Tendon injuries are common in occupations involving repetitive mechanical loading, and environmental heat exposure may further increase injury risk by altering tissue tolerance to strain. This study investigated cytoskeletal organization (F-actin), microRNA expression (miR-21-5p, miR-29a-3p, and miR-192-5p) and intracellular and extracellular heat shock protein 72 (HSP72) responses to combined heat under hyperthermic conditions (39 °C) and cyclic tensile strain in human achilles tendon-derived tenocytes from a single donor. Tenocytes were maintained at 37 °C without strain or exposed for 3 h to 39 °C without strain, 5% cyclic strain at 39 °C, or 10% cyclic strain at 39 °C. Moderate strain at 39 °C was associated with partially preserved F-actin organization, while higher strain at 39 °C produced greater qualitative disruption of the cytoskeleton. miR-21-5p and miR-29a-3p exhibited condition-dependent regulation, while miR-192-5p did not show a statistically significant change. In addition, intracellular and extracellular HSP72 levels varied across the experimental conditions, indicating activation of cellular stress responses under combined thermal and mechanical exposure. These preliminary findings demonstrate acute cytoskeletal remodeling, molecular and biochemical responses of human tenocytes to combined mild hyperthermia and cyclic mechanical strain providing a basis for further investigation of potential biological indicators of tendon stress in occupational settings.

The MHC class II transactivator (CIITA) is the master regulator of MHC class II (MHC-II) genes expression. In fact, it promotes both constitutive and interferon gamma-inducible expression of all MHC-II genes and thus regulates the optimal activation of adaptive immunity, which depends on the proper activation of CD4[+] T helper cells. In this review we will briefly discuss the role and regulation of CIITA and the downstream expression of MHC-II in adaptive immunity, followed by a discussion of how the induction of CIITA and MHC-II expression in tumor cells of different histological origins can make them surrogate antigen presenting cells for their own tumor antigens, thereby enabling them to elicit a protective adaptive anti-tumor immune response. Based on this concept, we will discuss possible strategies to use CIITA-driven MHC-II-positive tumor cells to unveil tumor antigens and develop different types of anti-tumor vaccines, including CIITA-based vaccination strategies. We will also address the major challenges associated with in vivo CIITA delivery and critically discuss the apparently conflicting preclinical evidence obtained in glioblastoma models. Finally, we will highlight the potential of epigenetic drugs to directly induce CIITA-driven MHC-II-positive tumor cells capable of stimulating the immune system and discuss the potential of CIITA and MHC-II expression as predictive markers of response to immunotherapy, thereby providing a rationale for combining CIITA-inducing strategies with immune checkpoint inhibitors for future human clinical setting.

Cisplatin is an effective chemotherapeutic agent with dose-limiting toxicity in non-target tissues. Hyperbaric oxygen therapy (HBOT) and mitochondrial transplantation (MitoTr) have been explored as supportive strategies in preclinical models. We examined whether HBOT and/or MitoTr modulate cisplatin-associated reductions in MTT metabolic activity and oxidative balance in healthy human mesenchymal stem cells (hMSCs) under incubator versus hyperbaric conditions. Bone marrow-derived hMSCs were seeded at 5 × 10[3] cells/well and exposed to cisplatin at an experimentally determined MTT-based LD50 (148 μM; 150 μM used). A delayed adjunctive intervention protocol was applied after LD50-level exposure: cisplatin or vehicle for 24 h, followed by 4 h MitoTr co-incubation (10× or 100× cell-number-scaled dose), then three HBOT sessions (90 min, 2.1 ATA) or matched incubator control. Endpoints were MTT absorbance (NAD(P)H-dependent oxidoreductase activity), total antioxidant status, total oxidant status, and oxidative stress index, analyzed by two-way ANOVA with Tukey post hoc. At 150 μM cisplatin, MTT absorbance was markedly reduced in all cisplatin-containing groups under both conditions (all p < 0.001 vs controls), and neither HBOT nor MitoTr restored MTT signal to control levels. Among cisplatin-free groups, M10 under HBOT exceeded control HB MTT absorbance (p = 0.009), whereas M10 under incubator conditions did not differ from control. HBOT reduced baseline MTT absorbance in cisplatin-free groups relative to matched incubator controls (p < 0.001). MitoTr lowered oxidant status under incubator conditions; HBOT improved redox indices in mitochondria-free groups. Under the LD50-level conditions tested, neither HBOT nor MitoTr restored NAD(P)H-dependent oxidoreductase (MTT) activity in hMSCs. Under non-toxic conditions, both modalities produced context-dependent redox changes. Whether lower, clinically relevant cisplatin exposures would yield partial protection remains an open question.

