Search Results (12,552)

Background/Objectives: Systemic lupus erythematosus (SLE) is characterized by diverse clinical presentations and complex immunological mechanisms. This study aimed to characterize patient serology associated with disease activity scored using the systemic lupus erythematosus disease activity index (SLEDAI) and investigate the molecular signature of complement activation (measured through C3dg, a complement breakdown product) in SLE patients utilizing high-throughput mass spectrometry and autoantibody profiling. Methods: Plasma samples from 39 SLE patients in four mutually exclusive groups based on either disease activity scores (high/low SLEDAI) or complement activation levels (high/low C3dg) were analyzed using rapid LC-MS/MS, followed by unsupervised and supervised protein expression analysis. Complement activation was evaluated by measuring C3dg levels, and disease activity was scored using SLEDAI. Autoantibody reactivities were profiled using global autoantibody protein microarrays. Data are available via ProteomeXchange with identifier PXD066214. Results: Differential proteomic analyses revealed 25 proteins associated with SLE disease activity (high vs. low SLEDAI scores) and 25 proteins linked to complement activation levels (high vs. low C3dg). Enriched pathways indicated that adaptive immune response, classical complement activation, and immunoglobulin production correlated with disease activity, while complement activation and coagulation cascades were primarily associated with complement activation levels. Autoantibody profiling highlighted distinct reactivity patterns between subgroups, suggesting varying degrees of immune-mediated tissue damage. Conclusions: In this study, disease activity and complement activation markers were associated with overlapping yet non-identical plasma proteomic patterns in SLE. These findings support the feasibility of rapid mass spectrometry-based proteomics and autoantibody profiling for generating candidate molecular signatures in SLE. These findings serve as exploratory signatures that require validation in larger independent cohorts before they can be considered for clinical stratification and decision-making. Full article

Background/Objectives: Postoperative monitoring of microvascular free flaps is critical for early detection of vascular complications and flap survival. Nurses play a central role in this process; however, qualitative evidence on their experiences and challenges remains limited. This study explored nurses’ experiences in free tissue flap care to identify clinical practices, challenges, and improvement needs. Methods: A phenomenological qualitative design was used. Data were collected through semi-structured interviews with nine nurses experienced in free tissue flap care, recruited via purposive and snowball sampling. Interviews were conducted online and lasted 30–45 min. Data were analyzed using content analysis with MAXQDA 2025. Inter-researcher reliability was 97%. Results: The findings were categorized into four main themes and seventeen subthemes: (1) clinical monitoring and evaluation in the care process, (2) challenges and difficulties, (3) emotional and professional reflections, and (4) suggestions for improving care. Nurses reported that flap care requires intensive monitoring, rapid decision-making, and close collaboration with physicians, especially within the first 24–48 h. Monitoring was largely based on observation and experience due to the lack of standardized protocols. Major challenges included high workload, frequent assessments, and donor site management. Emotional burden, stress, and responsibility were also prominent. Conclusions: Free flap care is a complex and demanding process for nurses. The lack of standardized monitoring tools and protocols is a key gap. Developing structured tools, improving training, and strengthening multidisciplinary collaboration may enhance patient safety and care quality. Full article

