Precise surface engineering is essential for robust characterization of extracellular vesicles and their interactions with biologically relevant interfaces. In this study, Au substrates were modified with C6 self-assembled monolayers (SAMs) bearing different terminal groups after multiple cleaning protocols to optimize monolayer compactness and interfacial quality. Surface area and functionality were further enhanced by gold nanoparticle (AuNP) deposition and secondary SAM formation, enabling layer-by-layer assembly of glycan-based polysaccharide nanostructures. Interface fabrication and growth were monitored by electrochemical methods and atomic force microscopy (AFM). Extracellular vesicles (EVs) from MDA-MB-231 breast cancer cells and benign RWPE-1 (control) cells were affinity isolated using magnetic microparticles, chromatographically preconcentrated, and subjected to protein-corona removal prior to analysis. The impact of the protein corona on antibody-mediated recognition was evaluated by ELISA and revealed markedly improved accessibility of EV surface markers following corona removal. Interactions of EVs with extracellular-matrix-mimicking glycan interfaces were investigated using surface plasmon resonance (SPR). Sensorgrams were analyzed using a physics-informed neural-network-assisted model to extract dissociation constants while reducing the influence of bulk refractive-index contributions typical of vesicle samples. The developed nanobiointerfaces enabled sensitive characterization of EV binding behavior and revealed distinct interaction phenotypes of malignant-derived and non-malignant-derived EVs toward glycan-based surfaces. These findings demonstrate that protein-corona composition and glycan-mediated interactions significantly influence EV recognition and highlight the potential of glycan-based nanobiointerfaces as tools for studying extracellular-vesicle biology, which may contribute to future liquid-biopsy development.

Artificial intelligence, particularly machine learning and deep learning, is a rapidly evolving field that is increasingly permeating all areas of modern society, including public healthcare. In the present work, we provide a comprehensive introduction to the application of machine learning in routine clinical practice and biomedical research, including biomarker discovery. The most widely used machine learning models are introduced together with their key characteristics, strengths, and limitations. Furthermore, the fundamental principles governing the interpretation of model outputs and communication with regulatory authorities are discussed and illustrated using both real-world and synthetic datasets through modeling in the Python programming environment. Particular emphasis is placed on maintaining data quality and ensuring robust validation procedures in the context of dynamic, continuously updated AI systems intended for diagnostic software registered as medical devices (SaMDs). The importance of data governance, external validation, interpretability, and clinical oversight is highlighted as a prerequisite for safe and effective implementation of AI technologies in healthcare - throughout both the development and commercialization phases. Finally, current trends in medical artificial intelligence are reviewed, together with their potential benefits and associated risks for patients. The presented overview aims to facilitate a deeper understanding of the opportunities and challenges associated with the integration of AI-driven solutions into modern healthcare systems.

Oxygen-deficient nanomaterials have emerged as a distinct class of functional materials that continue to attract extensive research interest due tothe presence of oxygen vacancies (OVs) and associated electronic structure modulation and broad technological relevance in catalysis, sensing, energy storage and conversion, and environmental remediation. This review provides a comprehensive examination of how different OVs preparation methods govern their formation, distribution, and stability, thereby dictating fundamental mechanisms in diverse nanostructured systems, with emphasis on the resulting structural distortions, electronic rearrangements, and defect–property correlations. Particular attention is devoted to advanced synthesis strategies designed to precisely and efficiently tailor vacancy concentration and distribution, as well as to state-of-the-art spectroscopic and microscopic methodologies enabling their quantitative and qualitative characterization. Furthermore, recent progress in the deployment of oxygen-deficient nanomaterials is evaluated in detail. Thermal and chemical reduction methods significantly increase the concentration of the vacancy and mixed-valence metal states, which enable enhanced electrical conductivity, redox properties and structural stability of the nanomaterials via excessive bulk defects.Plasma, laser, and electrochemical-assisted routes generate surface-localized OVs preferentially to serve as highly active catalytic sites during a wide range of catalytic reactions. However, non-stoichiometric type strategies, including high-energy particle bombardment and chemical vapour deposition, offer thermodynamically stable OVs engineering, compensating for the materials’ activity and durability.By integrating insights from synthesis, characterization, and application perspectives, this review seeks to provide a view that all the adopted methods regulate the charge-transfer kinetics and adsorption energetics by promoting metal-oxygen covalency and introducing donor states of the evolving landscape of oxygen-deficient nanomaterials in their multifunctional role in dictating performance, and their potential to inform the rational design of next-generation materials for sustainable technologies.