Airway epithelial dysfunction is a hallmark of asthma, contributing to chronic inflammation and structural remodeling. Recent research suggests the role of post-translational modifications in regulating epithelial immune responses. However, the role of specific deubiquitinating enzymes (DUBs) remains unclear. In this study, we identified OTU deubiquitinase 7a (OTUD7a) as a gene that is significantly downregulated in a murine model of allergic asthma. Transcriptomic analysis of two independent datasets (GSE71822 and GSE9383) consistently revealed the suppression of OTUD7a in asthmatic lungs. Furthermore, immunofluorescence analysis suggested that OTUD7a was detected in CC10-positive bronchiolar epithelial cells, non-ciliated epithelial cells known for their anti-inflammatory functions. To explore the molecular associations of OTUD7a, we established a club cell line overexpressing Flag-tagged OTUD7a and performed co-immunoprecipitation-mass spectrometry (Co-IP/MS). We identified 1135 candidate interacting proteins. In parallel, LC-MS/MS-based ubiquitinome profiling in ovalbumin (OVA)-induced asthmatic lungs revealed widespread changes in protein ubiquitination, with more than 3000 differentially modified ubiquitination sites. These changes were enriched in pathways associated with immune signaling, cytoskeletal dynamics, and epithelial barrier function. Collectively, our findings suggest that OTUD7a, as a DUB detected in CC10-positive bronchiolar epithelial cells, is markedly decreased in asthma and they present a comprehensive landscape of ubiquitination changes in allergic airway inflammation. Therefore, our study offers new insight into post-translational regulation in the pathogenesis of asthma.

Diabetic nephropathy (DN) is a major complication of diabetes and a leading cause of dialysis initiation. However, treatment options for DN are currently limited, highlighting the urgent need for novel therapies. The pathogenesis of DN involves persistent chronic hyperglycemia and advanced glycation end products, leading to inflammation, and oxidative stress. SMTP-44D, a compound derived from Stachybotrys microspora, exhibits anti-inflammatory and antioxidant properties. This study aimed to elucidate the renoprotective mechanism of SMTP-44D in DN. To establish a DN model, 6-week-old db/db mice underwent unilateral nephrectomy, and the model animals were treated with SMTP-44D for 10 weeks. Key physiological parameters, blood glucose levels, soluble epoxide hydrolase (sEH) substrates [epoxyeicosatrienoic acids (EETs) and lysophosphatidic acids (LPAs)], and the critical signaling molecule nuclear factor-kappa B (NF-κB) and Peroxisome Proliferator-Activated Receptor Gamma (PPARγ), were analyzed. Our results demonstrated that SMTP-44D increased the levels of serum EETs and LPAs in DN model animals. Following SMTP-44D treatment, the fraction of nuclear NF-κB decreased and cytoplasmic PPARγ level increased in the renal tissues. These results are consistent with the hypothesis that SMTP-44D inhibits sEH, causing an accumulation of substrates that act as signaling molecules controlling inflammatory processes via NF-κB. Furthermore, SMTP-44D improved insulin resistance, as evidenced by reduced blood glucose levels. In conclusion, SMTP-44D represents a promising novel therapeutic candidate for DN, offering a multifaceted approach that addresses inflammation, oxidative stress, and insulin resistance.

