Search Results (140)

Recent investigations into C-reactive protein (CRP) dynamics have shown that by evaluating the change in CRP level in a patient over time, it is possible to distinguish between bacterial and viral infections more accurately thereby guiding antimicrobial prescription practices. Consequently, a biosensor targeted towards CRP was developed using a nano- and microfiber mesh-based transducer. The produced transducers were functionalized with streptavidin, after which a biorecognition element, anti-CRP antibodies, could be bound to the sensor. Confirmation of the sensor production phases was obtained using fluorescence microscopy. The sensors were evaluated using Electrochemical Impedance Spectroscopy (EIS) and showed increasing changes in the impedance modulus corresponding to increasing concentrations of CRP in solution following a parabolic trend line. Full article

Micro(nano)plastics are important emerging contaminants and a current research hotspot in the environmental field. Micro(nano)plastics widely exist in various organic wastes such as waste sludge, food waste (FW) and livestock manure and often enter into digesters along with anaerobic digestion (AD) treatment of these wastes, thereby exerting extensive and profound influences on anaerobic process performance. This study reviews sources of micro(nano)plastics and their pathways entering the anaerobic system and summarizes the quantities, sizes, shapes and micromorphology of various micro(nano)plastics in waste sludge, FW, livestock manure, yard waste and municipal solid waste. The current advances on the effects of multiple micro(nano)plastics mainly polyvinyl chloride (PVC), polystyrene (PS) and polyethylene (PE) with different sizes and quantities (or concentrations) on AD of organic wastes in terms of methane production, organic acid degradation and process stability are comprehensively overviewed and mechanisms of micro(nano)plastics affecting AD involved in microbial cells, key enzymes, microbial communities and antibiotic resistance genes are analyzed. Meanwhile, coupling effects of micro(nano)plastics with some typical pollutants such as antibiotics and heavy metals on AD are also reviewed. Due to the extreme complexity of the anaerobic system, current research still lacks full understanding concerning composite influences of different types, sizes and concentrations of micro(nano)plastics on AD under various operating modes. Future research should focus on elucidating mechanisms of micro(nano)plastics affecting organic metabolic pathways and the expression of specific functional genes of microorganisms, exploring the fate and transformation of micro(nano)plastics along waste streams including but not limited to AD, investigating the interaction between micro(nano)plastics and other emerging contaminants (such as perfluorooctanoic acid and perfluorooctane sulphonate) and their coupling effects on anaerobic systems, and developing accurate detection and quantification methods for micro(nano)plastics and technologies for eliminating the negative impacts of micro(nano)plastics on AD. Full article

Long non-coding RNAs (lncRNAs) are increasingly recognized as key regulators of gene expression and cellular signaling in cancer. Their functions are primarily mediated through interactions with specific protein partners that modulate chromatin structure, epigenetic remodeling, transcription, and signal transduction. In this review, we explore reports and strategies for the proteomic characterization of lncRNA-associated proteins, particularly emphasizing high-throughput liquid chromatography–mass spectrometry (LC-MS)-based techniques. Affinity-based methods such as RNA pull-down, ChIRP MS, RAP-MS, BioID-MS, and SILAC-MS enable sensitive and specific mapping of lncRNA and protein complexes. These approaches reveal cancer-specific proteomic signatures, post-translational modifications, and mechanistic insights into tumor biology. The use of label-free quantification, bituminization, and crosslinking strategies further enhances the resolution of dynamic RNA–protein networks. Validation tools following bioinformatic analyses, such as Western blotting, immunohistochemistry, immunofluorescence, and ELISA, are used to prioritize and confirm findings. Candidate biomarkers from hepatocellular carcinoma to colorectal and prostate cancers, profiling lncRNA-associated proteins, hold promise for identifying clinically actionable biomarkers and therapeutic targets. This review highlights the translational relevance of lncRNA protein studies and advocates for their broader adoption in oncological research. In LC-MS workflows, proteins bound to lncRNAs are enzymatically digested into peptides, separated via nano-LC, and analyzed using high-resolution tandem MS. Label-free or isotope-labeled methods quantify differential enrichment, followed by bioinformatics-driven pathway annotation. Full article

