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.

Breast cancer remains the most prevalent malignancy among women worldwide. When suspicious lesions are detected, standard clinical workflows rely on histopathological evaluation of excised tissue (biopsy) and the choice of preoperative systemic therapies or primary surgical excision. This study explores the potential of X-ray grating-interferometry (GI)-based micro-computed tomography (μCT) as a complementary virtual tool to the gold-standard conventional histopathology Methods: Small (up to 2 cm) residual samples from 20 formalin-fixed mastectomy specimens (from 19 patients) underwent imaging using GI-based μCT and were subsequently processed for routine histology. Diagnostic concordance between gold-standard histological sections and μCT images was established with input from an expert pathologist and an expert radiologist Results: GI-based μCT enabled differentiation of neoplastic and benign findings from surrounding tissue based on morphology and gray-value contrast. While μCT lacks the cellular resolution of histology, its 3D volumetric data provided valuable spatial context for small structures. Imaging a 2 cm specimen required approximately 13.4 h, with negligible degradation in diagnostic image quality when the acquisition time was reduced to 4.4 h Conclusions: GI-based μCT demonstrates strong potential as an adjunct to gold-standard histopathology, providing high-quality soft-tissue contrast comparable to conventional μCT and aiding in the identification of regions of interest for further tissue-based analysis. The possible clinical utility of GI-based μCT diagnostics needs further clinical evaluation, also on smaller samples.

Tumor-infiltrating immune cells play a fundamental role in shaping patient prognosis across many cancers. Retroperitoneal dedifferentiated liposarcomas (DDLPS) and leiomyosarcomas (LMS) are associated with poor prognosis, therefore, a deeper understanding of the immune landscape in these tumors is warranted to identify novel prognostic biomarkers and immunotherapeutic targets. In this exploratory study, we analyzed spatial organization, frequency, and proportions of CD8[+] T cell (including activated, proliferating, and exhausted phenotypes), T helper, and macrophage subsets, and B cells in primary treatment-naive retroperitoneal DDLPS (n = 22) and LMS tissues (n = 10) using classical and multiplex immunohistochemistry. CD8[+]LAG-3[+] T cells comprised the highest proportion of all analyzed CD8[+] subsets. Their elevated density was associated with improved overall survival (OS). Moreover, higher density of CD8[+]Ki-67[+] T cells was linked to better OS and tended to independently predict OS. Interestingly, enhanced density of regulatory T cells was associated with favorable OS. Densities and proportions of T cell subsets did not significantly differ between DDLPS and LMS. A lower density of M2-like macrophages (CD68[+]CD163[+]) was linked to better progression-free survival (PFS) and tended to serve as an independent prognostic factor. Invasive margin revealed distinct prognostic patterns compared to tumor core, where high levels of CD8[+]Ki-67[+]TCF1[+] were linked to better PFS and elevated proportions of CD8[+]GrzB[+] and CD3[+]T-Bet[+] T cells were associated with inferior PFS. These findings highlight the clinical relevance of immune infiltration in retroperitoneal DDLPS and LMS. Moreover, they support the rationale for further exploration of the immune architecture for prognostic biomarkers and development of targeted immunotherapeutic strategies to improve the clinical outcomes of the patients.

Cholangiocarcinoma (CCA) is a highly lethal malignancy and often exhibits histological overlap with non-malignant bile duct lesions, including cholangitis and precancerous lesions. The lack of reliable markers to distinguish malignant from non-malignant biliary lesions underscores the need for improved diagnostic approaches. The cGAS-STING pathway, a key regulator of innate immune responses, may provide insight into inflammation-associated carcinogenesis. This study aimed to compare the expression of cGAS, STING, IRF3, and IFN-α among CCA, precancerous lesions, and cholangitis, and to evaluate their potential diagnostic relevance. A total of 198 formalin-fixed paraffin-embedded (FFPE) tissue samples were analyzed, including 164 cases of cholangiocarcinoma, 26 precancerous lesions, and 8 cases of cholangitis. Immunohistochemistry for cGAS, STING, IRF3, and IFN-α was performed, and protein expression was quantitatively assessed using H-scores. Diagnostic performance was evaluated using receiver operating characteristic (ROC) curve analysis. ROC-derived H-score cut-off values were applied in logistic regression models to assess association with CCA. IRF3 and IFN-α expression were markedly increased in precancerous lesions compared with cholangitis. High cGAS and IRF3 expression at ROC-derived cut-off values showed association with CCA. Logistic regression analysis demonstrated that elevated cGAS (H-score ≥ 202; p = 0.026) and IRF3 (H-score ≥ 201; p = 0.032) expression were associated with increased odds of CCA. Although cGAS and IRF3 showed limited standalone diagnostic accuracy, their ROC-derived H-score thresholds were modestly associated with CCA. This work could provide adjunctive diagnostic information when integrated with routine histopathological assessment.