Medicine and Pharmacology

Abstract: Glioblastoma multiforme (GBM) remains one of the most aggressive and treatment-resistant brain tumors, characterized by rapid proliferation and poor patient prognosis. Novel therapeutic strategies are urgently needed to improve clinical outcomes. In this study, we investigated the cytotoxic and pro-apoptotic effects of bortezomib (BORT), a proteasome inhibitor, and graphene oxide (GO), a nanomaterial with known anticancer potential, on human glioblastoma cell lines. Treatment with BORT and GO, both individually and in combination, significantly reduced cell viability in a dose-dependent manner, as determined by MTT. In this study we observed revealed enhanced apoptotic cell death, accompanied by increased activation of both caspase-8 and caspase-9, indicating simultaneous engagement of extrinsic and intrinsic apoptotic pathways. Western blot analysis demonstrated downregulation of anti-apoptotic proteins Bcl-2 and upregulation of pro-apoptotic markers (NOXA, cleaved PARP). A central finding of this work is the pronounced increase in intracellular reactive oxygen species (ROS) levels following BORT–GO treatment. The elevated ROS levels observed in BORT–GO–treated cells compared with free bortezomib therefore suggest a synergistic mechanism in which GO-mediated oxidative stress amplifies proteasome inhibition–induced apoptosis, which is particularly visible in the LN-229 line. Notably, the combination of BORT and GO exhibited synergistic effects, suggesting a potential cooperative mechanism through proteasome inhibition and oxidative stress induction. These findings indicate that graphene oxide may potentiate the antitumor efficacy of bortezomib and support further investigation of this combination as a promising therapeutic approach for glioblastoma.

Abstract: Triple-negative breast cancer (TNBC) is the most aggressive subtype and accounts for 15-20% of breast cancer cases. TNBC lacks expression of the estrogen, progesterone, and human epidermal growth factor receptor 2/neu (HER2/neu) receptors. Due to the lack of these receptors, targeted therapies are virtually ineffective. In addition, due to their aggressive nature, standard therapy options are limited by the development of resistance making TNBC very challenging to treat, highlighting the need for new therapeutic approaches. TNBCs undergo metabolic alterations to support growth and survival, one of which is glutamine addiction. TNBCs have been shown to demonstrate increased levels of GLS mRNA, which correlates with their dependence on exogenous glutamine for growth and survival. This study examined whether inhibiting glutamine metabolism enhances radiotherapy (RT) efficacy against TNBC. In two TNBC cell lines (MDA-MB-231 and 4T1), glutamine deprivation and the glutaminase (GLS) inhibitor CB-839 combined with ionizing radiation (IR) reduced colony formation in the combination treatment was significantly more effective than either treatment alone. In a murine model of TNBC, this combination significantly decreased mammary fat pad tumor growth. These findings demonstrate that inhibiting glutamine metabolism combined with RT represents a promising therapeutic strategy that could improve treatment outcomes in TNBC patients who currently have limited effective treatment options.

Abstract: Liposomal irinotecan (nal-IRI) has emerged as a cornerstone in the treatment of metastatic pancreatic ductal adenocarcinoma (mPDAC), particularly in combination with 5-fluorouracil and leucovorin (5-FU/LV) following progression on gemcitabine-based therapy. Despite its demonstrated survival benefit, gastrointestinal toxicity—most notably diarrhea—remains a clinically significant adverse event that can compromise dose intensity, treatment adherence, and patient quality of life. Diarrhea associated with irinotecan-containing regimens is mechanistically complex, encompassing both acute cholinergic and delayed secretory components mediated by mucosal injury and enterohepatic recirculation of the active metabolite SN-38. The liposomal formulation alters the pharmacokinetic profile of the drug, prolonging systemic exposure while maintaining toxicity risks. This review provides a comprehensive and updated overview of the pathophysiology, incidence, and clinical implications of diarrhea in nal-IRI–based regimens. Evidence-based management strategies are discussed, including pharmacologic interventions, supportive care, dose modifications, and patient education. Emerging therapeutic approaches—including microbiome modulation and pharmacogenomic-guided therapy—are also explored. A multidisciplinary and proactive management approach is essential to optimize outcomes and minimize toxicity.

