Search Results (3,941)

Background: Major level escalation following distal amputation for diabetic foot—defined as subsequent below-knee amputation (BKA)—represents a clinically meaningful endpoint with substantial functional implications. Identifying reliable and readily available preoperative biomarkers capable of predicting major level escalation remains a clinical challenge. This study aimed to evaluate the independent prognostic value of the C-reactive protein-to-albumin ratio (CAR) and blood urea nitrogen-to-albumin ratio (BAR) in predicting postoperative major level escalation. Methods: We retrospectively analyzed 151 consecutive patients who underwent distal lower extremity amputation for diabetic foot between January 2020 and October 2025. The primary outcome was ipsilateral below-knee amputation within the follow-up period. Preoperative CAR and BAR were calculated using laboratory parameters obtained within 24 h prior to surgery. Given the shared albumin component, two separate multivariable logistic regression models were constructed to evaluate independent associations, adjusting for age, peripheral arterial disease (PAD), and index amputation level. Results: During follow-up, 46 patients (30.5%) required major level escalation (BKA). Both CAR and BAR were significantly higher in patients who developed BKA (p < 0.001 and p = 0.006, respectively). In receiver operating characteristic (ROC) analysis, CAR demonstrated acceptable discriminative ability (AUC = 0.745; 95% CI 0.653–0.827), whereas BAR showed modest performance (AUC = 0.640; 95% CI 0.536–0.738). The optimal cutoff values were 1.50 for CAR (sensitivity 76.1%, specificity 61.9%) and 0.61 for BAR (sensitivity 73.9%, specificity 44.8%), although these thresholds were considered exploratory. In the primary multivariable analysis, both CAR (OR 1.16; 95% CI 1.02–1.32; p = 0.024) and BAR (OR 4.02; 95% CI 1.85–8.73; p = 0.005) were independently associated with major level escalation. In sensitivity analyses, BAR retained independent significance, whereas CAR did not (p = 0.100). Conclusions: Preoperative BAR demonstrated robust independent association with major level escalation across both primary and sensitivity analyses, while CAR showed association in the primary model only. These composite biomarkers may provide hypothesis-generating insights into systemic risk profiling in diabetic foot surgery, pending external validation. Full article

Chimeric antigen receptor (CAR) T-cell therapy is an effective treatment for hematologic malignancies. However, it is limited by high costs, risk of severe toxicities such as cytokine release syndrome and neurotoxicity, and heterogeneous patient responses. The current therapy monitoring depends largely on subjective symptom assessment, routine laboratory tests, and basic vital signs, without real-time, quantitative evaluation of CAR T-cell expansion or activation in clinical practice. This lack of timely immune monitoring hampers individualized care and contributes to increased treatment costs. To address this need, we present a proof-of-concept, label-free rapid optical imaging (ROI) biosensor with automated machine learning analysis for direct quantification of CAR T-cells from whole blood. This microfluidic platform integrates red blood cell (RBC) removal, CAR T-cell capture, and imaging-based quantification on a single chip, eliminating the need for centrifugation, staining, and operator-dependent interpretation. For validation, 50 μL whole blood samples spiked with Jurkat cells expressing CD19 CARs underwent RBC depletion by agglutination and microfiltration. The remaining blood components were then incubated on a sensor chip functionalized with recombinant CD19 protein. Captured CAR T-cells were imaged by brightfield microscopy and automatically enumerated using a machine learning algorithm trained on fluorescence-validated cells. The CD-19 cells’ capture performance was validated by flow cytometry and fluorescence imaging. The trained machine learning model validated at 88% sensitivity and 96% specificity. Buffer and whole blood calibration curves were established across clinically relevant concentrations (1–1000 cells/µL) with triple replicates. The results showed high correlation (0.975 and 0.990 R²) between the spiked concentration and the detected CAR T-cells, with a 95% certainty limit of detection (LOD) and quantification (LOQ) of 0.6 and 1.1 cells/µL for spiked buffer, and 14 and 67 cells/µL for spiked whole-blood, respectively. Full article

