five independent cohorts, capturing tumor-infiltrating lymphocytes (TILs) heterogeneity and identifying four immune phenotypes linked to prognosis and TIL:myeloid networks driving malignant progression. We found that in preclinical mouse models, mirroring inflamed human OCs, the recurrent Brca1^mut tumors maintained activated TILs:dendritic cells (DCs) niches but evaded immune control through upregulation of COX/PGE₂ signaling. Conversely, recurrent Brca1^wt tumors displayed loss of TILs:DCs niches and accumulated immunosuppressive tumor microenvironment (TME) networks featuring Trem2/ApoE^high tumor associated macrophages (TAMs) and Nduf4l2^high/Galectin3^high malignant states. Recurrent tumors recapitulate the immunogenic landscapes of original cancers. Our findings reveal BRCA-dependent TIL:myeloid crosstalk as key to persistent immunogenicity in recurrent OC and propose new targets to enhance chemotherapy efficacy.

<h4>Background</h4>Psychiatric symptoms are increasingly recognized as early manifestations of Alzheimer's disease (AD). These symptoms may reflect or contribute to underlying neurobiological changes, including tau burden, which represents more advanced AD pathology. Understanding factors associated with tau burden may help identify individuals at elevated risk and improve early detection strategies.<h4>Objectives</h4>To investigate the temporal relationships between neuropsychiatric symptoms, demographic factors, and tau burden, by examining amyloid (Aβ)-dependent, -independent, and interactive associations.<h4>Design</h4>Retrospective cohort study.<h4>Setting</h4>Alzheimer's Disease Neuroimaging Initiative.<h4>Participants</h4>We included 681 participants without dementia (mean age = 71.2 years, 51.8 % female).<h4>Measurements</h4>The participants underwent tau PET scanning with prior amyloid PET and Neuropsychiatric Inventory (NPI) interview assessments clustered around three time periods relative to tau PET: closest (0 - 2 years), mid (3 - 5 years), and furthest (6 - 8 years). Linear regression analyses, adjusting for age and APOE ε4 alleles, examined associations of NPI scores, sex, education, and their Aβ-status interactions with tau burden.<h4>Results</h4>Higher total NPI scores up to 2 years prior to tau PET were associated with greater tau burden independently of Aβ status, whereas anxiety symptoms demonstrated an Aβ-dependent relationship with tau. (NPI: β=0.117, 95 % CI: 0.049 to 0.185, p=0.001, Anxiety: β=0.249, 95 % CI: 0.073 to 0.424, p=0.006). NPI measured up to 5 years prior to tau PET interacted with Aβ on tau burden (0-2 years: β=0.272, 95 % CI: 0.136 to 0.407, p<0.001, 3-5 years: β=0.336, 95 % CI: 0.127 to 0.544, p=0.002). Sex and education showed minimal associations with tau at uncorrected statistical levels.<h4>Conclusions</h4>Neuropsychiatric symptoms were associated with tau burden up to two years before tau sampling, independently of Aβ, and interacted with Aβ status up to five years prior, suggesting that neuropsychiatric symptoms are related to tau in the short term and may represent manifestations of advancing AD pathology. Demographic factors showed minimal associations. These findings highlight the importance of evaluating neuropsychiatric and anxiety symptoms as potential indicators of increased tau pathology.

Huntington's disease (HD) is an autosomal-dominant inherited neurological disorder caused by an unstable trinucleotide CAG repeat expansion at the N-terminus of the IT-15 gene, which encodes the ∼350 kDa huntingtin protein (Htt). This mutation confers toxic properties, promoting neuronal dysfunction and death through multiple mechanisms. Effective treatments for HD are still lacking. Therefore, early-stage interventions could represent a promising approach to addressing the pathological aspects of HD. Recently, our laboratory explored the potential neuroprotective effect of murtilla (Ugni molinae, Turcz) fruit extract (ETE 19-1) in an HD cellular model and observed a significant anti-aggregation effect on mutant Htt (mHtt). In this study, we determine the beneficial effects of ETE 19-1 on HD progression in both brain and gut tissues. Our results show that a ETE 19-1 reduces motor impairment, decreases mHtt levels, and mitigates microglia activation in the striatum brain region of R6/2 mice. Moreover, we observed a reduction in gastrointestinal inflammation and an increase the gut microbiota biodiversity in R6/2 mice treated with ETE 19-1 extract. These findings highlight the impact of polyphenol-enriched natural products on the brain and gut inflammatory process during the progression of neurodegenerative diseases.

Ebola virus (EBOV) is a highly virulent non-segmented, negative-sense RNA virus and a causative agent of severe, often fatal disease in humans. EBOV encodes 7 genes and viral protein 35 (VP35) plays a key role in EBOV pathogenesis. VP35 is an EBOV polymerase cofactor that facilitates host immune evasion, viral RNA synthesis, and nucleocapsid assembly. Over the past 25 years since the first recognition that VP35 can antagonize innate immune responses, considerable progress has been made to define the molecular mechanisms by which VP35 mediates its multiple functions and interactions with host factors. Much of this work is based on our understanding of the VP35 sequence and structure as it relates to function. Here, we will provide a comprehensive review of the VP35 structure and known insights into function. Given its significant role, VP35 has also emerged as a therapeutic target for the development of countermeasures against EBOV.

<h4>Introduction</h4>We recently developed a new therapy using effective-mononuclear cells (E-MNCs) and demonstrated its efficacy in treating radiation-damaged salivary glands (SGs). The activity of E-MNCs in part involves constituent immunoregulatory -CD11b/macrophage scavenger receptor 1(Msr1)-positive-M2 macrophages, which exert anti-inflammatory and tissue-regenerating effects via phagocytic clearance of extracellular high mobility group box 1 (HMGB1). Focusing on the phenomena, this study investigated significance of regulating the HMGB1/toll-like receptor 4 (TLR4)/receptor for advanced glycation end products (RAGE) signaling pathway in the treatment of SG dysfunction caused by radiation damage.<h4>Methods</h4>E-MNCs were transplanted into radiation-damaged mice SGs, and changes of TLR4/RAGE expression were observed. Furthermore, the activation of downstream signals was investigated in both intact SGs and cultured SG epithelial cells after irradiation. Subsequently, TLR4-knock-out (KO) mice were employed to examine how HMGB1/TLR4/RAGE signaling affected damage progression.<h4>Results</h4>Expression of both TLR4 and RAGE was diminished in ductal cells and macrophages/vascular endothelial cells of damaged SGs with E-MNC transplantation, respectively. Meanwhile, expression of TLR2/4 and RAGE in damaged SGs markedly increased in association with extracellular HMGB1 accumulation. Downstream signals were activated, and intranuclear localization of phospho-nuclear factor-kappa B (p-NF-KB) in ductal cells and production of IL-6, tumor necrosis factor-α (TNF-α), and interferon-γ (IFN-γ) were observed. Additionally, culture supernatant of irradiated cultured SG epithelial cells contained damaged associated molecular pattern (DAMP)/senescence-associated secretory phenotype (SASP) factors. Treatment of cultured SG epithelial cells with this supernatant activated TLR4 signaling pathway and induced cellular senescence. In TLR4-KO mice, onset of radiogenic SG dysfunction was markedly delayed. However, TLR2/RAGE signalings were alternatively activated, and SG function was impaired.<h4>Conclusions</h4>Clearance of DAMPs such as HMGB1 may attenuate sterile inflammation in damaged SGs via suppression of the TLR4/RAGE signaling pathway. This cellular mechanism may have significant implications for the development of future cell-based regenerative therapies.

Glucose is the main source of energy and the source of carbon for the biosynthesis of several molecules, such as neurotransmitters, for most mammalian cells. Therefore, the transport of glucose into cells is very important. There are described three distinct families of glucose transporters: facilitative glucose transporters (GLUTs), sodium-dependent glucose cotransporters (SGLTs), and a uniporter, the SWEET protein. Impaired function and/or expression of these transporters due to, for example, mutations in their genes, may cause severe diseases. Associations with the impaired function of glucose transporters have been described in the case of neurodegenerative diseases (NDs) such as Alzheimer's disease, Parkinson's disease, Huntington's disease, GLUT1-deficiency syndrome, stroke, and traumatic brain injury. Changes in the presence of glucose transporters may be a cause of NDs, and they may be the effect of NDs. On the other hand, in many cases of neurodegenerative diseases, changes in the expression of glucose transporters may be a targeted therapy in the treatment of patients with these diseases.

Accounting for 15-30% of colorectal cancer cases, the serrated pathway remains poorly characterized compared to the adenoma-carcinoma sequence. It involves sessile serrated lesions as precursors and is characterized by BRAF mutations (BRAF^V600E), CpG island hypermethylation, and microsatellite instability (MSI). Using label-free proteomics, we compared normal tissue margins from patients with diverticular disease, sessile serrated lesions, low-grade adenomas, and high-grade adenomas. We identified S100A14 as significantly overexpressed in sessile serrated lesions compared to low-grade adenomas, high-grade adenomas, and normal tissues. This overexpression was confirmed by immunohistochemical scoring in an independent cohort. Gene expression analyses of public datasets showed higher S100A14 expression in BRAF^V600E-mutated and MSI-H colorectal cancers compared to microsatellite stable BRAF^wt tumors. This finding was confirmed by immunohistochemical scoring in an independent colorectal cancer cohort. Furthermore, single-cell RNA sequencing analysis from the Human Colon Cancer Atlas revealed that S100A14 expression in tumor cells positively correlated with the abundance of tumoral CD8⁺ cytotoxic T cells, particularly the CD8⁺ CXCL13⁺ subset, known for its association with a favorable response to immunotherapy. Collectively, our results demonstrate for the first time that S100A14 is a potential biomarker of serrated neoplasia and further suggests its potential role in predicting immunotherapy responses in colorectal cancer.

The glucagon-like peptide-1 receptor (GLP-1R), which belongs to the class B1 G protein-coupled receptor (GPCR) family, is an important target for treatment of metabolic disorders, including type 2 diabetes and obesity. The growing interest in GLP-1R-based therapies is driven by the development of various functional agonists as well as the huge commercial market. Thus, understanding the structural details of ligand-induced signaling are important for developing improved GLP-1R drugs. Here, we investigated the conformational dynamics of the receptor in complex with a selection of prototypical functional agonists, including CHU-128 (small molecule-biased), danuglipron (small molecule balanced), and Peptide 19 (peptide balanced), which exhibit unique, distinct binding modes and induced helix packing. Furthermore, our all-atom molecular dynamics (MD) simulations revealed atomic feature how different those ligands led to signaling pathway preference. Our findings offer valuable insights into the mechanistic principle of GLP-1R activation, which are helpful for the rational design of next-generation GLP-1R drug molecules.

Phylum Annelida are ubiquitous metazoans found in almost every terrestrial and aquatic habitat on Earth. Historically, taxonomic studies on the phylum have been focused largely on its majorgroups, polychaetes, oligochaetes and leeches, so that while family-level keys for each group are available, no single-source identification guide exists to the world's annelid families. Here, the first illustrated linear key to annelid families is provided and family-level descriptions and diagnoses that distinguish individuals of each family from those of other families in the phylum are updated. This information is generated from an annelid DELTA database of 334 characters and 166 mostly family-level taxa. A link is provided to downloadable software (ANNiKEY Interactive) allowing the same data to be interrogated using the open-source DELTA program Intkey, which enables both interactive identification and taxonomic query functionality. For each family-level taxon, a diagnosis, full description, links to taxonomic data at the World Register of Marine Species, illustrations of diagnostic features, and a summary of the recent literature, including a list of published keys to genera and species are provided.

Although malaria has been effectively controlled, it still poses a threat to global health. Artemisinins are the first-line antimalarial drugs. However, their therapeutic efficacy is significantly limited by poor solubility and short biological half-life. To overcome these limitations and enhance drug accumulation in <i>Plasmodium</i>, we developed a glucose-functionalized redox-responsive dihydroartemisinin (DHA) prodrug nanosystem (D@GLU-PMs-SS). The nanosystem was prepared by using DHA-dithiodipropionic acid-octadecylamine prodrug and D-α-Tocopherol polyethylene glycol 1000 succinate-arbutin conjugate. The resultant D@GLU-PMs-SS exhibited excellent stability under conditions of storage and physiological environment. D@GLU-PMs-SS could be activated by glutathione (GSH), leading to the dissociation of nanoparticles and subsequent release of free DHA. <i>In vitro</i> experiments revealed that the host erythrocyte uptake of glucose-functionalized nanoparticles was significantly enhanced <i>via</i> GLUT-mediated transport. Cellular experiments illustrated that D@GLU-PMs-SS effectively reduced GSH concentrations in <i>Plasmodium</i>. Furthermore, D@GLU-PMs-SS displayed remarkable efficacy in inhibiting the growth of <i>Plasmodium</i> while maintaining biosafety. Overall, this study developed a strategy to enhance the targeting of nanoparticles to improve their therapeutic efficacy against malaria, warranting further investigation in clinical trials.

Chronic kidney disease (CKD) affects an estimated 15% of all adults in Central Europe. Those affected are at high risk of cardiovascular disease and death. Inflammatory rheumatic systemic diseases manifest themselves extra-articularly with varying frequency. This article summarized the prevalence and pathogenetic mechanisms of CKD in rheumatic systemic diseases. The following databases were searched for references: PubMed, Web of Science, Cochrane Library, Scopus. The search period spanned from 1975 to 2025. Kidney involvement is almost always present in systemic lupus erythematosus and certain types of systemic vasculitis. In the context of rheumatic diseases, there are additional mechanisms that can contribute to enhancing the functional and structural integrity of the kidneys. These mechanisms include inflammation and an increase in cardiovascular risk. The prevalence of CKD is disproportionately high in certain entities of the rheumatic form. Given the disproportionately high prevalence of CKD in relevant entities of the inflammatory rheumatic group and the associated increase in the risk of cardiovascular disease and death, CKD screening should be an integral part of the care of affected patients.

Arfs are small Ras-superfamily proteins important for regulating membrane trafficking including the recruitment of vesicular coats as well as a diverse range of other functions. There are five Arfs in humans: two Class I Arfs (Arf1 and Arf3), two Class II Arfs (Arf4 and Arf5) and one Class III Arf (Arf6), with Class I and Class II Arfs present on the Golgi apparatus among other locations. These Golgi Arfs (Arf1, Arf3, Arf4 and Arf5) are highly similar in sequence, and knockout studies have established a complex pattern of redundancy, with Arf4 alone able to support cell survival in tissue culture. Moreover, adding to the complexity, functions of Arfs on distinct membranes can involve non-overlapping sets of effectors (e.g., COPI on <i>cis</i>-Golgi membranes and clathrin adaptors on <i>trans</i>-Golgi network). The three classes of Arfs are found in most metazoans, suggesting biologically important specialization the details of which are beginning to emerge. This review examines recent studies using siRNA and CRISPR/Cas9 knockouts of mammalian Arfs combined with functional assays of the secretory pathway in the context of detailed localization of fluorescently-tagged Arfs by fluorescent and super-resolution microscopy and the existing literature using more conventional techniques. We suggest that specificity of effector recruitment involves additional membrane determinants which need to be considered in future studies.

Tumor immunosuppression remains a major barrier to effective cancer immunotherapy and is often driven by the immunoregulatory activities of innate immune cells, such as myeloid cells within the tumor microenvironment (TME). Myeloid populations-including tumor-associated macrophages (TAMs), dendritic cells, granulocytes, monocytes and myeloid-derived suppressor cells (MDSCs)-play pivotal roles in dampening anti-tumor immune responses and promoting tumor progression. Recent advances in our understanding of myeloid cell biology have unveiled new therapeutic opportunities to disrupt these immunosuppressive mechanisms associated with tumor inflammation. This review highlights key signaling pathways and surface molecules involved in myeloid-mediated immune suppression, including CSF1R, PI3Kγ, mTOR, Syk, MerTK/Axl, and immune checkpoints such as Trem2, LILRBs, VISTA, and CD40. We examine preclinical and clinical findings that support targeting these pathways to reprogram the TME and enhance anti-tumor immunity. By integrating insights from mechanistic studies and therapeutic development, this review underscores the potential of myeloid cell-targeting strategies as promising adjuncts to current cancer immunotherapies. Finally, we discuss future directions and challenges in translating these approaches into durable clinical benefit.

<h4>Introduction</h4>Intravenous immunoglobulin (IVIG) is a therapy that uses pooled immunoglobulins from thousands of different donors. While it is primarily used to treat immunodeficiency and autoimmune diseases due to its immunomodulatory properties, IVIG has also been used as an off-label therapy for respiratory infections, including COVID-19. Clinical data regarding the efficacy of IVIG for COVID-19 has been controversial, and although some smaller studies have shown beneficial effects, others including a large randomized trial found no significant clinical impact but noted detrimental secondary effects.<h4>Methods</h4>We describe the first proteomic analysis from the plasma of COVID-19 patients treated with IVIG, as well as clinical outcomes.<h4>Results</h4>Patients that received IVIG early upon hospitalization have faster clinical improvement. Proteomic analysis showed that serum from patients with COVID-19 has increased levels of proteins associated with inflammatory responses, activation of coagulation and complement pathways, and dysregulation of lipid metabolism. IVIG therapy significantly impacted pathways related to coagulation. Given known crosstalk between coagulation and complement pathways, we also analyzed complement-related proteins. Overall, treatment with IVIG appeared to modulate coagulation (KNG1, ACTB, FGA, F13B, and CPB2) and complement (C1RL, C8G and CFD) related proteins.<h4>Discussion</h4>Our data is supported by similar findings observed in disease states other than COVID-19, where IVIG can impact coagulation and complement proteins. However, early administration seems to be critical determinants to optimize responsiveness to IVIG therapy in COVID-19.

Recent advancements in genetic engineering have enabled the creation of extensive mutant libraries across various species, driving the need for efficient screening methods to identify mutants of interest. In this study, we developed and optimized two rapid and straightforward screening techniques to identify genes involved in iron metabolism. Iron is an essential element for almost all organisms, and in pathogens, the ability to acquire iron from the environment and mitigate the toxic effects of intracellular iron often plays a crucial role in virulence. The first screening method exploits the autofluorescence property of porphyrins, while the second one is an optimization of growth assay on solid-agar suitable for large scale analyses. To validate these methods, we applied them to a recently published protein kinase deletion mutant library in <i>Aspergillus fumigatus</i>, a fungal pathogen that causes severe diseases in immunocompromised individuals. Our iron-specific screening approaches successfully identified strains with altered iron metabolism, including both previously known and novel mutants, generating a small set of genes that can serve as new targets for antifungal therapies. These methodologies provide the first large-scale tool for exploring iron metabolism-related genes and can be adapted for other organisms with available mutant libraries.

Mineral-microbe interaction is a driving environmental changes, regulating the biogeochemical cycling of elements, and contributing to the formation of ore deposits. Microorganisms are fundamental to mineral transformation processes, exerting a profound influence on biogeochemical cycles and the bioavailability of critical nutrients required for plant growth. In this review, we delve into the various mechanisms by which microbes facilitate mineral dissolution, precipitation, and transformation, with a particular focus on how these processes regulate the availability of both macronutrients and micronutrients in soils. Essential microbial activities such as phosphate solubilization, iron chelation, and sulfur oxidation play a pivotal role in enhancing nutrient uptake in plants, thereby supporting sustainable agricultural practices and reducing dependence on chemical fertilizers. Furthermore, microbial-driven mineral transformations are vital for environmental remediation efforts, as they contribute to the immobilization of toxic metals and the detoxification of contaminated soils. By examining key microbial-mineral interactions-including nitrogen fixation, siderophore production, and metal precipitation-this review underscores the indispensable role of microorganisms in improving soil fertility, fostering plant growth, and bolstering ecosystem resilience. The exploration of these microbial processes reveals significant potential for advancing bioremediation strategies and the development of biofertilizers, offering promising solutions to enhance agricultural productivity and address environmental challenges.

<h4>Introduction</h4>Shenfu injection (SFI) is widely used in clinical severe conditions including sepsis due to its pharmacological effects of invigorating Qi and reviving Yang for resuscitation. SFI is known to alleviate sepsis-related intestinal injury. However, the underlying mechanism remains exclusive. This study aimed to investigate the regulatory role of SFI in the self-renewal of intestinal stem cells as well as gut microbiota during sepsis.<h4>Methods</h4>Plasma of septic patients was collected for detecting intestinal dysfunction markers. Mechanistic investigations were performed using immunofluorescent co-staining, Western Blotting, hematoxylin-eosin(H&E) staining, immunohistochemical (IHC) staining, TUNEL staining, Enzyme-linked immunosorbent assay (ELISA), quantitative reverse transcription-PCR (RT-qPCR) and 16S rRNA gene sequencing.<h4>Results</h4>Plasma levels of D-lactate and intestinal-type fatty acid-binding protein (I-FABP) in septic patients with intestinal dysfunction were significantly reduced following SFI administration. Compared to the control mice, sepsis induced severe mucosal damage in the intestine, including the decrease of the villus length and the number of crypts, which was significantly inhibited by SFI administration. In addition, SFI significantly reduced the levels of pro-inflammatory cytokines Interleukin 1β (IL-1β), Interleukin 6 (IL-6), and tumor necrosis factor-α (TNF-α) in septic mice. Cell proliferation and migration in the intestine were significantly increased in the intestine of septic mice upon SFI treatment. Mechanically, SFI facilitated the expression of activating transcription factor 4 (ATF4) to promote the transcription of SRY-Box Transcription Factor 9 (Sox9), thereby activating the stemness and proliferation of crypt stem cells. Moreover, the gut microbiota composition in septic mice was modulated by SFI administration. In particular, high dose of SFI (HSF) treatment was found to decrease the proportion of harmful bacteria including Proteobacteria and Escherichia-Shigella, while increase the abundance of beneficial bacteria such as Alloprevotella and Butyricimonas in septic mice.<h4>Discussion</h4>Our findings revealed that SFI protects against sepsis-induced intestinal injury via promoting the self-renewal of crypt stem cells and regulating the gut microbiota composition, providing strong evidence for the potential of SFI in treating sepsis-related intestinal dysfunction.

Autophagy is a crucial mechanism implicated in both aging and cardiovascular disease, which are two closely interconnected conditions. Modulation of autophagy is expected to have profound impacts on cellular aging and maintenance of cardiovascular functions under physiological or pathological conditions. Consequently, modulation of autophagy could be an effective strategy for counteracting age-induced vascular and cardiac remodelling as well as alleviating cardiovascular disease. The present review comprehensively elucidates the multifaceted impacts of autophagy on aging of the cardiovascular system. We comprehensively analyse both vascular and cardiac tissues, including vascular and cardiac malignancies, in distinct contexts. We also emphasize the significance of non-coding RNAs (ncRNAs) in the epigenetic regulation of gene expression and their roles as biomarkers of cardiovascular pathologies while maintaining clear distinctions between the vascular and cardiac tissues. Preclinical and clinical models are described herein to highlight the importance of ncRNAs in disease treatment by considering their involvement in the modulation of autophagy within the cardiocirculatory system. Finally, we conducted a comprehensive meta-analysis of transcriptomic data to underscore the paramount importance of autophagy while demonstrating it as a process that is frequently dysregulated in both cardiac and vascular cells under pathological conditions. The findings presented herein emphasize the importance of investigating novel strategies for modulating autophagy as a potential therapeutic approach to the management of age-related cardiovascular disorders.

CAG repeat expansions within the HTT gene cause Huntington's disease (HD), a devastating neurodegenerative disease characterized by progressive movement, cognitive, and behavioral symptoms. These expansions result in the expression and accumulation of neurotoxic poly-glutamine (polyQ). Disease initiation depends on the length of the expansion, with fewer than 35 repeats of polyQ typically not pathogenic, while 40 or greater repeats almost always result in HD. Longer expansions correlate with earlier onset of disease; however, there may be other factors that contribute to disease initiation or progression, particularly in individuals with repeat lengths close to or below the 40 repeat length pathogenic threshold. Aggregates of the protein tau are a frequent co-pathology in HD and may modify disease presentation. To examine relationships between tau and polyQ <i>in vivo</i> , we generated <i>C. elegans</i> co-expressing 40 repeats of polyQ (polyQ(40)) and human tau pan-neuronally. We found that co-expression of tau and polyQ(40) results in mild worsening of motility defects and increased accumulation of total and phosphorylated tau but not polyQ. These results suggest that co-morbid tau and polyQ can worsen neuronal dysfunction, and the presence of tau pathology may contribute to disease phenotypes in patients with HD, particularly individuals with repeat lengths close to the pathogenic threshold.

<h4>Background and objectives</h4>Cell lines serve as invaluable tools in studying lung cancer biology and developing new therapies to combat the disease. However, commercially available cell lines are typically of Caucasian origin and may be less representative of the local genetic background. To address this, our lab previously immortalized cells from pleural fluid of a Filipino non-small cell lung cancer (NSCLC) patient via CDK4 transduction. Copy number variations (CNVs) are a type of genetic variation which may affect physiology and disease by disrupting gene function or altering gene expression, and in cancer, these may be associated with patient outcomes. CNV profiling can be valuable for understanding the biology of our immortalized cells and identifying genes that could serve as potential targets for diagnostic, prognostic, and therapeutic interventions. This study aimed to characterize previously immortalized NSCLC-derived cells, GL01, in comparison with an established lung adenocarcinoma (LUAD) cell line, A549, through whole-genome microarray-based copy number profiling.<h4>Methods</h4>DNA was extracted from GL01 and A549 cells using a commercially-available silica-based DNA extraction kit. DNA extracts were quantified and normalized for microarray analysis. Whole-genome copy number profiling was done using the OncoScan CNV Plus Assay following the manufacturer's protocols, and data was analyzed using the Chromosome Analysis Suite software. Functional analysis of genes identified to be involved in copy number aberrations was done using the PANTHER Classification System.<h4>Results</h4>Copy number aberrations span 1,592,737,105 bp in GL01 and 1,715,708,552 bp in A549, with a high degree of concordance between the two. Largescale and focal copy number aberrations previously identified to be recurrent in various LUAD cohorts were present in both GL01 and A549. Focal copy number aberrations associated with previously described lung cancer-related genes involve the PDE4D gene in GL01 and the SKIL and CDKN2A/CDKN2B genes in both GL01 and A549. PANTHER Pathway analysis of genes positively correlated with mRNA expression showed that the ubiquitin proteasome pathway was significantly overrepresented in both GL01 (FDR p = 0.000074) and A549 (FDR p = 0.000075), with 20 genes involved. Additionally, the KRAS:p.G12C/S:c.34G>T/A somatic mutation variant was detected in both GL01 and A549.<h4>Conclusion</h4>This study provides a method for identifying potentially clinically-relevant genes associated with a sample's copy number aberrations and the pathways they represent, providing personalized mechanistic, prognostic, and therapeutic insights into the cancer biology of our cells.

Background Although <i>Helicobacter</i> <i>pylori </i>infection is a primary risk factor for gastric cancer (GC), the specific bacterial components that causally drive carcinogenesis remain poorly understood. Traditional epidemiological studies are limited by confounding variables and the potential for reverse causation. This study aimed to dissect the causal effects of host antibody responses to various <i>H. pylori</i> antigens on GC risk using Mendelian randomization (MR). Methodology We conducted a two-sample MR study using summary statistics from large-scale genome-wide association studies (GWAS) of European-ancestry populations. Genetic instruments were selected for general <i>H. pylori</i> seropositivity and antibody levels against six antigens: cytotoxin-associated gene A (CagA), Catalase, GroEL, outer membrane protein (OMP), urease subunit A (UreA), and vacuolating cytotoxin A (VacA). The primary outcome was GC. Inverse-variance weighted (IVW) MR served as the main analysis, with comprehensive sensitivity analyses to assess the robustness of results. Multivariable MR (MVMR) was used to estimate the direct effects of each serotype, and a two-step mediation analysis was performed to explore potential mediating pathways. Results Genetically predicted general <i>H. pylori</i> seropositivity was causally associated with an increased risk of GC (odds ratio (OR): 1.12, 95% confidence interval (CI): 1.01-1.24; <i>P </i>= 0.027). The host antibody response to OMP showed a stronger causal effect (OR: 1.19, 95% CI: 1.08-1.30; <i>P </i>< 0.001). In contrast, no causal effects were observed for antibody responses to the classic virulence factors CagA or VacA (<i>P</i> > 0.05). In multivariable analysis, the effect of the anti-OMP response remained robust (OR: 1.18, 95% CI: 1.07-1.30; <i>P </i>= 0.001), while the association for general seropositivity was attenuated to null. Mediation analysis implicated tumor necrosis factor ligand superfamily member 18 (TNFSF18) as a potential mediator of the <i>H. pylori</i>-GC pathway, accounting for a substantial portion of the total effect (estimated at 47.0%), though this finding did not reach statistical significance (<i>P </i>= 0.077). Conclusions This MR study provides genetic evidence that the host immune response to <i>H. pylori</i> OMPs, rather than to classic virulence factors like CagA, is a key contributor to gastric carcinogenesis. This effect appears to be partially mediated by the inflammatory TNFSF18 pathway, suggesting that the chronic host-bacterial interactions at the gastric epithelial surface are critical to malignant transformation.

No abstract available.

<h4> Abstract </h4> Huntington’s disease (HD) arises from the toxic gain of function caused by a CAG expansion in the coding region of the HTT gene. HD is increasingly appreciated to emerge from multiple pathogenic processes, including somatic instability in mutant HTT’s ( mHTT ) CAG repeat tract, which leads to diverse deleterious consequences. These include the alternative processing of HTT pre-mRNA to generate the HTT1a transcript that encodes the very toxic, mHTT isoform referred to as HTT1a. We set out to compare the efficacy and safety of allele-selective lowering of mHTT compared to non-allele-selective lowering using antisense oligonucleotides (ASOs) in heterozygous Htt Q111 (Q111) mice. We developed a mutant specific ASO (MutASO) targeting Htt intron-1 that selectively reduced mutant full-length HTT, as well as HTT1a, in the brains of Q111 mice. Compared to the rescue provided by a pan-allele-targeting ASO (PanASO) that lowers wild-type HTT and full-length mHTT (sparing HTT1a), the MutASO essentially eliminated aggregate formation, and provided marked protection from transcriptional dysregulation in HD knock-in mice. Thus, by targeting the ASO to the region upstream of the cryptic polyadenylation sites required to generate the HTT1a transcript, our allele-selective MutASO potently reduced HTT1a protein levels. Here, our findings advocate that HTT1a may have a disproportionate impact on aggregate formation and transcriptional dysregulation and that lowering the levels of HTT1a could provide benefit when designing HTT-lowering based therapeutic strategies for HD.

Fibroblast growth factor 21 (FGF21), a pleiotropic hormone, is a significant modulator of energy homeostasis. We evaluated serum FGF21 levels in patients with a deficiency of mitochondrial aminoacyl t-RNA synthetase (aARSs). Six patients with mitochondrial aminoacyl tRNA synthetase deficiency and twelve healthy volunteers were included in this study. Whole-exome sequencing was used for molecular diagnosis. Serum FGF21 levels in the case group and healthy volunteers were analyzed using the enzyme-linked immunosorbent assay. Exome sequencing test revealed nine different pathogenic variants in the AARS2, EARS2, DARS2, SARS2, and WARS2 genes. A statistically significant difference was found between the serum FGF21 levels of the case and control groups: case group (n = 6), 882.49 ± 923.60 pg/mL; control group (n = 12), 20.89 ± 2.63 pg/mL (p &lt; 0.001). The area under the ROC Curve for FGF21 in the differential diagnosis of mitochondrial aminoacyl-tRNA Synthetase Deficiency was 0.933 (0.786-0.962). Sensitivity and Specificity 100%, Positive and Negative Predicted Value 100% were found for FGF21 &gt;27.4 pg/mL cut-off value. Assessment of FGF 21 levels as an indicator of mitochondrial damage in mt-aARS deficiency may provide insight into the level of damage. Investigating the biochemical mechanisms underlying the varying levels of damage caused by different aminoacyl tRNA synthetases is crucial for elucidating clinical heterogeneity.

Upstream open reading frames (uORFs) are short translated regions that occur in the 5□ untranslated regions (5□ UTRs) of mRNA transcripts where they primarily serve to repress expression translation of the downstream primary open reading frame (pORF). Their widespread presence across mammalian transcriptomes suggests an important role in shaping the proteome, although the mechanistic basis of their regulatory effects remain incompletely understood. Here we present an integrated experimental and computational investigation into the features that govern uORF-mediated translation control. Using high-resolution proteomics data from 29 healthy human tissues and machine learning-based simulations, we have systematically dissected how features including uORF length, amino acid composition, start codon position, stop codon position, and Kozak context influence repressive activity, and performed experimental validation using reporter gene constructs. We also investigated how multiple uORFs within a single 5□ UTR can interact in synergistic or antagonistic ways, with the potential to produce counterintuitive effects on pORF translation. From these studies, we present a model of uORF function, suggesting a hierarchy of uORF feature importance, and proposing that a combination of uORF translation initiation probability, ribosome recycling rate, intercistronic ternary complex recharging requirements, and ribosome stalling mechanisms underlie uORF repressive activity. Together, these studies provide a comprehensive view of the molecular logic underlying uORF activity, offering new insights into their endogenous and highlighting their potential as targets for drug development.

<h4>Abstract</h4>Neuropathic pain is pervasive among people with diabetes. The integrated stress response (ISR) is a key mechanism of translational regulation implicated in diabetic pain. In this study, we demonstrate that a reactive glycolytic metabolite, methylglyoxal (MGO), which is strongly associated with painful diabetic neuropathy, activates the ISR through the kinase general control nonderepressible 2 (GCN2). Methylglyoxal disrupts elongating ribosomes, triggering the recruitment of ribosome quality control factors and collision sensors. GCN2 activation by MGO requires the ribosomal P-stalk, a critical sensor for elongation factors. Moreover, neuronal sensitization and mechanical allodynia produced by MGO are GCN2-dependent. Overall, this study links ribosomal elongation dysfunction to metabolic pain and identifies GCN2 as a novel analgesic target for diabetic neuropathy.

Type 1 diabetes is characterised by the autoimmune destruction of pancreatic β-cells, leading to an absolute or near-absolute insulin deficiency. Although traditionally associated with childhood onset, it can manifest at any age, and it is increasingly recognised that there is significant heterogeneity in its clinical presentation. This review examines the intricate interplay between genetic susceptibility, environmental factors, and autoimmune mechanisms that contribute to the pathogenesis of type 1 diabetes. The role of clinical phenotype, along with diagnostic and clinical measurements of autoantibodies and C-peptide, in the classification of type 1 diabetes is discussed, alongside the challenges in diagnosing and classifying the disease. Emerging insights from studies into the heterogeneity of type 1 diabetes phenotypes and mechanistic endotypes underscore the need for refined diagnostic criteria, particularly in identifying autoimmunity in individuals initially diagnosed with type 2 diabetes. The impact of obesity and insulin resistance on disease progression and clinical management is also examined. Overall, this review aims to provide a comprehensive understanding of the evolving landscape of type 1 diabetes, highlighting critical areas for future research and potential therapeutic approaches tailored to individual patient profiles. PLAIN LANGUAGE SUMMARY: Type 1 diabetes is an autoimmune disease where the body does not produce insulin, leading to high blood sugar levels. Traditionally thought to start in children and young adults, type 1 diabetes can occur at any age. However, many factors contribute to when someone develops type 1 diabetes and how rapidly the disease progresses, including a person's combination of genetic factors, including specific genes that can either be protective or high-risk for the development of type 1 diabetes. Although it is widely assumed that a triggering event or events initiate the autoimmune process, the trigger or triggers remain unknown. However, it is this autoimmune process that causes progressive destruction of insulin-producing Β-cells in the pancreas that eventually leads to high blood sugar and the diagnosis of diabetes. Although we have several tools to diagnose and classify diabetes, including measuring autoimmune markers (antibodies) in the blood, there is significant variation in how individuals with type 1 diabetes can present, which can make recognizing and appropriately treating type 1 diabetes more challenging. Finding better ways to characterize the unique characteristics of each subgroup of individuals may provide new insights into how we can best tailor treatment to each of these patient groups.