Abstract: The CLIA-CA-62 assay is an in vitro diagnostic device registered in Russia and Kazakhstan for measuring a marker specific to epithelial carcinomas. This pilot project aimed to assess CA-62 utility for primary cancer screening in an asymptomatic cohort in Kazakhstan. The trial was interrupted in January 2022 for reasons unrelated to the scientific program before clinical outcomes could be obtained. Available baseline data were therefore used to characterize the CA-62 value distribution and perform a scenario-based assessment of estimated assay specificity at a reference value of 5,000 U/mL. The analysis included 1,214 quantitative CA-62 measurements from asymptomatic healthcare workers aged 45–70 years, collected during annual preventive examinations between September and October 2021. The distribution was markedly right-skewed, with 92.5% of samples in the normal zone (median: 3,371 U/mL; IQR: 1,965–4,415 U/mL; 95th percentile: 6,309 U/mL). At the 5,000 U/mL cutoff, 7.5% of results (91/1,214) were elevated. Scenario-based modeling assuming cancer prevalence of 0.5–2.5% and assay sensitivity of 65–95% yielded an estimated specificity of 92.79–94.75%. These findings provide an analytical foundation for prospective verification of CA-62 in primary screening settings.

Abstract: Despite advances in multimodal treatment, head and neck squamous cell carcinoma (HNSCC) remains a major clinical problem owing to its high recurrence rate and frequent development of treatment resistance. Abnormal histone modifications, particularly lysine methylation regulated by methyltransferases (KMTs) and demethylases (KDMs), have emerged as key drivers of HNSCC initiation, progression, and cellular plasticity. This review aims to comprehensively evaluate the role of selected KMTs and KDMs in HNSCC biology, with a focus on their contribution to resistance to immunotherapy, radiotherapy, and cytotoxic chemotherapy. We summarize and critically analyze preclinical and clinical studies investigating histone methylation dynamics in HNSCC, with particular emphasis on enzymes such as KMT2C/D, EZH2, NSD1/NSD2, SMYD3, G9a/EHMT2, LSD1, KDM2A/B, KDM3, KDM4, KDM5, KDM6, KDM7, and KDM8. Particular attention is given to pharmacological approaches targeting these proteins: we discuss smallmolecule inhibitors of EZH2, LSD1, KDM4/5/6, and other KMT/KDMs that are currently in preclinical development or earlyphase clinical trials, and we highlight completed and ongoing studies testing EZH1/2 inhibitors and epigenetic combinations in patients with recurrent and metastatic HNSCC. Deregulation of specific KMTs and KDMs reshapes histone methylation at key residues, thereby controlling cell cycle progression, epithelial-mesenchymal transition (EMT), stem cell phenotypes, DNA damage responses, and interactions with the immune system in HNSCC. Targeting disrupted histone methylation pathways may partially reverse epigenetic reprogramming of HNSCC cells and represents a promising strategy to improve treatment efficacy in advanced disease. We also summarize preclinical evidence and limited clinical data on targeting histone methylation in HNSCC and discuss therapeutic implications.

Abstract: Leukocyte migration is a fundamental process in both innate and adaptive immune responses. This process is tightly regulated by chemokines and their cognate receptors. The bioavailability of chemokines is further modulated by atypical chemokine receptors (ACKRs), a subset of chemokine receptor–like molecules that lack coupling to canonical G protein–mediated signaling pathways. Among these, ACKR4 regulates dendritic cell migration through ligand scavenging and has been implicated in tumor progression in murine models. We previously established anti-mouse ACKR4 (mACKR4) mAbs, A4Mab-1, A4Mab-2, and A4Mab-3, by N-terminal peptide immunization. This study examined the binding epitopes of A4Mabs. Alanine (or glycine) scanning within the N-terminal region (amino acids 2–19) was performed using flow cytometry and Western blotting. The results demonstrated that Tyr11, Tyr12, Glu14, Glu15, and Glu17 are critical for recognition by A4Mab-1, while Tyr11, Tyr12, Tyr13, Glu15, and Asn16 are essential for recognition by A4Mab-2 in flow cytometry and Western blotting. Furthermore, Glu14, Asn16, and Glu17 are essential for recognition by A4Mab-3 in flow cytometry. These findings contribute to the understanding of mACKR4 recognition by A4Mabs.