A simple and efficient dispersive micro solid-phase extraction (DMSPE) method using nano-TiO₂ as a sorbent was developed for the separation and preconcentration of (ultra) trace levels of lead in water samples prior to quantification by electrothermal atomic absorption spectrometry (ETAAS). Key experimental parameters affecting the DMSPE process, including pH, ionic strength, sorbent dosage, and preconcentration factor, were optimized. The optimized method demonstrated a preconcentration factor of 20, a relative standard deviation below 4.5%, and a detection limit of 0.11 µg/L. The procedure was validated using certified reference material (CRM TM-25.5) and applied to real water samples from a lake, a residential well, and industrial wastewater. Satisfactory recoveries (89–103%) confirmed the reliability of the method for the determination of low lead concentrations in complex matrices. Full article

Electroanalysis of monoamine neurotransmitters is a useful tool for monitoring relevant neurodegenerative disorders and diseases. Electroanalysis of neurotransmitters using analytical devices consisting of electrodes modified with tailored and nanostructured composite materials is an active research topic nowadays. Nano- and microstructured composite materials composed of various organic conductive polymers, metal/metal oxide nanoparticles, and carbonaceous materials enable an increase in the performance of electroanalytical sensing devices. Synergistic properties resulting from the combination of various pristine nanomaterials have enabled faster kinetics and increased overall performance. Herein, recent results related to the design and elaboration of electroanalytical sensing devices based on cost-effective and reliable nano- and microstructured composite materials for the quantification of monoamine neurotransmitters are presented. The discussion focuses on the fabrication procedures and detection strategies, highlighting the capabilities of the analytical platforms used in the determination of relevant analytes. The review aims to present the main benefits of using composite nanostructured materials in the electroanalysis of monoamine neurotransmitters. Full article

Background: Previous proteomic studies provided valuable information about cataracts, but unclarified issues, such as sex and ethnicity-associated differences, remain. This study aimed to provide additional data on cataract-related proteins regarding age, sex, and cataract type. Methods: Twenty-six female and seven male Turkish cataract patients were screened for visual acuity and dysfunctional lens index. A nano-LC-MS/MS system and Progenesis QI software v3.0 were used for protein identification and quantification. The remaining data were evaluated with SPSS Version 29.0 software. Results: Proteins that showed age-associated changes were mainly involved in cytoskeletal organization. A glyoxalase enzyme, caveolin 1, and HS90B were lower, and RAB8B and ATP6V1B1 were higher in lenses in women. Proteins with lower levels in cataractous lenses than in transparent lenses included filensin and phakinin, concurrent with previous publications, and LCTL, GDI, HSPB1, and EIF4A2, not reported before. Corticonuclear cataracts constituted the only group showing depletions in putatively protective proteins, while the cortical type was the least influenced. ANXA1 and DNHD1 positively, and TCPD, SEC14L2, and PRPS1 proteins negatively correlated with visual acuity. Conclusions: This study revealed cataract-related proteins concurrent with earlier studies and new ones hitherto unreported. Despite the low number of patients investigated, the results merit further research, as these new proteins are highly likely to be involved in cataractogenesis. Full article

Background/Objectives: Carbohydrate antigen 19-9 (CA19-9) is a clinically established biomarker primarily used for monitoring disease progression and recurrence in pancreatic and gastrointestinal cancers. Accurate and continuous quantification of CA19-9 in patient samples is critical for effective clinical management. This study aimed to develop dual-emitting molecularly imprinted nanopolymers (dual@nanoMIPs) for ratiometric and reliable detection of CA19-9 in serum. Methods: Dual-emitting nanoMIPs were synthesized via a one-step molecular imprinting process, incorporating both blue-emitting carbon dots (b-CDs) as internal reference fluorophores and yellow-emitting quantum dots (y-QDs) as responsive probes. The CA19-9 template was embedded into the polymer matrix to create specific recognition sites. Fluorescence measurements were carried out under 365 nm excitation in 1% human serum diluted in phosphate-buffered saline (PBS). Results: The dual@nanoMIPs exhibited a ratiometric fluorescence response upon CA19-9 binding, characterized by the emission quenching of the y-QDs at 575 nm, while the b-CDs emission remained stable at 467 nm. The fluorescence shift observed in the RGB coordinates from yellow to green in the concentration range of CA19-9 tested, improved quantification accuracy by compensating for matrix effects in serum. A linear detection range was achieved from 4.98 × 10⁻³ to 8.39 × 10² U mL⁻¹ in serum samples, with high specificity and reproducibility. Conclusions: The dual@nanoMIPs developed in this work enable a stable, sensitive, and specific detection of CA19-9 in minimally processed serum, offering a promising tool for longitudinal monitoring of cancer patients. Its ratiometric fluorescence design enhances reliability, supporting clinical decision-making in the follow-up of pancreatic cancer. Full article