Long-read RNA sequencing has transformed transcriptome analysis by enabling comprehensive mapping of full-length transcripts, providing an unprecedented resolution of transcript diversity, alternative splicing and transcript-specific regulation. In this study, we employed nanopore long-read RNA sequencing to profile the transcriptomes of three cell types commonly used to model brain disorders, human fibroblasts, induced pluripotent stem cells and stem cell-derived cortical neurons, identifying extensive transcript diversity with 15 072 transcripts in stem cell-derived cortical neurons, 13 048 in fibroblasts and 12 759 in induced pluripotent stem cells. Our analyses uncovered 35 519 differential transcript expression events and 5135 differential transcript usage events, underscoring the complexity of transcriptomic regulation across these cell types. Importantly, by integrating differential transcript expression and usage analyses, we gained deeper insights into transcript dynamics that are not captured by gene-level expression analysis alone. Differential transcript usage analysis highlighted transcript-specific changes in disease-relevant genes such as <i>APP</i>, <i>KIF2A</i> and <i>BSCL2</i>, associated with Alzheimer's disease, neuronal migration disorders and degenerative axonopathies, respectively. This added resolution emphasizes the significance of transcript-level variations that often remain hidden in traditional differential gene expression analyses. Overall, our work provides a framework for understanding transcript diversity in both pluripotent and specialized cell types, which can be used to investigate transcriptomic changes in disease states in future work. Additionally, this study underscores the utility of differential transcript usage analysis in advancing our understanding of neurodevelopmental and neurodegenerative diseases, paving the way for identifying transcript-specific therapeutic targets.

Pulmonary Mycobacterium tuberculosis (Mtb) infection results in a variety of heterogeneous lesion structures, from necrotic granulomas to alveolitis, but the mechanisms regulating their development remain unclear. Using a mouse model of concomitant immunity and subsequent aerosol infection, we demonstrate that counter regulation between neutrophils and CD4 T cells occurs very early during infection and governs these distinct pathologies. In primary Mtb infection, a dysregulated feed-forward circuit of neutrophil recruitment occurs, in which neutrophils hinder CD4 T cell interactions with infected macrophages, cause granuloma necrosis, and establish a replicative niche that drives a two-log increase in lung bacterial burden. Conversely, the rapid recruitment and activation of T cells due to concomitant immunity promotes local macrophage activation and dampens detrimental neutrophil responses. Together, these studies uncover fundamental determinants of tuberculosis lung pathology, which have important implications for new strategies to prevent or treat tuberculosis.

Although synaptic evolution has been extensively studied, how axons first arose remains unexplored. Because evolution often occurs by coopting existing features, we review the evolutionary histories, biophysics, and cell biology of cytokinesis, cell crawling, and ciliogenesis to explore the origin of axons. Although we found that cilia and axons are outwardly similar, and growth cones strongly resemble the leading edge of crawling cells, the biophysical processes and the critical proteins that drive each seem weakly linked to axons as a structure. In contrast, the traction force machinery that pulls daughter cells apart during cytokinesis and the cytoskeletal organization of cytokinetic bridges appear to have a one-to-one correspondence to neuronal growth cones and axons. Based on these observations, we propose the hypothesis that axons evolved due to mutations that partially activated cytokinesis in an interphase cell. To rigorously test this hypothesis, we suggest conducting systematic phylogenetic analysis of the genes essential for each process, paired with molecular genetic studies in which critical genes are systematically disrupted. Doing so will provide a framework for understanding the relationship between diverse cellular processes, the early evolution of neurons, and insights that could potentially assist in treating cancer and promoting neuronal regeneration.

Parkinson's disease is characterized by loss of dopamine neurons that project to the dorsal striatum, and mutations in <i>LRRK2</i> and <i>GBA1</i> are the most common genetic causes of familial Parkinson's disease. Previously, we showed that pathogenic <i>LRRK2</i> mutations inhibit primary cilia formation in rare interneurons and astrocytes of the mouse and human dorsal striatum. This blocks Hedgehog signaling and reduces synthesis of neuroprotective GDNF and NRTN, which normally support dopamine neurons vulnerable in PD. Here, we show that <i>GBA1</i> mutations also impair Hedgehog signaling and Hedgehog-dependent neuroprotective factor production by a distinct mechanism. Loss of GBA1 activity increases lysosomal accessible cholesterol and thus decreases accessible cholesterol in primary cilia of cultured cells; this change in lipid composition blocks ciliary Hedgehog signaling that depends on accessible cholesterol. Consistent with defects in Hedgehog signaling in the mouse dorsal striatum, <i>GBA1</i> mutant mice show reduced Hedgehog-induced <i>Gdnf</i> RNA expression in striatal cholinergic interneurons, with no detectable impact on cilia formation. Also, both <i>LRRK2</i> and <i>GBA1</i> mutations suppress Hedgehog-induced <i>Bdnf</i> expression in striatal astrocytes. These findings underscore the role of Hedgehog signaling in the nigrostriatal circuit and reveal a convergent mechanism by which distinct <i>LRRK2</i> and <i>GBA1</i> mutations may contribute to PD pathogenesis.

Combination therapies enhance the therapeutic effect of cancer treatment; however, identifying effective interdependent doses, durations, and sequences of multidrug administration regimens is a time- and labor-intensive task. Here, we integrated machine learning, automation, and large microfluidic arrays of cancer spheroids or patient-derived organoids formed in a tissue-mimetic hydrogel to achieve notable acceleration of the discovery of effective multidrug administration regimens. For the clinically approved drug combination, we found a sequential administration regimen leading to a substantial reduction in the total drug dose, in comparison with concurrent drug supply, both at comparable drug efficacy. For the drugs that are currently under clinical development, we found a synergistic effect of concurrently administered drugs and showed that the synergy diminishes for the sequential drug supply. The developed strategy holds promise for the discovery of effective combination therapies for advanced cancer treatment, including personalized chemotherapies.

In situ tumor vaccines have immense potential for immunotherapy because they generate whole tumor-derived antigens (TDAs) to activate antitumor immune responses. However, the rapid degradation and clearance of the released TDAs severely hinder subsequent antigen presentation and the final efficacy of the in situ vaccine. Here, we synthesized gold nanoxanthium coated with polydopamine (AuNX-PDA) to mimic the morphological and biological adhesion properties of xanthium and mussels, respectively. AuNX-PDA facilitated effective absorption of released TDAs after near-infrared II photothermal treatment and delivery of the absorbed TDAs to the endoplasmic reticulum and Golgi apparatus of dendritic cells for cross-presentation, thereby activating CD8⁺ T cells for efficient tumor-specific immunity. The nanovaccine (NV) significantly inhibited irradiated primary tumors and nonirradiated distant tumors by producing robust antitumor immune responses in B16F10 melanoma and 4T1 breast cancer mouse models. These findings highlight the potency of morphology- and adhesion-dual biomimetic NVs in whole-tumor vaccine therapy.

Hirschsprung disease (HSCR) is a congenital condition characterized by the improper migration of enteric neural crest cells, leading to aganglionosis most commonly in the rectosigmoid colon. This severe and life-threatening disorder often results in the development of Hirschsprung-associated enterocolitis (HAEC), which can occur either before or after surgical resection of the affected bowel segment. Using colonic tissue from patients with HSCR alongside the well-established endothelin receptor B knockout mouse model, we investigated epithelial regeneration dynamics and stromal-epithelial cross-talk in the distal ganglionic colon, a critical site for HAEC development. In individuals with HSCR but without epithelial damage, the distal ganglionic colon displayed impaired epithelial regeneration and alteration of intestinal stem cell dynamics, characterized by the reduction of leucine-rich repeat-containing G protein-coupled receptor 5 (LGR5⁺) epithelial stem cells. This phenomenon was consistent in the mouse model, where impaired regenerative ability preceded HAEC when epithelial damage occurred on site. Patients with HSCR also exhibited remodeling in stromal cells in this distal ganglionic colon region, with fewer primary sources of Wingless-related integration site (Wnt) signal-releasing stromal cells and the exclusive presence of proinflammatory (matrix metalloproteinase 1⁺) stromal cells. Stromal cells from the HSCR distal ganglionic colon failed to sustain the growth of colonic organoids. However, ibuprofen suppressed the proinflammatory stromal cells, leading to effective restoration of epithelial organoid growth. These observations underscore the crucial role of impaired stromal-epithelial cross-talk in HSCR and the pathogenesis of HAEC and suggest potential therapeutic targets for the prevention or treatment of the condition.

<p>Introduction: Congenital dyserythropoietic anemia type I (CDA-I) is a rare disorder of erythropoiesis. All CDA-I patients are expected to have iron overload and chronic hemolysis. Patients with severe anemia may undergo splenectomy. Hemochromatosis, chronic hemolysis, and splenectomy are all found to increase risk for thromboembolism in thalassemic patients. As CDA-I patients have similar findings, we sought to evaluate prevalence of thromboembolic events (TEEs) in these patients.<h4>Methods</h4>A retrospective case-control study was conducted, including 110 CDA-I patients (study group) and 326 age- and sex-matched iron deficiency anemia patients of the same ethnicity (control group). Patients were risk-stratified using Risk Assessment Models for thromboembolism.<h4>Results</h4>We identified 3 cases (2.7%) with TEEs in the CDA group and 1 case (0.3%) in the control group. All patients were females. VTE risk scores were low to moderate for CDA patients and higher for IDA patient. When compared to control group, CDA-I patients were nine times more likely to develop TEE (OR 9.11, 95% CI = 1.15-185.27, p = 0.057). All 3 CDA patients had a history of remarkable hemolysis and iron overload. Two underwent splenectomy.<h4>Conclusion</h4>These findings show that CDA patients appear to be at increased risk for TEEs. </p>.

Astrocytes are not a uniform population but exhibit diverse morphological, molecular, and functional characteristics. However, how this diversity originates and becomes establishes during development, remains largely unknown. Here, using single-cell RNA sequencing and spatial transcriptomics, we identify five astrocyte subtypes with unique molecular features, spatial distributions and functions in the mouse neocortex and characterize essential regulators for their formation. Using TrackerSeq to trace clonally related astrocytes, we identify two distinct lineages that give rise to these five subtypes. One lineage derives from Emx1⁺ radial glial cells that initially generate neurons and later switch to astrocyte production. The other, with minimal neuronal output, predominantly produces a distinct subset of astrocytes marked by Olig2. Olig2 knockout disrupts lineage specification, leading to changes at molecular, morphological and functional levels. These findings shed light on the cellular mechanisms underlying astrocyte diversity, highlighting the presence of multiple radial glial cell subtypes responsible for generating cortical astrocyte subtypes.

Determining the neuromodulators driving brain activity is critical for understanding cognition and neuropathology. Neuromodulators act through neuroreceptors in a coordinated manner, yet their interconnected dynamics are often overlooked. We show that a neuroreceptor-based modeling framework using cortical density maps of 19 neuroreceptors and transporters from Positron Emission Tomography (PET) can model BOLD-derived brain activity. This framework reconstructs activity across four datasets (N = 314) identifying two neuroreceptor modules linked to higher-order associative or somatomotor and visual networks. Applying the framework to independent datasets, we recover the binding profiles of LSD and Modafinil, demonstrating consistency with known pharmacological and neurobiological associations. Additionally, it uncovered associations between neuroreceptors, transporters, and altered brain activity in neuropsychiatric disorders. These findings demonstrate the framework's potential to elucidate neuromodulatory mechanisms and advance our understanding of brain function across diverse states and conditions.

Peripheral immune dysregulation is frequently reported in autism spectrum disorder (ASD); however, the underlying molecular mechanisms remain unclear. We recruited a well-defined cohort of young Arab children with ASD, aged 2-4 years, along with matched controls in Qatar. Using a multimodal approach, we integrated transcriptomic, proteomic, and single-cell RNA-seq data analyses from this cohort. Targeted transcriptomic profiling identified differential expression of 50 immune-related genes in the circulating PBMCs of children with ASD, three of which (JAK3, CUL2, and CARD11) negatively correlated with ASD symptom severity. These gene signatures were validated in independent studies using blood and brain tissues from individuals with ASD. Enrichment analysis revealed involvement of these genes in immune function, particularly through TNF signaling pathway. Proteomic analysis highlighted disrupted TNF signaling and upregulated levels of TNFSF10 (TRAIL), TNFSF11 (RANKL), and TNFSF12 (TWEAK) in plasma of individuals with ASD. Single-cell RNA-seq revealed that B cells, CD4 T cells, and NK cells potentially contributed to these upregulations in ASD. Dysregulated TRAIL, RANKL, and TWEAK signaling pathways were specifically observed in CD8 T cells, CD4 T cells, and NK cells of individuals with ASD. These findings provide new insights into immune dysregulation mechanisms in ASD and highlight potential therapeutic targets.

<h4>Background</h4>Oat silage is a key feed source in animal husbandry due to its high nutritional value and adaptability. However, off-flavors caused by butyric acid fermentation significantly reduce its quality, with Clostridium bacteria being the main cause. The mechanisms underlying microbiota dynamics and volatile metabolite interactions remain unclear. In this study, we integrated microbial sequencing and metabolomics to analyze the key microorganisms and metabolic pathways involved in off-flavor formation, aiming to provide a theoretical basis for optimizing the fermentation process.<h4>Results</h4>The pH of the off-flavor group (UO) increased to 5.90, with reduced lactic acid content and elevated butyric acid and propionic acid. Nutritional analysis showed increased neutral detergent fiber and acid detergent fiber in the UO group, along with a decline in crude protein, indicating hindered fiber decomposition and increased nutrient loss. Microbial community analysis revealed decreased Lactobacillus spp. abundance, increased Clostridium spp. and Enterococcus spp., and a significant rise in the α-diversity index. Volatile metabolomics detected 734 metabolites, with a significant up-regulation of 454 compounds in the UO group of esters (19.71%) and terpenoids (19.86%), with an increase in the abundance of sweetness and fruity related substances such as methyl phenylpropionate and 3-methylphenylmethyl butyrate. Correlation analysis revealed that Clostridium (e.g. Clostridium tyrobutyricum) was positively correlated with butyric acid and off-flavor metabolites (e.g., 2-methoxy-4-vinylphenol) (p < 0.05), whereas Lactobacillus was strongly correlated with lactic acid content and indicators of high-quality fermentation (low pH, high DM).KEGG pathway analysis further revealed that secondary metabolite biosynthesis (e.g., terpenoids) and phenylpropane metabolic pathways are the core mechanisms of off-flavor formation.<h4>Conclusion</h4>Oat silage off-flavors result from Clostridium-driven butyric acid fermentation, which converts carbohydrates and lactic acid into butyric acid and volatile esters. This process raises pH, affects fiber degradation (31.1% rise in NDF), and causes nutrient loss (16.3% drop in CP). The imbalance between Clostridium and Enterococcus, worsened by a 11.2% decrease in lactic acid bacteria, further degrades fermentation quality. The study innovatively used multi-omics analysis to clarify the molecular mechanisms of microbial dynamics and metabolite interactions behind off-flavor formation. It showed that boosting Lactobacillus abundance can suppress Clostridium activity.

<h4>Background</h4>Traumatic brain injury (TBI) is a global public health problem. Pathophysiology of TBI remains unclear. Thus, methods of TBI treatment are limited. Phosphorylation plays a vital role in many neurological diseases, including TBI. As an emerging technique, phosphoproteomic has been widely applied in many fields.<h4>Methods</h4>Rats were subjected to controlled cortical impact (CCI) or divided to sham group. Protein was extracted from cortex, digested and labeled by iTRAQ. After undergoing mass spectrometry (MS), differential expressed phosphorylated sites (DEPSs) were identified, and proteins of these DEPSs were also listed. PSD95 was chosen as a target protein for further research. ZL006 was used to treat TBI rats. Nissl staining was applied to assess lesion volume, apoptosis was evaluated by TUNEL immunofluorescence staining, and PSD95 phosphorylation was further validated by Western blot.<h4>Results</h4>A total of 2753 phosphorylation sites across 1001 proteins were identified. A total of 221 DEPSs were identified. Phosphorylation of PSD95 at serine 417 and 418 were significantly upregulated after TBI. PSD95 had most interactions with other differential expressed phosphorylated proteins (DEPPs). Following ZL006 treatment, brain lesion volume and apoptotic rate were significantly reduced, and phosphorylation of PSD95 at Ser418 was decreased.<h4>Conclusions</h4>After TBI, PSD95 was significantly phosphorylated at Ser417 and Ser418. ZL006 markedly reduced brain lesion volume and apoptotic rate and suppressed the phosphorylation of PSD95 at Ser418.

Proteins that bind to intrinsically disordered proteins (IDPs) and intrinsically disordered regions (IDRs) with high affinity and specificity could be useful for therapeutic and diagnostic applications^1-4. However, a general methodology for targeting IDPs or IDRs has yet to be developed. Here we show that starting only from the target sequence of the input, and freely sampling both target and binding protein conformations, RFdiffusion⁵ can generate binders to IDPs and IDRs in a wide range of conformations. We used this approach to generate binders to the IDPs amylin, C-peptide, VP48 and BRCA1_ARATH in diverse conformations with a dissociation constant (K_d) ranging from 3 to 100 nM. For the IDRs G3BP1, common cytokine receptor γ-chain (IL-2RG) and prion protein, we diffused binders to β-strand conformations of the targets, obtaining K_d between 10 and 100 nM. Fluorescence imaging experiments show that the binders bind to their respective targets in cells. The G3BP1 binder disrupts stress granule formation in cells, and the amylin binder inhibits amyloid fibril formation and dissociates existing fibres, enables targeting of both monomeric and fibrillar amylin to lysosomes, and increases the sensitivity of mass spectrometry-based amylin detection. Our approach should be useful for creating binders to flexible IDPs or IDRs spanning a wide range of intrinsic conformational preferences.

Breast cancer is the second most common cancer globally, with most deaths caused by metastatic disease, often following long periods of clinical dormancy¹. Understanding the mechanisms that disrupt the quiescence of dormant disseminated cancer cells (DCCs) is crucial for addressing metastatic progression. Infections caused by respiratory viruses such as influenza and SARS-CoV-2 trigger both local and systemic inflammation^2,3. Here we demonstrate, in mice, that influenza and SARS-CoV-2 infections lead to loss of the pro-dormancy phenotype in breast DCCs in the lung, causing DCC proliferation within days of infection and a massive expansion of carcinoma cells into metastatic lesions within two weeks. These phenotypic transitions and expansions are interleukin-6 dependent. We show that DCCs impair lung T cell activation and that CD4⁺ T cells sustain the pulmonary metastatic burden after the influenza infection by inhibiting CD8⁺ T cell activation and cytotoxicity. Crucially, these experimental findings align with human observational data. Analyses of cancer survivors from the UK Biobank (all cancers) and Flatiron Health (breast cancer) databases reveal that SARS-CoV-2 infection substantially increases the risk of cancer-related mortality and lung metastasis compared with uninfected cancer survivors. These discoveries underscore the huge impact of respiratory viral infections on metastatic cancer resurgence, offering new insights into the connection between infectious diseases and cancer metastasis.

BACKGROUND: Systemic Autoinflammatory disorders (SAIDs) are a group of immunedysregulation disorders driven by aberrant activation of the innate immune system, often linked to single-gene mutations. While consanguinity is a known risk factor for inherited disorders, including autoimmune diseases, its impact in the prevalence and genetic variability of SAIDs remains insufficiently explored. OBJECTIVE: To assess the consanguinity rate among patients with SAIDs and to evaluate its impact on the prevalence and genetic variability of rare monogenic SAIDs. METHODS: We conducted a descriptive, longitudinal, observational study at the main tertiary center in Saudi Arabia. Medical records of patients with a confirmed SAIDs were retrospectively reviewed. Rare SAIDs were defined as those with ≤ 50 globally reported cases. Demographic, clinical, and genetic data of patients with rare SAIDs were collected and analyzed to assess consanguinity and its impact on prevalence and genetic variability. RESULTS: A total of 200 patients with SAIDs were included in this study, with data derived from the autoinflammatory disease clinic database. Consanguinity was observed in 78% of patients, while 52% had a positive family history of SAIDs. Genomic DNA sequencing was performed on 85% of the cohort, utilizing whole exome sequencing (75%), Sanger sequencing (14.7%), and targeted gene panel sequencing (9.4%). The diagnostic genetic yield revealed a confirmed monogenic etiology in 67% of the cases. The cohort encompassed various types of SAIDs, including Pyrin inflammasopathies (n = 48), other inflammasopathies (n = 20), type I interferonopathies (n = 29), NF-kB mediated SAIDs (n = 7), miscellaneous SAIDs (n = 15), undifferentiated SAIDs (n = 42), and polygenic SAIDs (n = 39). Notably, 38 patients were classified as rare monogenic SAIDs, linked to mutations in genes, including NLRC4, LPIN2, RIPK1, PLCG2, PIK3CD, ISG15, ZNFX1, STAT1, SOCS1, OTULIN, PTEN, GJB2, SLC29A3, and IL-17RA. Among these, consanguinity was reported in 78% of first-and second-degree relatives. Additionally, novel pathogenic or likely pathogenic variants were identified in 70% of patients with rare monogenic SAIDs, spanning 14 distinct genes. CONCLUSION: Our cohort demonstrates that rare monogenic SAIDs and novel pathogenic variants tend to cluster within consanguineous families. These findings highlight the importance of considering consanguinity and family history in the clinical assessment of SAIDs. Larger studies involving consanguineous populations are needed to further investigate their impact on prevelance and clinical outcome. CLINICAL TRIAL NUMBER: Not applicable.

<h4>Background</h4>PANoptosis has been identified as a robust inflammatory cell death pathway triggered upon host defense against invaded pathogens such as bacteria and viruses, however, pathogen-free tumor PANoptosis has not been achieved yet. Reactive oxygen and nitrogen species capable of inducing robust and diverse cell death pathways such as pyroptosis, apoptosis, and necroptosis are supposed to be the potential triggers for tumor PANoptosis by ultrasound (US)-controlled sono-piezodynamic therapy.<h4>Methods</h4>S-nitrosothiols (SNO)-zinc peroxide (ZnO₂)@cyclic dinucleotide (CDN)@mesoporous tetragonal barium titanate (mtBTO) nanoparticles (NZCB NPs) were synthesized by hydrothermal method with subsequent annealing, in situ growth, and finally surface functionalization. Scanning electron microscopy, transmission electron microscopy, X-ray diffraction, atomic force microscopy, Fourier transform infrared spectroscopy, and electron spin resonance were used for materials characterizations. Murine melanoma B16 cells are employed to investigate the in vitro US-initiated tumor PANoptosis by NZCB NPs. In vivo US-initiated tumor PANoptosis was investigated on B16 tumor-bearing C57BL/6J mice.<h4>Results</h4>A "boiling-bubbling" strategy is developed to endow the piezoelectric BTO nanocatalysts, with mesoporous architecture, which enables the encapsulation of the immune-agonist CDN (9.4 wt%) to initiate innate immunity of the host. Then, SNO-functionalized ZnO₂ was further employed to cap the mesoporous nanocatalysts, forming multifunctional piezocatalytic NZCB NPs. Under US irradiation, intracellular massive reactive oxygen and nitrogen species such as superoxide anion radicals, nitric oxide (NO), and peroxynitrite (ONOO^-) could be produced from the piezoelectric NZCB NPs, which, synergized with CDN-triggered antitumoral immunity, lead to highly immunogenic tumor PANoptosis by NZCB NPs through the tumor microenvironment remodeling. Intratumoral injection of NZCB NPs leads to substantial tumor PANoptosis with immune potentiation, ultimately destroying the tumor xenografts effectively.<h4>Conclusion</h4>The present work presents the mesostructure design of piezocatalytic nanomaterials and the crosstalk between oxidative stress and antitumor immunity within the tumor, facilitating promising tumor PANoptosis by nanocatalytic oxidation with high effectiveness and biocompatibility.

Bacterial populations can display different susceptibilities to antibiotics among individual cells, even though they originate from the same parent cell. This variability can lead to treatment failure and the emergence of resistant bacteria. Understanding the factors influencing this variability is crucial for developing effective antibiotic treatments. The underlying cause of variation in susceptibility distribution within bacterial populations remains unclear, necessitating the development of new tools for measurement. Here, we use a droplet microfluidic single-cell antibiotic susceptibility assay and focus on antibiotic resistance conveyed by the TEM β-lactamases family. We investigate how the catalytic activity of β-lactamase, and the genetic characteristics of the host strains affect the susceptibility distribution within bacterial populations at the single-cell level. For this purpose, we selected TEM-1 with the least catalytic activity against cefotaxime, followed by its two variants, R164S and G238S, exhibiting moderate and significant catalytic activity, respectively. The results showed that increasing the catalytic activity causes an increase in the population's mean level of antibiotic resistance. While the type of β-lactamase influences the susceptibility distribution of the strains, this effect is independent of the catalytic activity of the strains. Besides, the genetic characteristics of the strains receiving the β-lactam resistance gene is an important factor that plays a role in the distribution of susceptibility.

Direct cardiac compression (DCC) devices, under development as a new modality for mechanical cardiac support (MCS), offer several advantages over presently available forms of MCS. DCC devices avoid the blood contact obligatory with other implantable MCS devices, the complications associated with blood contact and hematologic incompatibility, such as thrombosis, stroke, and the need for anticoagulation are avoided, and DCC does not require vascular access eliminating challenges such as bleeding and extremity ischemia. Arterial pressure pulsatility is also maintained with DCC. Significant and underappreciated advancements in DCC technology have occurred over the last decades with notable dramatic improvements in cardiac performance and minimal tissue damage. One device has entered clinical trials with a second device anticipated to follow. DCC is poorly understood by most cardiologists and cardiac surgeons. This review summarizes DCC development and advances so that upcoming human clinical trials can be properly assessed.

The thalamus is an essential element for sensory information processing, serving as a link between peripheral sensory stimuli and cortical circuits. Consequently, the development of thalamocortical (TC) projections has been a central focus in systems neuroscience. Although substantial progress has been made in understanding the mechanisms guiding thalamic axon navigation from the diencephalon to the cortex, our understanding of the processes underlying sensory modality specificity in TC circuits remains incomplete. Modern genomic, physiological and imaging approaches have yielded exciting results, providing novel insights into the specialization of visual, somatosensory and auditory TC circuits. Recent findings have shed light on the genetic and spontaneous activity mechanisms involved in the formation of distinct sensory modalities, rekindling the interest in the thalamus and opening new research perspectives on the development of this diencephalic structure.

Senescence emerged as significant mechanism of aging and age-related diseases, offering an attractive target for clinical interventions. Senescent cells release a senescence-associated secretory phenotype (SASP), including exosomes that may act as signal transducers between distal tissues, and propagate secondary senescence. However, the composition of exosomal SASP components remains underexplored. We identified ~1,300 exosome proteins released by senescent primary human lung fibroblasts induced by three different senescence inducers. In parallel, a small human plasma cohort from young (20-26 years) and old (65-74 years) individuals revealed 1,350 exosome proteins and 171 plasma exosome proteins were altered in old individuals. Of the age-regulated plasma exosome proteins, we observed 52 exosomal SASP factors that were also regulated in exosomes from the senescent fibroblasts, SERPINs, Prothrombin, Coagulation factor V, Plasminogen, and Reelin. We identified 247 exosome lipids. Following senescence induction phosphatidylcholines, phosphatidylethanolamines, and sphingomyelins increased significantly indicating cellular membrane changes. Significantly changed proteins were related to extracellular matrix remodeling and inflammation, both potentially detrimental pathways that can damage surrounding tissues and even induce secondary senescence. Our proof-of-principle study - even though initially from a rather small human cohort - suggested potential senescence biomarker candidates, enabling future surveillance of senescence burden in the aging population.

<h4>Introduction</h4>Sepsis, a systemic inflammatory syndrome, is frequently associated with intestinal dysfunction, which in turn exacerbates disease severity. Intestinal epithelial Paneth cells exhibit increased susceptibility to necroptosis upon inflammatory stimulation. Nuclear receptor Nur77 has been implicated in multiple programmed cell death pathways. However, the precise role of Nur77 in regulating Paneth cell necroptosis during sepsis remains unclear.<h4>Objectives</h4>This study elucidates the role and underlying molecular mechanisms of nuclear receptor Nur77 in regulating Paneth cell necroptosis during sepsis.<h4>Methods</h4>We employed both systemic and Paneth cell-specific Nur77 knockout mouse models. Paneth cell necroptosis was assessed using TUNEL staining, immunofluorescence, and transmission electron microscopy. Endoplasmic reticulum (ER) homeostasis was evaluated based on ultrastructural integrity, ER-phagy, and ER stress. Protein modification and protein-protein interaction were validated by structure prediction and immunoprecipitation.<h4>Results</h4>Systemic or Paneth cell-specific Nur77 knockout exacerbated intestinal inflammation by enhancing Paneth cell necroptosis during sepsis. Nur77 deficiency in Paneth cells altered ileal microbiota rather than intestinal stem cell niche after LPS challenge. Nur77 deficiency-induced Paneth cell necroptosis was attributed to impaired ER homeostasis caused by defective ER-phagy. Mechanistically, LPS induced Nur77-PKCα interaction and their subsequent translocation to the ER, which promoted AMFR phosphorylation, FAM134B ubiquitination and subsequently ER-phagy. Treatment with Nur77 agonists (BTP and Csn-B) alleviated intestinal inflammation and restored Paneth cell homeostasis in sepsis mice.<h4>Conclusion</h4>This study demonstrates that Paneth cell necroptosis plays a critical role in intestinal inflammation. Our work also identifies Nur77 as a potential therapeutic target to protect Paneth cells and maintain intestinal homeostasis during sepsis, pointing out the therapeutic potential of Nur77 agonist in sepsis.

<h4>Objective</h4>Metabolic reprogramming emerges as a central driver of therapy resistance and survival disadvantage in ovarian cancer. We recently demonstrated that inhibiting the enzyme Deiodinase type 3 (DIO3) reduces ovarian cancer growth, although the underlying mechanism remains unclear.<h4>Methods</h4>We studied DIO3 role in metabolism in genetically manipulated ovarian cancer cells using protein expression analysis, integrative proteomics, endogenous and extracellular metabolomics, metabolic assays including lactate and glutamate secretion, reactive oxygen species (ROS) production and the Seahorse Cell Mito Stress test.<h4>Results</h4>We reveled that inhibiting DIO3 suppresses glycolysis while enhancing ATP production through oxidative phosphorylation (OXPHOS). We corroborated these findings using two models of ovarian cancer xenografts, demonstrating a marked reduction in glycolytic proteins upon silencing or inhibiting DIO3 using our first in class small molecule. Moreover, altered glutamine metabolism was also documented, favoring urea cycle and TCA cycle engagement over antioxidant production, accompanied by elevated ROS. Intriguingly, DIO3 depletion in fallopian tube cells, the precursor of HGSOC, displayed distinct metabolic adaptations, including enhanced glycolysis and lipid metabolism, suggesting tissue-specific roles for DIO3.<h4>Conclusions</h4>These collective findings position DIO3 as a potential regulator of ovarian cancer metabolism, with implications for targeting this enzyme to disrupt tumor energetics as a novel therapeutic approach.

Endometrial cancer is one of the most prevalent gynecologic malignancies in developed countries, with its incidence steadily increasing each year. Early diagnosis is crucial for a favorable prognosis; however, certain patients experience recurrence and distant metastasis after surgery, similar to advanced cancer patients, with limited treatment options. Therefore, effective strategies for early screening, diagnosis, predicting local recurrence, and guiding rapid treatment interventions are essential for improving survival rates and prognosis. Liquid biopsy, a method known for being non-invasive, safe, and effective, has attracted widespread attention for cancer diagnosis and treatment. Although its clinical application in endometrial cancer is less established than in other cancers, research on biomarkers using liquid biopsy in endometrial cancer patients is currently in progress. This review examines the latest advancements in non-invasive biomarkers identified through liquid biopsy and provides a comprehensive overview of their clinical applications in endometrial cancer. Additionally, it discusses the challenges and future prospects of liquid biopsy, offering valuable insights into the diagnosis and personalized treatment of endometrial cancer.

The human gut microbiota significantly influences host health through its metabolic products and interaction with immune, neural, and metabolic systems. Among these, short-chain fatty acids (SCFAs), especially butyrate, play key roles in maintaining gut barrier integrity, modulating inflammation, and supporting metabolic regulation. Dysbiosis is increasingly linked to diverse conditions such as gastrointestinal, metabolic, and neuropsychiatric disorders, cardiovascular diseases, and colorectal cancer (CRC). Probiotics offer therapeutic potential by restoring microbial balance, enhancing epithelial defenses, and modulating immune responses. This review highlights the physiological functions of gut microbiota and SCFAs, with a particular focus on butyrate's anti-inflammatory and anti-cancer effects in CRC. It also examines emerging microbial therapies like probiotics, synbiotics, postbiotics, and engineered microbes. Emphasis is placed on the need for precision microbiome medicine, tailored to individual host-microbiome interactions and metabolomic profiles. These insights underscore the promising role of gut microbiota modulation in advancing preventive and personalized healthcare.

Lymphoma continues to pose a serious challenge to global health, underscoring the urgent need for new therapeutic strategies. Recently, the gut microbiome has been shown to play a potential role in regulating immune responses and influencing cancer progression. However, its molecular mechanisms of action in lymphoma remain poorly understood. This study investigates the antiproliferative and apoptotic activities of gut microbiota-derived metabolites, specifically nisin (N) and urolithin B (UB), individually and in combination 7:3 (5750 μM), against the human lymphoma cell line HKB-11. Comprehensive evaluations were performed using Alamar Blue viability assays, combination index (CI) analyses, reactive oxygen species (ROS) quantification, flow cytometry for apoptosis detection, and advanced bottom-up proteomics analyses. N and UB exhibited potent antiproliferative activity, with the 7:3 combination demonstrating strong synergistic effects (CI < 1), significantly enhancing apoptosis (<i>p</i> < 0.01) and ROS production (<i>p</i> < 0.0001) compared to the untreated control. Proteomics analyses revealed substantial alterations in proteins crucial to ribosomal biogenesis, mitochondrial function, cell cycle control, and apoptosis regulation, including a marked downregulation of ribosomal proteins (RPS27; Log₂FC = -3.47) and UBE2N (Log₂FC = -0.60). These findings highlight the potential of N and UB combinations as a novel and practical therapeutic approach for lymphoma treatment, warranting further in vivo exploration and clinical validation.

