Search Results (1,621)

Background: Mast cells are integral components of the tumor microenvironment and have been implicated in the regulation of tumor progression in various malignancies. The association between inflammation and colorectal cancer (CRC) development has become increasingly recognized. Depending on the tumor microenvironment, mast cells may exert either pro-tumorigenic or antitumorigenic functions. Objective: This study aimed to evaluate the relationship between stromal mast cell density and prognostic factors in patients with CRC. Methods: In this retrospective cohort study, 81 patients who underwent curative surgical resection for CRC were analyzed. Immunohistochemical staining for mast cell tryptase (MCT) was performed on paraffin-embedded tumor specimens. Mast cells were quantified in regions of hot spots within the tumor stroma. Patients were categorized as high mast cell density (MCC-H, ≥22 cells/HPF) or low mast cell density (MCC-L, <22 cells/HPF). Associations with clinicopathological parameters were assessed using chi-square or Fisher’s exact tests. Progression-free survival (PFS) and overall survival (OS) were analyzed using Kaplan–Meier estimates and log-rank tests. Independent prognostic factors were identified using multivariate Cox proportional hazards regression, with hazard ratios (HRs) and 95% confidence intervals (CIs) reported. Results: ROC analysis identified an MCC cut-off of 22 cells/HPF (AUC = 0.61; sensitivity = 0.67, specificity = 0.52) for mortality prediction. Multivariate analysis revealed lymph node involvement (HR: 1.41, 95% CI: 1.03–1.94, p = 0.033) and macroscopic tumor perforation (HR: 0.15, 95% CI: 0.04–0.55, p = 0.004) as independent predictors of PFS. High MCC (≥22) independently predicted improved OS (HR: 0.07, 95% CI: 0.006–0.87, p = 0.039). A significant association was observed between OS, MCC, and lymph node stage. Conclusions: Stromal mast cell count is an independent prognostic factor for overall survival in patients with CRC. Our findings suggest that MCC may serve as a reliable prognostic biomarker following surgical resection and could aid in postoperative risk stratification. Full article

Background: Folate receptor alpha (FRα) is a glycosylphosphatidylinositol-anchored membrane protein encoded by the FOLR1 gene. Its overexpression in various cancers, including ovarian carcinoma, makes it a promising target for antibody-drug conjugates (ADC). Mirvetuximab soravtansine-gynx, an FRα-targeting ADC, has been approved by the FDA and EMA for the treatment of FRα-positive, platinum-resistant ovarian cancer. In the United States, patient selection is tied to the VENTANA FOLR1 (FOLR1-2.1) RxDx Assay, an immunohistochemical (IHC) test that identifies tumors with ≥75% moderate-to-strong membrane staining. However, in the European Union, no specific IHC test is mandated, and alternative antibodies are frequently used in routine pathology, necessitating validation of their diagnostic performance. Methods and Results: We report the results of the first interlaboratory proficiency trial on FRα testing conducted by the German Quality Assurance Initiative in Pathology (QuIP^®). Sixty-eight pathology institutes participated across internal and open trials using a variety of antibodies and staining platforms. The VENTANA FOLR1 RxDx Assay demonstrated the highest reliability, with 83% of participating laboratories achieving a successful result. In contrast, alternative clones such as BN3.2 (Leica/Novocastra) and EPR20277 (Abcam) showed substantially weaker staining intensity, lower concordance with reference values, and success rates of only 22–25%, while other antibodies failed entirely. Problem analysis revealed that failures with the VENTANA FOLR1 (FOLR1-2.1) RxDx Assay were mainly due to interpretative challenges, whereas weak staining was the predominant issue with alternative clones. Participation in a preparatory online seminar improved pass rates, underscoring the importance of training. Conclusions: These findings highlight the critical importance of standardized, validated assays for FRα detection to ensure accurate patient selection for targeted therapies. The study emphasizes the need for further optimization of alternative antibodies before clinical implementation. Full article