The tissue regenerative potential of the liquid component of mesenchymal stem cells has gained significant attention. Stem cells from human exfoliated deciduous teeth-conditioned medium (SHED-CM), which is often extracted during orthodontic treatment, promotes bone regeneration. However, further investigation is warranted to determine which component of SHED-CM affects bone regeneration. Therefore, we focused on the extracellular vesicles contained in SHED-CM (SHED-EVs) and aimed to study their effects on osteoblasts. SHED-EVs were isolated using a pellet-down EV extraction kit and identified using transmission electron microscopy and NanoSight. SHED-EVs were added to human calvarial osteoblasts (HCOs), and cell proliferation and migration ability were examined with Incucyte^® and BrdU. Alkaline phosphatase (ALP) expression was confirmed using real-time PCR and ALP quantification. The bone differentiation potential was examined using Alizarin Red S (ARS) staining. SHED-EVs promoted proliferation and migration of HCOs. Real-time PCR and ALP quantification results demonstrated that HCOs cultured with SHED-EVs exhibited increased ALP expression. ARS staining revealed that SHED-EVs promoted bone differentiation of HCOs. These results suggest that SHED-EVs promote cell proliferation and migration and bone regeneration of osteoblasts, highlighting their potential in the development of bone regeneration therapies. Full article

Upon engagement of one of the nineteen secreted Wnt signaling proteins with one of the ten Frizzled transmembrane Wnt receptors (FZD_1–10), a wide variety of cellular Wnt signaling responses can be elicited, the selectivity of which depends on the following: (1) the specific Wnt-FZD pairing, (2) the participation of Wnt co-receptors and (3) the cellular context. Co-receptors play a pivotal role in guiding the specificity of Wnt signaling, most notably between β-catenin-dependent and -independent pathways, where co-receptors such as LRP5/6 and ROR1/2/PTK7 play major roles, respectively. It remains less understood how specific Wnt/FZD combinations contribute to the selectivity of downstream Wnt signaling, and we lack accurate comparative data on their binding properties under physiological conditions. Here, using fluorescently tagged Wnt3a, Wnt5a and Wnt16 proteins and cell lines expressing HiBiT-tagged Frizzled, we build on our ongoing efforts to provide a complete overview of the biophysical properties of all Wnt/FZD interactions using full-length proteins. Our real-time NanoBRET analysis using living cells expressing low receptor levels provides more accurate quantification of binding and will help us understand how these binary engagements control Wnt signaling outputs. We also provide evidence that LRP6 regulates the binding affinity of Wnt/FZD interactions in the trimeric Wnt-FZD-LRP6 complex. Full article

Albuminuria is a sensitive biomarker of kidney dysfunction, and the albumin/creatinine ratio (ACR) is an essential measure for monitoring diabetic kidney disease (DKD). Abnormal levels can indicate a propensity for progressive renal failure and other complications such as cardiovascular diseases. This study employed UV/Visible spectroscopy to analyze aqueous urine samples spiked with bovine serum albumin (BSA) and creatinine at clinically relevant concentrations (0–30 mg/L for albumin and 600–1800 mg/L for creatinine) using a multivariate method. UV/Visible spectra of co-spiked samples recorded in triplicate revealed distinct bands at 229 nm and 249 nm, corresponding to BSA and creatinine, respectively, alongside other amino acid bands. Partial Least Squares Regression (PLS-R) analysis for BSA yielded a Root Mean Square Error of Calibration (RMSEC) and Cross-Validation (RMSECV) values of 66.93 and 73.92 mg/L, respectively. For creatinine, RMSEC and RMSECV values were 244.32 and 275.65 mg/L, respectively. Prediction models for both BSA and creatinine compared to ELISA demonstrated a robust performance with R²_PRED values of 0.96 and 0.95, respectively, indicating strong model reliability. The Limit of Detection (LOD) for co-spiked samples was 19.82 mg/L for BSA and 58.43 mg/L for creatinine. The significance of the achieved Limit of Detection (LOD) lies in its ability to measure concentrations well below the normal physiological ranges of 0–30 mg/L for albumin and 600–1800 mg/L for creatinine. These results demonstrate the proof of concept of applying an UV/Visible-spectroscopy-based method as a rapid, cost-effective point-of-care (PoC) tool for ACR measurements, offering promising applications in the early diagnosis, monitoring, and prognosis of diabetic kidney disease and associated cardiovascular complications. The next stage will involve a pilot trial to evaluate the technology’s potential using clinical patients. Full article