Background/Objectives: Early detection through minimally invasive approaches is critical for timely patient stratification and optimal therapeutic decision-making in colorectal cancer (CRC). Liquid biopsy, based on the analysis of tumor-derived components in blood and other body fluids, has emerged as a promising strategy to overcome current limitations in CRC diagnosis and follow-up. This review evaluates the current landscape of liquid biopsy clinical trials in CRC, focusing on predictive biomarker detection, prognostic assessment, and disease monitoring. Methods: ClinicalTrials.gov was searched using the terms “colorectal cancer” and “liquid biopsy” yielding 153 registered trials. After manual screening, 44 trials were excluded for not using liquid biopsy for CRC management, leaving 109 trials for analysis. Of these, 25 were completed, and 13 had publicly available results related to liquid biopsy. Results: The included trials were conducted across 27 countries on four continents. Overall, 119 biomolecules assessments and 167 different endpoints were reported across 109 clinical trials. Because individual trials could evaluate multiple biomolecules and endpoints, counts exceed the total number of trials. Cell-free DNA (cfDNA) was evaluated in 92/109 trials (84%) and accounting for 77% of all biomolecule assessments. Circulatingtumor cells (CTCs) were analyzed in 9/109 trials (8%, representing 8% of all the biomolecules analyzed), and microRNAs (miRNAs) in 8/109 (7%, representing 7% of all the biomolecules analyzed). Treatment sensitivity was the most common endpoint (57/109, 52% of the clinical trials; representing 34% of all the 167 different endpoints analyzed), followed by disease progression (28/109, 26%; representing 17% of all the different endpoints analyzed) and diagnostic applications (21/109, 19%; representing 12% of all the different endpoints analyzed). Among the 25 completed studies, 10/25 (40%) were interventional and 15/25 (60%) observational, spanning 14 countries. The majority of completed trials (21/25, 84%) used cfDNA. Interventional studies were predominantly phase II (5/10), with fewer phase III trials (2/10), primarily evaluating treatment response, particularly in relation to EGFR inhibitors and RAS/BRAF mutation status. Four observational studies (4/15) investigated emerging biomarkers, including long noncoding RNAs and miRNAs. Conclusions: Current clinical trials highlight cfDNA as the dominant and most clinically advanced liquid biopsy biomarker in CRC, primarily used for treatment guidance and disease monitoring. In contrast, CTCs and RNA-based biomarkers remain underrepresented. The limited number of randomized late-phase trials, heterogeneity in study design, and technical challenges associated with emerging biomarkers underscore the need for standardized methodologies and robust validation before routine clinical implementation. Full article

Background/Objectives: Blood donation is a critical component of healthcare systems worldwide, yet donor recruitment remains challenging. This study evaluates the knowledge, attitudes, motivations, and practices (KAP) of blood donation among the general population in Saudi Arabia to identify key barriers and propose targeted interventions. Methods: A cross-sectional study was conducted using a structured, validated questionnaire distributed over five months (December 2022 to April 2023) via social media and in-person recruitment at the Central Blood Bank in Riyadh. A total of 1150 participants aged 18–60 years residing in Saudi Arabia were included in the final analysis. Statistical analysis was performed using SPSS version 22, with p < 0.05 considered significant. Results: Participants demonstrated moderate knowledge (mean score 5.43 ± 1.81 out of 9), with significantly higher scores among males, individuals aged 21–30 years, and those holding a bachelor’s degree. Attitudes toward donation were highly positive (mean score 15.46 ± 2.74 out of 20) and correlated with age, gender, marital status, and occupation. Despite this positive outlook, only 34.96% of participants had donated blood previously, although 95.25% expressed willingness to do so. Primary motivators included mobile donation units (89.22%) and paid leave (89.22%), whereas 51.22% of respondents considered current media campaigns ineffective. Common barriers to donation included health concerns (25.30%), time constraints (12.87%), and fear of needles (7.74%). Conclusions: This study reveals a critical disparity between positive public attitudes and actual donation practices in Saudi Arabia. To enhance donor participation, we recommend implementing convenient donation strategies such as mobile blood drives, workplace incentives, and more effective, culturally tailored educational campaigns. Addressing these factors could help Saudi Arabia improve its voluntary donation rates and ensure a sustainable, safe blood supply. Full article

Background: Multi-center resting-state functional magnetic resonance imaging (rs-fMRI) is important for neurodevelopmental disorder diagnosis, but cross-site differences in repetition time (TR) can cause temporal feature misalignment. In addition, blood-oxygen-level-dependent (BOLD) signals are non-stationary, so disease-related information may be distributed across multiple time scales. Existing methods usually do not explicitly model physical sampling intervals or coordinate temporal and spectral information across scales, which may limit cross-site generalization in heterogeneous multi-center settings. Methods: We propose Spec-RWKV, a spectrum-guided linear recurrent framework for multi-site rs-fMRI diagnosis. It includes three components: PrismTimeMix, which models temporal dynamics using decay rates derived from physical half-lives and converts them adaptively across TRs; a TR-adaptive continuous wavelet transform, which aligns spectral representations across sites by adjusting frequency coverage; and spectrum-guided adaptive temporal aggregation, which uses spectral context to weight temporal features. Results: On ABIDE-I and ADHD-200, Spec-RWKV achieved AUCs of 75.86% and 76.31%, respectively. Under leave-one-site-out validation, it achieved the best mean AUC on ABIDE-I and the best mean accuracy and AUC on ADHD-200. Conclusions: Spec-RWKV explicitly models sampling-rate differences and multi-scale spectral structure, with results supporting strong cross-site generalizability. Full article