Spontaneous coronary artery dissection (SCAD) is a non-atherosclerotic cause of acute coronary syndrome, characterized by the development of a false lumen within the coronary arterial wall, leading to narrowing or complete occlusion of the true lumen. This underrecognized condition accounts for a substantial proportion of sudden cardiac death (SCD), particularly among young, otherwise healthy women. Macroscopically, SCAD is defined by intramural hematoma and focal thickening of the arterial wall, while histological examination demonstrates separation of the tunica media, elastic fiber degeneration, and variable inflammatory infiltrates. Proposed pathogenic mechanisms include primary intimal tear and primary intramural hematoma, frequently associated with predisposing conditions such as fibromuscular dysplasia, connective tissue disorders, and specific hormonal states. In cases of myocardial infarction, the myocardium exhibits acute ischemic necrosis and early hypoperfusion injury. Postmortem diagnosis requires meticulous coronary dissection, adjunctive histochemical and immunohistochemical staining, and, when indicated, molecular autopsy (MA). The purpose of this review is to provide an updated synthesis of current knowledge on SCAD as a cause of SCD, integrating pathogenetic, morphological, and genetic perspectives, and to emphasize the role of MA as both a diagnostic and preventive tool. Full article

Background/Objectives: Despite the introduction of targeted therapies such as Nivolumab, survival outcomes for patients with esophageal adenocarcinoma remain poor. During tumorigenesis, some tumors develop auxotrophy by downregulation of Argininosuccinate Synthetase-1 (ASS1), making them reliant on external arginine supply and thus potentially susceptible to arginine deprivation therapy. Arginine deprivation therapy with agents such as pegargiminase has shown improved survival in patients with pleural mesothelioma exhibiting ASS1 loss in tumor cells. Therefore, we investigated the prevalence of ASS1 loss in esophageal adenocarcinoma. Methods: First, we compared the staining patterns of three antibodies for ASS1 with RNA in situ Scope analysis results to identify the most reliable antibody for ASS1 immunohistochemical staining in esophageal adenocarcinoma. Subsequently, we performed ASS1 immunohistochemical staining on samples from 97 patients who underwent curative resection. The staining results were classified into three categories based on expression levels: negative, low-positive, and positive. Results: Among all included patients, 6.2% exhibited an ASS1 loss, and 6.2% showed low ASS1 expression. Notably, patients with an ASS1 loss did not demonstrate a response to neoadjuvant therapy. Patients with ASS1 loss or low expression were significantly younger. Conclusions: Our findings indicate that approximately 12.4% of patients with esophageal adenocarcinoma may be eligible and could potentially benefit from arginine deprivation therapy. This underscores the urgent need for clinical trials evaluating the efficacy of pegargiminase in this patient population. Additionally, incorporating ASS1 immunohistochemical staining into pre-neoadjuvant biopsy assessments should be considered to optimize neoadjuvant treatment strategies and advance the implementation of personalized cancer therapy. Full article

Background: Epicardial adipose tissue (EAT) is a visceral fat depot surrounding the myocardium. It contributes to coronary artery disease (CAD) through local inflammation, while its metabolic activity, including the expression of uncoupling protein-1 (UCP-1) and incretin receptors (GLP-1R, GIPR), may exert protective effects. The relationship between EAT immunohistochemical features and imaging-derived volume remains unclear. Methods: We prospectively studied 50 patients undergoing cardiac surgery: 25 with CAD undergoing coronary artery bypass grafting and 25 without CAD undergoing valve replacement. EAT samples were immunohistochemically stained for CD3, CD68, MPO, UCP-1, GLP-1R, and GIPR. Preoperative CT was used to quantify EAT volume. Results: Patients with CAD more frequently had higher CD3 immunopositivity compared to the control group (84.0 vs. 58.3%, p = 0.047), with no difference in MPO and CD68 immunoexpression. UCP-1 expression was elevated in CAD patients (p = 0.004), whereas GLP-1R and GIPR immunopositivity were similar. EAT volume did not differ between CAD and non-CAD patients (102.87 cm³ vs. 99.38 cm³, p = 0.964) but correlated modestly with BMI (r_s = 0.325, p = 0.021). UCP-1 and GLP-1R immunopositivity, as well as larger LVEDD (left ventricular end-diastolic diameter), were positively associated with greater EAT volume. Conclusions: EAT in CAD exhibits increased T-cell infiltration and elevated UCP-1 expression, indicating an inflammatory yet metabolically active profile. Larger EAT volume was associated with UCP-1 and GLP-1R expression, underscoring the immunometabolic role of EAT in CAD. Full article