<h4>Introduction</h4>Visceral leishmaniasis (VL) occurs more frequently in immunosuppressed individuals, especially those undergoing immunosuppressive drug therapy for an autoimmune disease. In those receiving TNF antagonist therapy (anti-TNF), the course of VL is more severe and the response to traditional leishmanicidal treatments, such as antimonials (Sb), is often reduced. This effect of anti-TNF treatment is observed in our immunosuppressed-mouse model of VL. In this model, we compared anti-TNF immunosuppression with no immunosuppression before and after VL treatment with Sb.<h4>Methods</h4>Serum-derived extracellular vesicles (EVs) were analyzed through label-free quantitative proteomics to identify proteins involved in both VL severity and the impact of anti-TNF immunosuppression on treatment outcome.<h4>Results</h4>In total, 223 dysregulated proteins were found in the pre-treatment groups, the majority of which, such as vitronectin, haemopexin or caveolin-1, were downregulated in the anti-TNF samples. In contrast, 173 proteins were identified in the Sb-treatment groups, most of which were found enriched in the anti-TNF plus treatment samples (anti-TNF+Sb) including fibronectin, transferrin, vitronectin and dipeptidyl peptidase-4. These differentially-expressed proteins were associated with pathways related to the immune system, liver regeneration, and ion transport.<h4>Conclusion</h4>Our findings have useful implications for the clinical management of VL patients under anti-TNF immunosuppression.

Malignant tumors of the urinary system, such as kidney cancer, bladder cancer, and prostate cancer, remain a significant challenge despite the various treatment options available. Identifying therapeutic targets for urological tumors is crucial due to the potential for recurrence and metastasis. Recent research has highlighted the importance of RNA modifications in post-transcriptional regulation, impacting various biological functions in urological tumors, including tumorigenesis, progression, metastasis, and drug resistance. However, the specific mechanisms underlying these interactions are not fully understood. This review will focus on exploring the regulatory role of RNA modifications like m1A, m5C, and m7G in urological tumors, shedding light on the pathways and molecular mechanisms involved. This analysis aims to provide new insights for the treatment of urological tumors.

Gold nanoparticles (<b>AuNPs</b>) are renowned for their unique optical, electronic, and biocompatible properties, making them ideal for applications in drug delivery, diagnostics, imaging, and materials science. With green chemistry-based biological synthesis methods gaining popularity, eco-friendly alternatives to traditional chemical and physical methods techniques have emerged. Plant extract-derived AuNPs stand out for their remarkable medicinal properties, stability, and low reactivity, enhancing their biological applicability. This review explores the different synthesis methodschemical, physical, and biologicalhighlighting their advantages and challenges. We summarize the latest research (Updated until <b>March, 2025</b>), focusing on the most recent developments in the synthesis and multifield application of AuNPs, offering a comprehensive perspective on their potential.

Capacitive sensors are platforms that enable label-free, real-time detection at low non-perturbing voltages. These sensors do not rely on Faradaic processes, thereby eliminating the need for redox-active species and simplifying system integration for point-of-care diagnostics. However, their sensitivity in high-ionic-strength solutions, such as bodily fluids, is limited due to a reduced Debye length and non-specific interactions. The present review highlights advances in material integration, surface modification, and signal enhancement techniques to mitigate the challenges of deploying capacitive sensors in biofluids (sweat, saliva, blood, serum). This work further expands on the promise of such sensors for advancing liquid biopsies and highlights key technical challenges in translating capacitive systems to clinics.

<i>Background and Objectives</i>: This study investigated whether the interaction between heavy alcohol use and depression amplifies the risk of hyperglycemia in psychiatric patients. <i>Materials and Methods</i>: We conducted a cross-sectional study on 172 patients (aged 18-65) hospitalized at the "Elisabeta Doamna" Clinical Psychiatric Hospital, Romania. The data included fasting blood glucose, gamma-glutamyl transferase (GGT), Beck Depression Inventory (BDI), and Alcohol Use Disorders Identification Test (AUDIT) scores. <i>Results</i>: Moderate positive correlations were observed between depression scores and blood glucose (r = 0.44) and between alcohol consumption and blood glucose (r = 0.43). The interaction term (BDI × AUDIT) was statistically significant in multiple regression (β = 0.012, <i>p</i> = 0.001), and the model explained 39.1% of glucose variability. Logistic regression analysis revealed that neither high alcohol consumption (OR = 1.38, <i>p</i> = 0.441) nor severe depression alone (OR = 1.30, <i>p</i> = 0.582) were significantly associated with hyperglycemia. However, their interaction demonstrated a strong and statistically significant effect (OR = 19.3, 95% CI: 3.22-115.81, <i>p</i> = 0.001). The prevalence of hyperglycemia reached 95.8% in patients with both risk factors. <i>Conclusions</i>: The combined presence of high alcohol consumption and severe depression significantly increases the risk of hyperglycemia. These findings highlight the importance of integrated screening and interventions in psychiatric settings.

Multivalent lectin-glycan interactions (MLGIs) are vital for viral infection, cell-cell communication and regulation of immune responses. Their structural and biophysical data are thus important, not only for providing insights into their underlying mechanisms but also for designing potent glycoconjugate therapeutics against target MLGIs. However, such information remains to be limited for some important MLGIs, significantly restricting the research progress. We have recently demonstrated that functional nanoparticles, including ∼4 nm quantum dots and varying sized gold nanoparticles (GNPs), densely glycosylated with various natural mono- and oligo- saccharides, are powerful biophysical probes for MLGIs. Using two important viral receptors, DC-SIGN and DC-SIGNR (together denoted as DC-SIGN/R hereafter), as model multimeric lectins, we have shown that α-mannose and α-manno-α-1,2-biose (abbreviated as Man and DiMan, respectively) coated GNPs not only can provide sensitive measurement of MLGI affinities but also reveal critical structural information (e.g., binding site orientation and mode) which are important for MLGI targeting. In this study, we produced mannuronic acid (ManA) coated GNPs (GNP-ManA) of two different sizes to probe the effect of glycan modification on their MLGI affinity and antiviral property. Using our recently developed GNP fluorescence quenching assay, we find that GNP-ManA binds effectively to both DC-SIGN/R and increasing the size of GNP significantly enhances their MLGI affinity. Consistent with this, increasing the GNP size also significantly enhances their ability to block DC-SIGN/R-augmented virus entry into host cells. Particularly, ManA coated 13 nm GNP potently block Ebola virus glycoprotein-driven entry into DC-SIGN/R-expressing cells with sub-nM levels of <i>EC</i>₅₀. Our findings suggest that GNP-ManA probes can act as a useful tool to quantify the characteristics of MLGIs, where increasing the GNP scaffold size substantially enhances their MLGI affinity and antiviral potency.

Tissue fibrosis is a pathological feature of many diseases including muscular dystrophies such as Duchenne muscular dystrophy (DMD). Fibrosis may limit the effectiveness of gene therapy in muscle impacting on viral dosing but direct evidence is lacking. Strategies to reduce skeletal muscle fibrosis are limited. The fibrosis <i>Fap</i> gene is over-expressed in the skeletal muscles of a severe mouse model of DMD, suggesting that cells expressing membrane fibroblast activation protein (FAP) could be targeted by chimeric antigen receptor (CAR)-T cells. Two consecutive administrations of FAP-specific CAR-T cells in the severe DMD model reduced collagen deposits and fibrotic biomarkers and also reduced the number of FAP-positive cells in muscle. Single cell transcriptomics revealed that FAP-CAR-T cells triggered cellular interactions with otherwise inactive muscle resident macrophages and depleted specific subsets of FAP-highly-expressing fibro-adipogenic progenitor cells, pointing to their importance in the fibrosis process. Reducing fibrosis with FAP-CAR-T cells enhanced adeno-associated virus (AAV) microdystrophin gene transfer in the model by increasing vector copies, demonstrating that fibrosis is a restriction factor for AAV gene delivery in skeletal muscle. These results provide novel insights into therapeutic strategies for DMD or other fibrotic diseases.

<h4>Background</h4>Alanine aminotransferase (ALT) has a variable normal range according to race and ethnicity. So, the upper normal level of ALT localized for the Iranian population was determined in the Ravansar Non-Communicable Disease (RaNCD) cohort population by re-evaluation of high-risk people.<h4>Methods</h4>A cohort population with normal ALT results based on the current kit was checked for a history of liver diseases. After excluding them, the remaining population was included in the distribution diagram of individuals with apparently healthy livers. Participants whose ALT values were in the 90th to 100th percentile were re-evaluated by ultrasonography (US) and a checklist of liver disease. Patients identified as having liver disease or those with other abnormal liver enzymes were excluded, and the 95th percentile was extracted from the distribution diagram of the remaining population.<h4>Results</h4>After excluding liver disease, among 8046 participants of RaNCD, US and re-evaluation were performed in 543 high-risk individuals. Liver disease was diagnosed in 74.6% by US. The most common liver disease was metabolic dysfunction-associated steatotic liver disease (MASLD), accounting for 69.7%. Grade 2 or 3 of MASLD was found in 23.2%. After excluding patients with abnormal liver enzymes and liver disease, the 95th percentile of ALT was 29 U/L in women (sensitivity: 53%, specificity: 82%) and 36 U/L in men (sensitivity: 28%, specificity: 90%).<h4>Conclusion</h4>The calculated 95th percentile was lower than the routine cut-off value of the current kit in both sexes. Generalizability is a significant advantage of our results, provided by the lack of exclusion of patients with metabolic risk factors and the use of US to exclude MASLD.

Human midbrain organoids (hMOs) derived from induced pluripotent stem cells provide a powerful system to model disorders involving dopamine (DA) dysfunction, including Parkinson’s disease (PD) and neuropsychiatric conditions. However, current differentiation protocols still fall short in recapitulating early specification, substantia nigra pars compacta (SNpc)-like identity, and the functional maturation of vulnerable DA neurons. Here, we established a differentiation strategy that combines tri-phasic WNT modulation with dynamic bioreactor culture to generate hMOs enriched in SNpc-like DA neurons. This approach significantly increases the yield of TH⁺/GIRK2⁺ and TH⁺/ALDH1A1⁺ DA neurons and promotes enhanced synaptic maturation, robust electrophysiological activity, and elevated DA release. Single-cell transcriptomics revealed that this strategy drives the emergence of SOX6 + / GIRK2 + SNpc-like neurons, accompanied by upregulation of synaptic, metabolic, and maturation programs, alongside reduced cell stress and apoptotic signaling. Importantly, hMOs demonstrated vulnerability upon exposure to α-synuclein preformed fibrils, resulting in aggregate formation and DA neuron degeneration, supporting their use as a human model of PD-relevant pathology. Overall, this system provides a scalable and physiologically relevant approach to investigate molecular mechanisms underlying neurodegeneration and DA-related disorders.

Microbial interactions are fundamental to global ecological and evolutionary processes, exemplified by endosymbiosis between prokaryotes and single-cell eukaryotes that gave rise to organelles. While such associations remain widespread and ecologically important, the diversity and evolutionary dynamics of intracellular symbioses in many microbial ecosystems remain poorly understood. Here, we uncover a hidden layer of microbial complexity in the rumen ecosystem by identifying multiple endosymbiotic associations between ciliate protozoa and bacteria. Using genome-resolved metagenomics on protozoa enriched rumen fractions, we reveal diverse bacterial genomes exhibiting hallmarks of an obligate intracellular lifestyle. These candidate symbionts span several bacterial phyla and include close relatives of known endosymbionts and parasites of protists as well as previously unclassified or presumed free-living bacterial lineages that likely represent overlooked symbiont specialists. Our findings therefore expand the known distribution of bacterial endosymbiosis, establish the rumen – a key site of global carbon and nitrogen cycling – as a promising model for symbiosis research, and demonstrate the power of our approach to uncover hidden symbiotic associations across complex microbial communities. Overall, our results highlight the ubiquity and evolutionary significance of intracellular symbiosis as a shaping force in microbial ecosystems.

D-amino acid oxidase (DAO) catalyzes oxidative deamination of D-amino acids, producing 2-oxo acids, hydrogen peroxide, and ammonia. In mammals, DAO is essential for metabolizing both endogenous and exogenous D-amino acids. Previous studies on human DAO identify two promoter regions (P1 and P2), a negative regulatory element in intron 1, and several transcription factor (TF) binding sites. In this study, the regulatory mechanism of mouse DAO gene expression is investigated to compare with the human system. To determine the promoter activity in the upstream region of the initiation site, plasmids containing mouse DAO gene fragments inserted into the pGL4 [luc2P/Hygro] vector and assessed luciferase activity in LLC-PK1 cells are constructed. A series of deletion constructs is analyzed, revealing promoter activity in all tested fragments. The highest promoter activity is detected in the -333/-87 subregion, with residual activities in the -87/ + 111 region. Bioinformatics analysis identifies TFs, including NEUR, EGRF, ZF07, ZF11, KLFS, SP1F, and ZF02, which bind to both the human and mouse DAO genes at conserved positions, suggesting their critical role in regulating DAO promoter activity.

<h4>Background</h4>Lymph node (LN) metastasis in extrahepatic cholangiocarcinoma (eCCA) is associated with poor prognosis, but the impact of specific metastatic sites is unclear. This study investigated the clinical significance of LN metastasis around the common hepatic artery (N [CHA]) in eCCA.<h4>Methods</h4>A total of 291 patients who underwent curative resection for eCCA between 2002 and 2022 were retrospectively reviewed. Patients were classified as N1 (CHA), N1 (other, regional LN metastasis without CHA), or N0. Clinical characteristics and long-term outcomes were compared. The short-to-long axis ratio (SLR) of CHA nodes on preoperative CT was evaluated for diagnostic value.<h4>Results</h4>Of 291 patients, 164 had perihilar and 127 had distal cholangiocarcinoma. The N1 (CHA), N1 (other), and N0 groups included 33, 103, and 155 patients, respectively. Five-year cancer-specific survival (CSS) rates were 6.9% (N1 [CHA]), 24.7% (N1 [other]), and 60.3% (N0). N1 (CHA) and N1 (other) had CSS hazard ratios of 3.34 and 1.86, respectively (p < 0.01). The area under the receiver operating characteristics curve for SLR in predicting N1 (CHA) was 0.779.<h4>Conclusions</h4>N1 (CHA) is a strong negative prognostic factor in eCCA. CHA node status may serve as a useful imaging-based marker of biological resectability.

The critical bone defect is a common clinical challenge worldwide, characterized by long recovery times, a substantial risk of infection, and high disability rates. There remains a significant demand for synthetic bone-repairing materials to address this issue. In this study, porous monetite was synthesized through the reaction between monocalcium phosphate monohydrate and β-tricalcium phosphate, using paraffin microspheres as pore-forming agents. An injectable biphasic cement was then developed by blending the monetite granules with hemihydrate calcium sulfate, designed for minimally invasive surgical applications. The cement demonstrated excellent biocompatibility and osteogenic properties in vitro. Furthermore, in vivo studies confirmed the cement's superior bone repair capabilities, indicating its promising potential for the treatment of critical bone defects.

Endothelial cells form the inner layer of blood vessels and play key roles in circulatory system development and function. A variety of endothelial cell types have been described through gene expression and transcriptome studies; nonetheless, the transcriptional programs that specify endothelial cell fate and maintenance are not well understood. To uncover such regulatory programs, we studied the C. elegans head mesodermal cell (HMC), a non-contractile mesodermal cell bearing molecular and functional similarities to vertebrate endothelial cells. Here, we demonstrate that a Forkhead transcription factor, LET-381, is required for HMC fate specification and maintenance of HMC gene expression. DMD-4, a DMRT transcription factor, acts downstream of and in conjunction with LET-381 to mediate these functions. Independently of LET-381, DMD-4 also represses the expression of genes associated with a different, non-HMC, mesodermal fate. Our studies uncover essential roles for FoxF transcriptional regulators in endothelial cell development and suggest that FoxF co-functioning target transcription factors promote specific non-contractile mesodermal fates.

<h4>Background</h4>Genetic haemochromatosis (GH) is a long-term genetic condition which results in increased iron absorption into the blood and accumulation of iron into certain organs overtime. Increased absorption and accumulation can be fatal. GH can cause many symptoms including arthritis/joint pain, chronic fatigue, and cognitive difficulties. The aim of this study was to measure quality of life (QoL) in people diagnosed with GH (GH-diagnosed) compared to a healthy sample and identify possible explanations for this.<h4>Methodology</h4>QoL was measured in 535 healthy people and 1039 GH-diagnosed, through completion of the World Health Organisation Quality of Life-100 survey (WHOQOL-100). 985 GH-diagnosed respondents completed a GH-focussed survey, which was developed to get further details of the impact of GH.<h4>Results</h4>Comparison of the WHOQOL-100 overall QoL score between GH-diagnosed and the healthy sample found a significantly lower score in the GH-diagnosed. Physical, psychological, level of independence, and spiritual domains were significantly lower in the GH-diagnosed group. The GH-focussed survey found a high incidence of physical and mental symptoms, and some impact on social and work life. Areas in which participants suggest would improve their QoL included: improved healthcare especially with increased understanding of GH in medical professionals, increased access to appointments, in-person appointments, regular checks for organ damage, more nutrition or dietary advice, and local support groups.<h4>Conclusions</h4>Based on the WHOQOL-100 scores and GH-focussed survey, overall QoL is worse in people diagnosed with GH due to worse physical and psychological symptoms. Improved healthcare may aid in reducing the difference in QoL.

Accurate prediction of aqueous solubility for organic molecules is of great importance across a range of fields, from the design and manufacturing of energy materials, to assessing the environmental impact of potential pollutants. It is of particular significance to the pharmaceutical industry, in which problems with low aqueous solubility frequently hamper the development of new drugs. Experimental measurements of solubility are used extensively, but are often time-consuming, resource intensive and only applicable to already synthesized molecules. As such, there is a need for the development of computational approaches to predict solubility. In recent years, there have been considerable advances in physics-based methods, with several contrasting techniques able to give accurate predictions of solubility and a wealth of thermodynamic data for structural optimization. Here, we provide the reader with a thorough understanding of the theoretical background and practical applications of these physics-based methods to predict solubility. This includes discussions of the various advantages and disadvantages of each approach, and an indication of areas of continuing research. Experimental and data-driven methods to assess solubility are also discussed to provide context.

<h4>Purpose</h4>To identify ferroptosis-related genes associated with the development of Ulcerative colitis (UC), through bioinformatics and basic experiments.<h4>Methods</h4>Ferroptosis-related genes were identified from UC microarray data extracted from the GEO database and FerrDb. GO and KEGG pathway enrichment analyses were performed. Hub genes were identified through PPI analysis, leading to TF-hub gene and miRNA-hub gene regulatory network, predicting potential drug candidates by DSigDB. RT-qPCR, WB and IHC were employed to validate hub gene expression in animal samples. Clinical samples were gathered from Normal examiners and UC patients and IHC was performed to verify SLC7A5.<h4>Results</h4>Eleven ferroptosis-related DEGs were identified (nine upregulated and two downregulated genes) in UC, with eight genes chosen from the PPI network. MCC algorithm demonstrated that SLC7A11, PSAT1, SLC7A5, ACSF2, and ACSL4 were hub genes, predicting TFs, miRNAs and drugs. RT-qPCR confirmed significant differential expression of SLC7A5, ACSL4, and ACSF2. WB and IHC of mouse samples, as well as IHC of clinical samples, revealed significantly elevated SLC7A5 expression in the UC group compared to controls.<h4>Conclusion</h4>SLC7A5 emerged as a potential focus for understanding UC pathogenesis, potentially influencing ferroptosis. FOXP3, STAT6, and hsa-miR-186-5p are implicated in UC and ferroptosis, with minocycline identified as a potential treatment by inhibiting ferroptosis.

Mammalian cells synthesise tetrahydrobiopterin de novo, an essential cofactor for hydroxylation of aromatic amino acids, cleavage of ether lipids and the synthesis of nitric oxide. In contrast, kinetoplastid parasites are pterin auxotrophs and none of the above metabolic functions can account for the essential requirement of an unconjugated pterin for growth. Here we investigate the pterin requirements for growth and survival of two medically important parasites (T. brucei and L. major) in comparison with the model insect parasite, Crithidia fasciculata. The pterin concentration required to support 50% of maximum growth of each parasite was determined in defined pterin-free media for a variety of naturally occurring pterins. T. brucei and C. fasciculata showed an identical order of preference with the most active being 6-biopterin, followed by dihydrobiopterin > tetrahydrobiopterin > L-neopterin > sepiapterin. In contrast, L. major showed a pronounced growth preference (>200-fold) for the reduced pterins over the the oxidised forms 6-biopterin and L-neopterin. The unnatural isomers 7-biopterin or D-neopterin supported growth poorly, or not at all, in these organisms. Other pterins were inactive. HPLC analysis of pterins supporting growth established that these were metabolised to the tetrahydro-forms (>95%) with no evidence of further interconversion. In the absence of pterins, the parasites failed to grow and lost viability with <1% surviving beyond 5-14 days. Relatively high concentrations of folate or dihydrofolate (>500 nM) could support growth in the absence of unconjugated pterin and HPLC analysis identified pteridoxamine and 6-hydroxymethylpterin (as tetrahydro-form) in cell extracts. A common feature of pterins that support growth is the presence of at least one or more linear carbon substituents at position 6 of the pteridine ring with at least one hydroxyl group, ideally in the 1S configuration. The possible essential roles of these important metabolites are discussed.

Blood perfusion delivers oxygen and nutrients to all cells, making it a fundamental feature of brain organization. How cerebral blood perfusion maps onto micro-, meso- and macro-scale brain structure and function is therefore a key question in neuroscience. Here we analyze pseudo-continuous arterial spin labeling (ASL) data from 1305 healthy individuals in the HCP Lifespan studies (5-22 and 36-100 years) to reconstruct a high-resolution normative cerebral blood perfusion map. At the cellular and molecular level, cerebral blood perfusion co-localizes with granular layer IV, biological pathways for maintenance of cellular relaxation potential and mitochondrial organization, and with neurotransmitter and neuropeptide receptors involved in vasomodulation. At the regional level, blood perfusion aligns with cortical arealization and is greatest in regions with high metabolic demand and resting-state functional hubs. Looking across individuals, blood perfusion is dynamic throughout the lifespan, follows micro-architectural changes in development, and maps onto individual differences in physiological changes in aging. In addition, we find that cortical atrophy in multiple neurodegenerative diseases (late-onset Alzheimer's disease, TDP-43C, and dementia with Lewy bodies) is most pronounced in regions with lower perfusion, highlighting the utility of perfusion topography as an indicator of transdiagnostic vulnerability. Finally, we show that ASL-derived perfusion can be used to delineate arterial territories in a data-driven manner, providing insights into how the vascular system is linked to human brain function. Collectively, this work highlights how cerebral blood perfusion is central to, and interlinked with, multiple structural and functional systems in the brain.

PROteolysis-Targeting Chimeras (PROTACs) are an emerging class of molecules capable of inducing a forced approximation between a protein of interest (POI) and an E3 ligase enzyme (e.g., Cereblon), leading to the degradation of the POI by the cell's own machinery. Although in early stages of development, PROTACs' unique mechanisms of action offer a novel therapeutic strategy, which has attracted growing interest worldwide. Cereblon-based PROTACs are the most studied class of PROTACs and have been actively researched in recent years for the treatment of different diseases, from cancer to neurological disorders, with some of them already in clinical trials. In this review, we provide a comprehensive and critical analysis covering the recent advances, potential challenges and future prospects regarding the design and synthesis, as well as pre- and clinical evaluation of cereblon-based PROTACs. By integrating insights from drug discovery and development, a broad yet in-depth discussion is given to guide future research on cereblon-based PROTACs.

<h4>Background</h4>Chronic liver disease (CLD) is a growing global public health issue and ranks as the seventh leading cause of death in Ethiopia. However, regional-level data particularly from the West Arsi Zone are scarce. This study aimed to fill this gap by assessing the burden and key determinants of CLD among adults attending liver clinics, providing essential evidence for targeted interventions.<h4>Objective</h4>To determine the magnitude and associated factors with medically confirmed CLD among adult patients (≥ 18 years) undergoing follow-up for suspected or confirmed CLD in three public hospitals in the West Arsi Zone.<h4>Methodology</h4>An institution-based cross-sectional study was conducted from February to July 2022. A total of 384 adult participants were selected using systematic random sampling. Data were collected through structured interviews and medical record reviews. Multivariable binary logistic regression was used to identify factors associated with CLD, with adjusted odds ratios (AOR) and 95% confidence intervals (CI) reported.<h4>Results</h4>All 384 selected participants completed the study, yielding a response rate of 100%. Among them, 60.2% (231/384) were clinically diagnosed with chronic liver disease (CLD), with a higher prevalence observed among males (62%).

<h4>Background</h4>Immune escape is a critical barrier to effective cancer immunotherapy for cancers such as hepatocellular carcinoma (HCC). The aim of this study was to identify prognostic genes associated with immune escape and to analyse immune infiltration in HCC.<h4>Methods</h4>The TCGA-LIHC cohort gene expression matrix and TCGA cohort were downloaded from the UCSC Xena and TCGA databases, respectively, for differential expression analysis, as well as for clinical data and survival information. Additionally, gene expression matrices from HCC tumor tissue samples were downloaded from the ICGC database to validate prognostic models. Subsequently, enrichment analysis utilizing the Kyoto Encyclopedia of Genes and Genomes (KEGG) and Gene Ontology (GO) were conducted. Risk modeling was subsequently performed, followed by univariate and multivariate Cox regression analyses, as well as LASSO regression analysis. Overall survival (OS) curves, receiver operating characteristic (ROC) curves, and nomograms were also generated. Finally, immune infiltration analysis was performed by single-sample genomic enrichment analysis (ssGSEA) and GeneMANIA to predict the functions and pathways of associated with prognostic genes.<h4>Results</h4>A total of 4489 differential expression genes were obtained, including 3259 up-regulated, and 1230 down-regulated. Among them, 2123 GO biological functions and 334 KEGG results were enriched. Subsequently, eight differential genes related to immune escape became candidate genes. Finally, we constructed a risk model using three genes, CEP55, GPAA1 and PIGU, and demonstrated better results. The results of immune infiltration showed that the prognostic genes affected the patient's condition through these immune cells. Subsequently, we performed drug sensitivity analysis and finally discovered that CEP55 and PIGU were positively associated with five drugs in the high-risk group. And these three key prognostic genes have high expression levels in HCC tumor tissues.<h4>Conclusion</h4>Our study found that three prognostic genes: CEP55, GPAA1 and PIGU have good prognostic value for HCC patients, and are the pivotal prognostic biomarkers.

<h4>Introduction</h4>Increasing evidence shows that hyperglycemia-induced glucotoxicity and lipotoxicity that usually accompany diabetes development damage the endoplasmic reticulum and mitochondria of the hepatocytes in diabetic patients. Clinical studies highlighted the association between type 2 diabetes mellitus, comorbidities, and medications with liver function. The objective of this study is to explore the association between liver function tests' abnormalities and comorbidities, medications, and other risk factors in type 2 diabetes patients registered in the Best-Care system of the Saudi Ministry of National Guard-Health Affairs.<h4>Methods</h4>This is a cross-sectional study employing a chart of patients diagnosed with type 2 diabetes mellitus. We drew a simple random sample of 523 T2DM patients who had a liver function test from the Best-Care database of the Ministry. We applied various statistical analyses, including Student's independent t-test, Pearson's chi-squared test, Fisher's exact test, and odd ratios, to measure associations between different variables and liver function tests' abnormalities.<h4>Results</h4>About 35% of patients included in this study showed an abnormal level of gamma-glutamyl transferase and prothrombin time. Abnormalities of serum albumin, prothrombin time, and total serum protein tests were significantly associated with age (P < 0.05). Gamma-glutamyl transferase test abnormalities were significantly associated with gender (P < 0.05). The study found associations between several comorbidities and the abnormalities of liver function tests. These tests include the total bilirubin, albumin, total serum protein, gamma-glutamyl trans, international normalized ratio, and alanine aminotransferase. The associations were at significant levels (P < 0.05). Liraglutide was significantly associated with aspartate aminotransferase (OR = 14.40, 95% CI = 2.8, 73.2), while allopurinol was significantly associated with international normalized ratios (OR = 24.67, 95% CI = 2.95, 206.58) and total serum protein (OR = 5.44, 95% CI = 1.43, 20.83).<h4>Discussion</h4>This study is the first to examine the association between type 2 diabetes mellitus and liver function tests' abnormalities in Saudi Arabia. Although the results have a limited generalizability due to inherent biases, the findings align with similar studies in other populations. The study stresses the need to monitor liver functions, especially of T2DM patients who suffer from other conditions.

<h4>Introduction</h4>Primary biliary cholangitis (PBC) is a rare autoimmune liver disease involving bile duct damage and fibrosis. This study explores the role of HLA-G, an immunomodulatory molecule crucial for immune tolerance, in PBC pathogenesis and treatment.<h4>Methods</h4>A cohort of 166 PBC patients from Sardinia was compared to 180 healthy controls and 205 autoimmune hepatitis type 1 (AIH-1) patients. Plasma soluble HLA-G (sHLA-G) levels, <i>HLA-G</i> alleles, and <i>3'UTR</i> haplotypes were analyzed alongside clinical data, including therapy response to ursodeoxycholic acid.<h4>Results</h4>The UTR-1 haplotype was significantly more frequent in PBC patients than in controls (48.2% vs 34.3%, Pc= 0.0018). The extended haplotype <i>HLA-G*01:01:01:08/UTR-1</i> was also strongly associated with PBC (23.2% vs 12.5% in controls, Pc = 0.008; 23.2% vs 6.6% in AIH-1, Pc= 2.6×10_-9). PBC patients exhibited lower sHLA-G levels compared to controls and AIH-1 (9.1 U/mL vs 24.03 U/mL and 13.9 U/mL, respectively). Among <i>UTR-1</i> carriers, sHLA-G levels were particularly reduced in PBC patients. The <i>HLA-G*01:01:01:08/UTR-1</i> haplotype correlated with the lowest sHLA-G levels and poorer therapy response (60% vs 24.1%, P = 0.0001).<h4>Discussion</h4>These findings suggest HLA-G variants, especially <i>HLA-G*01:01:01:08/UTR-1</i>, as potential biomarkers for PBC prognosis and treatment outcomes.

<h4>Introduction</h4><i>Brucella</i> bacteria are adept at evading the human immune system, leading to a chronic infectious disease known as brucellosis, which poses significant global health challenges. This study addresses a notable gap in bibliometric analyses concerning the immune response to brucellosis.<h4>Methods</h4>We conducted a comprehensive literature screening from the Web of Science Core Collection, covering publications from 1980 to 2024. Relevant publications were analyzed using R software, VOSviewer, and CiteSpace for bibliometric analysis.<h4>Results</h4>A total of 733 publications were included in this study, revealing an average annual growth rate of 5.91% in publications. The United States led in publication volume and citations, followed by China and France. Prominent research institutions included INSERM (France) and CONICET (Argentina). <i>Infection and Immunity</i> was identified as a leading journal in the field, publishing 97 papers with 5,184 citations. Keyword co-occurrence analysis delineated three main research clusters: <i>Brucella</i> vaccine development and immune protection, <i>Brucella</i> molecular mechanisms and intracellular survival strategies, and host innate immunity and <i>Brucella</i> interaction mechanisms. Burst analysis highlighted increasing attention on keywords such as "protection," "type IV secretion system," "pathogenesis," and "prediction" since 2018.<h4>Discussion</h4>This bibliometric analysis sheds light on the global research landscape regarding the immune response in human brucellosis, pinpointing trends and gaps, with a focus on immune escape mechanisms and the future development of safe, effective vaccines.

In this paper, we report a case of traumatic wound infection caused by dorsal fin puncture of live fish. A 69-year-old woman developed progressive swelling of her right pinky finger after being stabbed by the dorsal fin of a live fish. The infection was confirmed by bacterial culture as a <i>Vibrio vulnificus</i> infection of a traumatic wound. The patient underwent antibiotic treatment, surgical decompression, debridement, and excision of necrotic tissue. Finally, the right pinkie finger was amputated due to dry gangrene. Early intervention and combined antibiotic therapy led to a good prognosis. As global ocean temperatures rise, the infection rate of this bacterium increases, and vigilance is needed. Clinical practice suggests that such infections should be considered in patients with a history of contact with seafood or seawater; Early diagnosis, active antibiotic therapy and necessary surgical intervention are the keys to improving the prognosis.

<h4>Background</h4>Flecainide is a Class IC antiarrhythmic drug used to treat arrhythmias such as atrial fibrillation (AF), paroxysmal supraventricular tachycardia, and ventricular tachycardia (VT). Its mechanism involves blocking sodium channels, leading to QRS widening, especially at higher heart rates. This property increases the risk of pro-arrhythmic events, particularly in patients with structural heart disease or ischaemia.<h4>Case summary</h4>A 67-year-old woman receiving flecainide for AF suffered an ischaemic stroke and later developed VT resistant to amiodarone and direct current cardioversion. Her condition improved significantly after intravenous sodium bicarbonate was administered, which counteracted the effects of sodium channel blockade. Later, she was found to have hyponatremia and slightly elevated flecainide values above the upper limit. This case highlights the complexities of flecainide therapy and the importance of timely intervention in managing arrhythmias.<h4>Discussion</h4>This case underscores the arrhythmogenic potential of flecainide. The development of VT suggests that flecainide may have created a substrate for arrhythmias due to its effects on cardiac conduction. The successful use of sodium bicarbonate illustrates an effective treatment strategy for reversing flecainide-induced toxicity. Clinicians should remain vigilant and consider alternative therapies in managing patients at risk for arrhythmias while on flecainide.

Diabetic Nephropathy (DN), a leading cause of disability and mortality in patients with diabetes, has become a complex global clinical issue that poses a severe challenge to public health. Research indicates that Non-coding RNAs (ncRNAs) participate in cell death and fibrosis through an endogenous competitive RNA (ceRNA) network. This network regulates kidney-specific cells such as podocytes, mesangial cells, and renal tubular epithelial cells, thereby establishing a multifaceted regulatory mechanism in DN progression. Furthermore, exosomal ncRNAs and their ceRNA networks, stem cell-derived exosomal ncRNAs, related biomolecules, and the targeted regulation of ncRNAs and ceRNA networks by traditional Chinese medicine all play significant roles in the advancement of DN. This review systematically summarizes the content of ncRNAs, ceRNA networks and DN, exosome ncRNA intervention in DN progression, and targeted regulation of ncRNA intervention in DN progression. Concurrently, it discusses the research progress and therapeutic status of ncRNAs as clinical biomarkers, challenges facing ncRNA-targeted therapy, therapeutic efficacy of exosomal ncRNAs and stem cell-derived exosomal ncRNAs, pharmacokinetic limitations of Chinese medicine components in regulating DN progression through ncRNA intervention, and analyses the bottlenecks in ncRNA-based diagnosis and cross-species conservation of circRNAs/lncRNAs. This study aimed to provide new insights for the in-depth exploration of the molecular mechanisms underlying DN and the development of targeted therapeutic strategies.