Abstract: Pancreatic ductal adenocarcinoma (PDAC) remains highly lethal and is largely refractory to immune checkpoint inhibition, reflecting limited antigen-specific priming, dominant myeloid suppression, dense desmoplasia, and abnormal vasculature that together enforce immune exclusion. CD40 is a central costimulatory receptor that links CD4⁺ T cell help (via CD40L/CD154) to antigen-presenting cell (APC) licensing and effective CD8⁺ T cell priming, positioning CD40 agonism as a rational strategy to enhance antitumor immunity in PDAC. In addition to robust expression on APC populations (DCs, macrophages, and B cells), CD40 has been reported on subsets of PDAC tumor cells and stromal compartments (including fibroblasts and endothelial cells), implying that CD40-directed therapies may engage multiple cellular nodes and influence both immune activation and microenvironmental remodeling. Mechanistically, CD40 signaling integrates TRAF-dependent pathways (canonical/non-canonical NF-κB, MAPK, and PI3K/AKT) that can promote APC maturation, IL-12-associated Th1 programming, macrophage repolarization, and matrix remodeling that may reduce physical barriers to immune infiltration; however, tumor-intrinsic CD40 signaling can be context-dependent and has been linked to divergent survival or apoptotic outcomes in different settings. Clinically, multiple CD40 agonists are in development, predominantly in combination regimens with chemotherapy, checkpoint blockade, and/or vaccine platforms, with evidence of pharmacodynamic immune engagement but variable efficacy and incomplete randomized validation. Baseline CD40 expression has not consistently predicted benefit, underscoring the need for spatially resolved profiling and on-treatment pharmacodynamic biomarkers to guide patient selection, sequencing, and regimen optimization. Ongoing studies and next-generation, tumor-localized agonist formats will define whether CD40 agonism can deliver consistent, durable clinical benefit in PDAC.

Abstract: Cancers are thought to result from the interaction of a small set of mutations in genes that regulate cell growth. A sixty-five year series of incidence data is available for cancer of the pancreas, spanning the period over which incidence increased threefold. These data were analysed using a multi-hit mutation model. The analysis yielded as parameters the averages of the number of interacting genes (with the number four being within the range found), the rate of their mutation, the age at which mutations began to accumulate and the number of persons at risk. Only this last parameter showed a significant increase over the years, a period which coincided with increased consumption of sugars. The effect of glucose on the proliferation of cells of the pancreas is suggested as a basis for these findings.

Abstract: Renal cell carcinoma encompasses a heterogeneous group of kidney tumors with wide variations in biological behavior, histologic subtype, and clinical aggressiveness. Accurate preoperative characterization is essential for management; however, it remains challenging due to overlapping imaging features and tumor complexity. CT is the most widely used imaging technique for renal mass evaluation, providing broad availability, high spatial resolution, and multiphasic acquisition capabilities. However, its ability to distinguish histologic subtypes and predict tumor aggressiveness remains limited. This review provides an updated overview of renal cell carcinoma epidemiology and evidence supporting CT as an essential imaging modality. It outlines key radiologic features of main histologic subtypes, highlights markers of aggressive behavior, and discusses the relationship between CT findings and the International Society of Urological Pathology (ISUP) grading system. We explore radiomics, summarizing its methodological foundations and applications in characterizing solid renal masses, emphasizing the need for multicenter studies and standardized radiomic workflows to develop accurate, reproducible tools for improving diagnostic accuracy and risk stratification for renal cell carcinoma.