Pituitary apoplexy is an uncommon but clinically urgent complication that often involves intrasellar hemorrhage and tissue necrosis. The mechanisms linking acute tissue injury to the inflammatory tumor microenvironment remain incompletely defined. Here, we characterized the apoplexy-associated microenvironment and examined whether macrophage mechanosensitive signaling contributes to inflammatory amplification and tissue damage in pituitary neuroendocrine tumors (PitNETs). We combined single-cell RNA sequencing (scRNA-seq), histological validation, clinical stratification, and in vitro functional assays using apoplectic and non-apoplectic human PitNET specimens. Macrophage state transitions, intercellular communication, and transcriptional regulatory programs were analyzed, followed by an experimental assessment of the PIEZO1–Ca²⁺ axis and macrophage-conditioned medium-induced tumor cell death. Histological validation confirmed macrophage accumulation in apoplectic PitNETs, including a 1.67-fold increase in IBA-1-positive cells (p < 0.001). CellChat-inferred interaction metrics increased descriptively in apoplectic samples. Apoplectic tissues showed higher TNF-α expression (3.00-fold; p < 0.0001) and higher PIEZO1 fluorescence in IBA-1-positive regions (1.39-fold; p = 0.001). Yoda1 increased Calcium 520 fluorescence in macrophages (1.72-fold; p = 0.002), whereas Piezo1 knockdown reduced the Yoda1-associated response (p = 0.003). Conditioned medium from activated macrophages increased total Annexin V/PI-positive death in AtT-20 cells (0.53 ± 0.53% to 32.48 ± 1.14%; p < 0.001) and GH3 cells (0.82 ± 0.50% to 30.92 ± 1.11%; p < 0.001); Piezo1 knockdown or TNF-α neutralization attenuated this effect. Clinically, pathological necrosis was associated with higher symptom frequencies and a greater adjusted likelihood of two or more clinical symptoms. Together, these findings indicate that PIEZO1-related macrophage signaling may participate in TNF-α-associated tumor cell necroptosis in pituitary apoplexy. Pathological necrosis was linked to greater acute symptom burden and perioperative hormonal abnormalities, suggesting that it may identify a clinically severe apoplexy subtype. Full article

Arrhythmogenic cardiomyopathy (ACM/ARVC) is an inherited myocardial disease characterized by progressive fibro-fatty replacement, ventricular arrhythmias, and an increased risk of sudden cardiac death. In addition to mutations in desmosomal genes, growing evidence suggests that microRNAs (miRNAs) actively contribute to disease pathogenesis by regulating key processes such as fibrosis, cell adhesion, and cardiac remodeling. This systematic review analyzed the main miRNAs identified in studies of human cardiac tissue and animal models of ARVC. Materials and Methods: Studies based on human myocardial tissue analysis (including autopsy and biopsy samples) and animal models of arrhythmogenic cardiomyopathy were included, using RNA sequencing, small RNA sequencing, miRNA arrays, and RT-qPCR. Studies on circulating miRNAs and narrative reviews were excluded. miRNAs were analyzed in relation to their functional pathways and their role in disease pathogenesis. Results: The synthesis of studies on human and animal cardiac tissue reveals a consistent miRNA signature associated with arrhythmogenic cardiomyopathy. MiR-21-5p and miR-29b-3p are associated with fibrosis and extracellular matrix remodeling, whereas miR-133a-b and miR-130a are linked to cardiomyocyte integrity loss and desmosomal dysfunction. A second group of miRNAs, including miR-217-5p, miR-708-5p, and miR-135b, regulates key pathways such as Wnt/β-catenin and Hippo signaling, contributing to structural remodeling and loss of cellular identity. Furthermore, downregulation of miR-499-5p is associated with mitochondrial dysfunction and cellular vulnerability, while the miR-142-3p, miR-182-5p, and miR-183-5p clusters contribute to differential molecular signatures compared with other cardiomyopathies. Overall, miRNAs converge on three main pathogenic axes: myocardial fibrosis, desmosomal impairment, and remodeling of cellular signaling pathways. Conclusions: The available evidence indicates that arrhythmogenic cardiomyopathy is regulated by a coordinated network of miRNAs that actively drives myocardial damage progression. These miRNAs represent not only biomarkers but also functional mediators of disease, suggesting potential diagnostic and therapeutic applications based on tissue-specific molecular signatures, including in post-mortem settings. Full article