<h4>Introduction</h4>The Population Health Management (PHM) Genomic Newborn Screens (GNBS) and Multi-Omics Intercepts for Human Phenotype Ontology (HPO) using Federated Data Platforms (FDP) represent a groundbreaking innovation in global health. This reform, supported by the UK's Genomic Medical Services (GMS) through "The Generation Study," aims to significantly reduce infant mortality by identifying and managing over 200 rare diseases from birth, paving the way for personalised health planning.<h4>Methods</h4>Using an ecosystem approach, this study evaluates a diverse pangenome to predict health outcomes or confirm diagnoses prior to symptomatic manifestations. GNBS standardises care by integrating diagnostic techniques such as blood spot analysis and full blood cell diagnostics to stratify risk. The approach enhances the understanding of rare diseases in primary care medicine, with biomedical and haematology diagnoses re-evaluated. Scientific proof of concept and fit-for-purpose technology align multi-omics in pre-eXams (X = Gen AI).<h4>Recommendations</h4>The Digital Regulation Service (DRS) assembles an agile group of experts to enhance medical science through human phenotype ontology (HPO) for precise disease segmentation, scheduling accurate eXam intercepts where needed. This team strategically plans regulation services for digital HPO eXam assurance and implements Higher Expert Medical Science Safety (HEMSS) frameworks. The DRS is responsible for overseeing gene, oligonucleotide, and recombinant protein intercepts; commissioning blood pathology HPO eXam intercepts; and monitoring preliminary eXams with advanced imaging techniques.<h4>Discussion</h4>In pursuit of excellence in PHM of HPO, HEMSS with Agile Group Development leverages the Genomic Newborn Screens (GNBS) and multi-omics to create personalised health plans integrated with NHS England Genomics and AI-driven DRS. The discourse extends to examining GNBS predictors and intercepts, focusing on their impact on public health and patient safety. Discussions encompass structured HPO knowledge addressing newborn health, ethical considerations, family privacy, and the benefits and limitations of pre-eXam screenings and life eXam intercepts. These debates involve stakeholders in adopting HPO-enhanced clinical pathways through Alliances for Health Systems Networking-Genomic Enterprise Partnerships (AHSN-GEP).<h4>Conclusion</h4>"The Generation Study" represents a paradigm in digital child health management using an HPO-X-Gen-AI framework, transitioning from trusted research to evidence-based discovery. This approach sets a standard for personalised healthcare practices, incorporating ontology risk stratification and future-ready analytics as outlined in the NHS Constitution. The discourse on higher expert medical science safety governance will continue in the forthcoming manuscript, "PHM Fit Lifecycles in Future Analytics," which will further explore developing localised health solutions for "Our Future Health."

<b>Background/Objective:</b> The hepatitis B virus (HBV) can cause chronic hepatitis B (CHB), which can rapidly progress into fatal liver cirrhosis (CHB-LC) and hepatocellular carcinoma (CHB-HCC). <b>Methods:</b> In this study, we investigated metabolites associated with distinct clinical stages of HBV infection for the identification of stage-specific serum metabolite biomarkers using ¹H-NMR-based metabolomics. <b>Results:</b> A total of 64 serum metabolites were identified, among which six core discriminatory metabolites, namely isoleucine, tryptophan, histamine (for CHB), and pyruvate, TMAO, lactate (for CHB-HCC), were consistently significant across univariate and multivariate statistical analyses, including ANOVA with FDR, OPLS-DA, and VIP scoring. These metabolites were closely linked to key metabolic pathways, such as propanoate metabolism, pyruvate metabolism, and the Warburg effect. <b>Conclusions:</b> The findings suggest that these six core metabolites serve as potential stage-specific biomarkers for CHB, CHB-LC, and CHB-HCC, respectively, and offer a foundation for the future development of metabolomics-based diagnostic and therapeutic strategies.

<h4>Background</h4>In recent years, comprehensive analyses using a genome-wide association study (GWAS) have been conducted to identify genetic factors related to athletic performance. In this study, we investigated the association between genetic variants and elite wrestling status across multiple ethnic groups using a genome-wide genotyping approach.<h4>Methods</h4>This study included 168 elite wrestlers (64 Japanese, 67 Turkish, and 36 Russian), all of whom had competed in international tournaments, including the Olympic Games. Control groups consisted of 306 Japanese, 137 Turkish, and 173 Russian individuals without elite athletic backgrounds. We performed a GWAS comparing allele frequencies of single-nucleotide polymorphisms (SNPs) between elite wrestlers and controls in each ethnic cohort. Cross-population analysis comprised (1) identifying SNPs with nominal significance (<i>p</i> < 0.05) in all three groups, then (2) meta-analyzing overlapped SNPs to assess effect consistency and combined significance. Finally, we investigated whether the most significant SNPs were associated with gene expression in skeletal muscle in 23 physically active men.<h4>Results</h4>The GWAS identified 328,388 (Japanese), 23,932 (Turkish), and 30,385 (Russian) SNPs reaching nominal significance. Meta-analysis revealed that the <i>ATP2A3</i> rs6502758 and <i>UNC5C</i> rs265061 polymorphisms were associated (<i>p</i> < 0.0001) with elite wrestling status across all three populations. Both variants are located in intronic regions and influence the expression of their respective genes in skeletal muscle.<h4>Conclusions</h4>This is the first study to investigate gene polymorphisms associated with elite wrestling status in a multi-ethnic cohort. <i>ATP2A3</i> rs6502758 and <i>UNC5C</i> rs265061 polymorphisms may represent important genetic factors associated with achieving an elite status in wrestling, irrespective of ethnicity.

The yak (<i>Bos grunniens</i>) is a key species in high-altitude rangelands of Asia. Despite their ecological and economic importance, yak production faces persistent challenges, including low milk yields, vulnerability to climate changes, emerging diseases, and a lack of systematic breeding programs. This review presents the genomic, physiological, and environmental dimensions of yak biology and husbandry. Genes such as <i>EPAS1</i>, which encodes hypoxia-inducible transcription factors, underpin physiological adaptations, including enlarged cardiopulmonary structures, elevated erythrocyte concentrations, and specialized thermoregulatory mechanisms that enable their survival at elevations of 3000 m and above. Copy number variations (CNVs) and single nucleotide polymorphisms (SNPs) present promising markers for improving milk and meat production, disease resistance, and metabolic efficiency. F1 and F2 generations of yak-cattle hybrids show superior growth and milk yields, but reproductive barriers, such as natural mating or artificial insemination, and environmental factors limit the success of these hybrids beyond second generation. Infectious diseases, such as bovine viral diarrhea and antimicrobial-resistant and biofilm-forming <i>Enterococcus</i> and <i>E. coli</i>, pose risks to herd health and food safety. Rising ambient temperatures, declining forage biomass, and increased disease prevalence due to climate changes risk yak economic performance and welfare. Addressing these challenges by nutritional, environmental, and genetic interventions will safeguard yak pastoralism. This review describes the genes associated with different yak traits and provides an overview of the genetic adaptations of yaks (<i>Bos grunniens</i>) to environmental stresses at high altitudes and emphasizes the need for conservation and improvement strategies for sustainable husbandry of these yaks.

<h4>Background</h4>Over 20 % of adults with insomnia disorder also experience chronic pain, termed insomnia disorder comorbid with chronic pain (ICCP), increasing risks for physical and mental diseases. Current treatments like cognitive behavioral therapy for insomnia show inconsistent pain relief, and non-opioid analgesics may exacerbate insomnia, underscoring the need for alternative approaches. Chinese herbal medicine (CHM) and acupuncture, guided by traditional Chinese medicine, may offer transdiagnostic benefits for ICCP, but a comprehensive review is lacking. This scoping review evaluates their therapeutic effects and mechanisms for ICCP.<h4>Methods</h4>PubMed, Wanfang, ClinicalTrials.gov and Google Scholar were searched up to December 31, 2024, for randomized controlled trials (RCTs) involving adults (≥18 years) with Diagnostic and Statistical Manual of Mental Disorders, Fifth Edition-defined insomnia and the International Association for the Study of Pain-defined chronic pain, treated with CHM or acupuncture. Effect sizes (modified Cohen's d) assessed efficacy of interventions, and the Cochrane Risk of Bias 2 tool evaluated the risk of bias.<h4>Results</h4>Six RCTs (487 participants) were included. CHM (modified Guipi decoction) showed medium to large effects for insomnia (<i>d</i> = 0.70-1.17) and pain (<i>d</i> = 0.67-1.42) versus diazepam/estazolam. Acupuncture had medium to large effects for insomnia (<i>d</i> = 0.64-0.99) and pain (<i>d</i> = 0.80-1.33) compared to treatment as usual. Combined CHM (Da Huoluo capsules) and acupuncture showed medium effects (<i>d</i> = 0.72 for insomnia; <i>d</i> = 0.57 for pain) versus multi-medications/traction. Most studies (83.33 %) had high risk of bias.<h4>Conclusion</h4>CHM and acupuncture show promise for ICCP management, but high risk of bias warrants cautious interpretation and further high-quality RCTs.

Excessive fructose intake drives intestinal aging and impairs intestinal stem cell (ISC) function, yet effective therapeutic interventions remain elusive. Astragaloside IV (AS-IV), a natural saponin from <i>Astragalus membranaceus</i>, has been widely recognized for its antiaging, anti-inflammatory, and gut-protective properties. Here, we revealed that AS-IV alleviates fructose-induced intestinal metabolic senescence via direct inhibition of ketohexokinase (KHK), the key rate-limiting enzyme in fructose metabolism. Molecular docking and site-directed mutagenesis identified Asn261 and Ala226 as distinct binding sites for AS-IV on KHK, with Asn261 also serving as a critical catalytic residue that is essential for KHK activity. Mutation at Asn261 abolished KHK enzymatic function, reduced the accumulation of fructose-derived metabolites such as palmitic acid and ceramide, and thereby prevented fructose-induced ISC cycle arrest. AS-IV's therapeutic efficacy was validated across <i>Drosophila</i>, murine intestinal organoids, and mice, where treatment consistently reversed high-fructose-induced intestinal metabolic senescence phenotypes, restored ISC proliferation, and preserved ISC homeostasis. These findings indicate that KHK is a previously unrecognized molecular target of AS-IV and reveal a conserved mechanism by which AS-IV modulates fructose metabolism to interfere with gut aging. Our results highlight its therapeutic potential in treating fructose-driven intestinal aging and associated metabolic disorders.

Actively identifying the risk factors and predictive indicators associated with portal vein thrombosis (PVT) in liver cirrhosis (LC) can enable early diagnosis and treatment, which is of great significance for prolonging the survival of patients with LC. Hemodynamic disturbances, advanced LC, vascular endothelial injury, and mutations in thrombophilic genetic factors are established risk factors for PVT-LC. Venous dilatation and decreased blood flow velocity contribute to hemodynamic disturbances. The severity of LC can be assessed by the degree of portal hypertension, liver metabolic function biomarkers, and validated liver scoring systems. Iatrogenic interventions, endotoxemia, and metabolic syndrome may induce vascular endothelial injury and hypercoagulability, the latter of which can be quantified via coagulation-anticoagulation-fibrinolysis biomarkers. Mutations in thrombophilic genetic factors, such as Factor V Leiden, MTHFR C667T, and JAK2 V617F, disrupt coagulation-anticoagulation homeostasis and predispose patients to PVT-LC. This review specifically focuses on comprehensively delineating established risk factors and predictive indicators for PVT-LC, thereby providing a theoretical foundation for the construction of clinically applicable PVT predictive models to guide early interventions and improve the prognosis. Future research should further validate the associations between recently proposed risk factors and PVT-LC, while simultaneously establishing cutoff values for indicators with robust predictive value to construct a clinically applicable PVT prediction framework.

Single-subject morphological brain networks derived from cross-feature correlation of macroscopic MRI-derived morphological measures provide an important means for studying the brain connectome. However, the validity of this approach remains to be confirmed at the microscopic level. Here, we constructed morphological brain networks at the single-cell level by extending features from macroscopic morphological measures to microscopic descriptions of neuronal morphology. We demonstrated the feasibility and generalizability of the method using neurons in the somatosensory cortex of a rat, neurons over the whole brain of a mouse, and neurons in the middle temporal gyrus (MTG) of a human. We found that interneuron morphological similarity was higher for intra- than interclass connections, depended on cytoarchitectonic, chemoarchitectonic, and laminar classification of neurons (rat), differed between regions with different evolutionary timelines (mouse), and correlated with neuronal axonal projections (mouse). Furthermore, highly connected hub neurons were disproportionately from superficial layers (rat), inhibitory neurons (rat), and subcortical regions (mouse), and exhibited unique morphology. Finally, we demonstrated a more segregated, less integrated, and economic network architecture with worse resistance to targeted attacks for neurons in human MTG than neurons in a mouse's primary visual cortex. Overall, our method provides an alternative avenue to study neuronal wiring diagrams in brains.

The proliferation and phenotypic modulation of smooth muscle cells (SMCs) to alternative mesenchymal states is a key process by which atherosclerotic lesions grow. The underlying mechanisms can be studied in mouse and pig atherosclerosis, but it remains unclear to what extent the mesenchymal plaque cell types in these species recapitulate human disease. Here, we integrate published and new single-cell RNA sequencing data of plaque mesenchymal cells from human carotid and coronary arteries, pig aorta and coronary arteries, and mouse brachiocephalic arteries. By applying consensus across multiple integration and gene homology-matching strategies, we identify a conserved core continuum of mesenchymal plaque cells, ranging from SMCs to extracellular matrix-producing fibroblast-like cells, which is stable across species and vascular beds. Notably, several other populations differed between human and experimental lesions. Subpopulations of SMCs marked by DLX5 and RERGL expression were specific to human carotid and coronary plaques, respectively. Mesenchymal cell states with strong pro-angiogenic and inflammation-associated gene signatures were identified in pig, but not human, coronary plaque datasets, with the pro-angiogenic phenotype associated with early stages of necrotic core development. Pericytes were solely present in pig and human plaques, while chondrocyte-like cells were unique to mouse lesions. The presented interspecies maps of mesenchymal cell diversity, and their markers may inform translational research into the role of SMCs and their derived progeny in atherosclerosis.

Protein-directed dynamic combinatorial chemistry (P-D DCC) is a powerful strategy for identifying ligands to protein targets of pharmacological significance. It leverages a thermodynamic templated effect, where proteins selectively amplify high-affinity binders. In contrast, although nucleic acids play critical roles in gene regulation and disease and offer significant therapeutic potential, they remain underexplored in drug discovery. While P-D DCC has been widely applied, the use of nucleic acid-directed dynamic combinatorial chemistry (NA-D DCC) is relatively limited. Expanding these methodologies is essential for tackling emerging infectious diseases and advancing therapeutic development. This review examines the applications, experimental design considerations, recent advancements, and P-D DCC and NA-D DCC perspectives.

pH-responsive nanocarriers have gain significant attention due to their ability to provide controlled cargo delivery with high precision in response to specific stimuli. However, the polymers used in the self-assembly of these nanocarriers must be carefully designed to meet the requirements of bio-relevant delivery. Here, we present an optimized synthesis of poly(2-methyl-2-oxazoline)-block-poly(2-(diisopropylamino)ethyl methacrylate) (PMOXA-b-PDPA) block copolymers tailored for obtaining carriers with vesicle architecture and thin membranes for an improved release behavior. By systematically modifying the synthesis conditions, we obtain a small library of copolymers, focusing on low molecular weight (MW) variants to reduce the membrane thickness of the resulting vesicles. We investigate the impact of membrane thickness on the kinetics and efficiency of cargo release in response to a pH shift from neutral to slightly acidic conditions that are particularly relevant in pathological environments like tumors. Model cargos of varying MWs, including doxorubicin hydrochloride, exhibited differential release profiles under these pH conditions. Together with no cytotoxicity, the thin membrane represents key aspects that support further development of such carriers for therapeutic applications.

<h4>Purpose</h4>In the current study, we have screened the OPTN gene in a cohort of unrelated juvenile open-angle glaucoma (JOAG) patients negative for possible pathogenic variants in CYP1B1 and MYOC genes.<h4>Methods</h4>Polymerase chain reaction (PCR) amplification and sequencing were employed to identify nucleotide variants within the coding sequence and intron-exon boundaries of the OPTN gene in 85 JOAG patients and 100 controls. A pathogenicity prediction of identified variants was performed by six distinct online algorithms. Possible structural alterations caused by pathogenic variants were investigated using GOR IV, PyMol, ChimeraX, and molecular dynamics simulations using Gromacs software.<h4>Results and discussion</h4>PCR amplification and sequencing revealed a total of 27 variations, encompassing eight missense, six synonymous, and 13 intronic changes. Out of the 85 patients, three JOAG individuals exhibited possible pathogenic variants. A novel missense variant p.(Q518L) was also observed and registered at NCBI with accession number PP898303. Computational algorithms identified three potential pathogenic variants. These variants induce disruptions and structural alterations which in turn compromise their functionality. This ultimately leads to retinal ganglion cells death and the eventual manifestation of glaucomatous damage resulting in vision loss.<h4>Conclusion</h4>This is the first report showing the involvement of pathogenic OPTN gene variants in JOAG cases from North Indian population which was unknown till now. This study provides population-specific data on genetic contribution of OPTN genes in JOAG pathogenesis. Genetic investigations like this may help in the understanding of disease pathogenesis and development of therapy for glaucoma management/treatment in the near future.

Huntington´s Disease (HD) is an autosomal dominant, neurodegenerative disorder with characteristic motor, behavioural, and cognitive impairment. Mutant Huntingtin may also affect peripheral tissue. We aimed to assess skeletal muscle (SMM) and fat mass, sarcopenia (EWGSOP2), and malnutrition in HD patients in early disease stages compared to age- and sex-matched healthy subjects. Body composition was evaluated by bioelectrical impedance analysis. The Unified HD Rating Scale and cognitive assessments were used for clinical characterization. Twenty early-stage HD patients (45% females) with a median age of 57 years, body mass index of 22 kg/m², Total Motor Score of 17 points, and Total Functional Capacity of 10 points were included consecutively and prospectively. Confirmed sarcopenia (15%, n = 3) was uncommon. Appendicular SMM index was reduced in 60% (n = 12) and body fat mass in 35% (n = 7). SMM reduction was significantly associated with low weight (p = 0.049) and body fat (p = 0.048). Patients in disease-stage2 (n = 11) had a lower weight (p = 0.009) and body fat (p = 0.008) than patients in disease-stage1 (n = 9; i.e., patients without functional decline). Weight was also lower (p = 0.011) when compared to 20 healthy controls (45% females; median age 56 years). Fifty-five % of HD patients were at risk for malnutrition or malnourished (Mini Nutritional Assessment). The latter correlated with weight (r_s = 0.724, p < 0.001), SMM (r_s = 0.473, p = 0.035), body fat mass (r_s = 0.611, p = 0.004), motor symptoms (r_s = - 0.519, p = 0.019), independence (r_s = 0.450, p = 0.046), and executive function (r_s = 0.526, p = 0.017). Reduction of muscle or fat mass and malnutrition are common even in early-stage HD, which may contribute to progressive wasting and dependence.

Innate-like T cells (ILTCs) have recently emerged as a new target of several cancer immunotherapies. Some unique features, such as rapid MHC-independent recognition of antigens, performing heterogeneous anti-tumor activities, and being less susceptible to tumor-induced suppression, suggest promising roles of ILTCs in cancer immunology. On the other hand, these cells exhibit a dualistic nature in cancer, which includes both pro-tumor and anti-tumor effects, highlighting the importance of the complex regulatory environment of their functioning and activation. The functions and evolution of ILTC are greatly influenced by microRNAs (miRNAs), small non-coding RNA molecules that mediate post-transcriptional regulation. Considering the kind of ILTCs, miRNA, and cancer type, this interaction resembles both a tumor promoter and suppressor. ILTC functions and evolution are closely associated with microRNAs (miRNAs), small noncoding RNA molecules that play posttranscriptional regulatory roles. Depending on the type of ILTCs, miRNA, and cancer, this interaction resembles both a Tumor promoter and a tumor Suppressor. This review addresses the complicated relationship between ILTCs and different miRNAs, such as miR-155, let-7 s, and miR-181a, expressed in tumor cells, ILTCs, or packed in tumor-derived exosomes. We will underscore the synergetic effects of the expression of miRNA in tumor and immune cells, influencing ILTC's function and cancer progression. We emphasized the recent and innovative therapeutic approaches, novel delivery systems, and CAR-T cell-based strategy, and the therapeutic potential of modifying the expression of miRNA to regulate the miRNA expression to modulate ILTC activity and improve cancer treatment.

<h4>Purpose of review</h4>This review examines the role of adipokines and gene polymorphisms in the development of depression and obesity. It is of great importance to understand the mechanisms that may be effective in the development of obesity and depression as their incidence increases.<h4>Recent findings</h4>Adipokines are released from adipose tissues and primarily regulate the connection between the metabolic and inflammatory effects of obesity and the brain cells and adipose tissue. Adipokines may potentially contribute to the pathophysiology of depression by influencing the HPA axis and neurotransmitters. According to some estimates, the genetic overlap between obesity and depression is as high as 12 percent. Furthermore, these genes may be linked to significant interconnected signaling networks that have a role in the etiology of both disorders. Obesity and depression are both on the rise globally, and it is thought that there is a bidirectional relationship between these two conditions. Obesity and obesity-induced depression seriously limit the psychosocial functionality of individuals and impair their quality of life. Having a high body mass index (BMI) raises the likelihood of developing depression. On the other hand, as the BMI elevates in people suffering from depression, the possibility of developing obesity also rises.

Lung adenocarcinoma (LUAD), the most common subtype of non-small-cell lung cancer (NSCLC), is often driven by mutations, particularly in epidermal growth factor receptor (EGFR), that guide targeted therapy choices. However, resistance to these treatments remains a major clinical challenge. Raf kinase inhibitory protein (RKIP), encoded by the PEBP1 gene, a metastasis suppressor, modulates key oncogenic pathways and may influence tumor aggressiveness and therapy response. Yet, its specific role in NSCLC remains unclear. This study investigates the influence of RKIP expression on NSCLC aggressiveness and explores its impact on therapy response, particularly to EGFR-targeted therapies. In silico analyses revealed that lower RKIP mRNA expression correlates with poorer survival outcomes in LUAD patients but not in other NSCLC subtypes. Genetic modulation of RKIP expression in LUAD cell lines demonstrated that its overexpression reduced migration, spheroid integrity, and suppressed tumor growth, whereas RKIP knockout had opposite effects, particularly in vivo. Expression profiling showed that RKIP overexpression impacts the activation of mitogen-activated protein kinase (MAPK), RAC serine/threonine-protein kinase (AKT), and signal transducer and activator of transcription 3 (STAT3) pathways, as well as processes related to extracellular matrix regulation and inflammatory responses. Importantly, in vitro and in vivo experiments demonstrated that RKIP overexpression sensitizes cells to anti-EGFR treatments, whereas RKIP knockout diminished their sensitivity. Overall, our findings indicate that RKIP modulates LUAD progression and response to EGFR-targeted therapies, although its clinical value as a biomarker requires further validation. These findings highlight RKIP's potential in overcoming therapeutic resistance and the need for further investigation into its regulatory mechanisms.

<h4>Background</h4>Chronic rhinosinusitis (CRS) with nasal polyps is a heterogeneous chronic inflammatory disease generally divided into two phenotypes including eosinophilic CRS with nasal polyps (eCRSwNP) and non-eosinophilic CRS with nasal polyps (neCRSwNP). However, its pathogenesis remains largely unknown. The aim of this study was to explore mechanistic differences between eCRSwNP and neCRSwNP using a bioinformatics approach.<h4>Methods</h4>We comprehensively analyzed single-cell RNA sequencing data from 3 healthy controls and 6 patients with CRSwNP (including 3 with eCRSwNP and 3 with neCRSwNP) to explore the heterogeneity and potential mechanisms of CRSwNP.<h4>Results</h4>Cluster analysis based on differential gene expression delineated 14 cell clusters. The eCRSwNP group exhibited a markedly higher prevalence of glandular cells and a notable reduction in fibroblasts, myoepithelial cells, and secretory cells compared to patients with neCRSwNP. Functional enrichment analysis of differentially expressed genes revealed the activation of pathways such as IL2-STAT5 signaling and the inhibition of apoptotic pathways in eCRSwNP compared to neCRSwNP. Significant differences in the metabolic profiles of epithelial cell subpopulations were observed between eCRSwNP and neCRSwNP. Furthermore, there were notable discrepancies in the numbers and functionality of immune cells between eCRSwNP and neCRSwNP. The CD4+Th2 cell subsets were found to be significantly enriched in eCRSwNP. The highest number of cellular communications from type 2 conventional dendritic cells (cDC2) to CD4+Th2 cells was found in CRSwNP, where the ICAM1-CD226 pathway from cDC2 to CD4+Th2 was significantly upregulated in eCRSwNP. In addition, eCRSwNP was mainly infiltrated with tissue-resident macrophages, whereas neCRSwNP was mainly infiltrated with monocyte-derived macrophages.<h4>Conclusions</h4>Our study provides new insights into the heterogeneity, molecular mechanisms, and biomarkers of CRSwNP, contributing to improved diagnostic and therapeutic options for this condition.

Geomagnetic storms represent a critical yet sometimes overlooked factor affecting the reliability of modern power systems. This study examines the relationship between geomagnetic storm activity-characterized by the Dst index and categorized into weak, moderate, strong, severe, and extreme intensities-and reported power outages of unknown or unusual origin in the United States from 2006 to 2023. Outage data come from the DOE OE-417 Annual Summaries, while heliospheric and solar wind parameters (including proton density, plasma speed, and the interplanetary magnetic field) were obtained from NASA's OMNIWeb database. Results indicate that years with a higher total count of geomagnetic storms, especially those featuring multiple strong or severe events, exhibit elevated incidences of unexplained power interruptions. Correlation analyses further reveal that increasingly negative Dst values, enhanced solar wind velocity, and higher alpha/proton ratios align with greater numbers of outages attributed to unknown causes, underscoring the pivotal role of solar wind-magnetosphere coupling. A simple regression model confirms that storm intensity and average magnetic field strength are statistically significant predictors of unexplained outages, more so than broad indicators such as sunspot number alone. These findings highlight the importance of monitoring high-intensity geomagnetic storms and associated heliospheric variables to mitigate potential risks. Greater attention to space weather impacts and improved reporting of outage causes could bolster grid resilience, helping operators anticipate and manage disruptions linked to geomagnetic disturbances.

Extracellular vesicles (EVs) play a crucial role in intercellular communication, signaling pathways, and disease pathogenesis by transporting biomolecules such as DNA, RNA, proteins, and lipids derived from their cells of origin, and they have demonstrated substantial potential in clinical applications. Their clinical significance underscores the need for sensitive methods to fully harness their diagnostic potential. In this comprehensive review, we explore EV heterogeneity related to biogenesis, structure, content, origin, sample type, and function roles; the use of EVs as disease biomarkers; and the evolving landscape of EV measurement for clinical diagnostics, highlighting the progression from bulk measurement to single vesicle analysis. This review covers emerging technologies such as single-particle tracking microscopy, single-vesicle RNA sequencing, and various nanopore-, nanoplasmonic-, immuno-digital droplet-, microfluidic-, and nanomaterial-based techniques. Unlike traditional bulk analysis methods, these methods contribute uniquely to EV characterization. Techniques like droplet-based single EV-counting enzyme-linked immunosorbent assays (ELISA), proximity-dependent barcoding assays, and surface-enhanced Raman spectroscopy further enhance our ability to precisely identify biomarkers, detect diseases earlier, and significantly improve clinical outcomes. These innovations provide access to intricate molecular details that expand our understanding of EV composition, with profound diagnostic implications. This review also examines key research challenges in the field, including the complexities of sample analysis, technique sensitivity and specificity, the level of detail provided by analytical methods, and practical applications, and we identify directions for future research. This review underscores the value of advanced EV analysis methods, which contribute to deep insights into EV-mediated pathological diversity and enhanced clinical diagnostics.

Through international gene-matching efforts, we identified 10 individuals with ultrarare heterozygous variants, including 5 de novo variants, in <i>BMAL1</i>, a core component of the molecular clock. Instead of an isolated circadian phenotype seen with disease-causing variants in other molecular clock genes, all individuals carrying <i>BMAL1</i> variants surprisingly share a clinical syndrome manifest as developmental delay and autism spectrum disorder, with variably penetrant sleep disturbances, seizures, and marfanoid habitus. Variants were functionally tested in cultured cells using a <i>Per2</i>-promoter driven luciferase reporter and revealed both loss-of-function and gain-of-function changes in circadian rhythms. The tested <i>BMAL1</i> variants disrupted <i>PER2</i> mRNA cycling, but did not cause significant shifts in cellular localization or binding with CLOCK. Conserved variants were further tested in <i>Drosophila</i>, which confirmed variant-dependent effects on behavioral rhythms. Remarkably, flies expressing variant <i>cycle</i>, the ortholog of <i>BMAL1</i>, also demonstrated deficits in short- and long-term memory, reminiscent of the highly prevalent developmental delay observed in our cohort. We suggest that ultrarare variants in the <i>BMAL1</i> core clock gene contribute to a neurodevelopmental disorder.

Inflammation plays a key role in the pathogenesis of various diseases, including inflammatory bowel diseases such as ulcerative colitis and Crohn's disease. Similar to animals, plant cells produce exosome-like nanovesicles (ELNs) that play a role in transmitting biological signals from specific types of cells or tissues to other cells or tissues. Here we show that ELNs derived from Peucedanum japonicum (PjELNs) exhibit potent anti-inflammatory effects on macrophages and are effective in a mouse colitis model induced by dextran sodium sulfate (DSS). RNA sequence analysis reveals that PjELNs suppress multiple inflammatory cytokine-mediated signaling pathways in LPS-stimulated macrophage cell lines. We further show that PjENLs promote the differentiation of goblet cells, thereby enhancing mucin production and providing a protective effect on mucosal damage in a DSS-induced murine model of colitis. We also demonstrate that PjELNs inhibit the DSS-induced reduction in the number of transit amplifying cells. PjELNs exhibit anti-inflammatory properties, partly due to the presence of miRNA-like small RNAs that directly regulate the expression of the inflammatory cytokine IL6. Importantly, these small RNAs exhibit cross-species effects in both humans and mice. These findings reveal the mechanism by which plant-derived ELNs modulate inflammatory responses, suggesting their potential as a preventive and therapeutic strategy for inflammatory diseases of the colon.

Developmental stuttering is a highly heritable, common speech condition characterized by prolongations, blocks and repetitions of speech. Although stuttering is highly heritable and enriched within families, the genetic architecture is largely understudied. We reasoned that there are both shared and distinct genetic variants impacting stuttering risk within sex and ancestry groups. To test this idea, we performed eight primary genome-wide association analyses of self-reported stuttering that were stratified by sex and ancestry, as well as secondary meta-analyses of more than one million individuals (99,776 cases and 1,023,243 controls), identifying 57 unique loci. We validated the genetic risk of self-reported stuttering in two independent datasets. We further show genetic similarity of stuttering with autism, depression and impaired musical rhythm across sexes, with follow-up analyses highlighting potentially causal relationships among these traits. Our findings provide well-powered insights into genetic factors underlying stuttering.

Brain cancer represents a complex disease influenced by a multitude of genetic and epigenetic factors. This study aims to elucidate the role of specific single nucleotide polymorphisms (SNPs) and long non-coding RNAs (lncRNAs) in the pathogenesis of brain cancer, employing a multi-omics approach. We conducted extensive eQTL, mQTL, haQTL, sQTL, and caQTL analyses to identify genetic variants and lncRNAs associated with brain cancer. Integration with GWAS-GWAS colocalization analysis provided insights into shared genetic mechanisms with other diseases. We further investigated copy number variation (CNV) and methylation status in relation to gene expression, and their prognostic implications in different brain cancer subtypes. SNP rs615552_AL359922.1 exhibited significant colocalization with key genes CDKN2A, CDKN2B, and CDKN2B-AS1, implicating its role in the regulation of gene expression. The long non-coding RNA CDKN2B-AS1 demonstrated both co-occurrence and co-expression with CDKN2A and CDKN2B, suggesting a coordinated regulatory mechanism among these genes. TERT emerged as a gene with shared susceptibility across brain cancer and other diseases, indicating a common genetic pathway. Methylation sites associated with mQTL, such as cg03935379 (TERT) and cg14069088 (CDKN2A), were identified as independent prognostic factors for lower-grade glioma (LGG), but not for glioblastoma multiforme (GBM). Bulk RNA-seq, spatial, and single-cell transcriptomic analyses revealed that CDKN2B-AS1 is predominantly expressed in malignant and dendritic cells (DCs), and is associated with DNA repair pathways in malignant cells and antigen presentation genes in DCs. This study offers a comprehensive perspective on the genetic and molecular factors influencing brain cancer. The results highlight the intricate nature of gene regulation in cancer and suggest the potential of CDKN2B-AS1 as a key regulator of immune responses and tumor suppressor genes. The SNP rs615552_AL359922.1 represents a significant pathogenic SNP for brain cancer. The identification of shared genetic mechanisms, particularly involving TERT, with other diseases points to new opportunities for developing therapeutic targets and diagnostic tools. Future research should focus on functional validation and the investigation of environmental interactions to fully leverage these findings for advancing brain cancer management.

The incidence of breast cancer continues to increase annually, posing a significant challenge for countries worldwide in terms of its prevention and treatment. Therefore, identifying novel therapeutic targets for breast cancer is urgently needed. The peroxiredoxin (PRDX) family is regarded as a good diagnostic marker for various tumors. However, the expression and prognostic significance of PRDX family members in breast cancer remain unclear and require systematic investigation. By using bioinformatic tools such as UALCAN, TIMER2.0, Human Protein Atlas Project (HPA), Gene Set Cancer Analysis (GSCA), and the cBioportal database, we systematically analyzed the expression pattern, prognostic value, methylation status and immune infiltrating association of PRDX gene family members in breast cancer. Through comprehensive analysis, we found that PRDX4 has good prognostic value and is closely related to immune infiltration, and further exploration of its oncogenic function in breast cancer is warranted. Subsequently, we performed a series of cellular assays to explore the potential role of PRDX4 in the progression of breast cancer. We demonstrated that PRDX4 promoted the proliferation, invasion, metastasis, and inhibited the apoptosis of breast cancer cells. In addition, PRDX4 expression was associated with the half maximal inhibitory concentration (IC₅₀) of neratinib which primarily targets human epidermal growth factor receptor 2 (HER2) and showed good binding in molecular docking. Our subsequent experiments showed that the PRDX4-HER2 axis may serve as a potential combined target for neratinib therapy. Our findings suggest that PRDX4 may be a potential diagnostic and prognostic marker for breast cancer, and targeting PRDX4 could represent a novel strategy to improve the efficacy of targeted therapy for patients with HER2-positive breast cancer.

The dynamic nature of supramolecular networks of telechelic polymers offers new avenues for the design of novel materials with enhanced melt strength and extensibility, increased energy at break, or self-healing properties. However, monitoring the kinetics of the underlying molecular-level scission-reaggregation events remains challenging, particularly in high-molar-mass polymers in the bulk state. Here, we employ solid-state ¹H NMR spectroscopy relaxation dispersion experiments to investigate the aggregation-scission dynamics in poly(ε-caprolactone) modified with oligopeptide end groups that form one-dimensional hydrogen-bonded aggregates. We have successfully determined the timescale of end-group dissociation directly and independently of any relaxation of the polymer segments at different temperatures in the bulk semi-crystalline and melt state. This site-specific, non-destructive method is applicable to entangled, high-molar-mass polymers without chemical modifications or modeling, provides critical insight into the dynamics of supramolecular networks in the bulk state, and promises to be a valuable tool for the directed development of next-generation functional materials.