Abstract: Background/Objectives: Clear cell renal cell carcinoma (ccRCC), the predominant malignant subtype of kidney cancer, is the leading cause of death among renal cell carcinoma patients. Although a subset of ccRCC patients benefit from select immune checkpoint inhibitors (ICIs), prognosis remains poor. While PD-1 and PD-L1 have been extensively studied, the prevalence and distribution of other immune checkpoints (ICs) and their relationship with epithelial-to-mesenchymal transition (EMT) remain poorly characterized. Here, we investigated the interplay between twenty ICs and EMT markers and assessed their combined prognostic relevance in ccRCC patients. Methods: Transcriptomic profiling and integrated bioinformatic analyses were performed, including differential expression, correlation analyses, survival analyses, forest plot analyses, ROC curve evaluation, and OncoPrint visualisation, complemented by analysis of single-cell RNA sequencing data, immunohistochemistry, and multiplex secretory IC (LegendPlex) assays. Results: Transcriptomic profiling of over 500 ccRCC tumors versus normal kidney tissue revealed dysregulation of ICs, particularly LAG3 and NT5E. Notably, expression of ICs, including LAG3 and NT5E, was associated with poor overall survival in 415 ccRCC patients. ICs that synergised with EMT phenotype provided improved prognostic discrimination compared to individual ICs. Correlation analyses, single-cell RNA sequencing and immunohistochemistry demonstrated that EMT-associated tumor cells exhibit coordinated expression of LAG3 and NT5E. Receiver operating characteristic analysis highlighted the potential clinical utility of LAG3 and NT5E. Conclusions: Collectively, this study defines an EMT-IC axis in ccRCC and demonstrates its relevance to tumor biology and patient outcomes, highlighting LAG3 and NT5E as potential prognostic markers and therapeutic targets.

Abstract: Cancer is increasingly recognized as a metabolic disease influenced by nutritional factors, with multi-omics technologies and artificial intelligence (AI), particularly machine learning (ML), enabling integrative analyses of diet, metabolism, and tumor biology interactions. This study aimed to synthesize evidence on these approaches for understanding the nutrition–metabolism–cancer axis and assess their translational potential in oncology, especially in low-resource settings. A PRISMA-compliant systematic review and meta-analysis searched PubMed, EMBASE, and Cochrane databases from 2018 to 2025, including studies on human cancers using ≥2 omics layers integrated via AI/ML and addressing nutritional/metabolic exposures. Random-effects pooling evaluated area under the curve (AUC), odds ratios (OR), and clinical endpoints, with subgroup analyses and quality assessments via QUADAS-2, ROBINS-I, TRIPOD, and PRISMA-AI. From 4812 records, 42 studies were included, yielding a pooled AUC of 0.88 (95% CI: 0.86–0.91) and OR of 2.4 (95% CI: 1.2–3.5), demonstrating encouraging but early-stage exploratory evidence of predictive performance. Cancer-specific signatures emerged in colorectal, breast, pancreatic, liver, and hematologic malignancies. A conceptual translational framework was proposed, integrating nutrition, omics, AI/ML, and oncology to illustrate a potential implementation pathway for developing countries like Saudi Arabia. These findings represent preliminary, hypothesis-generating evidence; the proposed framework requires prospective validation before clinical deployment, particularly in resource-limited settings.

Abstract: Renal cell carcinoma (RCC) remains a biologically heterogeneous disease with variable clinical outcomes, underscoring the need for robust biomarkers to guide risk stratification and therapeutic decision-making. Despite advances in immune checkpoint inhibitors and targeted therapies, clinically validated biomarkers are lacking, particularly in the perioperative setting. Kidney injury molecule-1 (KIM-1; also known as HAVCR1 or TIM-1) has emerged as a promising candidate with strong biological and clinical rationale. KIM-1 is a transmembrane glycoprotein minimally expressed in normal kidney tissue but markedly upregulated in injured and dedifferentiated proximal tubular epithelial cells, the cell of origin for clear cell RCC. Its extracellular domain is shed into circulation and urine, enabling non-invasive quantification. Beyond its role as a marker of renal injury, KIM-1 is implicated in immune modulation, chronic inflammation, and tumor biology, supporting its role as a dynamic indicator of tumor burden and disease aggressiveness. This review presents the current evidence supporting KIM-1 as a circulating biomarker and therapeutic target in RCC and discusses emerging strategies to address disease heterogeneity through biomarker-driven approaches. We examine its biological role, clinical utility in early detection and postoperative risk stratification, integration with other emerging biomarkers, and its development as a target for antibody–drug conjugates. The review concludes with a summary of the evolving landscape of KIM-1–directed biomarker strategies in RCC, which hold promise to refine patient selection, improve risk-adapted management, and advance precision oncology in this complex disease.