To characterize the distinct expression profiles of microRNAs (miRNAs) in serum and tissue and to delineate the heterogeneity of their regulatory mechanisms in early-stage ovarian cancer (EOC), thereby identifying candidate biomarkers for non-invasive early diagnosis. Differentially expressed miRNAs were identified by integrating publicly available datasets of EOC tissues and serum samples from the Gene Expression Omnibus (GEO) and The Cancer Genome Atlas (TCGA). Core miRNAs were subsequently screened through integrated differential expression analysis, weighted gene co-expression network analysis (WGCNA), and feature importance ranking derived from optimized machine learning models. Protein–protein interaction (PPI) networks and functional enrichment analyses (GO and KEGG) were performed on predicted target genes to systematically compare the functional discrepancies between serum- and tissue-derived miRNAs. No overlapping core miRNAs were observed between the two compartments. Serum miRNAs exhibited an overall up-regulated trend, whereas tissue miRNAs were predominantly down-regulated. Although the regulatory pathways demonstrated significant heterogeneity, they ultimately converged on the cell cycle and the PI3K-Akt signaling pathway, indicating high functional homology. Furthermore, serum miRNAs are not merely passive leakage products from tissues; current evidence suggests they may be selectively packaged into exosomes to participate in tumor regulation. Despite divergent expression profiles, serum and tissue miRNAs share homologous regulatory functions in EOC. These findings suggest that serum miRNAs accurately reflect the core molecular status of tumor tissues, providing a robust molecular foundation for liquid biopsy-based early detection strategies. Full article

Immunohistochemistry (IHC) is a widely used method for detecting specific proteins in tissue samples, helping diagnose diseases such as cancer. Traditional analysis methods rely heavily on human interpretation, which can lead to inconsistencies. In this study, we propose DeepFlare, a weakly supervised deep learning framework for cross-modality translation and segmentation of immunofluorescence and immunohistochemistry images. The proposed method utilizes multiplex immunofluorescence (mpIF) and co-registered IHC images, combined with preprocessing techniques such as affine transformation, stain normalization, noise reduction, and artifact removal. Multiple imaging channels, including hematoxylin, DAPI, Lap2, and nuclear envelope signals, are leveraged to generate segmentation masks using a U-Net++ architecture. The final segmentation mask is obtained through weighted fusion of modality-specific outputs. A generative adversarial network (GAN) is employed to measure translation fidelity between generated and real images. Weakly supervised learning techniques, including image-level supervision and consistency constraints, are applied to enhance performance under limited annotation scenarios. Pretrained pathology foundation encoders such as UNI and Virchow are integrated to extract multi-scale morphological and contextual features. Explainable AI techniques are incorporated to highlight critical regions and refine model attention. Experimental results demonstrate strong performance, achieving an SSIM of 0.7077 for image translation and a Dice score of 0.7424 for segmentation. The integration of the UNI encoder provides marginal improvement over the baseline (0.72 Dice score), indicating limited domain adaptation without fine-tuning on the dataset of 1264 training samples. Full article

In this study, we conducted a molecular investigation of hemotropic Mycoplasma spp. in bat species captured in the northern region of the Pantanal biome, Mato Grosso State, Brazil. Tissue samples were screened by qPCR targeting the 16S rRNA gene. Positive samples were subsequently subjected to conventional PCR assays targeting partial fragments of the 16S rRNA (~900 bp) and 23S rRNA (~800 bp) genes. Hemoplasma DNA was detected in four bat species: Glossophaga soricina, Molossops temminckii, Molossus rufus, and Desmodus rotundus. Phylogenetic analyses based on partial 16S and 23S rRNA gene sequences demonstrated that the detected hemoplasmas clustered predominantly with previously described bat-associated hemoplasmas from Brazil and other countries in the Americas. Notably, the detection in M. temminckii represents, to our knowledge, the first molecular evidence of hemotropic Mycoplasma infection in this bat species. These findings expand current knowledge regarding the occurrence, host range, and genetic diversity of hemotropic Mycoplasma spp. in bats from the Pantanal biome and contribute to wildlife surveillance efforts in this ecologically important region. Full article