Cancer stem cells (CSCs) are a specific subset of cancer cells that possess the ability to self-renew, resist therapies, and promote metastasis, making them a crucial target in cancer treatment. This study investigates the therapeutic potential of natural compounds in targeting CSCs, particularly their ability to inhibit key signaling pathways, induce apoptosis, and alter the tumor microenvironment. This article reviews the molecular mechanisms that maintain CSCs and contribute to their resistance, focusing on the roles of the WNT/β-catenin, Hedgehog, Notch, and PI3K/ATK/mTOR pathways. Several natural compounds, including curcumin, resveratrol, epigallocatechin gallate, and sulforaphane, were assessed for their effectiveness in targeting CSCs. The finding revealed that these natural compounds can inhibit CSC proliferation, enhance sensitivity to chemotherapy, and suppress the tumor-supportive microenvironment. Notably, compounds such as berberine and piperine were found to reverse drug resistance by downregulating efflux transporters, while quercetin and salinomycin selectively induced apoptosis in CSCs. Overall, natural compounds show promising potential for targeting CSCs in therapy. However, challenges related to bioavailability and metabolic stability must be addressed through advanced drug delivery systems and combination therapy.

Parkinson's disease (PD) is a progressive neurodegenerative disorder primarily characterized by the progressive loss of dopaminergic neurons. Mutations in the PLA2G6 gene, which encodes calcium-independent phospholipase A2 (iPLA2β), have been associated with autosomal recessive early-onset parkinsonism, a subtype of neurodegeneration marked by brain iron accumulation. Although the pathogenic mechanisms underlying PLA2G6-related neurodegeneration remains unclear, disturbances in iron metabolism, neuroinflammation, and mitochondrial dysfunction are thought to play key roles. Ferroptosis, an iron-dependent form of regulated cell death driven by lipid peroxidation, has recently been implicated in PD. Here, we demonstrate that the iPLA2β deficiency in dopaminergic neurons induces ferroptosis, aggravating neurodegeneration and motor deficits in a mouse model of PLA2G6-associated neurodegeneration (PLAN). This ferroptotic phenotype is characterized by increased iron accumulation, elevated lipid peroxidation, and impaired antioxidant defenses, including downregulation of ferritin heavy chain 1 (FTH1) and glutathione peroxidase 4 (GPX4). We further identify peroxiredoxin 6 (PRDX6) as a direct binding partner of iPLA2β and a critical regulator of ferroptosis. Loss of iPLA2β destabilizes PRDX6, promoting its degradation and thereby enhancing ferroptotic susceptibility. Notably, restoration of PRDX6 expression alleviates ferroptosis in iPLA2β-deficient cells, highlighting the protective role for the PRDX6/FTH1/GPX4 axis in maintaining redox homeostasis. Furthermore, treatment with the ferroptosis inhibitor Liproxstatin-1 (Lip-1) attenuated motor dysfunction and dopaminergic neuron loss in PLA2G6 knockout (KO) mice. Collectively, our findings uncover a novel mechanism linking iPLA2β deficiency to ferroptosis in PD and suggest ferroptosis inhibition as a promising therapeutic strategy for PD patients with PLA2G6 mutations.

<h4>Background</h4>Epigenetic acetylation plays an essential role in the development and drug resistance of luminal breast cancer. However, the acetylation regulatory network in luminal breast cancer remains underexplored.<h4>Methods</h4>We used the TCGA-BRCA database to explore the acetylation regulatory network in luminal breast cancer. Spearman correlation coefficients, Cox proportional hazards, and the STRING database were used to identify genes that were correlated with acetylation regulatory molecules in luminal breast cancer and could predict patient outcomes. An acetylation regulatory risk model was constructed via Consensus Cluster Plus and the LASSO risk model. GSEA, K‒M survival analysis, and receiver operating characteristic (ROC) curve analysis were used to analyze survival and possible regulatory pathways of the risk model. TIDE, Microenvironment Cell Populations-counter, and CIBERSORT algorithms were used to analyze the immune landscape of the risk model population. Patients' tumor specimens were used to detect the expression of KAT2B and TAF1L. The luminal breast cancer cell lines MCF-7 and T47D were used in cell viability, Transwell, western blotting, and RT‒qPCR experiments to confirm the risk model. Mouse model was constructed for in vivo validation of KAT2B and TAF1L function.<h4>Results</h4>In our study, we utilized the TCGA-BRCA database to conduct a comprehensive analysis of the acetylation regulatory pattern in luminal breast cancer. Using Consensus Cluster Plus and the LASSO risk model, we screened 6 acetylation-related genes (KAT2B, TAF1L, CDC37, CCDC107, C17orf106, and ASPSCR1) and constructed a 6-gene risk model of luminal breast cancer. Based on this model, luminal breast cancer patients were classified into high- and low-risk subgroups. The high-risk subgroup had a poor prognosis. Further analysis revealed that the high-risk subgroup was associated with lower CD8 + T-cell infiltration and greater responsiveness to immune checkpoint inhibitor therapy. In vitro and in vivo experiments revealed that knockdown of KAT2B and TAF1L dramatically inhibited tumor cell proliferation. In vitro experiments also showed knockdown of KAT2B and TAF1L dramatically inhibited tumor cell migration, increased lymphocyte infiltration, and significantly upregulated the expression of CD8 + T-cell-associated chemokines in luminal breast cancer cells.<h4>Conclusions</h4>In this study, we successfully constructed a 6-gene acetylation-associated risk model for luminal breast cancer, providing a new direction and evidence for personalized treatment. Our results also suggested that KAT2B and TAF1L might serve as potential therapeutic targets in luminal breast cancer.

<h4>Background</h4>The quantity and quality of diet are directly associated with the development of obesity and diseases. This study aims to investigate the relationship between the Healthy Eating Index-2015 (HEI-2015) and anthropometric measurements, including body mass index (BMI), body adiposity index (BAI), body roundness index (BRI), visceral adiposity index (VAI), a body shape index (ABSI), lipid accumulation product (LAP) index, and triglyceride glucose index (TyG) index in non-alcoholic fatty liver disease (NAFLD) patients with diabetes.<h4>Methods</h4>This is a cross-sectional study that was conducted on the baseline data from Fasa cohort study. The socio-demographic, anthropometric, and dietary data of 384 individuals with diabetes and NAFLD were extracted. Multiple linear regression was used in various models adjusted for potential confounders to determine the associations.<h4>Results</h4>Out of the 384 eligible individuals, 74.5% were women. The mean age of men was 55.74 ± 7.85 years, whereas that of women was 54.68 ± 8.54 years. After adjusting for possible confounders, including age, marital status, occupation, education status, smoking status, physical activity, and total energy intake, there was no significant association between the quartiles of HEI score and any of the anthropometric measurements, including BMI, ABSI, BAI, BRI, VAI, LAP index, and TyG index among both men and women.<h4>Conclusion</h4>Higher HEI score was not associated with anthropometric indices in NAFLD patients with diabetes, according to our findings. It is recommended that further observational studies should be conducted to clarify the current findings.

<h4>Objectives</h4>The objective of this research is to investigate the pathophysiological progression of HIV from acute infection to chronic immunodeficiency (AIDS) and to understand the host's immunological responses, which are pivotal for elucidating disease aetiology and optimizing antiretroviral therapy (ART). Additionally, the study aims to explore the role of exosomes (40-130 nm bilipid-layered vesicles released by nearly all cell types) as key mediators of intercellular communication in the context of HIV infection.<h4>Materials and methods</h4>Recent research has uncovered that cells infected with HIV-1 release exosomes carrying a mix of viral and host components such as proteins, nucleic acids, lipids, and other metabolites. To decipher the plausible role of exosome-derived proteins in HIV disease progression, the exosomes isolated from HIV patient's serum were subjected to LC-MS/MS analysis to identify exosome-derived human and viral protein sequences. The identified proteins were then investigated, annotated, and explored for protein-protein interaction (PPI) network between HIV and the human host's proteins. Earlier experimental efforts focused on identifying PPI networks in host cells or only within the virus.<h4>Results</h4>The analysis showed that out of twelve exosome-derived host proteins identified from HIV-1 patient's samples, only five of the proteins were associated with Toll-like receptors (TLR), inflammasome-activation, inhibition of apoptosis, innate immune response modulation, and autophagy pathways. In the TLR pathway, CDH5, ENO1, OGT, TJP1, and TRAF6 exosome-derived host proteins participated in regulation. Notably, CDH5, ENO1, OGT, and TRAF6 were shared among these pathways, BioGRID version 4.4 showed that HIV-1 Gag, Gag-pol, Env, and Nef proteins interact with 196, 162, 158, and 80 human proteins, respectively, associated with different innate immune response pathways.<h4>Conclusion</h4>These findings are a step ahead in comprehending the pathophysiology of HIV1 and the innate immune response pathways, providing excellent opportunities to explore further exosome-based biomarkers for theranostic approaches.

The constitutive centromere-associated network (CCAN) of the inner kinetochore links CENP-A-containing nucleosomes of the centromere to the outer kinetochore, ensuring accurate chromosome segregation during mitosis. CCAN binding at the centromere is stabilized upon mitotic entry, but the underlying mechanisms remain unclear. Here, we demonstrate that cohesin is essential for CCAN stability. The chromosomal passenger complex (CPC), independently of its kinase subunit Aurora B, regulates cohesin-mediated CCAN stability via heterochromatin protein-1 (HP1), Haspin kinase, and phosphorylation of the cohesin-release factor WAPL, which weakens WAPL's affinity for PDS5B. While cohesin depletion disrupts CCAN stability, neither separase-mediated cohesin cleavage nor depletion of the cohesion-essential Esco2 acetyltransferase affects CCAN stability, indicating that cohesin stabilizes the CCAN independently of sister chromatid cohesion. Furthermore, we show that WAPL phosphorylation maintains a centromere-proximal pool of cohesin and promotes the formation of the primary constriction. These findings establish a non-cohesive function of cohesin in stabilizing the CCAN during mitosis and suggest that cohesin-mediated organization of centromeric chromatin strengthens kinetochore engagement to ensure faithful chromosome segregation.

The efficacy of neoadjuvant radiotherapy (RT) in patients with rectal cancer (RC) is hindered by the plasticity and heterogeneity of cancer-associated fibroblasts (CAFs). However, the underlying mechanisms remain poorly understood. In this study, single-cell RNA sequencing of patients with RC samples revealed a CAF subpopulation characterized by high interferon (IFN) regulatory factor 1 (IRF1) expression. These IFN-licensed CAFs (ilCAFs) are enriched in tumors with enhanced RT responses across various solid tumors, including RC. Mechanistically, IFN gamma (IFN-γ) signaling drives the polarization of ilCAFs, leading to the recruitment of T cells and dendritic cells via CCL4/CCL5 secretion. Activation of IFN-γ/stimulator of IFN genes (STING) signaling reprograms the stroma and augments anti-tumor immunity in both RT-sensitive and RT-resistant colorectal cancer. Silencing STING in CAFs impairs ilCAF enrichment and diminishes tumor sensitivity to RT. Combining STING agonists with RT results in robust tumor control, providing a compelling rationale for clinical translation.

<h4>Introduction</h4>Chronic low back pain (CLBP) and depressive symptoms (DS) are highly prevalent, burdensome, costly and comorbid health conditions. Osteopathic manipulative treatment (OMT) was shown to improve pain and disability in patients with CLBP; however, the effect on comorbid DS remains less certain. Interestingly, CLBP and DS seem to be associated with changes in interoception, which may be reversed by OMT.<h4>Methods and analysis</h4>The study protocol proposes a single-blinded, parallel-group, randomised controlled trial to investigate the effect of OMT on clinical symptoms (depression, pain and disability) and interoceptive functions (interoceptive accuracy, sensibility and awareness) in patients with CLBP and comorbid DS. A sample of 60 adult subjects with CLBP and comorbid DS shall be recruited from osteopathic, orthopaedic and physiotherapeutic practices and educational institutes for osteopathy, sports science, psychology and medicine in Hamburg, Germany. Participants will be randomly allocated (1:1 ratio) to receive six 45 min treatment sessions of either OMT (standard-OMT group) or sham treatment imitating OMT (sham-OMT group). Primarily, symptoms of depression, pain and disability will be assessed with the Beck's Depression Inventory, Second Edition (BDI-II), Numeric Rating Scale (NRS) and Oswestry Disability Index (ODI). Secondarily, interoceptive accuracy, sensibility and awareness will be evaluated using the Heartbeat Tracking Task (HTT), Multidimensional Assessment of Interoceptive Awareness (MAIA-2) and confidence-accuracy correspondence (CAC). Ancillary, the therapeutic alliance will be investigated with the Helping Alliance Questionnaire. Data will be collected at baseline (t₀), the first, third and sixth treatment sessions (t₁, t₃, t₆) and at 3 months follow-up (t₇). The findings will be analysed for between-group differences using descriptive (mean and SD) and inductive statistics (mixed analysis of variance). It is hypothesised that standard-OMT, compared with sham-OMT, will reduce depression, pain and disability (BDI-II, NRS, ODI) and increase interoceptive accuracy, sensibility and awareness (HTT, MAIA-2, CAC) in patients with CLBP and comorbid DS.<h4>Ethics and dissemination</h4>The study was approved by the ethics committee of the Medical School Hamburg (MSH-2023/288). The anonymised dataset will be published in an online repository, and the results will be published in peer-reviewed scientific journals.<h4>Trial registration number</h4>DRKS00031694.

Poly(ADP-ribose) polymerase 2 (PARP2) plays a crucial role in DNA repair. A common single-nucleotide polymorphism (SNP) in the PARP2 gene, rs3093921, has been associated with pancreatic cancer and marginal zone lymphoma. This SNP results in a missense mutation, D235G, in the PARP2 protein. PARP2 is also reported to undergo posttranslational modifications (PTMs), particularly phosphorylation at serine residues 226, 232, and 353. The C-terminal region of PARP2 includes the Trp-Gly-Arg (WGR) domain, the ADP-ribosyl transferase (ART) domain, and the helical subdomain (HD). The latter two, spanning residues 220-583, comprise the catalytic region of PARP2. The DNA-induced enzymatic activation of PARP2 is regulated by local destabilization of the HD domain. We used molecular dynamics simulations to investigate the impact of these three PTMs on both the wild-type (WT) and the mutant (D235G) forms of PARP2. Our simulations suggest that the mutation leads to a change in structure and residual flexibility in specific regions of the HD domain compared with the WT. The presence of the PTMs also shows altered structure of the HD domain, although the residual flexibility remains unchanged compared with the unmodified WT. This is further supported by the destabilizing interactions between the mutation site and the HD domain observed in the presence of both the mutation and the PTMs. Importantly, our results suggest that the three PTMs mitigate the changes in structure and residual flexibility caused by the mutation, helping the PTM-modified mutant protein retain WT-like features. We observe that, while the PTMs alter the dominant mode of motion in the native protein, this shift is not observed in the mutant, further indicating that PTMs mitigate the structural and dynamic consequences of the mutation, preserving WT-like behavior. The mutation and PTMs cause notable changes in correlated motions within the WGR and ART domains, with the PTM-modified mutant structure showing the strongest correlations. Interestingly, a similar trend is observed for anticorrelated motions, particularly between the WGR and ART domains, where the PTM-modified mutant structure again exhibits the highest level of anticorrelation. Additionally the mutation in PARP2 markedly weakens interdomain interactions between the HD and ART domains, as well as between the HD and WGR domains. By contrast, the PTMs alone show no change on the HD-ART interaction, although they weaken the HD-WGR interaction. For the PTM-modified mutant structure the HD-ART interaction remains disrupted compared with the WT, but the extent of disruption is less than that caused by the mutation alone-suggesting a partial compensatory effect of the PTMs. Additionally, the HD-WGR interaction remains consistently weakened in all modified systems.

We aimed to investigate the association of brain morphology with behavioral and emotional problems in early adolescence using a brain atlas of the serotonin system. This pre-registered study used data from the Generation R Study, a large birth cohort in Rotterdam, the Netherlands. A total of 2492 children were included at age 10 years, with neuroimaging data and self-reported behavioral and emotional problems. Cortical surface area and thickness were measured in ten serotonin-coupled brain regions. Our primary analysis revealed that higher total, attention, and externalizing problem scores were associated with smaller cortical surface area in Regions 8, 9, and 10 (covering cingulate, orbitofrontal, temporal, and parietal areas) after adjusting for intracranial volume, highlighting region-specific effects less confounded by overall head size. Among these regions, only Region 9 showed relative enrichment for 5-HT_1A receptors, suggesting potential serotonergic involvement in externalizing problems. Secondary analyses showed that greater cortical thickness in Region 2, enriched with serotonin transporters (5-HTT) and involving parts of the temporal cortex and insula, was associated with higher total, attention, and internalizing problem scores (β = 0.07, PFDR = 0.003). A follow-up analysis in an independent adult sample (n = 100), with the same-subject structural MRI and molecular 5-HTT imaging, revealed a specific negative association between 5-HTT availability and cortical thickness in Region 2 (β = -0.22, P = 0.02). These findings suggest selective serotonergic contributions to cortical morphology related to behavioral problems.

Netrin-1, a central axonal guidance molecule discovered for its role in neuronal development, is also essential in neurodegenerative diseases. Netrin-1 inhibits leukocyte migration and inflammation-related tissue damage outside the central nervous system. Therefore, it can be viewed as a potential biomarker for inflammatory activity in neurodegenerative diseases. Recent studies highlight the dual roles of Netrin-1 in neuroinflammation and neurodegenerative diseases. In the context of neurodegeneration, Netrin-1 demonstrates both protective and harmful effects. This review highlights recent advancements in research regarding the dual roles of Netrin-1 in neuroinflammation and neurodegeneration. We discuss its involvement in protecting the blood-brain barrier (BBB) and regulating immune cell migration and its effects on various neurodegenerative diseases. A greater understanding of the multifunctionality of Netrin-1 could potentially be employed in developing new treatment modalities.

The 2002 movie <i>Catch Me If You Can</i> is a cat-and-mouse story in which Frank Abagnale Jr. successfully conned his way into several high-profile jobs while evading capture by FBI agent Carl Hanratty. Similarly, after entering host cells, viruses interact with or hijack host cellular machinery to replicate their genetical materials and assemble themselves for the next round of infection. Analogous to an FBI agent, host cells have numerous molecular "detectives" that recognize viral nucleic acids (NAs). These include RIG-I, MDA5, LGP2, TLR3, TLR7, TLR8, DHX36, DICER1, PKR, OAS1, ZAP, and NLRP1/6 for viral RNA, as well as cGAS, TLR9, AIM2, IFI16, IFIX, Ku70, MRE11, RNA polymerase III, hnRNPA2B1, LRRFIP1, DAI, DHX9 and DDX41 for viral DNA. However, much like the brilliant Frank Abagnale Jr., viruses have developed various strategies to evade host cellular surveillance-for example, by sequestering or modifying viral NAs and inhibiting or degrading host sensors. In this review, we will summarize the host sensors identified so far, discuss the latest understandings of the various strategies employed by viruses, and highlight the challenges associated with drug development to target virus or host factors. Considering recent global health challenges such as the COVID-19 pandemic and undergoing measles outbreak, understanding virus-host interactions at the molecular and cellular levels remains essential for the development of novel therapeutic strategies.

<h4>Background</h4>Hepatocellular carcinoma (HCC) is the second leading cause of cancer-related deaths in China. It has a high rate of postoperative recurrence and lacks prognostic markers. In this study, we first analyzed mitochondrial permeability transition (MPT) necrosis-associated long non-coding RNAs (lncRNAs), integrated multi-omics, and constructed a prognostic model. We also revealed the mechanism by which it regulates the immune microenvironment. This provides a new target for targeted therapy in HCC.<h4>Objective</h4>Screening and construction of a prognostic risk score model for MPT-driven necrosis-associated lncRNAs in HCC and exploration of their potential role in HCC.<h4>Methods</h4>Pearson's correlation analysis, in conjunction with The Cancer Genome Atlas (TCGA) and gene set enrichment analysis (GSEA) databases, was utilized for the identification of lncRNAs associated with mitochondrial permeability transition-driven necrosis. The development of a risk prognostic score for mitochondrial permeability transition-driven necrosis-associated lncRNAs was accomplished through the implementation of one-way regression analysis and Least Absolute Shrinkage and Selection Operator (LASSO) regression analysis. Bioinformatics analysis was performed to validate the prognostic ability and clinical application efficacy of the risk score model and prognostic genes and to explore their biological significance.<h4>Results</h4>MPT-driven necrosis-related lncRNAs (MPTDNRlncRNAs) strongly correlated with HCC were obtained through Pearson's correlation analysis. Additionally, MPT-driven necrosis-related prognostic lncRNAs were obtained through univariate Cox regression analysis. A new prognostic risk model consisting of three MPTDNRlncRNAs was constructed using LASSO-Cox regression. The model was tested using multiple bioinformatics methods, which suggested that it could significantly differentiate between high- and low-risk groups (p < 0.05) and demonstrated good survival prediction efficacy [area under the curve (AUC) = 0.725]. Differential genes in the high- and low-risk groups were enriched in pathways related to the cell cycle and cellular composition. Combined with immune cell infiltration and immune function scores, these results showed that the patients in the low-risk group had a more significant clinical response to immunotherapy (p < 0.05). Furthermore, the expression level of prognostic genes was verified using the RT-qPCR method on cancerous and paracancerous tissues from HCC patients who underwent HCC resection at our hospital.<h4>Conclusion</h4>The risk scoring model and prognostic genes in this study have been shown to possess satisfactory predictive values, which may prove beneficial for the assessment of risk and the selection of individualized chemotherapy regimens for patients with HCC. A preliminary discussion is presented on the potential biological significance of risk scores in HCC.

<h4>Background</h4>Peritoneal dissemination in colorectal cancer (CRC) is associated with poor prognosis due to limited efficacy of current therapeutic strategies. The cholinergic receptor nicotinic beta 2 subunit (<i>CHRNB2</i>), a component of the acetylcholine receptor, has been implicated in other malignancies, but its role in CRC remains unknown.<h4>Methods</h4>This study evaluated the expression and function of CHRNB2 in CRC. CHRNB2 mRNA levels were quantified by qRT-PCR in cell lines and clinical specimens. Functional assays were conducted using CRC cell lines with high CHRNB2 expression, in which CHRNB2 was knocked down by shRNA. Cell proliferation, migration, and invasion were assessed in vitro. In vivo effects were evaluated using subcutaneous and peritoneal xenograft models. The impact of CHRNB2 monoclonal antibody (mAb) treatment on CRC cell proliferation was also examined. Clinical correlations were assessed between CHRNB2 expression and clinicopathological features, including recurrence patterns.<h4>Results</h4>CHRNB2 expression varied among CRC cell lines, with the highest levels observed in LOVO cells. CHRNB2 knockdown significantly inhibited proliferation, migration, and invasion in vitro and suppressed tumor growth in vivo. CHRNB2 mAb treatment reduced cell proliferation. Clinically, high CHRNB2 expression correlated with a significantly higher cumulative rate of peritoneal recurrence, but not with recurrence in the liver, lungs, or lymph nodes. Multivariate analysis identified high CHRNB2 expression and T4 tumor depth as independent predictors of peritoneal recurrence.<h4>Conclusions</h4>CHRNB2 promotes the malignant phenotype of CRC, particularly in peritoneal dissemination. These findings suggest that CHRNB2 may serve as a novel diagnostic biomarker and therapeutic target for CRC with peritoneal metastasis.

About one billion people worldwide are affected by neurologic disorders. Among the various neurologic disorders, one of the most common is Alzheimer's disease (AD). AD is a neurodegenerative disorder that progressively affects cognitive functions, disrupting the daily lives of millions of individuals. Mild cognitive impairment (MCI) is often considered a prodromal stage of Alzheimer's disease. In this article, we retrieved data from the online available dataset GSE63060, which includes transcriptomic data of 329 blood samples, of which there are 104 cognitively normal controls, 80 MCI patients, and 145 AD patients. We used transcriptomic data related to all three groups to perform an over-representation analysis of the gene ontologies followed by a network analysis. The aim of our study is to pinpoint alterations, detectable through a non-invasive method, in biological processes affected in MCI that persist during AD. Our goal is to uncover transcriptomic changes that could support earlier diagnosis and the development of more effective therapeutic strategies, starting from the early stages of the disease, to slow down or mitigate its progression. Our work provides a consistent picture of the transcriptomic unbalance of many genes strongly involved in ribosomal formation and biogenesis and splicing processes both in patients with MCI and with AD.

Huntington's disease (HD) is characterized by progressive neurodegeneration, but increasing evidence points to multisystemic involvement, including early hepatic steatosis in pediatric HD. Therefore, it is important to consider systemic alterations, particularly in liver lipid metabolism. In this study, we analyzed fatty acid (FA) profiles in two symptomatic HD mouse models: 2-month-old <i>R6/2</i> mice representing early-onset HD and 22-month-old <i>Hdh^Q150/Q150</i> (<i>Hdh</i>) mice representing late-onset HD, along with age-matched wild-type (<i>WT</i>) controls. FA composition in liver tissue was assessed by gas chromatography-mass spectrometry (GC-MS). In <i>R6/2</i> mice, we observed increased levels of total iso-branched chain, monounsaturated, and n-6 polyunsaturated FAs compared to WT. In contrast, only a few FA species showed reduced concentrations in <i>Hdh</i> mice. Overall, our results indicate that <i>R6/2</i> mice exhibit more pronounced alterations in hepatic FA profiles than <i>Hdh</i> mice, suggesting that early-onset HD may be associated with more severe peripheral metabolic dysregulation.

The epigenetic landscape plays a pivotal role in regulating the functions of both germ and somatic cells (Sertoli and Leydig cells) within the testis, which are essential for male fertility. While somatic cells support germ cell maturation and testosterone synthesis, the epigenetic regulation of germ cells is critical for proper spermatogenesis and function. Epigenetic modifications such as DNA methylation, histone modifications, chromatin remodeling, and non-coding RNAs (ncRNAs) are crucial for regulating gene expression that is essential for spermatogenesis and reproductive function. Although numerous studies have highlighted the significance of the epigenome and its implications for male reproductive health, a comprehensive overview of the existing literature and knowledge is lacking. This review aims to provide an in-depth analysis of the role of epigenetics in spermatogenesis and reproductive health, with a specific focus on DNA methylation, histone remodeling, and small noncoding RNAs (sncRNAs). Additionally, we examine the impact of lifestyle and environmental factors, such as diet, smoking, physical activity, and exposure to endocrine-disrupting chemicals, on the sperm epigenome. We emphasize how these factors influence fertility, embryonic development, and potential transgenerational inheritance. This review underscores how recent advances in the understanding of the epigenetic modulation of testicular function can inform the pathophysiology of male infertility, thereby paving the way for the development of targeted diagnostic and therapeutic strategies.

Neurodegenerative diseases such as Alzheimer's disease, Parkinson's disease, Huntington's disease, and amyotrophic lateral sclerosis remain incurable. Current therapeutic strategies primarily focus on slowing disease progression, alleviating symptoms, and improving patients' quality of life, including the management of comorbid conditions. Over the past few decades, the incidence of diagnosed neurodegenerative disorders has risen significantly. As the number of affected individuals continues to grow, so does the urgent need for effective treatments that can halt or mitigate the progression of these diseases. Among the most promising therapeutic resources are bioactive compounds derived from fungi. The high quality of proteins, polysaccharides, unsaturated fatty acids, triterpenoids, sterols, and secondary metabolites found in fungi have attracted growing interest from researchers across multiple disciplines. One intensively studied direction involves the use of naturally occurring fungi-derived nutraceuticals in the treatment of various diseases, including neurodegenerative conditions. This article provides an overview of recent findings on fungal compounds-such as phenolic compounds, carbohydrates, peptides and proteins, and lipids-that may have potential applications in the treatment of neurodegenerative diseases and the alleviation of their symptoms.

Amyotrophic lateral sclerosis (ALS) is a rare motor neurodegenerative disease affecting multiple cellular proteins during the progression of the disease. ALS was first discovered by Charcot in 1869, and since then, scientists have been unable to identify a singular cause of the disease. Further, there are no effective treatments available to cure ALS. The benchmark discovery of humanized preclinical SOD1 mouse models, which recapitulates the clinical and pathological phenotypes of human ALS, gives hope to medicinal chemists and neuroscientists around the globe that a suitable drug-like molecule can be discovered and translated into human beings as a means to slow down the progression of the disease. However, little success has been achieved until now in terms of finding an effective treatment for heterogenic and incurable ALS. One area marked for improvement is the use of semiquantitative, antibody-based targeted Western blotting (WB) experiments, which lack the power to analyze multiple cellular events within the entire dysregulated proteomic system. With the inconsistency of WB experiments, unexpected cellular pathways go undiscovered, and hence, loss of translation with no target engagement is seen from preclinical to human clinical ALS. Recent advancements in discovery-based quantitative proteomics have many advantages over WB. These innovative techniques could help solve the inherent problem in WB and their inability to discover multiple altered proteins with the added capability of longitudinal analysis in preclinical SOD1 models, further validating the findings in human ALS. Herein, we applied a holistic approach to summarize various reports on the use of proteomics in ALS from the published literature, and importantly, we found that using a discovery-based proteomics approach in SOD1 preclinical ALS models has revealed a more diverse and global picture of pathological proteins that affect multiple pathways during different stages of disease progression. Furthermore, we found that the proteomic profiling of the humanized SOD1 mouse model provided a proof of principle for translating the diverse pathological biomarker proteins identified in clinical human ALS cases. Moreover, we believe that advancements in the proteomics approach toward ALS biomarkers could bridge the gap between preclinical and clinical studies, enabling scientists worldwide to discover novel biomarkers and treatments that modify the progression of ALS.

Conventional approaches to analyzing electrocardiograms (ECG) in discrete parameters (such as the PR interval) ignored the high dimensionality of data omitted subtle but relevant information. We applied a variational auto-encoder to learn the underlying distributions of the ECG of 41,927 UK Biobank participants, generating 32-dimensional representation (latent factors). The latent factors showed correlations to conventional ECG parameters and strong associations to cardiac phenotypes estimated from magnetic resonance imaging. We found definitive associations of the latent factors to conduction, rhythm, and structural disorders (all <i>p</i> < 4.51 × 10^-308) and additionally value in mortality prediction. Genome wide association study (GWAS) of the latent factors, revealed 170 genetic loci with 29 not previously associated with electrocardiographic phenotypes. Further characterization of the genetic signals suggested involvement in cardiac development, contractility, and electrophysiology. Our results supported that the deep representation learning of 12-lead ECG could provide clinically meaningful and interpretable insights into cardiovascular biology and health.

Platinum-based chemotherapy is the standard treatment for advanced non-small cell lung cancer (NSCLC); however, innate and acquired resistance is a major obstacle. To determine the transcriptional regulators of resistance, we first classified three-dimensional tumor spheroids derived from 11 NSCLC cell lines into cisplatin-sensitive or -resistant groups based on their cisplatin sensitivity and selected signature genes that were differentially altered between the groups. Using reverse engineering methods and functional validation, cAMP response element-binding protein 1 (CREB) was identified as a major regulator of cisplatin resistance. Among the putative target genes of CREB responsible for cisplatin resistance, cisplatin treatment significantly decreased the occupancy of CREB in the regulatory regions of <i>TNKS</i> and <i>KDM6A</i> in cisplatin-sensitive cells, but not in resistant cells, resulting in decreased expression of these protein in the sensitive group. Furthermore, CREB knockdown led to increased sensitivity to cisplatin with reduced levels of TNKS and KDM6A in both cisplatin-resistant tumor spheroids and tumors in a xenograft mouse model. In conclusion, our study delineates the role of CREB in cisplatin resistance and suggests that CREB inhibition is a potential therapeutic strategy for cisplatin-resistant NSCLCs.

The acquisition of resistance to anoikis is a critical driver of metastasis in various tumor types. However, the combined role of anoikis apoptosis in the progression and prognosis of hepatocellular carcinoma (HCC) remains largely unexplored. This study integrates known anoikis genes with single-cell datasets to identify differentially expressed Anoikis (DE-Anoikis) through unsupervised clustering, enabling the classification of samples from The Cancer Genome Atlas (TCGA). A prognostic risk model was constructed using univariate Cox proportional hazards regression and validated with external datasets from the International Cancer Genome Consortium (ICGC) and the Gene Expression Omnibus (GEO). The results revealed significant prognostic differences among DE-Anoikis-based HCC molecular subtypes, with functional enrichment analyses highlighting metabolic reprogramming differences. Furthermore, the anoikis-related prognostic model demonstrated robust predictive accuracy across multiple validation datasets. Two potential therapeutic drugs exhibited sensitivity in low-risk patients, offering novel insights into HCC treatment. Overall, this study identifies a unique subgroup of apoptosis-associated HCC and a prognostic model, providing further biological insights into the molecular mechanisms and therapeutic strategies for HCC.

<b>Background:</b> While timely blood transfusion is critical for restoring oxygen-carrying capacity after coronary artery bypass grafting (CABG), allogeneic blood product transfusions are independently associated with increased long-term mortality, necessitating a risk-stratified approach to balance oxygen delivery against immunological complications and infection risks. <b>Methods:</b> We retrospectively analyzed 3376 patients undergoing isolated CABG between 2005 and 2023 at a single tertiary center. Patients who died during their perioperative hospital stay within 30 days were excluded. Transfusion burden was assessed both as the absolute number of blood product units (packed red blood cells, platelet transfusion, fresh frozen plasma) and as a percentage of calculated patient blood volume. The primary outcome was all-cause mortality at 5 years. Flexible Cox regression with penalized smoothing splines, adjusted for EuroSCORE II, was used to model dose-response relationships. <b>Results:</b> From our cohort of 3376 patients, a total of 137 patients (4.05%) received >10 units of packed red blood cells (PRBC) perioperatively. These patients were older (median 71 vs. 68 years, <i>p</i> < 0.001), more often female (29% vs. 15%, <i>p</i> < 0.001), and had higher preoperative risk (EuroSCORE II: 2.53 vs. 1.41, <i>p</i> < 0.001). After 5 years, mortality was 42% in the massive transfusion group versus 10% in controls. Spline regression revealed an exponential increase in mortality with transfused units: 14 units yielded a 1.5-fold higher hazard of death (HR 1.46, 95% CI 1.31-1.64), rising to HR 2.71 (95% CI 2.12-3.47) at 30 units. When transfusion was indexed to blood volume, this relationship became linear and more tightly correlated with mortality, with lower maximum hazard ratios and narrower confidence intervals. <b>Conclusions:</b> Indexing transfusion burden to the percentage of patient blood volume replaced provides a more accurate and clinically actionable predictor of 5-year mortality after CABG than absolute unit counts. Our findings support a shift toward individualized, volume-based transfusion strategies to optimize patient outcomes and resource stewardship in a time of limited availability of blood products.

A series of rivastigmine hybrids, incorporating rivastigmine fragments (RIV) and a set of different antioxidant scaffolds, were designed, synthesized, and evaluated as multifunctional agents for the potential therapy of Alzheimer's disease (AD). In vitro bioactivity assays indicated that some compounds have very good antioxidant (radical-scavenging) activity. The compounds also displayed good inhibitory activity against cholinesterases, though the bigger-sized hybrids showed higher inhibitory ability for butyrylcholinesterase (BChE) than for acetylcholinesterase (AChE), due to the larger active site cavity of BChE. All the hybrids exhibited an inhibition capacity for self-induced amyloid-β (Aβ_1-42) aggregation. Furthermore, cell assays demonstrated that some compounds showed capacity for rescuing neuroblastoma cells from toxicity induced by reactive oxygen species (ROS). Among these RIV hybrids, the best in vitro multifunctional capacity was found for the caffeic acid derivatives enclosing catechol moieties (<b>4AY5</b>, <b>4AY6</b>), though the Trolox derivatives (<b>4AY2</b>, <b>4BY2</b>) presented the best cell neuroprotective activity against oxidative damage. Molecular-docking studies provided structural insights into the binding modes of RIV-based hybrids to the cholinesterases, revealing key interaction patterns despite some lack of correlation with inhibitory potency. Overall, the balanced multifunctional profiles of these hybrids render them potentially promising candidates for treating AD, thus deserving further investigation.