Cardiac amyloidosis is a rare and progressive condition characterized by the extracellular deposition of amyloid fibrils in multiple organs. Wild-type transthyretin amyloidosis (ATTR-wt) is the most common type affecting subjects above 60 years old. Recent and growing evidence suggests a potential link between GM and cardiac amyloidosis. In this scenario, the aim of the present study is to characterize the gut microbiota (GM), related metabolites and inflammatory biomarkers in ATTR-wt patients. In the ATTR patients we identified Prevotella_9 as the core OTUs (Operational Taxonomic Unit) of this group, alongside Prevotella 7, Prevotellaceae_UCG-003 and Prevotellaceae_NK3B31. In addition, there were increased levels of long fatty acids, including tetradecanoic, hexadecanoic and octadecanoic acids, in the ATTR group. The data obtained suggest that ATTR patients have an altered gut microbiota that could be used as a potential biomarker in metabolic and cardiovascular diseases, as well as a potential predictor of adverse prognosis in ATTR patients. In addition, the intestinal dysbiosis in ATTR patients could be associated with low-grade endotoxemia promoting a pro-inflammatory state due to the translocation of bacterial components, such as LPS (lipopolysaccharide), into blood circulation. Full article

Chemotherapy-induced gonadotoxicity severely compromises male fertility, yet effective interventions remain limited. Building on our previous finding that yam protein (YP) modulates the gut-microbiota axis, this study investigated its direct protective role against cyclophosphamide (CTX)-induced testicular injury. Spectral analysis revealed a protein fraction (L-YP) with strong intrinsic fluorescence and optimal cytoprotection against oxidative stress. Proteomic characterization revealed six dominant proteins (YP1–YP6). In vivo experiments demonstrated that L-YP upregulates the expression of tight junction proteins Occludin and ZO-1, restores hormone levels, and modulates inflammatory factors, thereby enhancing the integrity of the blood–testis barrier. Network pharmacology analysis and molecular docking predicted a potential binding affinity between key components such as YP2 and NF-κB p65, which may provide a structural basis for their regulatory role. Further validation at the gene level indicated that YP can improve the local testicular immune microenvironment by modulating the classical TLR4/MyD88/NF-κB inflammatory signaling pathway. These findings suggest that yam protein alleviates chemotherapy-induced testicular damage, potentially through barrier protection and anti-inflammatory mechanisms, indicating its promise as a dietary protective agent. Full article

Blood is a key component of organisms, serving numerous functions, including metabolism, innate and humoral responses, and hemostasis. Variations in hematological parameters can indicate the presence of infectious and non-infectious diseases, chronic stress, and other pathological or physiological conditions. Complete blood count testing is common in human and veterinary medicine and, when combined with clinical examination, contributes to disease diagnosis and prognosis and the monitoring of therapeutic progression. Nevertheless, hematological analysis is not routinely performed in sheep due to the lack of case-specific reference intervals, complicating the interpretation of the results. Indeed, hematological parameters may be affected by various non-pathological (environmental, genetic, physiological) and pathological factors, and they require further understanding and relevant adjustments to be universally applicable. Therefore, the objective of this paper is to summarize the existing literature and describe how various pathological and non-pathological factors affect hematological parameters in sheep, thereby supporting their incorporation into health management practices. Full article

Positive mental health (PMH) has recently become a key topic in biomedical research. Previous studies have explored the correlation between biological and psychological measures, but only a few have focused on the relationship between PMH and aging. This study aimed: (i) to explore the association between PMH and biological aging; (ii) to determine if and to what extent the observed association could be explained by shared genetic and environmental effects. A total of 401 twins (age 19–81 years, 32% male) from the Italian Twin Registry were recruited, and the twin study design was applied. A self-report psychological test battery was used to evaluate several PMH components. Blood samples were collected from participants to determine telomere length (TL) and mitochondrial DNA copy number (mtDNAcn). TL was negatively associated with attachment anxiety (r = −0.11, p = 0.037). A bivariate twin model provided heritability estimates of 0.14 (95% CI 0.001–0.43) for TL and 0.32 (0.16–0.45) for attachment anxiety, and a substantial negative genetic correlation [r_g = −0.55 (−1.00–0.00)] between them. Under the limitations of a cross-sectional study with a self-report wellbeing assessment, these results suggest that anxiety in a relationship with a partner may contribute to accelerated TL shortening, and shared genetic factors may underlie this link. Full article