This review paper summarizes the pathophysiology of rheumatoid arthritis (RA), its incidence and etiology, and conventional RA protein-based biomarkers. A brief overview of novel protein-based biomarkers and biomarkers with potential to be used in the diagnostics of seronegative RA is also provided. The core of the review resides in the comprehensive characterization of the glycosylation of immunoglobulins and their utility as novel biomarkers for the diagnostics of RA and seropositive RA. Two main approaches to the analysis of glycans present on immunoglobulins and other proteins are provided, including instrument-based and lectin-based approaches showing their clinical performance as RA biomarkers.

The current study shows a direct electron transfer (DET) and direct bioelectrocatalysis of the sarcosine oxidase (SOx) on the screen-printed carbon electrode (SPCE) modified by a hybrid bionanocomposite composed of chitosan (CS) and Ti3C2Tx MXene for the first time. A detailed electrochemical investigation revealed a pair of redox peaks at SPCE/CS-MXene/SOx, i.e. an anodic peak at a potential value of approx. -0.7 V and a cathodic peak at a potential value of approx. -1.0 V at pH 7.0, displaying direct electron transfer of SOx. Further experiments showed homogeneous DET with SOx not to be strongly adsorbed on the interface; this might be a prerequisite for keeping the enzyme active towards catalysis. The DET of SOx is not reversible with the maximal current observed at pH 7.0. It can also be concluded that scan rate significantly influences the redox behaviour of SOx and that at scan rates above 0.3 V s−1 the redox behaviour of SOx is quite stable with Ipc/Ipa achieving a stable value of -1.55. In addition, detailed analysis revealed that the enzyme exhibited E1/2 of -(0.781 ± 0.003) V at pH 7.0. Direct bioelectrocatalysis is more effective at pH 7.0 than at pH 9.0, achieving a high maximal current of -7.57 × 10−5 A at a potential value of -663 mV upon the addition of 1 mM sarcosine. Furthermore, direct bioelectrocatalysis of SOx is highly reproducible when examined using six different SPCE electrodes.

N-glycome assays, utilizing a combination of matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) with linkage-specific N-glycan modifications, enabled the identification of specific N-glycans present in the plasma samples obtained from both colorectal cancer (CRC) patients and healthy controls. These assays determined two different types of sialic acid linkages: alpha-2,6-linked sialic acids and alpha-2,3-linked sialic acids, to expand the pool of potential glycan biomarkers. Biostatistical data evaluation was carried out using seven different approaches, selecting the top glycan discriminators, i.e. in a form of box plots revealing p values (Mann-Whitney U test); using principal component analysis, simulated annealing search for the best markers (maximization of Log-likelihood function (LogLik) or Area Under Curve (AUC) from Receiver Operating Curve (ROC)) and logistic regression with L1 regularization (Lasso). There are two glycans that rank among the top discriminators across all seven data evaluation methods. A more detailed data evaluation from the ROC was utilized to extract clinical parameters such as sensitivity, specificity, and AUC values. It is noteworthy that the combination of two biomarkers yields a remarkably high AUC value of 0.967 to discriminate CRC patients from healthy controls. Furthermore, the combination of three glycan biomarkers only marginally increased the AUC to a value of 0.993.

Direct methanol fuel cells are promising energy conversion devices, which traditionally rely on platinum on carbon black (Pt/C) as electrocatalysts to perform electro-oxidation of methanol at the anode. Still, classical Pt/C catalysts suffer from several issues, such as sluggish reaction kinetics, surface poisoning, insufficient durability, and high cost, as they use up to 20 wt% of precious Pt metal as active catalytic sites. To address these issues, several alternative Pt-based electrocatalysts have been suggested as alternatives for methanol oxidation reaction. In this review, we consider selection of constituting materials of the Pt-based electrocatalysts for methanol oxidation, with a focus on their influence on the performance of Pt-active sites, whetheras supports or active co-catalysts. Among different chemical elements from the periodic table, s-block elements primarily modulate the electrode/electrolyte interfaces, while p-, d- and f-block elements tune the electronic structure of Pt and Pt‒Pt bond length through coupling effects, which is beneficial in terms of anti-poisoning electrocatalyst behaviour. We specify the role of those elements in methanol oxidation kinetics and outline contemporary strategies to achieve enhanced performance, durability, and economic viability of the direct methanol fuel cells.