Osteosarcoma (OS), the most common primary bone cancer of mesenchymal origin in children and young adolescents, remains a challenge due to metastasis and resistance to chemotherapy. It displays severe aneuploidy and a high mutation frequency which drive tumor initiation and progression; however, recent studies have highlighted the role of epigenetic modifications as a key driver of OS pathogenesis, independent of genetic mutations. DNA and RNA methylation, histone modifications and non-coding RNAs are among the major epigenetic modifications which can modulate the expression of oncogenes. Abnormal activity of these mechanisms contributes to gene dysregulation, metastasis and immune evasion. Therapeutic targeting against these epigenetic mechanisms, including inhibitors of DNA and RNA methylation as well as regulators of RNA modifications, can enhance tumor suppressor gene activity. In this review, we examine recent studies elucidating the role of epigenetic regulation in OS pathogenesis and discuss emerging drugs or interventions with potential clinical utility. Understanding of tumor- specific epigenetic alterations, coupled with innovative therapeutic strategies and AI-driven biomarker discovery, could pave the way for personalized therapies based on the molecular profile of each tumor and improve the management of patients with OS.

Bronchial asthma is a complex and heterogeneous disease, with ferroptosis, a form of non-apoptotic cell death, contributing to its pathogenesis by inducing airway epithelial damage, inflammatory infiltration, and airway remodeling. Investigating ferroptosis-related characteristic genes and potential therapeutic compounds may enhance asthma management. This study employed differential analysis and machine learning to identify ferroptosis-related characteristic genes in asthma using the GSE179156 dataset and FerrDb V2 database. Immune infiltration analysis explored the associations between these genes and immune cells, while potential small-molecule drugs were screened through the Connectivity Map (CMap) database and evaluated via molecular docking and molecular dynamics simulations. Two ferroptosis-related characteristic genes, AGPS and APELA, were identified, with AGPS upregulated and APELA downregulated in asthma, both significantly correlated with various immune cells. A diagnostic model based on these genes demonstrated high predictive accuracy. Additionally, KU-55933 was identified as a potential small-molecule inhibitor of AGPS, with stable binding confirmed through computational simulations. These findings emphasize the role of ferroptosis-related genes in asthma and propose promising therapeutic candidates, providing novel insights into its diagnosis and treatment.

<h4>Background</h4>Although recent research has highlighted lactylation, a post-translational modification driven by elevated lactate levels, as a critical regulator of key cellular pathways in hepatocellular carcinoma (HCC), its contribution to the poor prognosis of Clonorchis sinensis (Cs)-infected HCC remains poorly understood.<h4>Methods</h4>We first identified the significant upregulation of the lactate metabolism enzyme LDH in Cs-infected HCC patients through clinical retrospective analysis. We then conducted a multi-omics analysis (RNA-Seq, ATAC-Seq, WGBS-Seq, oxWGBS-Seq, and ChIP-Seq) to examine the differences in 392 lactate metabolism-related genes (LMRGs) between Cs-infected and Cs-noninfected HCC tumors. Six key differentially expressed LMRGs were further validated using RT-qPCR assays to confirm their expression and potential role in HCC progression.<h4>Results</h4>The differential expression levels of 8 LMRGs, along with 71 accessible regions and 42 CpG sites in the promoters of LMRGs, were identified. Notably, we also demonstrated that histone modifications, including H3K9ac, H3K79me2, H3K4me2, H3K4me3, H3K27ac, and H3K4me1, were associated with chromatin accessibility in the promoters of LMRGs. Finally, the TCGA-LIHC cohort confirmed that the differential expression of LMRGs between Cs-infected and Cs-noninfected HCC tumors significantly affects the survival outcomes of HCC.<h4>Conclusions</h4>Our findings revealed that lactylation plays an important role in reshaping the characteristics of HCC during Cs infection, expanding our understanding of the unique features of Cs-infected HCC.

Deletions within the chromosomal locus 1p31.1 are rare, with only a limited number of documented cases. The typical clinical presentation includes intellectual disability, failure to thrive, and craniofacial abnormalities. Some cases may also present with cardiac, gastrointestinal, and genitourinary malformations. Variability in deletion size contributes to a broad spectrum of clinical phenotypes, and a comprehensive understanding of the syndrome's manifestations is still evolving. This case study aims to provide additional insights into 1p31.1 microdeletion syndrome, enhancing knowledge of its genetic and phenotypic characteristics to improve recognition by clinicians. Here, we report a case featuring a 14.385 Mb deletion isolated to the 1p31.1 region, encompassing 41 genes. The deletion manifested with microcephaly, distinctive facial morphology, hypotonia, developmental delay, bilateral cryptorchidism, and flat feet. Notably, our case also exhibited congenital thickening of the lingual and labial frenulum, a trait not typically associated with this deletion.

Objective Metabolic dysfunction-associated steatotic liver disease (MASLD) can rapidly progress to steatohepatitis, liver fibrosis, and hepatocellular carcinoma. However, it is a multisystem disease, and most of the MASLD-related morbidity and mortality is due to its extrahepatic complications. In light of the growing impact of MASLD on extrahepatic organs, we aimed to evaluate spleen fat deposition and compare it with liver fat deposition by vibration-controlled transient elastography using its attenuation parameter. Design Eighty patients from the outpatient department, who met the criteria of MASLD, were included in the study. Both liver and spleen elastography were performed via FibroScan at the same time before treatment. Results Out of the 80 people studied, 69 (86.3%) were male patients. Their age ranged from 22 to 86 years (mean 42.44± 11.92), and BMI ranged between 20.5 to 39.8 kg/m² (28.12± 3.99). Hypertension was present in 45 (56.3%) patients. Sixteen (20%) patients had mild liver steatosis, 26 (32.5%) had moderate, and 28 (35%) patients recorded the presence of severe fatty liver. Using the same cut-off values for spleen, 28 (35%) patients had normal attenuation (steatosis), four (5%) had mild spleen steatosis, 17 (21.3%) had moderate, and 31 (38.8%) patients had severe fatty spleen. A bivariate correlation analysis revealed a significant correlation between hepatic and splenic steatosis (p=0.01). Furthermore, patients with elevated spleen steatosis demonstrated higher liver fat content as indicated by raised liver controlled attenuation parameter (CAP) values (289.79±30.10 dB/m) compared to those without spleen steatosis (264.67±41.85 dB/m). Conclusion This study shows correlation of fatty liver with that of steatosis of the spleen, highlighting the extrahepatic multiorgan effects of the MASLD.

BACKGROUND: Aggregation of misfolded mutant Huntingtin (mHTT) is a pathological characteristic in Huntington’s disease (HD), implying clearance of mHTT is a therapeutical direction for this neurodegenerative disorder. Based on previous studies, Insulin-like growth factor 2 (IGF2) enhances microfilament polymerization in HD models; however, the role of IGF2 against mHTT aggregates is still unclear. RESULTS: Here, we demonstrate that IGF2 expression is significantly lower in symptomatic HD patients compared to presymptomatic individuals, and IGF2 activation mechanistically enhances phosphorylation of Protein Kinase B(AKT; serine/threonine kinase), which subsequently reduces mHTT aggregates in vitro. Furthermore, IGF2 stimulates Nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) signaling, promoting the secretion of mHTT within extracellular vesicles, thereby aiding cellular clearance. In vivo studies in R6/2 HD transgenic mice reveal that IGF2 administration improves motor functions and decreases mHTT levels. CONCLUSIONS: Collectively, our findings elucidate the multifaceted role of IGF2 in HD, highlighting its therapeutic potential through modulation of AKT and NF-κB signaling pathways.

Damage-associated molecular patterns (DAMPs) are a cause of Crohn's disease (CD). Peroxiredoxin 1 (Prdx1), a newly identified DAMP, plays a critical role in organ injury with its potent proinflammatory properties. However, its specific role in CD remains unclear. Here, we identify serum Prdx1 as a DAMP involved in CD. Serum Prdx1 levels were significantly increased and positively correlated with the severity of intestinal inflammation in both CD patients and mice with experimental colitis. Genetic knockout of Prdx1 or administration of a Prdx1-neutralizing antibody attenuated colitis in mice, as evidenced by restoration of the colonic epithelium, improved disease activity, and reduced colonic inflammation. These protective effects were impaired by introduction of recombinant Prdx1 (rPrdx1). Mechanistically, Prdx1 exacerbated intestinal inflammation by promoting macrophage infiltration and subsequent cytokine production. Depletion of macrophages abolished the rPrdx1-mediated exacerbation of colitis. Further, rPrdx1 was internalized by macrophages, leading to lysosomal disruption and subsequent activation of the NLRP3 inflammasome. Pharmacological inhibition of NLRP3 effectively abrogated rPrdx1-induced exacerbation of colitis. In conclusion, serum Prdx1 promotes intestinal inflammation in CD at least in part by activating the NLRP3 inflammasome through lysosomal disruption in macrophages. These findings highlight the pathogenic role of Prdx1 in CD and reveal therapeutic potential of managing CD via neutralization of circulating Prdx1.

For over a century it has been believed that the vertebrate kidney arises exclusively from the intermediate mesoderm. Here, we overturn this paradigm by demonstrating that some nephrons, the functional units of the kidney, originate from the somites- blocks of paraxial mesoderm best known for their contribution to muscle and connective tissues. Using a combination of the GESTALT technique to assign developmental ancestry, somite transplantation experiments and Cre-lox fate-mapping, we show that somites can contribute to the nephrons in the adult zebrafish kidney. Our findings uncover an unexpected developmental connection between the somites and kidneys, potentially offering new pathways for developing regenerative treatments for kidney diseases.

Effective management of osteoporosis requires individuals to take responsibility for following medication, exercise, and dietary guidelines. The aim of this study was to provide important insights into self-care behaviors and their determinants, especially dietary patterns related to vitamin D and calcium intake among elderly women, both healthy and those with osteoporosis. This cross-sectional descriptive-comparative study included 250 postmenopausal women aged 60 and above, consisting of 125 healthy women and 125 women with osteoporosis which conducted in Tabriz, Iran. The data collection instruments comprised a demographic questionnaire, Menopausal Self-Care Questionnaire, and Vitamin D and Calcium Food Frequency Questionnaire. Multivariate linear regression models were employed to identify predictors of self-care behaviors and dietary intakes. The mean (Standard deviation: SD) total self-care score (33-165) in healthy women was higher 118.97 ± 19.92 compared to osteoporotic women 84.7 ± 14.98 (p < 0.001). Healthy women also exhibited significantly higher daily dietary intakes of calcium (850.52 ± 147.92 mg vs. 546.71 ± 60.28 mg, p < 0.001) and vitamin D (3.38 ± 0.65 mg vs. 2.0 ± 0.34 mg, p < 0.001) than osteoporotic women. Multivariate analysis identified household income, age, education, exercise, and BMI as key predictors of self-care behaviors and dietary intakes (p < 0.05) in healthy and osteoporotic elderly women. Postmenopausal women with osteoporosis exhibited poorer self-care behaviors and lower calcium/vitamin D intake compared to healthy peers, with socioeconomic factors (income, education), exercise, age, and BMI as key predictors.

The application of vibrational microspectroscopy to the study of in vitro mesenchymal stem cells (MSC) osteogenic differentiation is a promising approach towards the understanding of the molecular processes involved in bone fabrication. Both infrared (IR) and Raman microspectroscopies have been applied, with a clear predominance of the latter. Bone marrow MSC have been the target of most studies, followed by those originating from dental/oral and adipose tissues. Interests have increasingly addressed single cell and extracellular matrix characterization at the molecular level. Most studies have focused on the characteristics and maturity of time-course mineralization, attempting to localize mineral aggregates formed onto the evolving collagen strands. Some reports have focused on time-dependent changes in protein structure and other components of extracellular matrix components. Besides spectral band examination through position, linewidth and shape, selected band ratios have proved largely informative to assess mineral species evolution and mineral-to-organic matrix interactions over time. The increasing use of multivariate analysis (or chemometrics) and machine learning strategies to detect finer spectral variations is evident, as is the promise of more recent IR and Raman variations to provide higher sensitivity and spatial resolution conditions. The label-free non-invasive nature of vibrational microspectroscopy makes it particularly promising for rapid and effective selection of suitable MSC donors, to support scale-up procedures for translation to the clinic. Some of the challenges to be faced are briefly discussed.

<h4>Background</h4>The pathogenesis of chronic thromboembolic pulmonary hypertension (CTEPH) is poorly understood. Studies of the genetic risk factors for CTEPH are likely to improve our understanding of CTEPH pathogenesis and may lead to novel treatment and prevention strategies. Genetic analysis focused on shared gene variants in high-risk disease pedigrees can aid in the identification of rare variants with a strong effect on disease risk.<h4>Methods</h4>We identified 13 CTEPH high-risk pedigrees and performed whole-exome sequencing in 22 CTEPH cases from these pedigrees, focusing on rare and deleterious variants that were shared between related CTEPH cases. We validated CTEPH candidate gene variants in two independent CTEPH cohorts, one from Utah (n=78) and one from the University of California San Diego (n=238), and compared them to controls from the UK Biobank.<h4>Results</h4>A rare and predicted deleterious missense variant in <i>STAB2</i> was identified in two related CTEPH cases. Qualifying <i>STAB2</i> variant alleles were observed more frequently in both CTEPH cohorts (pooled allele frequency 4.6%) than in subjects from the UK Biobank with a history of pulmonary embolism (PE) (allele frequency 2.2%, p=0.0002) or without a history of PE (allele frequency 1.9%, p<0.0001). CTEPH subjects with qualifying <i>STAB2</i> variants had elevated levels of plasma von Willebrand Factor (vWF) and factor VIII, consistent with the known role of <i>STAB2</i> as a genetic regulator of circulating vWF levels.<h4>Conclusions</h4>Rare variants in <i>STAB2</i> are identified in a CTEPH high-risk pedigree and are over-represented in nonrelated CTEPH cases compared to controls with PE. These data suggest that <i>STAB2</i> is a CTEPH risk gene.

Asthma is a chronic, complex, and heterogeneous respiratory condition marked by airway hyperresponsiveness and reversible airflow limitation. Recent evidence highlights the significant role of metabolic changes, particularly glycolytic reprogramming, in the pathogenesis of asthma. Glycolysis, a fundamental pathway for both anaerobic and aerobic oxidation, not only generates adenosine triphosphate (ATP) but also supplies substrates for lipid-based ATP storage. While initially studied in the context of cancer, glycolysis has been associated with asthma through its interplay with programmed cell death, which is crucial in asthma pathophysiology. This study offers a comprehensive overview of current research on glycolytic reprogramming in asthma, emphasizing its potential implications and significance. The findings aim to inform future studies and contribute to the development of asthma-related precision medicine. A deeper understanding of the interplay between glycolysis and the development and progression of asthma may facilitate the development of innovative therapeutic approaches for this complex condition.

<i>Tubastraea coccinea</i> and <i>T. tagusensis</i>, commonly known as sun corals, are two species of stony corals (Scleractinia, Dendrophylliidae) native to the Indo-Pacific region (<i>T. coccinea</i>) and the Galapagos Islands (<i>T. tagusensis</i>), respectively. They are considered highly invasive species, particularly in the Western Atlantic Ocean, due to high adaptability to various ecological conditions and notable resilience. Given their demonstrated invasiveness, it is important to delve into their physiology and the molecular bases supporting their resilience. However, to date, only a few molecular tools are available for the study of these organisms. The primary objective of the present study was the development of an efficient RNA extraction protocol for <i>Tubastraea coccinea</i> and <i>T.a tagusensis</i> samples collected off Ilha Grande Bay, Rio de Janeiro (Brazil). The quantity of isolated RNA was evaluated using NanoDrop, while its purity and quality were determined by evaluating the A260/A280 and A260/230 ratios. Subsequently, based on genes known for <i>T. coccinea</i>, two housekeeping genes and seven stress response-related genes were isolated and characterized, for the first time for both species, using a molecular approach. An interactomic analysis was also conducted, which revealed functional interactions among these genes. This study represents the first report on gene networks in <i>Tubastraea</i> spp., opening new perspectives for understanding the chemical ecology and the cellular mechanisms underlying the invasiveness of these species. The results obtained will be useful for ecological conservation purposes, contributing to the formulation of strategies to limit their further expansion.

We present a case series of six children who were infected with different species of Nontuberculous mycobacteria (NTM) and <i>Mycobacterium riyadhense</i> was the most prevalent isolate representing 50% of the total pathogens. Four of the reported cases were immunocompromised with disseminated NTM diseases and two were infected with <i>M. avium</i>, <i>M. abscessus</i>, and the other two infected with <i>M. riyadhense</i>. Most patients responded to medical therapy, except for the <i>M. avium</i> case, which was fatal despite combination therapy. Due to the presence of mycolic acid in the cell wall of NTM isolates, prolonged combination therapy is required for treatment, and in some cases, natural resistance may also emerge. Most of the patients reported in our study were immunocompetent. This suggests that NTM can infect children at various body sites regardless of immune status. We highlighted our experience in diagnosing and treating these patients, with special attention to <i>M. riyadhense</i>.

<b>Background and Aims</b>: Sodium-glucose cotransporter-2 (SGLT2) inhibitors have shown promise in metabolic dysfunction-associated steatotic liver disease (MASLD). This large real-world study aimed to evaluate the effects of SGLT2 inhibitors on MASLD patients' clinical outcomes and liver-related complications over extended follow-up. <b>Patients and Method</b>: Data were sourced from TriNetX, a global health research platform with de-identified electronic medical records spanning 135 million patients across 112 healthcare organizations worldwide. We included MASLD adults diagnosed according to ICD9/10 criteria. Following propensity score matching based on 34 variables (demographics, comorbidities, laboratory tests and medication history), SGLT2 inhibitor-treated (n = 19,922) patients were compared with non-SGLT2 inhibitor (n = 19,922) cases. Exclusion criteria included baseline improved alanine aminotransferase (ALT) and alkaline phosphatase (ALP) levels > 4 upper normal limit (UNL), baseline advanced liver disease, liver transplant and cancer, past anticoagulation and non-MASLD etiologies. Assessed outcomes included survival, biochemical, hematologic, AFP, metabolic and cardiovascular parameters, progression to advanced liver disease (ALD), synthetic function, and metabolic markers over 1, 5, and 10 years. <b>Results</b>: Following matching, both cohorts were well-balanced across baseline characteristics. After one year, the SGLT2 inhibitor group demonstrated significantly reduced BMI (33.2 ± 6.2 vs. 34.1 ± 6.5 kg/m², <i>p</i> < 0.001), improved ALT (40.3 ± 31.5 vs. 48.3 ± 41.2 U/L, <i>p</i> < 0.001), and better glycemic control (HbA1c 7.35 ± 1.51% vs. 7.93 ± 1.72%, <i>p</i> < 0.001). The SGLT2 inhibitor group showed higher 10-year survival rates (95.00% vs. 88.69%, <i>p</i> < 0.001), fewer cardiovascular events (10.19% vs. 11.80%, <i>p</i> < 0.001), and markedly reduced progression to advanced liver disease (6.90% vs. 14.15%, <i>p</i> < 0.001). These benefits were consistent across clinical, laboratory, and medication-defined ALD categories. Notably, rates of hepatic decompensation events were significantly lower with SGLT2 inhibitor therapy. <b>Conclusions</b>: In this large real-world cohort, SGLT2 inhibitor use in MASLD patients was associated with significantly improved long-term survival, cardiovascular, and liver-related outcomes over 10 years of follow-up. These benefits likely result from combined metabolic improvements, anti-inflammatory effects, and direct hepatoprotective mechanisms. SGLT2 inhibitors represent a promising therapeutic strategy for improving outcomes in MASLD.

Glycerophospholipids (GPLs) play important roles in cellular compartmentalization and signaling. Among them, phosphatidic acids (PA) exist as many distinct species depending on acyl chain composition, each one potentially displaying unique signaling function. Although the signaling functions of PA have already been demonstrated in multiple cellular processes, the specific roles of individual PA species remain obscure due to a lack of appropriate tools. Indeed, current synthetic PA analogues fail to preserve all the functions of natural PA. To circumvent these limitations, we developed a novel synthetic approach to produce PA analogues without compromising structural integrity of acyl chains. Moreover, addition of a clickable moiety allowed flexible grafting of different molecules to PA analogues for various biological applications. Hence, this innovation also provides powerful tools to investigate specific biological activities of individual PA species, with potential applications in unraveling complex GPL-mediated signaling pathways.

Huntington's disease is an autosomal dominant neurodegenerative disorder of variable progression. Its major features are motor dysfunction, cognitive decline, and psychiatric disturbances. The onset of HD in a patient occurs because of a polyglutamine-expanding mutation within the HTT gene, which leads to the formation of mutant huntingtin protein that aggregates and disrupts neuronal function. Epidemiologically, HD afflicts about 5-10 people per 100,000 throughout the world. However, among populations of European descent, its prevalence is increased. Even after much study into the disorder, myths prevail relating to onset and inheritance of this disorder; including myths such as non-genetic transmission, along with myths such as variation in symptoms, the myths feed on stigma, contributing to a delay in diagnosis and management. Neurodegenerative level in HD affects the basal ganglia especially the striatum leading to impaired motor coordination, chorea, and cognitive deficits. Pathophysiology encompasses excitotoxicity, mitochondrial dysfunction, oxidative stress, and impaired protein clearance mechanisms that end in neuronal loss. The future research areas in the management of HD include gene silencing techniques, stem cell therapy, and even advanced neuroprotective agents acting through a disease-modifying mechanism. The hope of CRISPR-Cas9 gene editing is correction at the source level, and ASOs target reduction in the expression of the mutant huntingtin protein. The introduction of personalized medicine for discovery based on biomarkers could further buttress early diagnosis and effectiveness of treatment. The most revolutionary approach towards the treatment of HD can be a multi-disciplinary approach encompassing conventional therapies and novel genetic techniques.

Osteoarthritis (OA) is a multifactorial disease characterized by joint inflammation and cartilage degeneration, with no disease-modifying drugs available. The vicious cycle between the inflammatory microenvironment (inflamed soil) and dysfunctional chondrocytes (degeneration-related seeds) drives the chronic progressive deterioration of OA. Here, we report a genetically engineered chondrocyte-mimetic nanoplatform (termed HKL-GECM@MPNPs) comprising a honokiol (HKL)-loaded mitochondrion-targeting nanoparticle core coated with an interleukin-1 receptor type 2 (IL-1R2)-overexpressing chondrocyte membrane. HKL-GECM@MPNPs fuse with OA chondrocytes, transferring IL-1R2 onto the plasma membrane and reprogramming the inflamed microenvironment through IL-1β blockade. Mitochondrion-targeting cores then directly deliver HKL to restore mitochondrial sirtuin-3 in OA chondrocytes, reprogramming the cells' pathological phenotype. Intra-articular injection of HKL-GECM@MPNPs in OA mice reduces inflammation, alleviates joint pain, and mitigates cartilage damage through a synergistic effect. Moreover, HKL-GECM@MPNPs effectively reverse cartilage degeneration in human OA cartilage explants. This approach highlights the potential of HKL-GECM@MPNPs to combine IL-1β blockade and mitochondrial sirtuin-3 restoration as a promising strategy for OA treatment.

The rixosome is a large multisubunit complex that initiates RNA decay during critical nuclear transactions including ribosome assembly and heterochromatin maintenance. The overall architecture of the complex remains undefined because several subunits contain intrinsically disordered regions (IDRs). Here, we combined structural and functional approaches to establish PELP1 as the central scaffold of the rixosome upon which the enzymatic subunits modularly assemble. The C-terminal half of PELP1 is composed of a proline-rich IDR that mediates association with the AAA-ATPase MDN1, histones, and the SUMO-specific protease SENP3. The PELP1 IDR contains a glutamic acid-rich region that we establish can chaperone the histone octamer in vitro. Last, the x-ray structure of a small linear motif (SLiM) from the PELP IDR bound to SENP3 reveals how PELP1 allosterically activates SUMO protease activity. This work provides an integrated structural model for understanding the rixosome's dynamic architecture and how it modularly coordinates several cellular functions.

Microglia, the innate immune cells of the central nervous system (CNS), act as first responders to brain injury. Their ability to switch between different neuroprotective and neurotoxic phenotypes, plays a central role in maintaining brain homeostasis. Recently, the P2Y12 receptor (P2Y12R) has been identified as a promising molecular biomarker for microglia activity, as its expression level is dependent on microglia phenotype and function. P2Y12R positron emission tomography (PET) might be a valuable diagnostic tool, however, tracers with sufficient brain retention have not been reported so far. Herein, we report a brain-permeable P2Y12R PET tracer for <i>in vivo</i> imaging of P2Y12R-positive microglia. Nicotinate [¹⁸F]<b>12</b> exhibited nanomolar affinity and specificity for the target receptor and showed a reduced uptake in microglia-depleted (PLX) mice, in comparison to WT and Trem2 knockout (Trem2^-/-) mice. <i>Ex vivo</i> immunohistochemistry (IHC) and PET data revealed a strong correlation between microglia abundance, P2Y12R expression levels and tracer uptake.

<h4>Background</h4>Obtaining human neurons and astrocytes for in vitro studies presents a significant challenge owing to the complexity of replicating their development and functionality outside the human brain. The Ntera-2 cell line is a valuable source of human neurons and astrocytes in neuroscience research. However, differentiating Ntera-2 cells into neurons and astrocytes with all-trans retinoic acid is complicated by the lack of reliable markers to monitor differentiation stages effectively. This study aimed to characterize neuron-enriched and pure astrocyte cultures at two maturation stages and to compare these with the original Ntera-2 cells. Ntera-2 cells and NT2 cells are used interchangeably in this publication.<h4>Methods</h4>Using an advanced proteomic approach, we assessed the protein composition and abundance of neuron and astrocyte co-cultures and discovered that the astrocytic protein profile in co-culture with neurons was more representative compared with that in pure astrocyte cultures. Additionally, electrophysiological studies were conducted to investigate the best astrocyte content for neuronal functionality.<h4>Results</h4>Mass spectrometry-based analysis provided insights into over 9000 proteins, covering well-known protein markers, proteins unique to specific cell types, and differentially expressed proteins. Notably, differences in transcription factors, regulatory proteins, intermediate filaments, and proteins unique to early and mature astrocytes highlighted the distinct maturation, activation, and functional profiles of the various cells. These findings offer a straightforward tool for characterization and monitoring the differentiation process. Three weeks of maturation in pure culture yielded immature astrocytes; however, extending the maturation period to 6 weeks significantly altered the composition of the cellular proteome, indicating increased astrocyte maturity. Studies revealed a broader repertoire of astrocytic proteins in co-culture with neurons. Meanwhile, electrophysiological analyses demonstrated that a high content of astrocytes is essential for neuronal functional maturity.<h4>Conclusions</h4>Astrocyte-neuron co-cultures offer a more accurate model of neural tissue than pure cultures, highlighting the complexity of cell maturation and providing insights for improving in vitro modeling of human neural development.

BACKGROUND: Circular RNA (circRNA) plays a pivotal role in regulating nucleus pulposus cells (NPCs) function, making it a promising therapeutic target for intervertebral disc degeneration (IDD). This study aimed to identify circRNA closely associated with IDD and assess their potential as therapeutic target. METHODS: Circ_0003251 was identified via circRNA sequencing and validated in degenerated human nucleus pulposus (NP) tissues. Its properties were characterized using Sanger sequencing, oligo(dT) primers, RNase R treatment, and fluorescence in situ hybridization. Functional experiments assessed its role in vitro, while molecular mechanisms were explored through luciferase reporter assays, RNA-binding protein immunoprecipitation, and immunofluorescence. To assess therapeutic potential, circ_0003251 was encapsulated in PLGA (poly(lactic-co-glycolic acid)) microspheres (MS) and evaluated in vitro and in vivo. RESULTS: Circ_0003251 expression was significantly downregulated in degenerated NPCs and NP tissues. Its upregulation enhanced NPCs proliferation, extracellular matrix synthesis, and reduced apoptosis. Mechanistically, circ_0003251 acted as a miR-637 sponge, inhibiting AKT1 suppression and alleviating NPCs degeneration. PLGA MS successfully delivered circ_0003251, enhancing its therapeutic effect and delaying IDD in vivo. CONCLUSION: Circ_0003251 was a promising biomarker and therapeutic target for IDD. PLGA MS delivery ensured effective application, offering a novel strategy for IDD treatment.

<h4>Background</h4>Sarcomas are frequently infiltrated with immunosuppressive myeloid cells. Vimseltinib is an inhibitor of the colony-stimulating factor 1 receptor kinase and has been shown to decrease tumor-infiltrating myeloid cells in preclinical models. We hypothesized that vimseltinib combined with the Programmed death-ligand 1 (PD-L1) inhibitor avelumab would be safe, tolerable, and clinically effective in patients with advanced sarcoma.<h4>Methods</h4>This was a phase I study of vimseltinib plus avelumab in patients with unresectable or metastatic sarcoma. The study used a standard 3 + 3 dose-escalation design, followed by dose expansion in patients with select histological subtypes. Vimseltinib was administered daily by mouth in 28-day cycles; avelumab was administered intravenously every 2 weeks. The primary objectives of the dose-escalation and dose-expansion phases were to determine the recommended phase II dose and to estimate the best objective response rate by RECIST version 1.1.<h4>Results</h4>Thirteen patients were treated in the dose-escalation phase, and 19 patients were treated in the dose-expansion phase. The most common treatment-related adverse events were asymptomatic increases in serum levels of amylase, lipase, creatine phosphokinase, aspartate aminotransferase, and alanine aminotransferase. One of six patients treated at the highest dose level had a dose-limiting toxicity (grade 4 increase in aspartate aminotransferase). The highest dose level was determined to be the recommended phase II dose. There were no objective responses. The median progression-free survival of patients treated at the recommended phase II dose was 1.55 months (95% confidence interval 1.25-1.78 months). Flow cytometric analysis of peripheral blood mononuclear cells revealed a decrease in myeloid-derived suppressor cells and regulatory T cells after treatment. RNA sequencing of paired tumor samples revealed an increase in tumor-infiltrating T cells and a decrease in macrophages after treatment.<h4>Conclusions</h4>Vimseltinib plus avelumab was generally safe and well tolerated. This combination had minimal clinical efficacy in our population of heavily pretreated patients with sarcoma.

Epstein-Barr virus-associated hemophagocytic lymphohistiocytosis (EBV-HLH) is a fatal hyperinflammatory disorder distinct from self-limiting EBV-induced infectious mononucleosis (IM). However, the immunological mechanisms underlying the divergence between benign EBV infection and fulminant HLH-particularly in the absence of inherited immunodeficiency-remains unclear, and systematic comparisons of immune landscapes across EBV-associated disease spectra are lacking. In this study, by enrolling children with IM and healthy volunteers as controls, we utilize single-cell RNA sequencing to identify unique immunological characteristics of EBV-HLH. Our analysis indicates that patients with EBV-HLH exhibite widespread activation of NF-κB signaling pathway. Furthermore, excessive cytokine secretion by T and NK cells is observed, along with a shift in monocyte differentiation towards an inflammatory phenotype, and the aggregation of IDO1⁺ monocytes. Metabolic pathway analysis reveals that L-kynurenine, a downstream metabolite of IDO1, is specifically elevated in EBV-HLH and mediates the production of multiple pro-inflammatory cytokines. Collectively, our study maps the immune landscape in pediatric EBV-HLH at single-cell resolution, uncovering potential role of IDO1⁺ monocytes and L-kynurenine as biomarkers.

Selective autophagy is a lysosomal degradation pathway that is critical for maintaining cellular homeostasis by disposing of harmful cellular material. Although the mechanisms by which soluble cargo receptors recruit the autophagy machinery are becoming increasingly clear, the principles governing how organelle-localized transmembrane cargo receptors initiate selective autophagy remain poorly understood. Here we demonstrate that the human transmembrane cargo receptors can initiate autophagosome biogenesis not only by recruiting the upstream FIP200/ULK1 complex but also via a WIPI-ATG13 complex. This latter pathway is employed by the BNIP3/NIX receptors to trigger mitophagy. Additionally, other transmembrane mitophagy receptors, including FUNDC1 and BCL2L13, exclusively use the FIP200/ULK1 complex, whereas FKBP8 and the ER-phagy receptor TEX264 are capable of utilizing both pathways to initiate autophagy. Our study defines the molecular rules for initiation by transmembrane cargo receptors, revealing remarkable flexibility in the assembly and activation of the autophagy machinery, with important implications for therapeutic interventions.

RNA molecules can populate ensembles of alternative structural conformations; however, comprehensively mapping RNA conformational landscapes within living cells presents notable challenges and has, as such, so far remained elusive. Here, we generate transcriptome-scale maps of RNA secondary structure ensembles in both Escherichia coli and human cells, uncovering features of structurally heterogeneous regions. By combining ensemble deconvolution and covariation analyses, we report the discovery of several bacterial RNA thermometers in the 5' untranslated regions (UTRs) of the cspG, cspI, cpxP and lpxP mRNAs of Escherichia coli. We mechanistically characterize how these thermometers switch structure in response to cold shock and reveal the CspE chaperone-mediated regulation of lpxP. Furthermore, we introduce a method for the transcriptome-scale mapping of 5' UTR structures in eukaryotes and leverage it to uncover RNA structural switches regulating the differential usage of open reading frames in the 5' UTRs of the CKS2 and TXNL4A mRNAs in HEK293 cells. Collectively, this work reveals the complexity of RNA structural dynamics in living cells and provides a resource to accelerate the discovery of regulatory RNA switches.

The Organ Donation and Transplantation Network (OPTN) will soon undergo a historic transformation to modernize the national organ transplant system. For the first time in 40 years, the OPTN contract will transition from 1 vendor to 5 to improve transparency, strengthen patient-centered communication, improve patient safety, better manage finances, and upgrade information technology (IT) for organ transplantation. This will be a remarkable change with bipartisan Congressional support. However, the Health Resources and Services Administration, which oversees OPTN, has recognized the complexity of updating the OPTN IT system. On January 17, 2025, it requested submissions for a multiple-award blanket purchase agreement called OPTN Next-Generation IT to help with modernization to improve the OPTN database for patient care and research. This perspective piece reviews the strengths and weaknesses of the current OPTN database using heart transplantation as an example and provides a blueprint to achieve the goals of a flexible database, more comprehensive data collection with clear definitions, and necessary changes to improve efficiency. It also reviews the involvement of the Health Resources and Services Administration and Congress in improving the OPTN, and of sharing OPTN data with the Scientific Registry of Transplant Recipients to generate Scientific Registry of Transplant Recipients national and program reports as well as research.