Retinitis pigmentosa (RP) is accompanied by retinal oxidative stress and neuroinflammation, but whether local retinal stress is associated with detectable circulating molecular changes remains unclear. Using Rho[P23H/+] mice at postnatal days 40, 90, and 240, we combined electroretinography, hematoxylin and eosin (H&E)-based outer nuclear layer (ONL) measurement, retinal qPCR, plasma redox assays, public transcriptomic reanalysis, and plasma CSF1 ELISA to characterize molecular changes during degeneration. Rho[P23H/+] mice showed progressive ERG attenuation and ONL thinning. Retinal Hmox1, Sod1, and Cat transcripts were increased, whereas plasma total oxidant status, total antioxidant status, oxidative stress index, glutathione peroxidase, total superoxide dismutase, catalase, and malondialdehyde showed no consistent genotype-dependent changes. Reanalysis of GSE152474 and GSE156533 identified immune- and glial-enriched modules containing Csf1 and Csf1r. qPCR validation showed a stage-dependent pattern of retinal Csf1 and Csf1r expression, with Csf1r increased at day 40 and both Csf1 and Csf1r increased at days 90 and 240. Plasma CSF1 concentration did not differ significantly between genotypes at day 90 but was higher in Rho[P23H/+] mice than in age-matched WT controls at day 240. These findings identify a stage-dependent pattern of retinal Csf1 and Csf1r expression and a late-stage genotype-associated difference in plasma CSF1 concentration, although the mechanistic relationship between the retinal and circulating findings remains undetermined.

Antiphospholipid syndrome (APS) is an autoimmune thrombotic disorder characterized by the presence of antiphospholipid antibodies, including anti-phosphatidylserine/prothrombin antibodies (aPS/PT). While neutrophil extracellular traps (NETs) are implicated in the pathogenesis of APS, the role of aPS/PT in NET induction and its contribution to thrombosis remain unclear. This study aimed to clarify the effects of NETs induced by a monoclonal aPS/PT antibody on platelet activation and their potential contribution to thrombo-inflammatory responses. NETs were induced by stimulating peripheral blood neutrophils from healthy donors with aPS/PT. Their morphology and platelet-activating capacity were compared with NETs induced by anti-neutrophil cytoplasmic antibodies (ANCAs). Proteomic analyses were conducted to comprehensively compare protein compositions of these NETs, and candidate proteins associated with platelet activation in aPS/PT-induced NETs were identified. Functional inhibition assays were then conducted to assess whether blocking these candidates would suppress aPS/PT-induced NET-mediated platelet activation. We found that binding of aPS/PT to neutrophils induced NET formation, with a larger and more fibrous morphology compared to ANCA-induced NETs. Platelets trapped in aPS/PT-induced NETs showed significantly higher activation compared to those trapped in ANCA-induced NETs. Proteomic analyses identified histone H3 as a potential mediator of platelet activation in aPS/PT-induced NETs. Correspondingly, plasma concentrations of H3.1 nucleosome were significantly higher in patients with APS than in healthy controls. Blockade of histone H3 using a neutralizing antibody significantly suppressed platelet activation mediated by aPS/PT-induced NETs. These findings suggest that aPS/PT-induced NETs contribute to platelet activation and may promote thrombo-inflammatory responses in APS. Targeting histone H3 within aPS/PT-induced NETs may provide a potential therapeutic strategy for thrombo-inflammatory processes in APS.

There is increasing evidence that translation is not limited to annotated protein-coding genes. Ribosome profiling sequencing, mass spectrometry-based proteomics, and immunopeptidomics have identified the productive translation of non-canonical open reading frames (ORFs). This suggests that the functional proteome includes not only conserved proteins but also proteins hidden in non-coding RNAs and de novo proteins. Some of these translated products are functional peptides, while others may be non-functional, potentially arising from evolutionary events. Several non-canonical ORF-encoded peptides have been found to regulate multiple physiological and pathological functions, particularly in cancer, immunity, and inflammation, indicating that they have potential as biomarkers and novel therapeutic targets. To better understand the diversity of functional peptides and translated non-canonical ORFs based on existing data, we summarize their classification according to transcriptional features and supporting evidence, including non-canonical ORFs located in ncRNAs and canonical mRNAs. This review provides a concise summary of the origin, discovery methods, and classification of non-canonical ORFs. It offers insights into the origins and functions of non-canonical ORF-encoded peptides from an evolutionary perspective, while also exploring the biological functions and regulatory mechanisms of these non-canonical ORF-encoded hidden proteins in tumorigenesis and progression.