Background: Despite advances in immunotherapy, non-small-cell lung carcinoma (NSCLC)’s clinical success is limited, possibly due to substantial immunological alterations in advanced cancer patients. This study examines the immunomodulatory effects of sEVs derived from lung adenocarcinoma (ADC) and squamous cell carcinoma (SCC) on T cells. Methods: SEVs were isolated from lung cancer cell lines and Jurkat-E6.1. SEV size and morphology were analyzed by NTA and TEM, respectively, while Western blotting confirmed sEV markers. SEV uptake was assessed, followed by resazurin assay, RNA isolation, quantification, cDNA preparation, RT-PCR, nano LC-MS, and bioinformatic analysis, before and after treating Jurkat-E6.1 cells with sEVs from A549 and SKMES1. Results: Cancer-derived sEVs were efficiently internalized by immune cells, reducing T-cell viability. The real-time PCR analysis showed downregulation of KI67, BCL2, BAX, TNFA, IL6, TGFβ, and IL10, suggesting reduced proliferation, dysregulated apoptosis, and impaired inflammatory and immunosuppressive signaling, and the upregulation of GZMB and IL2 suggests retained cytotoxic potential but possibly dysfunctional T-cell activation. Proteomic analysis revealed 39 differentially abundant proteins (DAPs) in ADC-treated T cells and 276 in SCC-treated T cells, with 19 shared DAPs. Gene Ontology (GO) analysis of these DAPs highlighted processes such as sEV biogenesis, metabolic pathways, and regulatory functions, with ADC sEVs influencing NAD metabolism, ECM binding, and oxidoreductase activity, while SCC sEVs affected mRNA stability, amino acid metabolism, and cadherin binding. The cytoplasmic colocalization suggests the presence of these proteins in the cellular and extracellular lumen, indicating the potential of further release of these proteins in the vesicles by T cells. Conclusion: Lung cancer-derived sEVs regulate T-cell activities through immunoregulatory signaling. The molecular interactions between sEVs and immune cells can reveal novel tumor immune regulatory mechanisms and therapeutic targets. Full article

The worldwide adoption of artificial turf in sports facilities and urban landscapes, alongside the systematic transition from natural grass and soil-based grounds, has raised growing concerns about its contribution to the significant source of nano- and microplastics in ecosystems. This review examines current knowledge on the mechanisms of nano- and microplastic generation from artificial turf systems and their environmental impacts. Combined mechanical stress, ultra-violet radiation, and weathering processes contribute to the breakdown of synthetic grass fibers and infill materials, generating particles ranging from nanometer to millimeter scales. These nano- and microplastics are detected in drainage systems and surrounding soils near sports facilities. Laboratory studies demonstrate that artificial turf-derived nano- and microplastics can adversely affect soil microbial communities, aquatic organisms, and potentially human health, through various exposure pathways. While current mitigation approaches include hybrid turf, particle retention systems, and improved maintenance protocols, emerging research focuses on developing novel, environmentally friendly materials as alternatives to conventional synthetic turf components. However, field data on emission rates and environmental fate remain limited, and standardized methods for particle characterization and quantification are lacking. This review identifies critical knowledge gaps, underscoring the need for comprehensive research on long-term ecological impacts and highlights the future goal of mitigating nano- and microplastic emissions from artificial turf systems into the ecosystem. Full article