Sickle cell anemia (SCA) is a genetic disorder characterized by chronic hemolysis, primarily driven by red blood cell lysis. Its pathophysiology is centered, though not exclusively, on the increased release of intracellular components, such as hemoglobin degradation products, which are known to stimulate innate immune responses and promote prothrombotic states. Current therapies alleviate symptoms, yet patients remain exposed to a chronic inflammatory milieu punctuated by episodes of acute pain. The recurrence of these crises can be life-threatening due to ischemia–reperfusion injury, hypercoagulability, and respiratory complications. Central mechanisms are marked by elevated hemolysis, heightened inflammatory signaling, and increased procoagulant activity, largely driven by soluble molecules released into the plasma, such as hemoglobin, nuclear molecules and other products. These compounds are recognized from sensors on immune and endothelial cells, named Pattern Recognition Receptors (PRRs), and constitute canonical pathways for intracellular activation. Four main types have been extensively studied in the literature over recent years in both infectious and sterile inflammatory contexts; still, only a few have elucidated the mechanisms underlying acute and chronic inflammation in patients with SCA. Although Toll receptors were shown to be major in triggering immunity, other receptors were found to be important regarding this function, which suggested a multifactorial mechanism for this triggering. Therefore, here, we propose a comprehensive review of previously published findings regarding the expression, activation, and dynamics of Toll-like, NOD-like, and RIG-I–like receptors in the progression of SCA and its associated inflammatory features. Full article

Background/Objectives: The growing prevalence of obesity necessitates innovative gut-targeted material strategies to modulate diet-associated metabolic dysfunction. This study investigates a spray-dried konjac glucomannan–montmorillonite (KGM-MMT) hybrid designed to integrate fermentable polysaccharide properties with luminal lipid-adsorptive clay functions within a single micro-engineered formulation. Methods: In HFD-fed mice treated for 42 days with 2% w/w KGM-MMT, cumulative body weight gain was attenuated by 7.6%, with an AUC of 5094 ± 52.95, compared to 5513 ± 81.35 in HFD controls (p < 0.0001). Results: Serum IL-6 concentrations were reduced by 97% (p = 0.0002), while blood glucose decreased by 46% (p < 0.0001); these effects were greater than those observed with MMT (24%, p = 0.0271) and KGM (16%, ns). Gut microbiota profiling demonstrated a significant 6.2-log₂-fold increase in Lactobacillaceae (p = 0.023) and a 2.4-log₂-fold increase in Enterococcaceae (p = 0.015) following KGM-MMT treatment. Functional shifts inferred from 16S rRNA gene-based prediction indicated a 1.9-fold increase in short-chain fatty acid-related pathways and a 5.4-fold increase in bile acid deconjugation pathways. Conclusions: Although the KGM-MMT hybrid did not consistently outperform its individual components across all endpoints, it consolidated complementary KGM- and MMT-associated effects within a single dosage form. These findings support spray-dried KGM-MMT as a gut-targeted biomaterial strategy that integrates multiple luminal and microbiota-associated functions within a single formulation. Future studies should define dose–response relationships, validate microbiota-derived functional predictions using higher-resolution approaches, and assess durability and safety under longer-term exposure. Full article

This study investigated the effects of chronic heat stress (HS) on brain injury in broilers and the associated molecular changes. A chronic HS model was established by exposing broilers to 35 °C from 08:00 to 20:00 daily from 21 to 42 days of age, and samples were collected at 28, 35, and 42 days of age. Chronic HS significantly impaired growth performance and was associated with histopathological and ultrastructural alterations in brain tissue. Serum antioxidant enzyme activities and the total antioxidant capacity were significantly reduced, whereas malondialdehyde levels were significantly increased, indicating sustained oxidative stress (OS). Blood–brain barrier (BBB) permeability, assessed by Evans blue extravasation, was significantly higher in HS birds and was accompanied by reduced mRNA expression of the tight junction-related genes ZO-1 and Claudin-5. In addition, chronic HS was associated with increased mRNA expression in genes related to cellular stress, oxidative stress, and inflammation, including key components of the TLR4/MyD88/NF-κB/NLRP3 pathway, as well as decreased expression of IL-4. These findings suggest that chronic HS is associated with enhanced OS, altered neuroinflammatory gene expression, and BBB impairment in the broiler brain. Overall, this study provides evidence that chronic HS is associated with brain injury in broilers and highlights a potential link among OS, inflammation-related transcriptional changes, and BBB dysfunction, thereby offering a basis for further mechanistic and interventional studies. Full article