Liposarcomas, the most common subtype of soft tissue sarcomas, show variable biological behavior and therapeutic response. Programmed death-ligand 1 (PD-L1) and cytotoxic T lymphocyte marker CD8 have been implicated in tumor immune evasion and prognosis in various malignancies, while Maspin, a tumor suppressor, has shown a negative prognostic impact in sarcomas. This study aimed to investigate the clinicopathological significance of PD-L1, CD8, and Maspin expression in liposarcomas. A retrospective analysis of 42 liposarcoma cases diagnosed between 2016 and 2023 was conducted. Immunohistochemical staining for PD-L1 (using DAKO 22C3 and 28-8 clones), CD8, and Maspin was performed. PD-L1 expression was assessed using the tumor proportion score (TPS) and tumor cell score (TC). CD8 expression was evaluated using an H-score, and Maspin positivity was assessed based on subcellular localization. Correlations with clinicopathological parameters were statistically analyzed using chi-squared and Fisher’s exact tests. Most liposarcomas exhibited low PD-L1 expression (<10%), but increased PD-L1 levels correlated with poor differentiation (G3), higher CD8 infiltration (H-score > 10%), and cytoplasmic Maspin positivity. Statistically significant associations were found between high PD-L1 expression and high CD8 infiltration (p = 0.007 for 22C3; p = 0.0331 for 28-8) and between PD-L1 positivity and Maspin expression (p = 0.003 for 22C3; p = 0.0113 for 28-8). CD8 infiltration was generally low across cases, and PD-L1 expression in inflammatory cells was noted predominantly in tumors with higher PD-L1 TPS/TC scores. High PD-L1 expression in liposarcomas is associated with poor tumor differentiation, increased CD8 infiltration, and Maspin positivity, suggesting an immune-evasive phenotype. Despite low overall expression rates, PD-L1 could serve as a prognostic biomarker and a potential target for immunotherapeutic strategies in liposarcomas. Further studies are necessary to standardize PD-L1 assessment and explore effective immunotherapy approaches for these tumors. Full article

A large number of regulatory proteins are found in both the cytoplasm and the nucleus. Changes in their nuclear abundance are important for cellular signalling, biological activity, and disease mechanisms. Accurate quantification of nuclear staining is therefore essential in studies of cellular function, therapeutic targeting, drug design, and drug resistance. However, manual scoring is time-consuming, unsuitable for high-throughput applications, and introduces potential bias. As expected, manual scoring by six observers with varying levels of expertise led to highly variable results. Moreover, it was far from achieving good interobserver reliability. To overcome these limitations, LoQANT (Localisation and Quantification of Antigen Nuclear sTaining), an open, freely available ImageJ plugin, was developed for reliable and efficient quantification of nuclear signals. LoQANT is a single cell-based approach to assess the proportion of cells with a positive nuclear signal, independent of cytoplasmic staining, in both immunohistochemically and fluorescently stained samples across various sample types. It also provides semiquantitative and quantitative measurements of nuclear staining intensity. The script, its version for batch analysis, and complete user guide are available at GitHub. Full article