Background/Objectives: Bitter taste receptors (T2Rs), specifically T2R38, are present in the respiratory epithelium and react with bacterial quorum-sensing molecules to induce an innate immunity response. Although TAS2R38 polymorphisms have been correlated with susceptibility to chronic rhinosinusitis (CRS), they have not yet been explored in odontogenic rhinosinusitis (ORS), a distinct form of CRS with particular microbial and inflammatory features. We aim to establish a proof-of-concept methodology for investigating TAS2R38 genetic variants in ORS using direct maxillary sinus tissue analysis and demonstrate the feasibility of this translational approach. Methods: We conducted a prospective pilot case–control study of 36 ORS patients and 37 controls undergoing septoplasty without sinonasal disease. Maxillary sinus mucosal biopsies were obtained intraoperatively with informed consent. Genomic DNA was extracted using the PureLink Genomic DNA Mini Kit and quantified via NanoDrop spectrophotometry. TAS2R38 haplotypes were determined and classified as taster (PAV/PAV), non-taster (AVI/AVI), or intermediate (PAV/AVI) phenotype. Results: Among fully classifiable canonical TAS2R38 phenotypes (32 ORS patients, 28 controls), distributions were: tasters 12.5% vs. 25.0%, non-tasters 31.3% vs. 25.0%, and intermediate 56.3% vs. 50.0%. AVI/AVI non-taster status was not significantly associated with ORS susceptibility (OR = 1.36, 95% CI: 0.44–4.25; Fisher’s exact p = 0.775). Conclusions: This proof-of-concept study demonstrates that genotyping-grade genomic DNA can be recovered from acutely inflamed maxillary sinus mucosa, validating this substrate for future tissue-based expression, functional, and microbiome analyses not obtainable from peripheral samples; germline genotyping itself does not require sinus tissue. The observed difference in non-taster prevalence (31.3% vs. 25.0%) did not reach statistical significance and is reported descriptively. This directional trend is hypothesis-generating only and, given the limited statistical power, does not constitute evidence for an association. The demonstrated feasibility, together with the established biological rationale, supports an adequately powered confirmatory study and lays the foundation for future investigation of taste receptor genetics in ORS pathogenesis, and potentially personalized therapeutic strategies. Full article

Introduction: The fish fauna in the Yucatan Peninsula exhibit a unique composition, shaped by the region’s complex geological and hydrological history. Thorichthys meeki is a widely distributed cichlid species that has been the subject of morphological, behavioral, and phylogenetic studies. Nevertheless, the historical and biogeographical processes that have shaped its diversification and genetic structure remain unexplored. Objective: The objective of this study was to identify phylogeographic patterns and assess the genetic diversity of T. meeki in southeastern Mexico. Methodology: Genomic DNA was extracted from the muscle tissue of specimens preserved in 90% ethanol. DNA was amplified using a polymerase chain reaction (PCR) and sequences were obtained from two molecular markers: cytochrome oxidase 1 (CO1) and S7 intron 1. Phylogeographic analyses included genetic diversity indices molecular variance analyses to assess population structure and haplotype networks. Results: A total of 60 CO1 and 40 S7 intron 1 sequences were obtained from 26 sampling sites. For CO1, 14 haplotypes were identified, with high haplotype diversity (Hd = 0.84), and low nucleotide diversity (π = 0.0048). For S7 intron 1, 16 haplotypes were recovered with lower haplotype diversity (Hd = 0.543) and π = 0.021. A hierarchical AMOVA (2 groups) showed the greatest variation; 72.3% in CO1 and 79.8% in S7 within population groups with significant ΦST values. Haplotype networks identified three haplogroups for each gene. A single CO1 haplotype was found in 26 sequences from 13 sampling sites. For S7 intron 1, one haplotype was identified in 22 sequences and was shered from all populations. Conclusions: A widely distributed haplotype for CO1 and S7 reflects historical connectivity among hydrological systems. These preliminary results provide an interesting insights into the processesthat have influenced the distribution of T. meeki on the Yucatán Peninsula. Full article

Polycyclic aromatic hydrocarbons (PAHs) are widespread, persistent pollutants that can be sequestered within human adipose tissue due to their lipophilic nature. While this accumulation poses toxicological risks depending on dose and individual susceptibility, the specific morphological impact of chronic PAH storage on tissue architecture remains poorly defined. Here, we performed a histopathological and morphometric analysis on human subcutaneous adipose tissue samples characterized by high pyrene levels. We evaluated tissue organization, collagen distribution, the presence of inflammatory, neural, and vascular alterations and adipocyte morphometry to assess the structural response to PAH sequestration. Despite high pyrene concentrations, PAH-positive tissues maintained preserved overall architecture with normal collagen distribution, absence of lymphocytic infiltration, low macrophages, unaltered nerve fiber patterns, without evidence of vascular remodeling. Morphometry revealed smaller adipocyte area in PAH-positive samples, although not statistically significant. Our experimental data indicate that high PAH accumulation does not necessarily induce subcutaneous adipose tissue remodeling, suggesting that biochemical or metabolic alterations might occur even in the absence of evident histological changes. Further studies, with a broadened cohort, are needed to define the threshold at which PAHs’ presence translates into permanent tissue damage. Full article