Therapeutics derived from donated blood or its constituents are classifiable into blood components and plasma derivatives. The latter are defined as medicines/drugs/pharmaceuticals produced from the industrial fractionation of thousands of pooled plasma donations and characterised with relative precision to a pre-defined specification through sampling of a homogenous pharmaceutical batch. The former are defined as components/biologicals produced using relatively simple (but increasingly complex) technologies in blood centres from single or small pools of isolated components from whole blood and are pre-specified through regulatory standards with relatively wide limits because of the inherent biologic variability of individual donors. This review discusses the evolution of technology to reduce the risk of pathogen transmission by blood-derived therapeutics, assess the state of the approved technologies for pathogen-reduced blood components, and examine the features of the blood-provider and regulatory framework globally that have shaped, and in some instances impeded, the implementation of component pathogen reduction to an extent equivalent to that achieved for plasma derivatives. The ensuing risks to the public’s confidence in the blood supply are discussed, and remedial actions are proposed. The features of a new paradigm for blood safety are outlined. Full article

Ten substituted quinobenzothiazinium salts were tested for their ability to inhibit acetylcholinesterase (AChE) and butyrylcholinesterase (BChE). All the compounds inhibited AChE in the IC₅₀ range of 0.03–0.658 µM, with 5,8,10-trimethyl-12H-quinolino[3,4-b][1,4]benzothiazin-5-ium chloride (3d) being the most potent inhibitor, with an IC₅₀ value significantly better than that of the clinically used rivastigmine and galantamine and comparable to that of tacrine and donepezil. The IC₅₀ values for BChE inhibition ranged from 0.34 to 4.25 µM; 5,9-dimethyl-12H-quinolino[3,4-b][1,4]benzothiazin-5-ium chloride (3b) exhibited the strongest BChE inhibitory activity and in general, all the investigated compounds were more potent inhibitors than rivastigmine and galantamine. Based on the calculated selectivity index values, they are rather preferential inhibitors of AChE. Cytotoxicity tests performed on normal human dermal fibroblasts (HFF-1) did not demonstrate any significant cytotoxicity under the tested conditions. The distance-oriented structure distribution for the studied molecules was related with the activity data using principal component analysis and hierarchical clustering analysis. (SAR)-based evaluation is reported to predict activity cliffs using a similarity–activity landscape index for the AChE inhibitory response values. Moreover, direct protein-mediated in silico methods were utilized to identify factors that may be relevant for quantitative (Q)SAR modeling. In practice, target-oriented molecular docking was used to organize the spatial distribution of the ligand property space for the anti-AChE system. In general, this series of alkylated quinobenzothiazinium salts with potent inhibitory activity against cholinesterases fulfills Lipinski’s rule of five based on in silico predictions and is also expected to have high absorption in the human gastrointestinal tract. All active derivatives are also expected to penetrate the blood–brain barrier, making them promising compounds for further research and possible use in Alzheimer’s disease therapy. Full article

Despite the identification of several mediators of arteriogenesis, the growth of natural bypass, the role of lymphocytes, particularly T cells, in this process remains poorly defined. Among these, γδ T cells, which express alternative T cell receptors, have emerged as a key immune component. This study examined the roles of αβ and γδ T cells in arteriogenesis using a murine hindlimb model. While the absence of αβ T cells did not affect arteriogenesis, γδ T cell depletion markedly reduced vascular cell proliferation and perfusion recovery. Early phase analyses revealed impaired mast cell activation, whereas platelet–neutrophil aggregates and neutrophil extravasation were unaffected. In the later proliferative phase, γδ T cell depletion hindered perivascular M2-like (MRC1⁺) macrophage accumulation. Flow cytometric analysis of whole blood in wildtype mice revealed a temporal shift in γδ T cell populations from a CD27⁺/CD39⁻ phenotype, commonly associated with pro-inflammatory functions and IFNγ production, to CD39⁺ phenotypes, which have been linked to anti-inflammatory properties and IL-10 production. In rescue experiments, administration of IFNγ to γδ T cell-depleted mice restored mast cell activation, whereas IL-10 treatment reestablished M2-like (MRC1⁺) macrophage accumulation. These findings collectively identify γδ T cells as critical regulators of both early and late phases of arteriogenesis through coordinated inflammatory and regenerative mechanisms. Full article