Introduction: Bladder cancer is associated with a high recurrence rate, and outcomes for muscle-invasive and metastatic disease remain poor. New targeted therapies, such as the FGFR inhibitor erdafitinib, have been introduced, but the progression from non-muscle-invasive to muscle-invasive disease remains a major clinical challenge. Methods: In this study, we performed immunohistochemical staining for FGFR1-FGFR4 on surgical specimens from 192 cases of urothelial carcinoma. We also conducted various functional assays on human bladder cancer cell lines to assess protein/gene expression, cell proliferation, migration, invasion, and colony formation. Results: FGFR2 and FGFR3 expressions were found to be significantly down-regulated in high-grade (0.014) and muscle-invasive (0.002) tumors, respectively. Functionally, the FGFR inhibitor erdafitinib suppressed cell proliferation and migration, and FGFR3 silencing also markedly reduced proliferation, migration, invasion, and colony formation in cancer cell lines. Conclusions: The down-regulation of FGFR3 in muscle-invasive bladder cancer, coupled with the inhibitory effect of its inactivation on cell growth, suggests a significant role for FGFR3 in bladder cancer progression. Full article

Background and Clinical Significance: Primary Cutaneous Cribriform Apocrine Carcinoma (PCCAC) is a rare, inert low-grade cutaneous malignancy that is diagnosed on histopathologic assessment. PCCAC usually presents in middle-aged adults as a solitary, subcutaneous nodule on the extremities. Characterized by anastomosing tubules and solid/cribriform nests of atypical epithelial cells generating a sieve-like display, the tumor is a histopathological variant of apocrine metaplasia of the skin. PCCAC also follows characteristic staining patterns. It is important to distinguish PCCAC from other similar histological variants, which may hold more grievous indications. Case Presentation: A 47-year-old female presented with an enlarging, itchy growth of several months on her back. On physical exam, an indurated pink, nontender papule of 8 mm on the left lateral side wall was noted. Histopathology demonstrated a well-circumscribed, pandermal tumor composed of anastomosing solid and cribriform nests, tubules, and cords of mildly atypical, eosinophilic epithelial cells forming a glandular lumina. An immunohistochemical study revealed the tumoral epithelium to express CK7, CK5/6, BER-EP4, CD117 (C-kit), and S100. Positive EMA and CEA staining highlighted intratumoral glandular ductal differentiation and apocrine secretion. Immunohistochemical stains for CK20, GATA-3, and p63 were negative. Conclusions: We present this case to distinguish the histological attributes of PCCAC and help differentiate it from more concerning visceral metastatic malignancies. We follow with a narrative review of the histopathologic differential for PCCAC and feature reconciliation of corresponding staining patterns reported in the literature. Full article

Background/Objectives: TP53 mutations in advanced epidermal growth factor receptor (EGFR)-mutated non-small cell lung cancer could worsen prognosis. Therefore, we aimed to investigate the clinical significance of TP53 mutations and p53 expression in these patients. Methods: Patients with advanced/metastatic EGFR-mutated lung adenocarcinoma treated with first-line tyrosine kinase inhibitors were retrospectively enrolled. Sanger sequencing was performed to detect TP53 mutations and immunohistochemical staining was used to verify p53 protein expression levels. Kaplan-Meier and Cox proportional hazards analyses were used to estimate survival and hazard ratio (HR) with 95% confidence interval (CI). Results: The study involved 83 patients with adequate tumor samples for TP53/p53 analysis. Patients with tumor p53 immunostaining ≥50% showed significantly better overall survival (OS) (HR: 0.49 [95% CI: 0.30–0.81], p < 0.001), but TP53 mutations were not associated with inferior progression-free survival (PFS) or OS (missense vs. wild-type [PFS, HR: 0.68 (95% CI: 0.40–1.15), p = 0.151; OS, HR: 0.88 (95% CI: 0.56–1.42), p = 0.599]). Areas under the receiver operating characteristic curves of TP53 mutations with different cut-off values for p53 positivity were 0.51–0.56. The Kaplan-Meier survival analysis revealed significant survival benefits in patients with EGFR L858R substitution and tumor p53 immunostaining ≥50% (median PFS: 8.0 vs. 5.3; median OS: 20.4 vs. 15.3 months; log-rank p = 0.025 and 0.049, respectively). Conclusions: Tumor p53 immunostaining (≥50%) was associated with better OS, especially in patients with TP53 mutations or L858R. Prospective clinical trials are required to explore the prognostic significance of p53 expression in the genomic era of TP53 mutations. Full article