Objective: Our objectives were to describe molecular profiling in a real-life cohort of patients with radioiodine-resistant (RAI-R) differentiated or poorly differentiated thyroid cancer (DTC or PDTC) treated with lenvatinib and to focus on factors potentially influencing the quality of tissue samples for molecular analysis, including the impact of storage time, defined as the interval between tissue collection and molecular testing. Design: We retrospectively included all lenvatinib-treated RAI-R DTC or PDTC patients tested with DNA- and/or RNA-based next-generation sequencing (NGS) in our center, also analyzing the results of fluorescence in situ hybridization (FISH) for RET fusions if the sample did not satisfy quality criteria for RNA-based NGS analysis. We investigated differences in terms of histotype, biopsy site, or storage time between adequate and inadequate samples for RNA-based NGS. Results: At least one gene alteration was detected in 50% of the cohort (18 out of 36 patients); RAS and BRAF were the most frequent mutations, while gene fusions accounted for 5.6% of cases. Tissue samples were more frequently adequate for DNA-based NGS compared to RNA-NGS analysis (93.9% vs. 58.3%, p < 0.001). The median storage time was significantly longer in the case of inadequate samples for RNA-based NGS compared with adequate specimens (41.5 vs. 9.5 months, p = 0.016); samples archived for ≥3 years led more frequently to an inadequate result. Conclusions: Advanced RAI-R TC candidates for systemic therapy often harbor gene alterations. An adequate result was less frequently achieved in cases of RNA-based NGS than in DNA-based NGS, especially if the interval between tissue collection and molecular analysis was longer; nevertheless, the limited cohort size precludes definitive conclusions. Full article

Accurate apple origin identification and non-destructive internal quality evaluation are important for fruit traceability, quality grading, and post-harvest management. Unlike previous studies mainly focusing on origin classification, this study established a dual-task near-infrared spectroscopy framework integrating geographical origin classification and soluble solid content (SSC, °Brix) prediction for Fuji apples. Samples were collected from three representative production regions in China: Alar in Xinjiang, Yantai in Shandong, and Luochuan in Shaanxi. Near-infrared diffuse reflectance spectra were acquired from 375 apples, generating 3000 spectral samples for origin classification and 750 SSC-calibrated samples for sugar content prediction. For classification, six deep learning models were evaluated using standardized full-spectrum input without chemometric spectral preprocessing, and the Transformer achieved the best performance, with a test accuracy of 96.22%. For SSC regression, spectra were preprocessed using standard normal variate and Savitzky–Golay filtering. The DNN model achieved the best prediction performance, with MAE = 0.5958 °Brix, RMSE = 0.7333 °Brix, R² = 0.8646, and Pearson r = 0.9338. These results indicate that near-infrared spectroscopy combined with deep learning can support both Fuji apple origin authentication and non-destructive local tissue SSC assessment. Full article