Sudden cardiac death (SCD) in individuals ≤35 years remains a major challenge for cardiology and forensic medicine, often affecting subjects without prior symptoms. At the same time, the consumption of energy drinks (EDs) and the medical or non-medical use of cognitive enhancers or "smart drugs" (SDs), including methylphenidate, amphetamine derivatives, and modafinil, has increased markedly among adolescents, students, and athletes. This review integrates mechanistic, clinical, epidemiological, and forensic evidence to clarify the role of EDs and SDs as arrhythmogenic triggers, identify at-risk populations, and outline diagnostic and regulatory implications. ED formulations consistently produce measurable cardiovascular effects. Experimental whole-heart models show that caffeine-taurine combinations shorten action potential duration and effective refractory period, facilitating re-entry. Clinical and forensic case reports describe SCD or near-fatal arrhythmias shortly after ED intake, particularly in individuals later diagnosed with long-QT syndrome, catecholaminergic polymorphic ventricular tachycardia, or other inherited channelopathies, supporting a trigger-on-substrate mechanism. Prescription SDs increase sympathetic tone, myocardial oxygen demand, and autonomic stress, especially under dehydration, sleep deprivation, strenuous exercise, or polysubstance use. Although population-level risk is low, adults with hypertension or occult cardiac disease and young individuals with latent genetic substrates exhibit substantially increased vulnerability. Stimulants seldom act as sole causes of SCD but can significantly lower the arrhythmic threshold in susceptible individuals, particularly under physiological or environmental stress. Integrating clinical screening, rigorous forensic investigation, including molecular autopsy, and improved regulatory measures are essential to reduce preventable stimulant-associated cardiac deaths while preserving legitimate therapeutic use.

Histological grading, using the Nottingham Grading System (NGS), is a major prognostic indicator for breast cancer. NGS involves the scoring of cancer cell-related morphological features, yet it overlooks tumor microenvironment (TME) components such as collagen. Collagen proteins, integral to the extracellular matrix (ECM), influence tumor architecture and progression but their relationship with histological grade is not fully characterized. Here, we assessed intratumoral collagen deposition using Masson Trichrome staining of whole slides (n = 166), proteomic profiling (n = 2) and transcriptomic analyses of the METABRIC (n = 1827) and TCGA-BRCA (n = 753) cohorts. We showed that low-grade tumors display significantly higher intratumoral collagen deposition compared to high-grade tumors. Moreover, we demonstrated that collagen expression at the transcript and protein levels (Masson Trichrome) could discriminate Grade II carcinomas into distinct prognostic groups, in which patients with Grade II carcinomas with elevated levels of collagen expression were associated with lower pTNM stage and better survival outcomes. Our results support the inclusion of TME features, such as collagen deposition, to enhance prognostic accuracy in breast cancer.