Abstract: Melanoma treatment has been transformed by immune checkpoint blockade, yet many patients still experience primary resistance, limited durability of response, or acquired resistance. These limitations underscore the need for additional targets that reflect melanoma biology while enabling new therapeutic strategies. The B7-H6/NKp30 axis has gained attention as a link between tumor cell stress, immune recognition, and therapy-related adaptation. B7-H6 (NCR3LG1), an inducible ligand for NKp30, has been detected in melanoma cell lines and tumor specimens, and soluble B7-H6 has been identified in a subset of patients. Membrane-bound B7-H6 may support NK-cell activation, whereas ligand shedding and accumulation of soluble B7-H6 may reduce effective antitumor recognition and promote immune evasion. Emerging evidence further suggests that B7-H6 expression may be linked to tumor-intrinsic programs relevant to melanoma cell survival, migration, and adaptation to therapeutic stress. In this review, we examine the role of the B7-H6/NKp30 axis in immune surveillance, tumor escape, biomarker development, and therapeutic targeting, and discuss its translational potential in melanoma.

Abstract: Background/Objectives: The EGFR T790M gatekeeper mutation drives resistance in non-small cell lung cancer by sterically hindering inhibitors and restoring ATP affinity. As emerging C797S mutations render covalent inhibitors obsolete, novel non-covalent strategies are critical. This study identifies inhibitors that redefine the mutant methionine sulfur atom as a primary stabilizing anchor rather than a liability. Methods: A reinforcement learning-based generative AI framework (DrugEx) sampled 100,000 molecules, prioritized through QSAR-based classification (mean ROC-AUC: 0.91 ± 0.01) and physicochemical filtering. Four lead candidates underwent 200 ns all-atom molecular dynamics (MD) simulations. Mechanistic stability and energetic convergence were quantified via Free Energy Landscape (FEL) analysis, post-simulation Ramachandran validation, and ensemble-averaged Molecular Mechanics Generalized Born Surface Area (MM-GBSA) binding free energy calculations. Results: Candidate 106 demonstrated high mutation tolerance by redistributing interactions toward the Met790 sulfur atom, contrasting with reference scaffolds 14 and 88 which suffered significant affinity loss. MD analysis revealed that potency is fundamentally dictated by successful recruitment of the thioether environment, locking the complex within a narrow, well-defined thermodynamic basin. Candidate 106 maintained stable binding (−11.0 kcal/mol by docking) corroborated by a robust MM-GBSA ΔGbind of −50.51 kcal/mol, primarily driven by persistent π-sulfur contacts (85% occupancy). Ramachandran analysis confirmed over 90% favored region occupancy, ensuring interactions occur without non-physical protein strain. Conclusions: Successful T790M/C797S resistance bypass is achievable by exploiting the gatekeeper methionine’s electronic environment. This mutation-aware blueprint provides a non-covalent strategy that avoids the metabolic and resistance-prone liabilities of covalent warheads.