<h4>Background</h4>Zagociguat (zag) is a CNS-penetrant, soluble guanylate cyclase (sGC) stimulator that has been evaluated in phase 2a, with phase 2b ongoing, clinical studies of primary mitochondrial disease (PMD) subjects with mitochondrial encephalomyopathy, lactic acidosis, and stroke-like episodes syndrome (MELAS). To explore its utility in a broader array of PMDs and secondary mitochondrial disorders, we performed prfeclinical modeling of zag across larval and adult zebrafish models with biochemical deficiencies in diverse respiratory chain (RC) complexes or dihydrolipoamide dehydrogenase (Dldh).<h4>Methods</h4>Zag was evaluated for tissue uptake, gross toxicity, protection from RC toxin-induced brain death, neuromuscular dysfunction, heartbeat loss, and biochemical dysfunction in transgenic or toxin-exposed zebrafish with mitochondrial enzyme deficiencies in complex I (<i>ndufs2</i> ^<i>-/-</i> or rotenone-exposed wild type (WT)), complex IV (<i>surf1</i> ^<i>-/-</i> or azide-exposed WT), multiple RC complexes (<i>fbxl4</i> ^<i>-/-</i> ), or pyruvate dehydrogenase complex (<i>dldh</i> ^<i>-/-</i> ). Zag effects were also studied on the whole-body oxygen consumption capacity (MO₂) and swimming activity of WT and complex IV disease adult zebrafish.<h4>Results</h4>Similar zag levels were observed in adult brains and tail muscle. No morphological or functional toxic effects of zag were observed on larvae viability. Zag provided neuromuscular protection in complex I deficient genetic and pharmacologic inhibitor models. In complex IV deficient models, prevention from brain death occurred at 100 nM zag in high-dose azide-exposed WT larvae; however, no rescue of swimming or neuromuscular phenotypes in low-dose azide-exposed <i>surf1</i> ^<i>-/-</i> larvae was observed. A total of 100 nM zag rescued MO₂ and maximum swimming speed in adult <i>surf1</i> ^<i>-/-</i> zebrafish. Larval swimming activity was also preserved with 10 nM zag treatment in azide-stressed <i>fbxl4</i> ^<i>-/-</i> larvae but not at 10 nM, 100 nM, or 1 µM zag in <i>dldh</i> ^<i>-/-</i> larvae. Zag (10 nM) enhanced complex I enzyme activity that is suggestive of mitochondrial biogenesis and key aspects of mitochondrial physiology in azide-exposed <i>surf1</i> ^<i>-/-</i> and <i>fbxl4</i> ^-/- larvae.<h4>Conclusion</h4>Preclinical evaluation of zag demonstrated its safety, significant protection of neuromuscular dysfunction and/or acute RC stressor-induced decompensation, and improved mitochondrial physiology across multiple different genetic and/or pharmacologic models of RC-deficient PMD. Thus, zag may yield therapeutic potential for an array of diseases with mitochondrial dysfunction beyond MELAS, potentially including Leigh syndrome spectrum disorder and primary mitochondrial myopathies.

Intervertebral disc degeneration (IDD) is a leading cause of spinal disorders worldwide. Current clinical therapies for IDD are often constrained by limited efficacy, notable adverse effects, and high treatment costs. Thus, there is a pressing need for safer and more effective treatment strategies. In recent years, natural product-based therapies have garnered increasing attention due to their multi-target mechanisms and relatively low toxicity. This review comprehensively summarizes recent advances in the application of natural products for IDD treatment, with a focus on flavonoids (e.g., quercetin, hyperoside), glycosides (e.g., ginsenosides, notoginsenosides), terpenoids (e.g., aucubin, celastrol), phenolic compounds (e.g., curcumin, resveratrol), and alkaloids (e.g., berberine, evodiamine). These compounds exert their therapeutic effects by modulating critical signaling pathways, including Sirtuin-1 (SIRT1), Nuclear Factor-kappa B (NF-κB), Mitogen-Activated Protein Kinase (MAPK), Phosphoinositide 3-Kinase/Protein Kinase B (PI3K/Akt), and Nuclear Factor Erythroid 2-Related Factor 2 (Nrf2). Collectively, they exhibit potent anti-inflammatory, antioxidant, anti-apoptotic, anti-senescence, and regenerative properties. The insights presented herein provide a robust theoretical foundation to support future preclinical and clinical investigations, highlighting the considerable promise of natural products in IDD management.

<h4>Background</h4>Spread through air spaces (STAS) represents a novel invasion mechanism in adenocarcinoma that considerably influences lung cancer clinical outcomes; however, studies of its mechanisms at the spatial level are lacking.<h4>Methods</h4>We used the NanoString GeoMx digital spatial profiling (DSP) technology to conduct a spatial transcriptomic analysis of surgically resected tissues from non-small-cell lung cancer (NSCLC) patients with or without STAS.<h4>Results</h4>Compared with tumor nests in non-STAS patients, <i>HLA-DRB5</i> and <i>RASGRF1</i> were significantly less expressed in compartments of STAS, suggesting their inhibitory roles in the occurrence of STAS. Meanwhile, an increase in CD4 T memory cells and a decrease in B cells were observed in the tumor immune microenvironment of STAS. Furthermore, distinct molecular profiles were observed between tumor cells in tumor nests and in air spaces in STAS patients, which was highlighted by the elevated <i>ITGA2</i> expression in the air spaces. These results were validated in an independent cohort by multiplex immunofluorescence stainings.<h4>Conclusion</h4>This study is the first to use DSP to analyze spatial transcriptomic profiles of NSCLC tumor nests and air space tumors, and it identifies potential module features that may be used for STAS identification and prognosis.

<h4>Introduction</h4>Protocols for tissue clearing have been established and optimized for the central nervous system. However, significant modifications are required for clearing different tissue types. Therefore, effective optical clearing for cardiovascular tissue remains a major challenge. The goal of this study is to better understand the responses of porcine left anterior descending artery (LADA) to label-free multiphoton imaging.<h4>Methods</h4>To this end, the effects of different clearing methods (i.e., benzyl alcohol benzyl benzoate-BABB and glycerol), formalin fixation, variations in formalin fixation times (0-240 min), and extended storage in BABB (up to 14 days) are investigated. We compare tissue characteristics under different conditions (e.g., tissue clearing reagent and/or tissue fixation), particularly with regard to tissue preservation and transparency across z-stacks (i.e., imaging depths).<h4>Results</h4>The glycerol clearing method exhibited relatively lower tissue transparency, whereas BABB increased mean AF-AUC from 0.0035 ± 0.0009 to 0.1205 ± 0.0168 and SHG-AUC from 0.0003 ± 0.0002 to 0.0072 ± 0.0040 (<i>p</i>< 0.001), enabling robust signal intensities at deeper layers of LADA tissue. In addition, we observed that BABB preserves fluorescent signals even after extended tissue storage with no significant loss in integrity over 14 days. Finally, we found that formalin fixation in combination with the glycerol clearing method significantly improved tissue preservation compared to the glycerol clearing method alone. However, in combination with the BABB clearing method, fixation reduced tissue transparency and signal intensity compared to BABB clearing without fixation.<h4>Discussion</h4>These findings establish BABB as the superior, label-free clearing agent for deep 3D multiphoton microscopy/second harmonic generation imaging of cardiovascular tissue and underscore the necessity of tailoring fixation parameters to the chosen clearing method.

<h4>Background</h4>Glioblastoma (GBM) was considered the most aggressive type of primary brain tumor, marked by poor clinical outcomes and a high tendency to relapse. The therapeutic efficacy of GBM was significantly compromised by tumor heterogeneity, dysregulated metabolic pathways, the formation of an immunosuppressive microenvironment, and treatment resistance. Therefore, multi-dimensional therapeutic strategies targeting GBM-specific molecular features, its intrinsic properties, and microenvironmental regulatory networks were considered to potentially provide new breakthroughs for overcoming treatment resistance in GBM.<h4>Methods</h4>We analyzed single-cell RNA sequencing (scRNA-seq) data processed with the Seurat package to accurately identify cell types. Spatial transcriptomics integrated Multimodal Intersection Analysis, TransferData, and Robust Cell Type Decomposition techniques to characterize the spatial distribution patterns of key cell subtypes. CellChat was employed to assess intercellular communication networks. Furthermore, <i>in vitro</i> experiments confirmed the main regulatory role of YEATS4 (key transcription factor of C2 <i>PCLAF</i>+ subtype) in GBM malignant progression.<h4>Results</h4>Through scRNA-seq, we identified the C2 <i>PCLAF</i>+ subtype in GBM and analyzed its molecular characteristics and functional role in tumor progression. This subtype exhibited a unique malignant phenotype, marked by significant proliferative activity, characteristic metabolic reprogramming, and dysregulated cell death regulation mechanisms. Spatial transcriptomics revealed its preferential localization within specific tumor niches. Furthermore, the C2 <i>PCLAF</i>+ subtype established a specific interaction with fibroblasts through the MDK-LRP1 ligand-receptor pair. Critically, silencing <i>YEATS4 in vitro</i> significantly inhibited GBM malignancy. Additionally, the prognostic risk score model based on the C2 <i>PCLAF</i>+ subtype demonstrated significant clinical translational value.<h4>Conclusion</h4>Our study systematically elucidated the malignant characteristics of the C2 <i>PCLAF</i>+ subtype and its molecular mechanisms driving GBM progression. This subtype promoted therapeutic resistance through unique metabolic reprogramming, MDK-LRP1-mediated microenvironmental interactions, and immunosuppressive properties. <i>YEATS4</i> knockdown effectively suppressed malignant tumor behaviors, highlighting its therapeutic potential. These findings provided novel targeted intervention strategies to address GBM heterogeneity and treatment resistance, offering promising avenues for overcoming current therapeutic limitations.

<h4>Background</h4>Lymph node metastasis markedly worsens prognosis in lung adenocarcinoma (LUAD); however, the evolutionary dynamics and regulatory mechanisms underlying the heterogeneity of malignant epithelial cells during this process remain poorly understood and warrant comprehensive investigation.<h4>Methods</h4>We performed a comprehensive single-cell transcriptomic analysis of epithelial cells from 18 samples comprising normal lung tissue and lymph node metastases. Malignant epithelial cells were identified via inferred copy number variation (CNV) profiles. Key malignant subpopulations were further characterized through trajectory inference, cell-cell communication mapping, gene set variation analysis (GSVA), and reconstruction of transcription factor regulatory networks. To assess clinical relevance, we developed and validated a prognostic model-termed the EAS score-based on the transcriptional signatures of malignant epithelial subsets, using integrated data from multiple TCGA and GEO cohorts. The functional role of the hub gene SELENBP1 was experimentally validated through quantitative PCR (qPCR), Western blotting, immunohistochemistry (IHC), Transwell migration assays, colony formation assays, flow cytometry, ROS quantification, and subcutaneous tumorigenesis assays <i>in vivo</i>.<h4>Results</h4>Single-cell transcriptomic analysis identified four distinct malignant epithelial subtypes (Clusters 0-3), each characterized by unique patterns of CNV. Leveraging these defined cellular subpopulations, we constructed a highly accurate model for prognostication in LUAD, enabling reliable classification of patients based on clinical outcomes. Through detailed comparisons between groups with divergent prognostic risks, the study revealed notable differences across the tumor microenvironment (TME), including alterations in pathway activity, gene enrichment distributions, mutation profiles, and anticipated responses to immune checkpoint blockade. In addition, functional validation experiments confirmed that SELENBP1 plays a tumor-suppressive role, further supporting its relevance as a potential intervention target in LUAD.<h4>Conclusion</h4>This research provides insights into the evolutionary complexity and heterogeneity of malignant epithelial populations in lymph node metastatic sites of LUAD. It also presents a scoring system based on prognostic indicators, which serves as a reliable tool for forecasting patient survival outcomes. Moreover, the discovery of SELENBP1 as a candidate tumor suppressor emphasizes its importance in guiding both clinical risk categorization and the design of personalized treatment strategies for individuals classified as high-risk LUAD cases.

<b>Background/Objectives:</b> Emerging studies have indicated the importance of intra-tumoral neuronal signals in tumor progression and immune modulation. However, there is limited insight into neuroimmune crosstalk, and the molecules involved are largely unknown. This study investigates the relationship between tumor-derived neuropeptides and immune modulation in head and neck squamous cell carcinoma (HNSC). <b>Methods:</b> By utilizing neuropeptide databases and web tools leveraging TCGA data, neuropeptides' expression and their associations with neurotrophic factors, immune cell infiltration, and immune checkpoints were analyzed, followed by survival analysis. <b>Results:</b> Over half of the neuropeptides were expressed in HNSC, with 16% exhibiting differential expression compared to normal counterparts. Notably, differentially expressed neuropeptides showed significant correlations with neurotrophic factors, immune cell infiltration, and checkpoint genes. Further, their expression was significantly different in responder and non-responder patient samples subjected to immune checkpoint therapy. Neuropeptide genes-PTHLH, NMB, GAST, APLN, and LYNX1-were identified and emerged as crucial mediators in neuroimmune crosstalk. Additionally, the neurotrophic gene NTRK1 exhibited extensive correlation with immune checkpoint genes, underscoring the prevalence of neuroimmune crosstalk in HNSC. <b>Conclusions:</b> These findings shed light on the role of tumor-derived neuropeptides in neuroimmune regulation in HNSC, offering valuable insights for future studies to decode the cancer neuroscience of HNSC progression and therapy.

The serrated pathway represents a significant route to colorectal cancer (CRC), accounting for approximately 15-30% of cases, yet the specific epithelial cell subpopulations driving this pathway remain poorly understood. This study explores the causal relationship between serrated epithelial cells and CRC risk using single-cell transcriptomics and Mendelian randomization (MR). Publicly available single-cell RNA sequencing data were utilized to analyze epithelial cell subpopulations in CRC, focusing on specific serrated cells (SSCs). By integrating genome-wide association study data, MR was employed to assess the causal relationship between gene expression patterns and CRC risk. The study found that an increase in SSCs is closely associated with CRC progression. MR analysis revealed a significant correlation between expression changes in specific genes, such as <i>IER3</i> in SSCs, and CRC risk (<i>p</i> < 0.05). Functional analyses indicated that <i>IER3</i> may promote malignancy by regulating cell proliferation, adhesion, and immune evasion. Several genetic loci related to SSC gene expression were identified and validated for CRC risk association. This study demonstrates the significant role of serrated epithelial cell subpopulations in CRC development, particularly through key genes such as <i>IER3</i>, providing new perspectives for understanding CRC pathogenesis and future therapeutic strategies.

Chinese hamster ovary (CHO) cells are the most common protein production platform for glycosylated biopharmaceuticals due to their relatively efficient secretion systems, post-translational modification (PTM) machinery, and quality control mechanisms. However, high productivity and titer demands can overburden these processes. In particular, the endoplasmic reticulum (ER) can become overwhelmed with misfolded proteins, triggering the unfolded protein response (UPR) as evidence of ER stress. The UPR increases the expression of multiple genes/proteins, which are beneficial to protein folding and secretion. However, if the stressed ER cannot return to a state of homeostasis, a prolonged UPR results in apoptosis. Because ER stress poses a substantial bottleneck for secreting protein therapeutics, CHO cells are both selected for and engineered to improve high-quality protein production through optimized UPR and ER stress management. This is vital for optimizing industrial CHO cell fermentation. This review begins with an overview of common ER-stress related markers. Next, the optimal UPR profile of high-producing CHO cells is discussed followed by the context-dependency of a UPR profile for any given recombinant CHO cell line. Recent efforts to control and engineer ER stress-related responses in CHO cell lines through the use of various bioprocess operations and activation/inhibition strategies are elucidated. Finally, this review concludes with a discussion on future directions for engineering the CHO cell UPR.

Inflammatory bowel disease (IBD), which includes ulcerative colitis (UC) and Crohn's disease (CD), is a significant global health issue characterized by a complex etiology and high rates of recurrence. IBD increases the risk of acquiring colorectal cancer (CRC). CRC, the third leading cause of cancer worldwide, is becoming increasingly prevalent each year. Treatments targeting IBD and associated CRC do not yield effective results. One of the cell death processes associated with IBD pathogenesis is apoptosis. Although apoptosis helps maintain the intestinal barrier of the gut, excessive apoptosis is linked to the development of IBD and contributes to the growth and progression of CRC. In IBD, pro-apoptotic molecules are elevated, while they decrease in CRC. Therefore, therapies that inhibit pro-apoptotic molecules in IBD and enhance apoptosis in CRC may help prevent IBD and the associated CRC. This article reviews the mechanisms of apoptosis and its involvement in IBD and CRC. It also examines possible therapies, including gene and combination therapies that target apoptosis molecules and their signaling pathways in IBD and CRC.

<b>Rationale:</b> Vascular dementia (VaD), driven by chronic cerebral hypoperfusion (CCH), leads to synaptic degeneration and cognitive decline, yet mechanisms linking vascular dysfunction to synaptic loss remain unclear. Intermittent fasting (IF) has emerged as a potential intervention, but its effects on synaptic integrity in VaD are unknown. This study aims to investigate the effects of IF against synaptic degeneration and cognitive impairment induced by CCH. Methods: Bilateral common carotid artery stenosis (BCAS) was employed to induce chronic CCH by placing 0.18 mm micro-coils around each common carotid artery in mice. To assess temporal differences, the coils remained in place for 1, 7, 14, or 30 days. IF was implemented for 16 hours daily over three months prior to BCAS induction. Cognitive impairment was evaluated using the Barnes maze test. White matter lesions (WMLs) and neuronal loss were assessed using Luxol fast blue and cresyl violet staining, respectively. Immunoblotting and immunohistochemistry were performed to quantify synaptic protein levels. Synaptic integrity was examined using transmission electron microscopy. Proteomic analysis of the hippocampus was conducted to investigate molecular adaptations to IF following CCH. <b>Results:</b> We demonstrate that a 16-hour IF regimen preserves cognitive function and synaptic density despite persistent hypoperfusion. Behavioral assays revealed that IF prevented spatial memory deficits in BCAS mice, while electron microscopy confirmed synaptic preservation without altering baseline architecture. Surprisingly, key synaptic protein levels remained unchanged, suggesting IF protects synaptic function rather than abundance. Proteomic profiling revealed dynamic hippocampal adaptations under IF, including upregulation of synaptic stabilizers, enhanced GABAergic signaling, and suppression of neuroinflammatory mediators. CCH induced microglial engulfment of synapses, suggesting a role in complement-mediated synaptic pruning. Temporal pathway analysis revealed IF's multi-phase neuroprotection: early synaptic reinforcement, mid-phase metabolic optimization, and late-phase suppression of chronic neuroinflammation. <b>Conclusion:</b> These findings establish IF as a potent modulator of synaptic resilience in VaD, acting through coordinated preservation of synaptic structure, inhibition of inflammatory synapse loss, and metabolic reprogramming. Our results highlight IF's potential as a non-pharmacological strategy to combat vascular cognitive impairment by targeting the synaptic vulnerability underlying dementia progression.

Cancer is an extremely complex disease characterized by abnormal cell growth due to genetic and environmental factors. With the rise of the field of epigenetic transcriptomics, 5-methylcytidine (m⁵C) modification has been identified as one of the most common chemical modifications occurring in various RNA types. The writers, erasers, and readers of m⁵C modification regulate cancer initiation, progression, and therapeutic responses, such as the proliferation, metastasis, angiogenesis, metabolic reprogramming, immune escape, and therapeutic resistance of tumour cells, by regulating RNA stability, translation, nuclear export, and splicing processes. In this review, we elucidate the biological process of m⁵C modification, summarize the abnormal expression of RNA-modifying proteins (RMPs) in common malignant tumours, explore their functional effects on malignant hallmarks of cancer and molecular mechanisms, and prospect the potential clinical application value of m⁵C.

Water buffalo (<i>Bubalus bubalis</i>) are a primary source of milk in Pakistan, where bovine mastitis is a significant health issue among cattle, leading to substantial economic losses. <i>Staphylococcus aureus</i> is a predominant pathogen associated with mastitis; however, a detailed molecular characterization of the strains in the country remains limited. We previously characterized mastitis strains from the Hazara division of Khyber Pakhtunkhwa, Pakistan. In this study, we investigated mastitis cases in the Peshawar division, including samples from both animals and human farm workers for comparison. Higher rates of mastitis (67.27% of animals) and sub-clinical mastitis (91.03% of positive animals) were identified in Peshawar than for those (34.55% and 75.31%, respectively) previously observed in Hazara. Methicillin-susceptible <i>S. aureus</i> (MSSA) belonging to clonal complex 9 (ST2454) were predominant. Methicillin-resistant <i>S. aureus</i> (MRSA) belonging to ST22 and ST8 were also detected in the Nowshera district. While no <i>S. aureus</i> colonization was observed among animal handlers, evidence of hand contamination suggests a potential route for pathogen spread. Low levels of antibiotic resistance were noted amongst isolates, but higher rates were seen in MRSA. This study presents only the second comprehensive molecular investigation of <i>S. aureus</i> isolated from buffalo mastitis in Pakistan and indicates a concerning rise in mastitis within the province.

Viral oncolysis is considered a promising cancer treatment method because of its good tolerability and durable anti-tumor effects. Compared with other oncolytic viruses, Newcastle disease virus (NDV) has some distinct advantages. As an RNA virus, NDV does not recombine with the host genome, making it safer compared with DNA viruses and retroviruses; NDV can induce syncytium formation, allowing the virus to spread among cells without exposure to host neutralizing antibodies; and its genome adheres to the hexamer genetic code rule (genome length as a multiple of six nucleotides), ensuring accurate replication, low recombination rates, and high genetic stability. Although wild-type NDV has a killing effect on various tumor cells, its oncolytic effect and working mechanism are diverse, increasing the complexity of generating engineered oncolytic viruses with NDV. This study aims to employ whole-genome CRISPR-Cas9 knockout screening and RNA sequencing to identify putative key regulatory factors involved in the interaction between NDV and human colon cancer HCT116 cells and map their global interaction networks. The results suggests that NDV infection disrupts cellular homeostasis, thereby exerting oncolytic effects by inhibiting cell metabolism and proliferation. Meanwhile, the antiviral immune response triggered by NDV infection, along with the activation of anti-apoptotic signaling pathways, may be responsible for the limited oncolytic efficacy of NDV against HCT116 cells. These findings not only enhance our understanding of the oncolytic mechanism of NDV against colonic carcinoma but also provide potential strategies and targets for the development of NDV-based engineered oncolytic viruses.

<h4>Background & aims</h4>Hepatocellular carcinoma (HCC) is the third deadliest cancer worldwide. Its high mortality is primarily attributed to late-stage diagnosis. While mutations in driver genes, such as those encoding β-catenin (CTNNB1), tumor protein p53 (TP53), and telomerase reverse transcriptase promoter (TERTp) are well-documented in the literature, dysregulation of microRNAs (miRNAs), small non-coding RNAs that serve as crucial translational regulators, remains poorly understood.<h4>Methods</h4>We conducted microRNA profiling by microarrays in 45 paired (tumor and non-tumor adjacent tissue) samples from non-viral HCC patients. We performed clinical correlation, ROC analysis and survival analysis of time to recurrence (TTR), disease-free survival (DFS) and overall survival.<h4>Results</h4>We identified 23 significantly dysregulated miRNAs (p ≤ 0.05, fold change ≥2). We investigated their differential expression and its relationship with clinical and pathological variables. Further, we found that miRNA-1972 may serve as an important positive prognostic marker because its high levels were associated with longer TTR and DFS. Significant positive results were obtained in receiver operating characteristic analysis for miR-1972, miR-3651 and miR-486-5p.<h4>Conclusion</h4>miRNA-1972 is a strong prognostic marker in non-viral HCC. Dysregulation of several other miRNAs relates to pathological variables such as amount of stroma within tumor, microvascular invasion and micronodularity.

Proto-oncogenes in the <i>RAS</i> superfamily play dual roles in maintaining cellular homeostasis, such as regulating growth signals and contributing to cancer development through proliferation and deregulation. Activating proto-oncogenes in vitro transforms cells, underscoring their centrality in gene regulation and cellular networks. Despite decades of research, poor outcomes in advanced cancers reveal gaps in understanding Ras-driven mechanisms or therapeutic strategies. This narrative review examines <i>RAS</i> genes and Ras proteins in both housekeeping functions, such as cell growth, apoptosis, and protein trafficking, as well as in tumorigenesis, integrating insights from human (<i>HRAS</i>, <i>KRAS</i>, <i>NRAS</i>), mouse (<i>Hras</i>, <i>Kras</i>, <i>Nras</i>), and <i>Drosophila melanogaster</i> (<i>ras</i>) models. While <i>RAS</i> mutations are tightly linked to human tumors, the interplay between their standard and oncogenic functions remains complex. Even within the same tissue, distinct cancer pathways-such as the mitogen-activated protein kinase (MAPK) and phosphoinositide 3-kinase (PI3K) pathways-can drive varied disease courses, complicating treatment. Advanced-stage cancers add further challenges, including heterogeneity, protective microenvironments, drug resistance, and adaptive progression. This synthesis organizes current knowledge of RAS gene regulation and Ras protein function from genomic alterations and intracellular signaling to membrane dynamics and extracellular interactions, offering a layered perspective on the Ras pathway's role in both housekeeping and tumorigenic contexts.

Given the key importance played by the redox-active metals iron (Fe), copper (Cu), and manganese (Mn) in vital cellular processes, such as DNA synthesis, oxidative phosphorylation, the detoxification of reactive oxygen species (ROS), and angiogenesis, it is not surprising that their dysregulation plays a causative role in many human diseases. The same applies to redox-inactive zinc (Zn), which is involved in numerous biological functions, and serves as a structural element, a catalyst, and a participant in both intracellular and intercellular signaling and in maintaining immune system function. An imbalance in redox active (Fe, Cu, Mn) or redox inactive (Zn) metal ions, whether in excess or deficiency, is harmful and may disrupt the structural, regulatory, and catalytic roles of various antioxidant enzymes (superoxide dismutases (SODs), catalase (CAT), glutathione peroxidases (GPxs)), proteins, receptors, transporters, alter sulfhydryl homeostasis, generate high levels of ROS (e.g., hydroxyl radicals by the Fenton reaction), initiate lipid peroxidation, cause DNA damage, and lead to cell death via mechanisms such as ferroptosis, cuproptosis, cellular senescence, or inflammation. Maintaining redox homeostasis is essential for regulating numerous cellular signaling pathways. Redox-sensitive signaling pathways, such as the nuclear factor kappa B (NF-κB), mitogen-activated protein kinase kinase (MAPK), and nuclear factor erythroid 2-related factor 2 (Nrf2) pathways, form an intricate network that governs cellular responses to redox metal-induced oxidative stress and inflammation. The Nrf2 pathway is primarily responsible for mediating antioxidant defenses, whereas the NF-κB and MAPK pathways play roles in proinflammatory and stress-related responses. Dysregulation of redox-active Fe, Cu, Mn, and redox-inactive Zn can alter epigenetic regulatory mechanisms such as DNA methylation, histone modification, and non-coding RNA expression. The dyshomeostasis of metal ions is closely related to the pathogenesis of lung, renal, and gastrointestinal diseases, neurodegenerative disorders (Alzheimer's disease, Parkinson's disease, and Huntington's disease), psychiatric conditions (schizophrenia), and various cancers. This review summarizes recent findings on the role of iron, copper, manganese, and zinc in maintaining physiological functions, redox homeostasis, and human diseases. See also the graphical abstract(Fig. 1).

Alzheimer's disease (AD) is a progressive neurodegenerative disorder characterized by cognitive decline and memory loss, with few effective treatments currently available. The multifactorial nature of AD, shaped by genetic, environmental, and biological factors, complicates both research and clinical management. Recent advances in artificial intelligence (AI) and multi-omics technologies provide new opportunities to elucidate the molecular mechanisms of AD and identify early biomarkers for diagnosis and prognosis. AI-driven approaches such as machine learning, deep learning, and network-based models have enabled the integration of large-scale genomic, transcriptomic, proteomic, metabolomic, and microbiomic datasets. These efforts have facilitated the discovery of novel molecular signatures and therapeutic targets. Methods including deep belief networks and joint deep semi-non-negative matrix factorization have contributed to improvements in disease classification and patient stratification. However, ongoing challenges remain. These include data heterogeneity, limited interpretability of complex models, a lack of large and diverse datasets, and insufficient clinical validation. The absence of standardized multi-omics data processing methods further restricts progress. This review systematically summarizes recent advances in AI-driven multi-omics research in AD, highlighting achievements in early diagnosis and biomarker discovery while discussing limitations and future directions needed to advance these approaches toward clinical application.

Skeletal muscle differentiation is a complex process regulated by a network of genes and transcription factors. Recent studies have revealed the roles of circular RNAs (circRNAs) and microRNAs (miRNAs) in modulating gene expression during myogenesis. In this study, we focused on the functional interplay between circAtxn10, miR-143-3p, and the nicotinic acetylcholine receptor subunit alpha 1 (Chrna1) in skeletal muscle differentiation. Our results demonstrate that circAtxn10 expression increases during myogenic differentiation and acts as a sponge for miR-143-3p through direct binding. We identified Chrna1 as a direct target of miR-143-3p through three binding sites in its 3'-UTR and showed that both miR-143-3p mimic and Chrna1 knockdown significantly impair myogenesis. Notably, Chrna1 overexpression dramatically enhanced myogenic marker expression and myotube formation. Our findings establish a regulatory axis involving circAtxn10, miR-143-3p, and Chrna1 that plays a critical role in modulating skeletal muscle differentiation, providing new insights into the complex molecular mechanisms regulating myogenesis.

Ferroptosis is closely associated with the various pathological manifestations of asthma. This study aimed to explore the role of the stimulator of interferon genes (STING) in modulating airway inflammation in asthma, with a particular focus on regulating ferroptosis in airway epithelial cells. Using an ovalbumin (OVA)-sensitized mouse model of asthma, the OVA group exhibited significant inflammatory cell infiltration in the airways, increased mucus secretion, and elevated levels of inflammatory cytokines compared with those noted in the normal group. Additionally, the ferroptosis-related protein acyl-CoA synthetase long-chain family member 4 (ACSL4) was upregulated, whereas glutathione peroxidase 4 (GPX4) was downregulated, accompanied by elevated malondialdehyde (MDA) levels and reduced superoxide dismutase (SOD) activity. Furthermore, both messenger ribonucleic acid and protein levels of STING were significantly increased in the lungs of OVA-sensitized mice, with predominant expression in airway epithelial cells. After intervention with STING inhibitor C-176, the OVA + C-176 group demonstrated reduced inflammatory cell infiltration and mucus hypersecretion in the airways, along with decreased serum levels of IgE and Th2-associated cytokines (IL-4 and IL-13), but increased levels of the Th1 cytokine IFN-γ. Moreover, ACSL4 protein and MDA levels were significantly decreased, whereas SOD activity was significantly restored following C-176 intervention. Double immunofluorescence staining revealed the colocalization of STING and ACSL4, with their expression levels significantly reduced following C-176 treatment. Co-immunoprecipitation confirms the interaction between STING and ACSL4. Collectively, these findings indicate that STING regulates airway inflammation in asthma by modulating ferroptosis and lipid peroxidation, highlighting STING as a potential therapeutic target for asthma.

Antibodies are critical for the immune response and serve as important tools due to their ability to recognize specific amino acid sequences, or epitopes. Based on the latter, they are utilized as diagnostic tools in biological and biomedical research. Huntington's disease (HD) is a neurodegenerative condition caused by CAG repeat expansions in the huntingtin (HTT) gene and characterized by amyloid-like protein deposits in patients. Multiple anti-HTT antibodies are used in HD research for their ability to recognize specific HTT inclusions in both post-mortem tissue and in laboratory conditions. Some of the antibodies are seen as detectors of distinct structural motifs. However, most knowledge of their binding mechanism stems from studies of soluble monomers or short fragments of the epitopes, rather than the aggregated, misfolded target protein. Here, we investigate how MW8 antibodies interact with HTT exon 1 (HTTex1) fibrils, using solid-state NMR, electron microscopy, and complementary techniques. Magic angle spinning (MAS) NMR revealed localized impacts of the antibody on exposed parts of the HTTex1 fibrils: the flanking segments that form its "fuzzy coat". Antibody binding affected the structure and dynamics of the fuzzy coat, but also modulated the propensity for forming supramolecular fibril clusters, which has important implications for (reducing) cytotoxicity.

LC3-interacting regions (LIRs), or Atg8-interacting motifs (AIMs), are short linear motifs found in unstructured loops or intrinsically disordered regions of many autophagy-related proteins. LIRs were initially identified for their role in binding to Atg8 family proteins on autophagosomal membranes. However, emerging evidence suggests that LIRs and their surrounding residues mediate interactions with a wide array of proteins beyond Atg8s. This broadens the biological significance of LIRs in autophagy, rendering them an organizing principle of the autophagy machinery. In this perspective, we explore recent advances highlighting the multifunctional roles of LIRs, including their capacity to mediate binding with diverse factors. We discuss insights into the mechanisms underlying LIR-mediated interactions and propose an updated model to explain Atg8 diversification in higher eukaryotes. We conclude by addressing key challenges and outlining future directions for understanding LIR biology and its broader implications for cellular homeostasis.

Transactive response DNA binding protein 43 kDa (TDP43) proteinopathy, characterized by the mislocalization and aggregation of TDP43, is a hallmark of several neurodegenerative diseases, including Amyotrophic Lateral Sclerosis (ALS). In this study, we describe the development of a new model of TDP43 proteinopathy using human induced pluripotent stem cell (iPSC)-derived neurons. Utilizing a genome engineering approach, we induced the mislocalization of endogenous TDP43 from the nucleus to the cytoplasm without mutating the TDP43 gene or using chemical stressors. Our model successfully recapitulates key early and late pathological features of TDP43 proteinopathy, including neuronal loss, reduced neurite complexity, and cytoplasmic accumulation and aggregation of TDP43. Concurrently, the loss of nuclear TDP43 leads to splicing defects, while its cytoplasmic gain adversely affects microRNA expression. Strikingly, our observations suggest that TDP43 is capable of sustaining its own mislocalization, thereby perpetuating and further aggravating the proteinopathy. This innovative model provides a valuable tool for the in-depth investigation of the consequences of TDP43 proteinopathy. It offers a clinically relevant platform that will accelerate the identification of potential therapeutic targets for the treatment of TDP43-associated neurodegenerative diseases, including sporadic ALS.

Molecular diversity is one of the most pursued objectives in drug discovery, and diversity-oriented synthesis (DOS) perfectly responds to the achievement of this goal. In this paper, we describe a DOS approach applied to the antitumor field with the aim of identifying new anticancer structures and their associated targets. To accomplish this ambitious project, after an initial stage of phenotypic evaluation, we set up an integrated platform of inverse virtual screening (IVS), bioinformatics, and omics to predict the biological targets of the most promising compounds <b>31</b> and <b>63</b>. Several proteins emerged from this study, and the most interesting ones were assessed by biophysical and in cellulo experiments, leading to the validation of six targets involved in calcium regulation, endoplasmic reticulum stress, and apoptosis. This work allowed us to identify two hit compounds with an interesting antitumor mechanism, but principally, to validate our platform as a fruitful tool for untargeted DOS campaigns.