Cancer is a leading cause of death worldwide, resulting in substantial economic costs. Because cancer is a complex, heterogeneous group of diseases affecting a variety of cells, its detection may sometimes be difficult. Herein we review a large group of the gastrointestinal cancers (oral, esophageal, stomach, pancreatic, liver, and bowel cancers) and the possibility of using glycans conjugated to protein backbones for less-invasive diagnoses than the commonly used endoscopic approaches. The reality of bacterial N-glycosylation and the effect of epithelial mucosa on gut microbiota are discussed. Current advantages, barriers, and advantages in the prospective use of selected glycomic approaches in clinical practice are also detailed.

The review article provides a short introduction to exosomes with the focus to use exosomes as disease markers itself (i.e. their concentration or presence of some specific receptors) or a source of disease biomarkers such as proteins and metabolites. In detail, we are discussing various methods of exosome isolation and the main focus of the review paper is on affinity capture of exosomes, since some of them can be applied to the isolation of specific sub-populations of exosomes produced by some specific organs. The article provides a comprehensive overview of magnetic (bio)affinity capture applied to the detection of exosomes or exosomal cargo using different (bio)affinity capture ligands such as antibodies, DNA aptamers, peptides, glycan-based recognition, transferrin-based approaches, affinity based on recognition of phospholipids of exosomes and other approaches including electrostatic interactions. The review in detail provides key analytical and clinical parameters of such approaches in a form of an extensive table summarising outcomes published in the last two years (2023–2024). Finally, the review paper also provides conclusions sections discussing pros and cons of magnetic (bio)affinity capture for exosome isolation and/or determination of exosomal content.

Herein, the basic properties, composition, and isolation methods for the separation of exosomes are described, since they can be a rich source of disease biomarkers. Then an introduction to MXenes, a novel class of 2D nanomaterials with interesting properties applicable in numerous fields including biosensing is provided. Also, aptamers are described as an alternative to antibodies for the robust biorecognition of analytes of interest. The final part of the article gives examples in which these three key components are integrated for sensitive isolation and/or electrochemical detection of exosomes. The conclusion provides a summary of these initial achievements and also an outlook for future discoveries in exosome aptasensing using advanced nanomaterials, i.e., MXenes. MXenes have a promising outlook for affinity‐based biosensing, being hydrophilic and rich in surface functional groups. In addition, the free plasmons present in MXenes can be used for covalent grafting of biorecognition elements using diazonium moieties. This is an especially interesting approach for the immobilization of DNA/RNA aptamers, which can be readily modified by diazonium moieties.

BACKGROUND:The medication used to treat benign prostate hyperplasia (BPH), a common condition in men over 50 years of age, can alter the levels of biomarkers used in prostate cancer detection. Commonly used medications for BPH include alpha-blockers, 5-alpha reductase inhibitors (5-ARIs), and muscarinic antagonists. We studied the impact of these drugs on total prostate-specific antigen (tPSA), free PSA (fPSA), [-2]proPSA, fPSA/tPSA ratio, and the Prostate Health Index (PHI), as well as novel potential biomarkers in the form of glycan composition of fPSA. PATIENTS AND METHODS:Serum samples were collected from 564 males with BPH, with a mean age of 68.5 years. The samples were used to measure levels of tPSA, fPSA, and [-2]proPSA. The fPSA/tPSA and PHI were then calculated. The glycan composition of fPSA was analyzed using lectin-based glycoprofiling. Pharmacotherapy data was collected from the patients' medical records. RESULTS:Alpha-blocker monotherapy was associated with higher fPSA and fPSA/tPSA ratio, and decreased PHI. Levels of tPSA were not impacted. Alpha-blocker and 5-ARI dual therapy was associated with reduced levels of fPSA, [-2]proPSA, and PHI. Therapy combining alpha-blockers and antimuscarinic agents did not significantly influence biomarker levels apart from an increase in a Maackia amurensis lectin-recognized glycan originating in fPSA. CONCLUSION:BPH pharmacotherapy notably affects prostate cancer biomarkers. Recognizing the impact of pharmacotherapy is crucial for achieving an accurate diagnosis of prostate cancer and for planning treatment.