Abstract: Background: Intertumoral differences in glioblastoma (GBM) malignancy pose significant challenges for clinical management. Distinct microvascular growth patterns contribute substantially to tumor heterogeneity. Ultrasound localization microscopy (ULM) enables microscale mapping of microvascular network remodeling by tracking individual microbubble trajectories in vivo. This study evaluated whether ULM-derived microvascular heterogeneity metrics can facilitate histopathology-based stratification of GBM malignancy. Methods: An orthotopic glioblastoma model was established in 113 Sprague–Dawley rats, and ULM-derived heterogeneity parameters were extracted from tumor regions of interest. Spearman’s rank correlation coefficients were used to assess associations between microvascular heterogeneity metrics and histopathological indices. The Kruskal–Wallis H and Mann–Whitney U tests were used to compare metrics across different levels of microvascular maturity and cell proliferation. A decision tree–based diagnostic model was developed using ULM-derived microvascular features. Results: Microvascular heterogeneity was significantly negatively correlated with the vascular maturity index (p < 0.001) and positively correlated with the cell proliferation index (p < 0.001), supporting the biological and pathological relevance of ULM-derived heterogeneity metrics. Compared with transitional microvessels, mature microvessels exhibited significantly lower tortuosity (p = 0.002). Orientation variance, fractal dimension, connectivity, local thickness, and the spatial distribution index also tended to decrease but did not reach statistical significance (p = 0.074–0.529). In contrast, all corresponding metrics were significantly higher in immature microvessels (p ≤ 0.007). Compared with the low-proliferation group, all heterogeneity-related parameters were significantly higher in the high-proliferation group (p < 0.001). The decision-tree model based on microvascular heterogeneity demonstrated high performance at the sample level in predicting microvascular maturity and cell proliferation status, achieving accuracies of 90.29% (p = 0.029) and 92.23% (p = 0.026), respectively. Conclusions: We developed a clinically implementable decision-tree diagnostic model to support GBM malignancy stratification. As super-resolution ultrasound advances toward clinical translation, our findings may help inform future clinical decision-making.

Abstract: Brain metastases remain a major cause of morbidity and mortality in patients with cancer, particularly melanoma and non-small cell lung cancer. Stereotactic radiosurgery (SRS) is a cornerstone of management for limited intracranial disease, offering high local control while minimizing the neurocognitive toxicity associated with whole-brain radiotherapy. Immune checkpoint inhibitors (ICIs) have also transformed systemic therapy for tumors with central nervous system involvement, creating increasing clinical need to define how best to integrate these modalities.The combined use of SRS and ICIs has raised an important question regarding optimal treatment timing. Retrospective evidence suggests that concurrent or near-concurrent administration, commonly defined as treatment within approximately 2–4 weeks, may improve local control and intracranial response. Several studies also suggest a potential survival advantage compared with sequential treatment, although these findings are limited by selection bias and require prospective validation. Most contemporary analyses do not show a significant increase in radionecrosis (RN) with concurrent single-agent ICI; however, emerging data suggest that dual checkpoint blockade may increase the risk of symptomatic RN.This narrative review synthesizes the biologic rationale, clinical evidence, and toxicity considerations for combining SRS and ICIs in patients with brain metastases. We emphasize differences between single agent and dual ICI strategies, highlight dosimetric predictors of RN such as V12 Gy, and propose a practical framework for treatment integration. Overall, concurrent SRS with single-agent ICI appears feasible and is associated with favorable intracranial outcomes in selected patients, whereas dual ICI warrants more cautious, individualized decision-making. Prospective studies are needed to define optimal sequencing, patient selection, and toxicity mitigation strategies.

Abstract: Background: Acute lymphoblastic leukemia (ALL) is the most common malignancy in the pediatric population. Despite significant therapeutic advances over the past four decades, a critical survival gap persists between North America and Latin America, indicating that non-biological determinants exert a decisive influence on clinical outcomes across the continent. Objective: To synthesize the evidence on the impact of socioeconomic determinants and access barriers on clinical outcomes in pediatric ALL across the Americas, published between 2015 and 2025. Methods: An integrative review was conducted following the five-stage Whittemore and Knafl framework and PRISMA 2020 reporting guidelines. The databases PubMed/MEDLINE, Scopus, SciELO, and LILACS were searched using structured Boolean queries combining MeSH and DeCS controlled vocabulary. Studies examining pediatric ALL (ages 0-18 years) in any country of the Americas, published between January 2015 and December 2025, were included. Results: A five-year overall survival (OS) gap of up to 40 percentage points was identified between high-income countries and Latin American settings (>90% vs. 50-75%). Low household income and lack of private insurance independently predicted inferior event-free survival (EFS), with differences of up to 30%. Treatment abandonment reached 30-40% in rural Latin American areas, driven by geographic distance exceeding 70-150 km to specialized centers and financial toxicity. Treatment-related mortality (TRM) of 15-25% was primarily attributable to sepsis and the absence of pediatric early warning systems (PEWS). A high prevalence of high-risk genomic alterations, including IGH::CRLF2 rearrangements and IKZF1 deletions linked to Amerindian and Hispanic ancestry, was documented. Conclusions: Survival outcomes in pediatric ALL across the Americas are primarily determined by biosocial factors. Closing the survival gap requires strengthening supportive care infrastructure, implementing ancestry-adapted precision medicine, and dismantling the socioeconomic barriers that limit equitable access to curative therapy.