Objective: To evaluate the efficacy of NADPH oxidase 4 and miR-146a-5p in current treatment planning for ascending aortic aneurysms, independent of aortic diameter, and to develop protocols that will ensure the treatment of ascending aortic aneurysms, which pose a risk for aortic dissection, without complications. Methods: Patients who met the inclusion criteria and underwent surgery at Dr. Siyami Ersek Chest, Heart, and Vascular Surgery Training and Research Hospital for ascending aortic aneurysms and coronary artery disease between 2023 and 2024 were included in the study. This study was designed as a prospective study. Demographic, biochemical, radiological, and echocardiographic data were collected, and NOX4 mRNA and miR-146a-5p expressions were examined and compared in tissue samples. Results: The study was conducted on a total of 50 patients, with 25 patients in the aneurysm group and 25 patients in the control group. miR-146a-5p expression levels were found to be significantly decreased in the patient group compared to the control group (p = 0.001). When NOX4 mRNA expression levels were examined, no significant difference was found between the control and aneurysm groups. No correlation was found between NOX4 mRNA and miR-146a-5p levels (p = 0.764). When the relationship between ascending aorta diameter and both NOX4 mRNA and miR-146a-5p was examined, it was found that miR-146a-5p expression was negatively correlated with ascending aorta diameter (p = 0.036) and did not show a significant correlation with NOX4 mRNA levels (p = 0.318). A similar correlation was also found with ascending aorta length. The correlation of NOX4 mRNA and miR-146a-5p expression levels with age, gender, and ejection fraction was investigated separately. No significant correlation was found for all three variables. The optimum cut-off value to be used to separate the patient group from the control group using miR-146a-5p expression levels, as well as the sensitivity and specificity of miR-146a-5p expression levels when this cut-off value was used, was calculated using an ROC curve. Specificity for miR-146a-5p expression was found to be 88%, and sensitivity was found to be 66%. Conclusions: The study found promising results indicating that NOX4, shown to be a determinant of vascular oxidative stress, is not involved in the development of ascending aortic aneurysms; however, miR-146a-5p, which functions in the regulation of many inflammatory responses, including the regulation of NOX4 expression, may help prevent the development of ascending aortic aneurysms. Further studies aimed at elucidating the genetic and biochemical processes involved in aneurysm development suggest that miR-146a-5p could be a therapeutic target for preventing aneurysms. Full article

Osteoarthritis (OA) causes chronic pain and impaired mobility in dogs. Since current therapies cannot restore damaged articular cartilage, tissue engineering approaches offer promising therapeutic strategies. This study aimed to investigate whether peri-ovarian adipose tissue (POAT) represents a biologically competent and functionally relevant alternative source of mesenchymal stromal cells (MSCs) compared to subcutaneous adipose tissue (SAT). Samples were collected from five healthy and normal-weight Labrador Retrievers undergoing routine ovariectomy. MSCs were characterized according to the International Society for Cellular Therapy, including doubling time, growth curves, colony-forming unit assays, immunophenotyping, and trilineage differentiation potential. Chondrogenic differentiation was assessed through Alcian Blue staining and qPCR analysis of COL2A1, COL1A1, COL10A1, and SOX9 expression at multiple timepoints. MSCs derived from both adipose depots showed comparable mesenchymal characteristics, proliferative capacity, immunophenotypic profiles, and multilineage differentiation potential. POAT-MSCs exhibited enhanced chondrogenic differentiation compared to SAT-MSCs, with stronger extracellular matrix deposition and significantly increased COL2A1 expression at later stages of differentiation than SAT-MSCs. SOX9 expression supported a more advanced chondrogenic commitment in POAT-derived cells, while COL10A1 expression remained low and stable in both groups. These preliminary findings suggest that POAT, routinely discarded after ovariectomy, may represent a promising and ethically advantageous source of canine MSCs for regenerative medicine. Full article

Although the Kazakh horse is a dual-purpose breed renowned for both milk and meat production, the extent to which surgical castration alters gene expression in its muscles has not yet been fully elucidated. In this study, left longissimus dorsi muscle (LDM) samples were obtained from six Kazakh stallions (W group) and six Kazakh geldings (S group) to comparatively evaluate meat quality parameters, examine histological characteristics in tissue sections, and apply transcriptomic profiling to comprehensively explore the principal regulatory pathways and candidate genes through which surgical castration modulates LDM growth. The results demonstrated that surgical castration did not induce significant alterations in meat color or pH-related parameters. However, cooking loss and shear force values were markedly diminished, accompanied by a marked decrease in muscle fiber cross-sectional area. Transcriptomic analysis identified 848 differentially expressed genes (DEGs) in total, comprising 415 upregulated and 433 markedly downregulated DEGs, which were predominantly enriched in key biological pathways, including actin cytoskeleton regulation. Moreover, eleven core candidate genes, including MYL2, MYL3, and TNNI1, were further screened and identified. Full article