Methyltransferase-like 13 (METTL13) is a member of the methyltransferase-like (METTL) family characterized by two conserved seven-beta-strand(7BS) catalytic domains. Current evidence identifies METTL13 primarily as a dual protein methyltransferase that modifies the translation elongation factor 1 A(eEF1A), including Lys55 dimethylation and N-terminal methylation. Through this best-established mechanism, METTL13 enhances eEF1A GTPase activity, reshapes translation elongation and codon-specific translational output, and promotes protein production in cells. In multiple cancer settings, METTL13 and the eEF1A-K55 methylation axis are associated with increased translational demand, tumor progression, and Ras-driven tumorigenesis. At the same time, accumulating studies indicate that METTL13 functions are strongly context dependent. Beyond its canonical eEF1A-centered role, METTL13 has been linked to additional regulatory programs, including the c-Cbl/SERCA2a axis, RNA-associated pathways, and disease-specific signaling networks, although these noncanonical mechanisms remain less well defined. Intriguingly, METTL13 may also exert protective or tumor-suppressive effects in selected contexts, such as ischemic heart failure, hereditary deafness, and clear cell renal cell carcinoma. In this narrative review, we summarize the structural and functional features of METTL13, its regulatory network, and current evidence for its roles in malignant and non-malignant diseases. By integrating structural features, molecular modification mechanisms, translational control, disease pathophysiology, and early clinical-translation evidence, this review reframes METTL13 as a context-dependent regulator of proteome adaptation and highlights unresolved mechanistic questions regarding substrate specificity, microenvironment-dependent functional outputs, and context-specific intervention strategies.

Congestion is the central pathophysiological mechanism and a major determinant of morbidity and mortality in acute heart failure (AHF). Natriuretic peptides (NPs), although guideline-recommended, provide only an indirect assessment of congestion and may have limited accuracy. This systematic review synthesized evidence on four biomarkers, including carbohydrate antigen 125 (CA-125), adrenomedullin (ADM) and its derivatives, soluble suppression of tumorigenicity-2 (sST2), and soluble cluster of differentiation 146 (sCD146), focusing on their associations with congestion and prognosis in AHF. A systematic search of PubMed and Embase was conducted in June 2025. Eligible studies included patients with AHF and evaluated at least one biomarker in relation to clinical, imaging, or laboratory markers of congestion and/or prognostic outcomes. After screening and full-text review, 39 studies met the inclusion criteria. CA-125 consistently correlated with systemic congestion, with higher admission levels predicting increased mortality and rehospitalization. Interventional studies suggested its potential role in guiding decongestive therapy. ADM derivatives were associated with congestion severity and residual congestion at discharge. Bio-adrenomedullin reflected rapid hemodynamic changes due to its short half-life, while mid-regional proadrenomedullin provided prognostic information. sST2, primarily reflecting myocardial stress and inflammation, demonstrated robust prognostic value and dynamic decline during hospitalization. sCD146 emerged as a novel marker strongly associated with systemic congestion, with additional diagnostic value in conditions where NPs underperform. Overall, these biomarkers provide mechanistically complementary information to NPs, improving congestion assessment and risk stratification in AHF. Larger prospective and interventional studies are required to define their role in routine clinical practice and in guiding decongestive strategies.

By a classical approach, the invasive margins of colorectal carcinomas can be typed as expansive or infiltrative, the latter portending a poor prognosis. To gain insight into the tumor biology behind these morphological features, we submitted 18 microsatellite-stable colorectal carcinomas that could be typed as having invasive margins of the infiltrative (N = 7) or expansive type (N = 11) with confidence to nanoString nCounter® analysis (Tumor Signaling 360™ Panel). Tissue microarray punch samples were obtained from the tumors and prior to RNA extraction histological sections were prepared. In an unsupervised cluster analysis of the gene expression data, 9 tumors, all of which expansive-types, were assigned to cluster 1, and all 7 cases with infiltrative-type margins to cluster 2 which also included 2 expansive-type cancers. Thus, invasive margin-types were mirrored in gene expressions; however, tumor budding, the second type of colorectal carcinoma invasion phenotype, was not. Nanostring Annotation Scores were significant (p < 0.05) for signaling pathways (TGFb, PDGF, MET, FGFR), extracellular matrix remodeling, and anti-tumor immunity processes, but any hopes that the tumor biology behind the two phenotypes of invasion could be pinpointed to differential expressions of a small set of genes were not fulfilled. Taken together, the data indeed give a molecular underpinning to the two invasion phenotypes, pointing out that matrix features and anti-tumor immunity are key. Nevertheless, we failed to gain a more detailed insight into the mechanics at work, and this may well be due to general limitations of the technology employed.