Prostate cancer (PCa) is the second most common cancer. In this paper, the isolation and properties of exosomes as potential novel liquid biopsy markers for early PCa liquid biopsy diagnosis are investigated using two prostate human cell lines, i.e., benign (control) cell line RWPE1 and carcinoma cell line 22Rv1. Exosomes produced by both cell lines are characterised by various methods including nanoparticle-tracking analysis, dynamic light scattering, scanning electron microscopy and atomic force microscopy. In addition, surface plasmon resonance (SPR) is used to study three different receptors on the exosomal surface (CD63, CD81 and prostate-specific membrane antigen-PMSA), implementing monoclonal antibodies and identifying the type of glycans present on the surface of exosomes using lectins (glycan-recognising proteins). Electrochemical analysis is used to understand the interfacial properties of exosomes. The results indicate that cancerous exosomes are smaller, are produced at higher concentrations, and exhibit more nega tive zeta potential than the control exosomes. The SPR experiments confirm that negatively charged α-2,3- and α-2,6-sialic acid-containing glycans are found in greater abundance on carcinoma exosomes, whereas bisecting and branched glycans are more abundant in the control exosomes. The SPR results also show that a sandwich antibody/exosomes/lectins configuration could be constructed for effective glycoprofiling of exosomes as a novel liquid biopsy marker.

We investigated the use of boron-doped diamond (BDD) with different surface morphologies for the enhanced detection of nine different peptides by matrix-assisted laser desorption/ionisation mass spectrometry (MALDI-MS). For the first time, we compared three different nanostructured BDD film morphologies (Continuous, Nanograss, and Nanotips) with differently terminated surfaces (-H, -O, and -F) to commercially available Ground Steel plates. All these surfaces were evaluated for their effectiveness in detecting the nine different peptides by MALDI-MS. Our results demonstrated that certain nanostructured BDD surfaces exhibited superior performance for the detection of especially hydrophobic peptides (e.g., bradykinin 1-7, substance P, and the renin substrate), with a limit of detection of down to 2.3 pM. Further investigation showed that hydrophobic peptides (e.g., bradykinin 1-7, substance P, and the renin substrate) were effectively detected on hydrogen-terminated BDD surfaces. On the other hand, the highly acidic negatively charged peptide adrenocorticotropic hormone fragment 18-39 was effectively identified on oxygen-/fluorine-terminated BDD surfaces. Furthermore, BDD surfaces reduced sodium adduct contamination significantly.

The initial part of the review provides an extensive overview about MXenes as novel and exciting 2D nanomaterials describing their basic physico-chemical features, methods of their synthesis, and possible interfacial modifications and techniques, which could be applied to the characterization of MXenes. Unique physico-chemical parameters of MXenes make them attractive for many practical applications, which are shortly discussed. Use of MXenes for healthcare applications is a hot scientific discipline which is discussed in detail. The article focuses on determination of low molecular weight analytes (metabolites), high molecular weight analytes (DNA/RNA and proteins), or even cells, exosomes, and viruses detected using electrochemical sensors and biosensors. Separate chapters are provided to show the potential of MXene-based devices for determination of cancer biomarkers and as wearable sensors and biosensors for monitoring of a wide range of human activities.

This review briefly introduces readers to an area where glycomics meets modern oncodiagnostics with a focus on the analysis of sialic acid (Neu5Ac)-terminated structures. We present the biochemical perspective of aberrant sialylation during tumourigenesis and its significance, as well as an analytical perspective on the detection of these structures using different approaches for diagnostic and therapeutic purposes. We also provide a comparison to other established liquid biopsy approaches, and we mathematically define an early-stage cancer based on the overall prognosis and effect of these approaches on the patient’s quality of life. Finally, some barriers including regulations and quality of clinical validations data are discussed, and a perspective and major challenges in this area are summarised.