Background: The aim of this study was to determine Ezrin and MMP-2 immunohistochemical expressions in the gingival tissue of patients with or without diabetes and to determine the role of the molecular pattern involvement in the evolution of periodontal disease. Material and Methods: In this histological study, we investigated 53 subjects with periodontal disease (test group—27 patients with type 2 DM; control—26 patients without diabetes). Samples from both groups were subjected to the immunohistochemistry (IHC) technique to evaluate the immunoreactivity (IR) intensity of Ezrin and MMP-2. Results: Among diabetic patients with periodontitis, 55.4% of patients exhibited intensely positive expression (+++) of Ezrin, and 44.6% of patients showed moderate expression (++) of Ezrin. All patients with diabetes and periodontitis showed intensely positive expression for MMP-2. In contrast, the control group showed negative expressions of Ezrin and MMP-2 (-) in 100% of cases. Significant statistical differences were found between Ezrin and MMP-2 expression in gingival samples of diabetic patients and non-diabetic patients with periodontal disease (p < 0.05). Conclusions: Ezrin and MMP-2 are significantly overexpressed in patients with diabetes and stage 2–3 periodontitis compared with non-diabetic patients with periodontal disease. Ezrin showed an exclusive pattern of moderate to strong positive staining in the diabetes–periodontitis group and complete absence in controls. MMP-2 displayed a broader range of staining intensities, with a predominance of strong positivity in all locations. Ezrin may represent a more consistent discriminative marker, whereas MMP-2 reflects a wider spectrum of tissue activation related to inflammation and tissue remodeling. Full article

Endometriosis is a common gynecological condition that affects fertility in many women of reproductive age worldwide. This multifaceted disease exhibits a pathogenesis characterized by hormonal and immune system dysregulations, alongside increased angiogenic activity within the peritoneum. The aberrant proliferation of endometrial tissue outside the uterus is associated with vascularization in ectopic endometriotic lesions. Consequently, RNA interference (RNAi)-based angiogenic therapies targeting the VEGFA gene present a promising strategy for the treatment of endometriosis. To ensure the efficacy of RNAi-based therapy, it is critical to develop carriers capable of precisely delivering small interfering RNA (siRNA) to target cells. Additionally, the instability of polyplexes in vivo must be regarded as a pivotal aspect influencing the success of non-viral delivery. In this study, we introduce ternary polyplexes comprising siRNA and a carrier derived from an arginine–histidine-rich peptide, which is further coated with a glutamate–histidine-rich polymer modified using an SDF-1 chemokine-derived ligand for targeting CXCR4-expressing cells. The physicochemical characteristics of the siRNA-polyplexes, along with cellular toxicity and GFP gene silencing efficacy, were assessed in vitro. The anti-angiogenic potential of anti-VEGFA siRNA-polyplexes was evaluated by measuring the size of endometrial lesions, conducting immunohistochemical staining, and analyzing VEGFA gene expression. For in vivo experiment, a rat model of endometriosis induced by subcutaneous auto-transplantation of uterine tissue was utilized. A significant reduction in the growth of endometriotic implants and silencing of VEGFA gene expression was observed when compared to the saline-treated control group. The results of this study strongly suggest that the developed ternary polyplexes have significant potential as an efficient tool for the development of anti-angiogenic RNAi-based therapies for endometriosis. Full article