Abstract: Mediastinal staging remains a critical step in the management of non-small-cell lung cancer (NSCLC), as it directly impacts treatment planning, surgical decision-making, and overall prognosis. For many years, mediastinoscopy was considered the standard approach; however, in routine practice, less invasive techniques have largely taken its place. Endobronchial Ultrasound (EBUS) and Endoscopic Ultrasound (EUS) have become widely adopted because they allow real-time sampling of lymph nodes with good accuracy and a low complication rate. In clinical seĴings, these techniques are often used together rather than separately, as each provides access to different nodal stations. This combined approach improves diagnostic yield and reduces the number of patients who require surgical staging. At the same time, recent updates in the IASLC TNM classification, including the proposed 9th edition, have introduced more detailed nodal categories, making accurate tissue confirmation even more important in daily practice. In this review, we summarize the current use of combined EBUS and EUS in mediastinal staging, focusing on their practical advantages, limitations, and roles across different clinical scenarios. We also discuss their relevance in the context of molecular testing and evolving treatment strategies. Despite their strengths, there are situations in which negative results should be interpreted with caution and confirmed surgically. Overall, these techniques have reshaped the approach to mediastinal staging and are now central to modern lung cancer care.

Abstract: Introduction: Conventional oncological pathology practice faces critical challenges stemming from interobserver variability and an ever-growing clinical workload. This review evaluates the technological maturity and clinical utility of artificial intelligence (AI) as a diagnostic and predictive support tool in histopathology. Methods: An integrative review of the literature (2019–2026) was conducted in PubMed, Scopus, and IEEE Xplore, following the methodology of Whittemore and Knafl. Studies on the diagnostic accuracy of deep learning algorithms in neoplasia histopathology were selected, with methodological quality assessed using QUADAS-2. Results: The evidence confirms that convolutional neural networks (CNNs) achieve diagnostic accuracy comparable to or exceeding that of pathologists in binary classification tasks, consistently reporting areas under the curve (AUC) > 0.98 in lung, breast, and prostate cancer. A disruptive finding is the validation of predictive computational histology, capable of inferring genotypic alterations—such as EGFR mutations or microsatellite instability—directly from standard hematoxylin and eosin (H&E) images, offering a cost-effective alternative for molecular screening. The evidence strongly supports the “augmented intelligence” model, in which the pathologist–AI synergy surpasses individual performance and mitigates visual fatigue. Conclusions: AI has transcended the experimental phase to become a robust technology for triage and digital phenotyping. Its definitive clinical adoption requires prioritizing multicenter external validation and the development of explainable AI (XAI) interfaces to overcome the “black box” barrier.

Abstract: Pancreatic cancer remains one of the most lethal malignancies worldwide, with pancreatic ductal adenocarcinoma accounting for the vast majority of cases and characterized by extensive desmoplasia, immune exclusion, and resistance to systemic therapies. Increasing evidence implicates lysosomal cathepsins as important regulators of these defining features of pancreatic tumor biology. Cathepsin-dependent proteolysis and lysosome-associated signaling pathways contribute to extracellular matrix remodeling, regulate immune cell trafficking, and influence antigen processing and presentation. Beyond their classical degradative functions, cathepsins participate in stress-adaptive cellular programs linked to autophagy, metabolic regulation, and proteostasis, supporting tumor cell survival under hypoxic, nutrient-limited, and therapy-induced stress conditions. Within the tumor microenvironment, dysregulated cathepsin activity promotes immune evasion by reshaping cytokine networks, impairing effective antigen presentation, and reinforcing physical and functional barriers to cytotoxic T-cell infiltration. Collectively, these mechanisms position the lysosome–cathepsin system as a central regulator of proteolytic remodeling, immune exclusion, and adaptive therapy resistance in pancreatic cancer, highlighting its potential relevance for emerging combinatorial therapeutic strategies.