Marine polysaccharides are widely available sustainable renewable macromolecules, which have attracted considerable attention owing to their enhanced biocompatibility, biodegradability, noncytotoxic, nonimmunogenic properties, and close similarity to the native cellular microenvironment of tissues and organs. Herein, a comprehensive overview of the main sources and properties of most studied cationic, anionic, and neutral marine-origin polysaccharides, their main chemical functionalization strategies, as well as their processing into advanced biofunctional materials/devices is provided. Several recent examples are given on the bottom-up processing of marine-origin polysaccharide-based biomaterials in the form of nano-/microparticles and capsules, nanofibers, thin films, membranes, hydrogels, cryogels, and (bio)inks to be used as high added-value antimicrobial coatings, adhesives, and wound dressings, or in food packaging, cosmetics, controlled drug delivery, <i>in vitro</i> disease modeling, or tissue engineering and regenerative medicine. The main challenges hampering the clinical translation and commercialization of most marine-origin polysaccharide-based biomaterials and devices, and future perspectives in the field are also discussed.

The process of protein aggregation, central to neurodegenerative diseases like Huntington's, is challenging to study due to its unpredictable nature and relatively rapid kinetics. Understanding its biomechanics is crucial for unraveling its role in disease progression and cellular toxicity. Brillouin microscopy offers unique advantages for studying biomechanical properties, yet is limited by slow imaging speed, complicating its use for rapid and dynamic processes like protein aggregation. To overcome these limitations, we developed a self-driving microscope that uses deep learning to predict the onset of aggregation from a single fluorescence image of soluble protein, achieving 91% accuracy. The system triggers optimized multimodal imaging when aggregation is imminent, enabling intelligent Brillouin microscopy of this dynamic biomechanical process. Furthermore, we demonstrate that by detecting mature aggregates in real time using brightfield images and a neural network, Brillouin microscopy can be used to study their biomechanical properties without the need for fluorescence labeling, minimizing phototoxicity and preserving sample health. This autonomous microscopy approach advances the study of aggregation kinetics and biomechanics in living cells, offering a powerful tool for investigating the role of protein misfolding and aggregation in neurodegeneration.

Pesticide exposure is increasingly recognized as a potential environmental factor in idiopathic Parkinson's disease, though the molecular mechanisms remain unclear. This study explores how pesticide exposure alters gene regulation in key brain regions using the rotenone rat model. We performed H3K27ac ChIP-sequencing to profile active regulatory elements in the substantia nigra and motor cortex. Despite uniform complex I inhibition across regions, we observed region-specific epigenomic changes associated with rotenone exposure. RNA-sequencing confirmed transcriptomic alterations. We identified a strong, rotenone-induced immune response in the substantia nigra, including increased activity in the C1q complement pathway, suggesting immune involvement driven by regulatory mechanisms. In contrast, the cortex showed dysregulation of synaptic function at the gene regulatory level. Our results highlight a role for gene regulatory mechanisms potentially mediating the effects of pesticide exposure, driving region-specific functional responses in the brain that may contribute to the pathology and selective vulnerability that characterise Parkinson's disease.

The coordinated development of cortical circuits composed of excitatory and inhibitory neurons is critical for proper brain function, and disruptions are linked to a spectrum of neuropsychiatric disorders. While excitatory neurons are generated locally in the cortical proliferative zones, inhibitory cortical interneurons (cINs) originate in the basal telencephalon and migrate tangentially into the cortex. Here, we show that DNA methyltransferase 1 (DNMT1) is essential for the migration and integration of somatostatin (SST)-expressing interneurons in mice. Dnmt1 deletion causes premature exit of SST⁺ cINs from the superficial migratory stream and alters the expression of key developmental genes. Unexpectedly, Dnmt1-deficient SST⁺ interneurons also exert non-cell-autonomous effects on cortical progenitor cells, resulting in subtle yet lasting alterations in cortical layering. These findings propose a role for DNMT1 in governing the migration of SST⁺ interneurons and mediating their instructive signaling to cortical progenitor cells, thereby shaping cortical architecture and influencing long-term network function.

<h4>Background</h4>Microbial lung infections may promote development of lung cancer through overlapping molecular mechanisms. This analysis aimed to identify a co-regulated peripheral blood gene signature in lung adenocarcinoma (LUAD) and microbial lung infections.<h4>Methods</h4>A total of 403 peripheral blood transcriptomic profiles from five GEO test datasets-two LUAD (GSE39345, GSE103527) and three infection-related (GSE40012, GSE65682, GSE103119)-were analyzed using the limma package. Differentially expressed genes (DEGs) were defined by|log₂FC| >1 and p < 0.05. Two additional GEO datasets (GSE42826 and GSE42830), comprising 30 blood samples (16 LUAD, 14 lung infection), served as validation sets. Shared DEGs were subjected to KEGG and GO enrichment analyses. Protein-protein interaction (PPI) networks were constructed in Cytoscape, and the top 10 hub genes were identified. Expression data of hub genes were compared between validation LUAD and lung infection samples using the Mann-Whitney U test, followed by linear regression and Pearson correlation to confirm co-regulation. Immune cell infiltration was assessed using xCell deconvolution algorithm.<h4>Results</h4>Ninety-three significant DEGs were shared between LUAD and infection datasets, including 40 upregulated and 53 downregulated genes. Eight hub genes showed consistent differential expression in both LUAD and lung infection: BCL6, CD163, S100A12 (upregulated); and FLT3LG, RPL13, RPL14, RPL22, RPS4X (downregulated), of which BCL6, S100A12, FLT3LG, RPL13, RPL14, RPL22 and RPS4X were significantly co-regulated (R² >0.8, p < 0.001) and correlated (p < 0.05). Immune profiling revealed that upregulated genes were associated with immunosuppressive cells such as Tregs and M2 macrophages, while downregulated genes were positively correlated with antitumor immune cell infiltration including CD8⁺ T cells and M1 macrophages. Consistent immune, stroma and microenvironment scores were observed between LUAD and lung infection.<h4>Conclusion</h4>This analysis identified a blood-based 7-gene signature shared between LUAD and microbial lung infections, associated with immunosuppressive microenvironment features, suggesting a potential link between infection-driven inflammation and tumor-promoting immune modulation.

Pediatric patients with fusion-driven leukemias frequently have a poor prognosis and need more effective therapies. Adoptive T-cell therapies, using expanded autologous T cells, have shown promise as an immunotherapeutic for patients with tumors characterized by high mutational burdens. However, this approach has not been shown to be effective in pediatric leukemias. In this study, we analyzed samples from pediatric patients with fusion-driven acute lymphoblastic, acute myeloid, and mixed phenotypic leukemias, including those with KMT2A-rearrangements. T cells were attained from bone marrow samples, expanded, and their reactivity against autologous leukemic blasts was tested. Strikingly, we observed leukemia-reactive T cells in nearly all patients (33 of 34) at diagnosis or relapse. Furthermore, some patients contained clones reactive to fusion neoantigens and other tumor-associated antigens, and candidate samples were further enriched by selecting for PD1^hi and CD39⁺ T-cell populations. These clones were only present at the initial diagnostic timepoint and could not be detected at later times after treatment, even with deep sequence profiling. Altogether, our data suggest that adoptive T cell therapy, using expanded leukemia-reactive T cells identified at diagnosis, has potential as a novel therapeutic for these patients.

There is significant variation in the reported estimates of Huntington's disease (HD) prevalence in different settings. This systematic review was undertaken to describe and assess the sources of heterogeneity in estimated prevalence values, and to consider the role of quantitative synthesis in the context of such heterogeneity. Observational studies from which a prevalence estimate (point or period) or cumulative incidence of HD could be calculated between 1993 and 2024 were sought from Medline and Embase databases. The study features are described and the sources of heterogeneity are discussed. A meta-regression was conducted including predictor variables: continent, median age of population, number of years since 1993, case ascertainment method, and Healthcare Access and Quality Index score. 43 studies met the inclusion criteria. Significant clinical and methodological heterogeneity between studies is described, including differences in case definitions and ascertainment methods, and in the estimates of disease burden calculated. There were differences in the estimated point prevalence between regions and populations within regions, while the estimated point prevalence was shown to be increasing since 1993. Wide prediction intervals in the overall pooled point prevalence (95% prediction interval: 0.32-37.55 cases per 100,000), and the European pooled point prevalence (95% prediction interval: 1.64-19.18 cases per 100,000), indicate the scale of heterogeneity between studies and settings. While such heterogeneity currently limits the validity and utility of quantitative synthesis, developing an accepted consensus on the minimum standards and reporting requirements for HD prevalence studies could reduce the methodological heterogeneity between future studies, enabling more valid and meaningful quantitative synthesis in future.

<h4>Background</h4>Lysosomes are monolayer membrane-encapsulated organelles containing acid hydrolases, crucial for intracellular substance breakdown and cellular homeostasis. They are also involved in autophagy. Although autophagy is linked to cancer, the role of lysosome-related genes in cervical cancer prognosis remains unclear. This study aimed to develop a prognostic model for cervical cancer based on lysosome-related genes and explore its applications in the tumor microenvironment, radiotherapy prognosis, and clinical pharmacology.<h4>Methods</h4>We identified differentially expressed lysosome-related genes in cervical cancer and normal tissues using the TCGA database. A prognostic model was constructed using LASSO-Cox regression, validated with ROC curves and PCA analysis, and further verified using the GEO dataset GSE63514. In vitro and in vivo experiments were conducted to explore key genes, and their biological significance and pharmacological potential were analyzed.<h4>Results</h4>A five-gene (AP1B1, DNASE2, LAMP3, NPC1, and LAPTM4A) lysosome-associated prognostic model was developed. LAMP3 was identified as the most differentially expressed gene. Knockdown of LAMP3 significantly reduced cervical cancer cell migration and invasion through lysosomal and autophagic pathways. Daidzein was found to have high binding affinity for LAMP3, suggesting its therapeutic potential.<h4>Conclusion</h4>Lysosome-related gene modeling has significant clinical value. LAMP3 knockdown inhibits cervical cancer progression by reducing autophagy and lysosomal function. Daidzein shows potential as a novel therapeutic agent. However, further validation in larger cohorts is needed due to the limited sample size in this study.

Advances in single-cell technology enable large-scale generation of omics data, promising for clarifying gene regulatory networks governing different cell type/states. Nonetheless, prevailing methods fail to account for universal and reusable regulatory modules in GRNs, which are fundamental underpinnings of cell type landscape. We introduce cRegulon to infer regulatory modules by modeling combinatorial regulation of transcription factors based on diverse GRNs from single-cell multi-omics data. Through benchmarking and applications using simulated datasets and real datasets, cRegulon outperforms existing approaches in identifying TF combinatorial modules as regulatory units and annotating cell types. cRegulon offers new insights and methodology into combinatorial regulation.

<h4>Background</h4>Cardioembolic stroke (CS) and atherosclerosis (AS) are closely related diseases. Ferroptosis, a novel form of programmed cell death, may play a key role in CS and AS. However, the pathophysiological mechanisms underlying their coexistence remain unclear. This study aims to identify the hub genes and pathways involved in developing both diseases.<h4>Methods</h4>CS (GSE58294) and AS (GSE20129) datasets were obtained from the Gene Expression Omnibus database, and a ferroptosis (FR)-related gene dataset was downloaded from the FR database. A study was conducted to examine differentially expressed genes (DEGs) in healthy individuals and patients diagnosed with CS and AS. Gene ontology and Kyoto encyclopedia of genes and genomes analyses were performed to explore the functions of common FR-related DEGs (FRDEGs). Two machine learning algorithms, Least Absolute Shrinkage and Selection Operator (LASSO) regression and Support Vector Machine Recursive Feature Elimination (SVM-RFE), were used to screen for overlapping FRDEGs in CS and AS. To validate the prediction results, blood samples were collected from healthy controls and patients with CS and AS for quantitative real-time PCR. The correlation between biomarkers and clinical features was also evaluated.<h4>Results</h4>A total of 69 and 39 FRDEGs were identified in CS and AS, respectively. The hub genes, CIRBP, CREB5, MAPK14, PEBP1, and PTGS2, were identified using multiple methods. The area under the curve was > 0.7 for both models constructed using CS and AS datasets. A strong correlation was observed between neutrophil levels and expression of the hub genes. Additionally, several types of cancer indicated elevated expression of these hub genes compared to normal tissues.<h4>Conclusions</h4>In summary, the diagnostic model based on the FR-related gene PTGS2 demonstrated significant and specific diagnostic value for CS and AS, reflecting the status of blood lymphocytes, monocytes, and neutrophils. A pan-cancer study suggested it could serve as a new clinical prognostic marker and therapeutic target across various cancer types. This model may aid in the diagnosis of CS and AS. The findings offer new insights into the pathogenesis of these diseases.

BackgroundFriedreich ataxia is a rare genetic disorder caused by mutations in the <i>FXN</i> gene, typically presenting with balance and coordination difficulties between ages 7 and 15 years. Neurologic symptoms are progressive and lead to loss of ambulation and especially in children other symptoms such as cardiomyopathy, scoliosis, and fatigue are common. The FACHILD natural history study aimed to expand knowledge about the disease course and evaluate clinical outcome assessments in children. We report on functional performance testing, clinical rating scales, and patient-reported outcomes as clinical outcome assessments for Friedreich ataxia. Over a 3-year period, all tests and assessments were conducted to evaluate their sensitivity to progression and correlate with established measures such as neurologic rating scales.MethodsIndividuals with genetically confirmed Friedreich ataxia, aged 7-18 years, were enrolled from October 2017 to November 2022. This analysis focused on ambulatory individuals, including timed walks (25-foot, 1 minute, and 6 minutes), the timed up and go, and the 9-hole pegboard test. Additionally, the Berg Balance Scale and FA-Activities of Daily Living were assessed. Progression data were analyzed using mixed models for repeated measures, with detailed analyses of intermittent missing data. Data from the Friedreich Ataxia Clinical Outcome Measures Study was used to augment analyses when available.Findings and InterpretationFunctional performance outcome measures are sensitive and clinically relevant tools for assessing disease progression in children with Friedreich ataxia. In early to moderately affected populations, the 1-Minute Walk demonstrated promising properties, showing comparable sensitivity to the modified Friedreich Ataxia Rating Scale and the Upright Stability Score.

Two-dimensional covalent organic frameworks (COFs) are a special kind of crystalline polymers produced by the polycondensation reaction of organic linkers. However, the polymerization is often uncontrolled and conventionally yields polycrystalline powders. For that reason, the development of new methods to obtain single-crystalline COFs is an urgent need. Herein, we report a modulated, room-temperature, and fast synthesis of a highly fluorinated COF where the reaction conditions were systematically analyzed to yield single-crystalline rods of around three microns. The increase in crystal size enabled the analysis of precise structural features such as the mechanical properties or the different aggregation states of the bidimensional framework. In addition, the reaction mechanism was computationally elucidated, and the roles of the modulator agent and the catalyst were analyzed, confirming the critical importance of trifluoroacetic acid and 2,3,5,6-tetrafluoroaniline in modulating Schiff base bond formation and, consequently, the crystallinity of the resulting COF. This study reveals new insights into the crystallization of high-quality 2D COFs.

Time-course monitoring of blood biomarkers with rapid turnaround has the potential to revolutionize the diagnosis, stratification of phenotypes, and therapeutic/prognostic approaches for various acute inflammatory diseases in both clinical and preclinical studies. Current approaches, however, are hampered by slow turnaround times and large sample volume requirements, limiting the exploration of disease mechanisms and therapeutic strategies. Here, we developed a microfluidic digital ELISA platform prototype, combining single-molecule counting with whole blood assay capability for the first time from small animal models. This platform is semi-automated and enables repeated, rapid biomarker monitoring with just 3.5 μL of whole blood collected from the tail. Our platform demonstrated high sensitivity and multiplexity, allowing real-time cytokine profiling within a 2-h turnaround. Using a murine sepsis model, we achieved precise temporal monitoring of cytokine levels, demonstrating prognostic capability by correlating early-stage cytokine levels with a liver-injury biomarker. This microfluidic platform enables high temporal resolution and rapid monitoring of biomarker dynamics in a single mouse using freshly collected whole blood, significantly reducing the number of animals needed for preclinical studies. This technology has strong potential to transform ICU therapeutic strategies and preclinical research, enabling personalized treatment based on real-time biomarker profiles.

NMDA receptors (NMDARs) are widely distributed throughout the central nervous system (CNS) and play pivotal roles in normal physiological processes such as synaptic plasticity, learning, and memory. Substantial evidence indicates that NMDAR dysfunction, particularly excessive calcium influx, critically contributes to the pathogenesis of major neurodegenerative diseases, including Alzheimer's disease (AD), Parkinson's disease (PD), Huntington's disease (HD), and amyotrophic lateral sclerosis (ALS). Dysregulated glutamatergic signaling synergizes with pathological protein aggregation (e.g., Aβ, <i>α</i>-synuclein, mutant huntingtin) to drive neuronal loss. We systematically delineate NMDAR-related mechanisms underlying neurodegeneration, highlighting spatial-specific roles (e.g., synaptic NMDAR-mediated neuroprotection versus extrasynaptic NMDAR-mediated excitotoxicity) and crosstalk with mitochondrial dysfunction and oxidative stress. We critically evaluate current therapeutic strategies targeting NMDARs, including subunit-selective modulators, downstream effector modulation, and glutamate transporter modulation designed to restore NMDAR homeostasis. Consequently, NMDARs and their modulators represent promising therapeutic targets for these refractory conditions. This review comprehensively summarizes current research on the involvement of NMDARs and the glutamatergic system in neurodegenerative diseases. Furthermore, we discuss the clinical application of NMDAR-targeting agents and explore emerging therapeutic strategies focused on modulating NMDAR-related pathways. This article aims to provide a reference for elucidating the molecular mechanisms underlying these neurodegenerative disorders and to highlight potential avenues for future drug development.

The blood-brain barrier (BBB) is the main obstacle preventing access to the central nervous system (CNS). It is therefore a major challenge in CNS studies, e.g., investigations of novel therapeutic agents for brain tumors, such as glioblastoma multiforme (GBM). Ensuring the structural and functional integrity of the BBB is essential for such studies. Therefore, the BBB and blood-brain-tumor barrier (BBTB) behaviors must be further investigated to enhance the treatment effectiveness in neurodegenerative diseases (NDDs). Researchers are striving to use nanoparticles (NPs) and/or develop nano delivery systems (NDSs) to efficiently overcome the barriers to transporting neurotherapeutics to the brain, focusing on targeting disease or tumor sites. In this regard, BBB disease modeling enables examination of the transport of these molecules and/or systems from the bloodstream to the brain. Facilitating their transport is likely to enhance their investigation in CNS studies and potentially lead to their use in treating various NDDs. This review describes the BBB, NPs, and/or NDSs used in BBB studies and evaluates the ability of existing BBB disease models to precisely forecast the in vivo efficacy of NPs or NDSs.

<b>Background and Objectives</b>: Accurate noninvasive differentiation between hepatocellular carcinoma (HCC) and intrahepatic cholangiocarcinoma (ICC) remains a clinical challenge. This study aimed to assess the dignostic performance of apparent diffusion coefficient (ADC) values from diffusion-weighted MRI in distinguishing between HCC and ICC, with histological confirmation as the gold standard. <b>Materials and Methods</b>: A retrospective analysis was performed on 61 patients (41 HCC, 20 ICC) who underwent liver MRI and percutaneous biopsy between 2019 and 2024. ADC values were measured from diffusion-weighted sequences (b-values of 0, 500, and 1000 s/mm²), and regions of interest were placed over solid tumor areas. Statistical analyses included <i>t</i>-tests, one-way ANOVA, and ROC curve analysis. <b>Results</b>: Mean ADC values did not differ significantly between HCC (1.09 ± 0.19 × 10^-3 mm²/s) and ICC (1.08 ± 0.11 × 10^-3 mm²/s). ROC analysis showed poor discriminative ability (AUC = 0.520; <i>p</i> = 0.806). In HCC, ADC values decreased with lower differentiation grades (<i>p</i> = 0.008, η² = 0.224). No significant trend was observed in ICC (<i>p</i> = 0.410, η² = 0.100). Immunohistochemical markers such as CK-7, Glypican 3, and TTF-1 showed significant diagnostic value between tumor subtypes. <b>Conclusions</b>: ADC values have limited utility for distinguishing HCC from ICC but may aid in HCC grading. Immunohistochemistry remains essential for accurate diagnosis, especially in poorly differentiated tumors. Further studies with larger cohorts are recommended to improve noninvasive diagnostic protocols.

RKIP and LKB1, encoded by <i>PEBP1</i> and <i>STK11</i>, respectively, have emerged as key regulators of cancer pathophysiology. However, their role in shaping tumor progression through modulation of the tumor microenvironment (TME) is not yet fully understood. To address this, we performed a comprehensive pan-cancer analysis using TCGA transcriptomic data across 33 cancer types, grouped by their tissue of origin. We investigated <i>PEBP1/STK11</i> co-expression and its association with transcriptomic reprogramming in major TME components, including immune, mechanical, metabolic, and hypoxic subtypes. Our results revealed both positive and inverse correlations between <i>PEBP1/STK11</i> co-expression and TME-related molecular signatures, which did not align with classical cancer categorizations. In a subset of tumors, <i>PEBP1/STK11</i> co-expression was significantly associated with improved overall survival and reduced mortality (HR < 1). Notably, we predominantly observed inverse correlations with pro-inflammatory and immunosuppressive chemokines, immune checkpoints, extracellular matrix components, and key regulators of epithelial-to-mesenchymal transition. In contrast, we found positive associations with anti-inflammatory chemokines and their receptors. Importantly, <i>PEBP1/STK11</i> co-expression was consistently linked to reduced expression of drug resistance genes and greater chemosensitivity across multiple tumor types. Our findings underscore the co-expression of <i>PEBP1</i> and <i>STK11</i> as a promising target for future studies aimed at elucidating its potential as a biomarker for prognosis and therapeutic response in precision oncology.

Radiotherapy is a standard treatment for advanced lung cancer, but resistance remains a significant cause of treatment failure. This study aimed to investigate lactate-associated genes to identify patients likely to benefit from radiotherapy. RNA-seq data from 99 patients with lung cancer who underwent radiotherapy were analyzed to identify differentially expressed genes (DEGs) between resistant and sensitive cases. Bioinformatics tools were used to assess the prognostic relevance of lactate-related genes, and a risk score model was develpoed based on these genes. Dysregulation of these genes in patients with lung cancer undergoing radiotherapy was validated through <i>in vitro</i> experiments. Molecular docking was used to explore potential radiosensitizers. The analysis identified 1482 DEGs, with enrichment analysis highlighting lactate metabolism pathways. A risk score model was constructed using the lactate-related genes ADAMTS3, FADS2, and RTBDN to classify patients into high- and low-risk subgroups. Functional enrichment analysis revealed the model's impact on DNA repair and tumor immunity. A nomogram was developed for clinical implementation. Wet lab experiments further confirmed these findings. In conclusion, a novel risk score model based on lactate-related genes was developed to predict radiotherapy outcomes in lung cancer. FADS2 was identified as a potential biomarker for predicting resistance to radiotherapy. This study is the first to examine the predictive value of lactate-related genes for radiotherapy efficacy in lung cancer, offering valuable insights for personalized treatment strategies to improve therapeutic outcomes.

3-Nitropropionic acid (3-NPA) is a deadly neurotoxic nitroalkane found in numerous fungi and leguminous plants. 3-NPA, known as an antimetabolite of succinate, irreversibly inhibits succinate dehydrogenase and disrupts mitochondrial oxidative phosphorylation. Its utility in modeling Huntington's disease (HD) and oxidative stress has garnered significant research interest. Derivatives of 3-NPA, formed through esterification, have a wide range of biological activities including neurotoxic, antiviral, insecticidal, antimicrobial and antioxidant properties. This review systematically summarizes the structural characteristics, biological activities, and chemical synthesis of 3-NPA-derived compounds, providing valuable insights for further research and therapeutic applications.

Metabolic dysfunction-associated steatotic liver disease (MASLD) encompasses a range of liver conditions, from simple hepatic steatosis to its more severe inflammatory form known as metabolic dysfunction-associated steatohepatitis (MASH). Despite its growing clinical significance and association with cirrhosis and cancer, there are currently few pharmacological treatments available for MASLD, highlighting the urgent need for new therapeutic strategies. This narrative review aims to elucidate the molecular mechanisms of lipophagy in MASLD progression, emphasizing how its dysfunction contributes to hepatic steatosis and lipotoxicity. We also explore the intersection of lipophagy failure with oxidative stress and inflammation in the liver, focusing on key signaling pathways, such as mTORC1 and AMPK, and discuss the therapeutic potential of targeting these pathways by systematically reviewing the literature from PubMed, Scopus, and Google Scholar databases. Recent studies suggest that lipophagy, the selective autophagic degradation of lipid droplets, is crucial for maintaining hepatic lipid homeostasis. Indeed, some vital components of the lipophagy machinery seem to be functionally inhibited in MASLD, resulting in the accumulation of intracellular triacylglycerol (TAG), lipotoxicity, and subsequent oxidative stress, all of which contribute to disease progression. In summary, impaired lipophagy is a central pathological mechanism in MASLD, making it an important therapeutic target. A deeper understanding of these mechanisms may offer new strategic insights for combating the progression of MASLD/MASH.

<b>Background</b>: Gastric cancer (GC) is a malignant tumor of the gastrointestinal tract, characterized by high mortality rates and responsible for about one million new cases each year globally. Surgery is the main treatment, but achieving radical resection remains a relevant intraoperative challenge. Fluorescence-guided surgery offers clinicians greater capabilities for real-time detection of tumor nodules and visualization of tumor margins. In this field, the main challenge remains the development of fluorescent dyes that can selectively target tumor tissues. <b>Methods</b>: we examined the expression of the most suitable GC markers, including carcinoembryonic antigen cell adhesion molecule-5 (CEACAM5) and Claudin-4 (CLDN4), in GC cell lines. To further evaluate their expression, we performed immunohistochemistry (IHC) on tumor and healthy tissue samples from 30 GC patients who underwent partial gastrectomy at the Digestive System Surgery Unit, AOU Careggi, Florence. Additionally, we validated anti-CEACAM5 expression on patient-derived organoids. Furthermore, we developed a fluorescent molecule targeting CEACAM5 on the surface of GC cells and assessed its binding properties on patient tissue slices and fragments. <b>Results</b>: in this work, we first identified CEACAM5 as an optimal GC biomarker, and then we developed a fluorescent antibody specific for CEACAM5. We also evaluated its binding specificity for GC cell lines and patient-derived tumor tissue, achieving an optimal ability to discriminate tumor tissue from healthy mucosa. <b>Conclusions</b>: Overall, our results support the development of our fluorescent antibody as a promising tumor-specific imaging agent that, after further in vivo validation, could improve the accuracy of complete tumor resection.

The pathogenesis of Huntington’s disease is still incompletely understood, despite the remarkable advances in identifying the molecular effects of the Htt mutation in this disease. When we focus on movement disorders, clinical studies offer us some hints about this issue. Human studies employing positron emission tomography have identified a reduction in phosphodiesterase 10A (PDE10A) as the earliest event in the brain of patients with Huntington’s disease, which occurs about 25 years before symptom onset. A PDE10A mutation is also known to cause childhood-onset chorea. GNAL encodes the olfactory type G-protein α subunit (Gα olf ), strongly expressed in the striatum, and its mutation causes familial dystonia. PDE10A and Gα olf are both critical regulators of cyclic AMP and are abundant in striatal spiny projection neurons. These findings suggest that maintaining cyclic AMP levels in the striatum might be an essential target for the pathogenesis of movement disorders such as chorea and dystonia. Why and how these changes in the striatum cause movement disorder are still a mystery. Here we suggest that a key might be evaluating these messenger systems in light of the circuit-level compartmental organization of the striatum, in which there is particular vulnerability of the striosome compartment. We developed machine learning algorithms to define with high precision and reproducibility the borders of striosomes in the brains of q175 Huntington’s disease model mice from 3-12 months of age. We demonstrate that multiple molecules including Gα olf , PDE10A, dopamine D1 and D2 receptors, adenosine 2A receptors, and mu-opioid receptors differentially change their expression patterns in striosomes across ages by comparison with their expression patterns in the matrix compartment. An early and pronounced differential vulnerability of striosomes has been demonstrated in studies of post-mortem Huntington’s brains. Our findings here, mapping the molecular distributions across age in a widely studied mouse model of Huntington’s disease, may help to pinpoint the pathogenic mechanisms of Huntington’s disease by demonstrating the differential molecular changes in the striosome compartment.

Hydrogel microparticles (HMPs) are powerful tools to study and manipulate cellular behavior in 3D cell culture systems and animal models. Here, fully DNA-based HMPs are presented, whose material properties can be precisely tuned by sequence-programmable design of self-assembling DNA nanostructures. These DNA-HMPs offer control over size, stiffness, viscoelasticity and ligand presentation. They are formed by microfluidic encapsulation of two types of orthogonal DNA nanostars and a sequence-complementary DNA linker in water-in-oil droplets. By varying the valency of the DNA nanostar designs, tunable mechanical properties are achieved – spanning three orders of magnitude in Young’s modulus from 30 Pa to 6.5 kPa with distinct viscoelastic behavior. Click-chemistry based functionalization with the small fibronectin-derived peptide cyclic-RGD (c[RGD]) enables integration into fibroblast spheroids. DNA-HMPs are stably retained within the spheroids for several days and undergo design- and stiffness-dependent remodeling, indicating active interactions between the cells and the DNA-HMPs. Combining tunable material properties and inherent biocompatibility of DNA with straightforward functionalization and stimuli-responsiveness, these DNA-HMPs represent a versatile tool to probe and manipulate tissue behaviors in 3D cell cultures and in vivo models. <h4>Table of Contents</h4> DNA hydrogel microparticles are designed to exhibit controllable viscoelasticity and stiffness across three orders of magnitude from 30 Pa to 6.5 kPa. They are uptaken into fibroblast spheroids where they are actively remodeled by cellular forces depending on their mechanical properties.

<h4>SUMMARY</h4> Huntingtin (HTT) function is enigmatic, as the native protein plays critical roles in neuronal health, while mutant HTT (mHTT), carrying an expanded polyglutamine stretch, triggers neurotoxicity and contributes to the pathogenesis of Huntington’s disease (HD). We recently found that HTT is part of a nuclear transcription-coupled DNA repair (TCR) complex with DNA repair enzymes including polynucleotide-kinase-3’-phosphatase (PNKP). This complex resolves DNA lesions during transcription to maintain genome integrity, while in HD, mHTT impairs the activity of this complex, resulting in accumulation of DNA lesions. Using molecular, cellular biology and computational methods, we find that HTT has a role in assembling a functional DNA repair complex in mitochondria. Together with mitochondrial RNA polymerase and transcription factors, HTT resolves mitochondrial DNA lesions to preserve mitochondrial genome integrity and function. Pathogenic mHTT impairs this activity, resulting in persistent DNA lesions and reduced mitochondrial function in HD. Importantly, restoring activity of this complex in a Drosophila HD model through ectopic HTT or PNKP expression significantly improves mitochondrial genome integrity and ameliorates motor deficits. <h4>HIGHLIGHTS</h4> HTT organizes a functional, multifactorial mitochondrial DNA repair complex Mutant HTT impairs the mitochondrial DNA repair complex causing DNA damage accumulation HTT-associated repair complex resolves mitochondrial DNA lesions and DNA integrity Restoring repair activity in HD flies rescues mitochondrial DNA integrity and motor defects

In vertebrates, two major cell types produce extensive pigmentation: neuroepithelium-derived retinal pigment epithelium (RPE) of the eye and neural crest-derived melanocytes. Both produce melanin, express opsins, and exhibit photosensory functions. However, the evolutionary relationship between these cells - whether pigmentation was coopted or they share a common ancestry - remains unclear. We explore these scenarios including the hypothesis of a shared origin from an ancestral pigmented photosensory structure. For this, we harness single cell transcriptomics, chromatin accessibility and spatial transcriptomics data, to connect the transcriptional programs in melanocytes, pinealocytes and RPE with that of the pigmented cells in the sensory vesicle of the tunicate Ciona. The results reveal common regulatory gene expression modules spanning beyond pigment production, including photoreception, metabolism and biosynthesis. This evidence does not favor a model where pigmentation was coopted into one of these cell types, and rather supports the homology of melanocytes and RPE. Further, phylotranscriptomics approach expose recently-evolved melanocyte-specific and RPE-specific functions, which diversified after these types split from the ancestral cell type. Overall, our results support that melanocytes and RPE evolved from ancestral pigmented photosensory structures in chordates, initiating the origin of the neural crest - a major evolutionary driver of the vertebrate lineage.

CoCrMo (Cobalt-Chromium-Molybdenum, CCM) alloys offer excellent wear resistance for the articulating surfaces of load-bearing implants. However, cancer-causing cobalt ions may be released <i>in vivo</i> during articulation. Bacterial infection and biofilm formation on the implant surface are also contributing factors to the failure of these implants. Systemic or localized antibiotics are often not effective against such bacterial infections. Naturally, there is a need to design alloys that show inherent bacterial resistance with minimal release of cobalt ions. This study uses a potential biomedical alloy with copper (Cu) to provide inherent bacterial resistance and hydroxyapatite (HA) ceramic to enhance wear resistance by forming a solid lubricating tribofilm. CCM, CCM-3Cu, CCM-3Cu-1HA, and CCM-3Cu-2HA were processed using a laser-based directed energy deposition (L-DED) additive manufacturing (AM) technique. Antibacterial efficacy was evaluated using <i>Pseudomonas aeruginosa</i>, a Gram-negative bacterium, over 48 and 72 h. An extensive tribocorrosion study was conducted in physiologically relevant Dulbecco's Modified Eagle Medium (DMEM) at loads of 5 and 10 N, accompanied by microstructural analysis. Copper exhibits enhanced bacterial resistance, and the addition of HA improves wear resistance while also decreasing cobalt ion release. The wear resistance of CCM-3Cu-2HA showed a 6-fold improvement compared to CCM. These results indicate that HA-reinforced CCM-3Cu alloys are promising for the articulating surfaces of load-bearing implants.

Defining the mechanisms that promote development and progression of myeloproliferative neoplasms (MPNs) is important for understanding the mechanisms of malignant hematopoiesis and critical development of new treatment approaches. We provide evidence for a key and essential role of the kinase ULK1 in MPN pathophysiology. Our studies demonstrate that genetic or pharmacological targeting of ULK1 delays substantially disease development in <i>Jak2</i>^V617F-mutant MPN models in vivo and establish that ULK1 activity is required for transcription of genes that control hematopoietic stem cell differentiation. Pharmacological targeting of ULK1 exhibits potent therapeutic effects, resulting in reduction of early stage erythroid progenitors in spleen and bone marrow, decreased levels of hemoglobin, and reduced spleen size in MPN mouse models in vivo. Taken together, these findings provide the first evidence for a novel protumorigenic role for ULK1 downstream of the hyperactive JAK2 signaling in MPNs and raise the potential of ULK1 as a new therapeutic target for the treatment of MPNs.

Over the past two decades, peptide drug discovery has experienced a remarkable renaissance, with organic chemistry and biotechnology emerging as pivotal tools for developing peptidomimetics that exhibit improved stability, specificity, and bioavailability compared to conventional peptides. This review systematically examines methodologies for modifying peptide backbones to achieve targeted properties, highlighting recent advances facilitated by modern biotechnological innovations for novel molecular transformations. Additionally, the review emphasizes the practical applications of peptides and peptidomimetics, showcasing their successful integration into medicine and pharmacology. This manuscript evaluates achievements and challenges in the field and identifies critical areas for further research. Its overarching aim is to synthesize current knowledge and propose strategic directions for advancing peptide-based therapeutics.