The glycoprofiling of two proteins, the free form of the prostate-specific antigen (fPSA) and zinc-α-2-glycoprotein (ZA2G), was assessed to determine their suitability as prostate cancer (PCa) biomarkers. The glycoprofiling of proteins was performed by analysing changes in the glycan composition on fPSA and ZA2G using lectins (proteins that recognise glycans, i.e. complex carbohydrates). The specific glycoprofiling of the proteins was performed using magnetic beads (MBs) modified with horseradish peroxidase (HRP) and antibodies that selectively enriched fPSA or ZA2G from human serum samples. Subsequently, the antibody-captured glycoproteins were incubated on lectin-coated ELISA plates. In addition, a novel glycoprotein standard (GPS) was used to normalise the assay. The glycoprofiling of fPSA and ZA2G was performed in human serum samples obtained from men undergoing a prostate biopsy after an elevated serum PSA, and prostate cancer patients with or without prior therapy. The results are presented in the form of an ROC (Receiver Operating Curve). A DCA (Decision Curve Analysis) to evaluate the clinical performance and net benefit of fPSA glycan-based biomarkers was also performed. While the glycoprofiling of ZA2G showed little promise as a potential PCa biomarker, the glycoprofiling of fPSA would appear to have significant clinical potential. Hence, the GIA (Glycobiopsy ImmunoAssay) test integrates the glycoprofiling of fPSA (i.e. two glycan forms of fPSA). The GIA test could be used for early diagnoses of PCa (AUC = 0.83; n = 559 samples) with a potential for use in therapy-monitoring (AUC = 0.90; n = 176 samples). Moreover, the analysis of a subset of serum samples (n = 215) revealed that the GIA test (AUC = 0.81) outperformed the PHI (Prostate Health Index) test (AUC = 0.69) in discriminating between men with prostate cancer and those with benign serum PSA elevation.

Post-translational modifications of proteins play an important role in their stability, solubility and in vivo function. Also, for several reasons, such as the Golgi fragmentation during cancerogenesis, glycosylation as the most common modification is especially promising in offering high cancer specificity which, in combination with tissue-specific biomarkers available in the case of prostate diseases (PSA, PSMA, PAP), may lead to the development of novel oncodiagnostic approaches. In this review, we present the importance of subterminal glycan structures based on the N-acetylated monosaccharides GlcNAc and GalNAc in N- and also O-glycans, structures of which they are a component (LacNAc, LacdiNAc, branched structures). We also discuss the importance and clinical performance of these structures in cases of prostate cancer diagnostics using lectin-based affinity methods, which could be implemented in clinical laboratory practice in the future.

In this study, a stable and highly hydrophobic perfluoroctylsilane grafted Ti3C2X (PFMXene) possessing repeatable and stable photothermal behavior was applied for fabrication of liquid marble (LM) and hydrogel-based LM. PFMXene-based hydrogel LMs demonstrate floating ability and self-assembly on the water's surface. Moreover, controlled non-contact movement of hydrogel marble through the immersed tool in the water surface reached a speed up to 2 cm s(-1). The structural character of the hydrogel matrix allows controlled light-induced disintegration of hydrogel liquid marble or application as a water evaporation system.

In this study, an assay for detection of the cancer biomarker Thomsen-nouvelle (Tn) antigen on the ELISA plates format was designed and developed. The effects of size and the interfacial density of the negative charge of magnetic beads (MBs) on the specific sensitivity of the bioaffinity interaction were studied. In particular, gly-conanoconjugate, i.e. glycan Tn antigen conjugated to bovine serum albumin (BSA) was covalently immobilised on MBs for the bioaffinity detection of anti-Tn antibodies as cancer biomarkers. Six different MBs were used in the study, i.e. carboxy-modified MBs of 250 nm, 500 nm, 1000 nm and 2800 nm and epoxy-modified MBs of 2800 nm and 4500 nm. In order to evaluate which MBs are the best suited for detection of the analyte anti-Tn antibodies, sensitivities of detection (slopes from calibration curves) were calculated. Next, specific sensitivities were calculated for each type of MBs as a ratio of sensitivity of detection to the mass of MBs. From zeta potential zeta for each type of MBs, the interfacial charge density on MBs was calculated, expressed as the density of zeta potential zeta d (ratio of zeta potential to surface area of MBs, i.e. zeta d = zeta/A). Then, we evaluated the effect of size and zeta d on the specific sensitivity of detection of anti-Tn antibodies in order to understand the immobilisation process on nanoscale. We also identified an optimal value of zeta d on MBs; this was essential to achieve highly sensitive detection of the analyte, which made it possible to attain limit of detection (LOD) of (0.31 +/- 0.01) ng mL-1 or (2.10 +/- 0.04) pM for analyte detection. In addition, the optimal assay configuration was highly selective and enabled reliable detection of the analyte in human serum with a recovery index in the range of 102-104%.