Background and Objectives: This pilot study examines the expression of MDM2 and NF-κB proteins in CD138-positive plasma cells in patients with multiple myeloma treated with bortezomib-based therapy, exploring their possible interplay and correlations with clinical and selected prognostic factors. Materials and Methods: We analyzed MDM2 and NF-κB protein expression via double immunohistochemical staining of bone marrow trephine biopsies obtained at diagnosis and after bortezomib-based therapy. Statistical analyses were performed to assess the significance of changes in protein expression before and after therapy and their association with clinical and prognostic parameters. Results: Key findings revealed a positive correlation between MDM2 and NF-κB, as well as a significant decrease in their expression following therapy. Decreased NF-κB expression seems to be an independent prognostic factor for improved renal function. Conclusions: The results demonstrated for the first time the in vivo protein expression of MDM2 in the bone marrow of patients with multiple myeloma, as well as the possible effect of bortezomib on the expression of this protein in the microenvironment of multiple myeloma. Full article

Background/Objectives: The use of dentin grafts in bone regeneration has gained increasing attention as an alternative to conventional grafting materials. Mesenchymal stem cells (MSCs), known for their osteogenic potential, have been combined with various biomaterials to enhance regenerative outcomes. This study aimed to evaluate the regenerative potential of dentin grafts and bovine-derived xenografts, with or without MSCs, in experimentally created bone defects in a rat model. Methods: A total of 25 male rats were randomly assigned to five groups: control, dentin graft, dentin graft and MSC, xenograft, and xenograft and MSC. Standardized 2-mm cortical defects were created bilaterally in the femoral shafts. Histological and immunohistochemical analyses were performed after a 90-day healing period. Statistical evaluation was carried out using the Kruskal–Wallis H test and Bonferroni-adjusted pairwise comparisons. Results: Complete healing was achieved in all groups without evidence of complications or inflammatory reactions. Immunohistochemical staining demonstrated no positive vascular endothelial growth factor (VEGF), collagen type I (COL1), or osteopontin (OPN) reactions in defect areas, consistent with complete maturation, although collagen type 3 (COL3) positivity was observed in residual xenograft material. Quantitative analysis showed that the dentin graft and MSC group achieved the highest degree of new bone formation (M = 92.88%, SD = 6.09), significantly greater than the control (p = 0.002) and xenograft groups (p = 0.013). Conclusions: Both dentin grafts and xenografts demonstrated enhanced bone defect healing when combined with MSCs. Nevertheless, dentin grafts in conjunction with MSCs yielded the most favorable regenerative outcomes, suggesting their clinical superiority over conventional xenografts. Full article

Background/Objectives: Proliferative verrucous leukoplakia (PVL) is an oral potentially malignant disorder characterized by a high risk for cancer development. Current evidence suggests that the evolution and malignant transformation of PVL is driven by a reciprocal crosstalk between the epithelial cells and the subepithelial immune microenvironment. The aim of the present study was to compare for the first time the immunohistochemical expression of the immune response-related proteins GATA-binding protein 3 (GATA3), c-KIT/cluster of differentiation (CD)117, CD56 and CD45 between PVL, PVL-associated oral squamous cell carcinoma (OSCC) and solitary (localized) oral leukoplakia (OL) cases. Methods: Thirty-six formalin-fixed and paraffin-embedded tissue specimens were used; sixteen from 8 patients with PVL, ten from 10 patients with PVL-OSCC and ten from 10 patients with OL. Immunohistochemistry was conducted using monoclonal primary antibodies against GATA3, c-KIT/CD117, CD56 and CD45. A semi-quantitative method was applied to score staining, and statistical analysis included Wilcoxon signed-rank test, Kruskal–Wallis test with Dunn’s post hoc test and Spearman’s correlation coefficient test. Results: A significantly decreased GATA3 expression was found in PVL-OSCC cases compared with PVL and OL cases. c-KIT/CD117 and CD56 proteins were consistently expressed in all study groups, while a significantly higher CD45 expression was noted in PVL than OL. No significant correlation between markers was found. Conclusions: These data collectively underscore an activated yet disturbed immune response that might be involved in the development and progression of malignancy in PVL that may also be considered as unique and interesting in vivo model of oral carcinogenesis. Full article