This study presents a digital surface-enhanced Raman scattering (SERS) method to enhance the sensitivity of SERS detection for low-concentration analytes. Conventional SERS analysis using average Raman intensity faces limitations in distinguishing low concentrations due to the substrate’s sparse distribution of target molecules. To overcome this challenge, we used a binary code-based data analysis approach. Gold nano popcorn substrates were utilized for high-sensitivity detection, with malachite green isothiocyanate (MGITC) as the target molecule. Raman mapping data were analyzed using both the conventional average Raman intensity method and the proposed digital SERS approach. In the digital SERS method, a threshold value was established based on the mean and standard deviation of Raman signals in the absence of target molecules. Pixels with Raman intensities exceeding this threshold were assigned a value of “1”, while those below were assigned “0”. Quantification was then performed based on these digital counts corresponding to MGITC concentrations. Our results demonstrate that the digital SERS method significantly improved the ability to distinguish and quantify analytes in low-concentration ranges that were indiscernible using the conventional approach. This analytical technique shows promise for ultrasensitive chemical analysis and expands the capabilities of SERS-based detection methods, potentially revolutionizing the field of trace analyte detection. Full article

Osteoarthritis, affecting over 8 million people in the UK, remains a debilitating condition with limited treatment options. Current therapies primarily address symptoms and can exacerbate joint damage over time. Developing disease-modifying drugs that alleviate inflammation and promote joint regeneration is crucial for long-term patient benefit. This study investigates the potential of exosome-like nano-vesicles isolated from grapefruit juice (GEVs) as a novel therapeutic approach for osteoarthritis. GEVs possess regenerative properties and present a promising avenue for clinical translation. In this study, nano-vesicles were isolated and characterized in terms of protein quantification, size, and morphology. In vitro studies demonstrated the safety and efficacy of GEVs, showing an enhancement in human chondrocyte migratory activity of over 13%. GEVs exhibited a dual mechanism of action, reducing inflammation and oxidative stress while promoting cellular regeneration. Specifically, they reduced the expression of COX2 and PTGS2, markers associated with inflammation and pain sensitization, and enhanced the expression of antioxidant genes SD2 and GPX in osteoarthritic-like chondrocytes. Additionally, GEVs downregulated the expression of ADAMTS-5 and hypertrophic COL10 while upregulating chondrogenic markers ACAN, COL2, and SOX9. This research signifies a significant advancement in osteoarthritis therapy, offering a natural, safe, and cost-effective treatment option with the potential for long-lasting benefits. Clinical translation of GEV therapy holds promise for improving patient outcomes and reducing the burden on healthcare systems. Full article

This study critically examines the limitations of the official Italian methodology used for detecting bovine adulteration milk in Protected Designation of Origin (PDO) Mozzarella di Bufala Campana (MdBC). This method focuses on the whey fraction of cheese samples, which comprises about 1% of total MdBC proteins, and is based on a high-performance liquid chromatography (HPLC) quantification of the bovine β-lactoglobulin A (β-Lg A) as a marker. Here, we have demonstrated that this official methodology suffers from measurement inconsistencies due to its reliance on raw bovine whey standards, which fail to account for β-Lg genetic polymorphisms in real MdBC samples and protein thermal modifications during cheesemaking. To overcome these limitations, we propose a dual proteomics-based approach using matrix-assisted laser desorption ionization (MALDI-TOF) mass spectrometry (MS) and nano-HPLC-electrospray (ESI)−tandem mass spectrometry (MS/MS) analysis of MdBC extracted whey. MALDI-TOF-MS focused on identifying proteotypic peptides specific to bovine and buffalo β-Lg and α-lactalbumin (α-La), enabling high specificity for distinguishing the two animal species at adulteration levels as low as 1%. Complementing this, nano-HPLC-ESI-MS/MS provided a comprehensive profile by identifying over 100 bovine-specific peptide markers from β-Lg, α-La, albumin, lactoferrin, and osteopontin. Both methods ensured precise detection and quantification of bovine milk adulteration in complex matrices like pasta filata cheeses, achieving high sensitivity even at minimal adulteration levels. Accordingly, the proposed dual proteomics-based approach overcomes challenges associated with whey protein polymorphism, heat treatment, and processing variability, and complements casein-based methodologies already validated under European standards. This integrated framework of analyses focused on whey and casein fraction enhances the reliability of adulteration detection and safeguards the authenticity of PDO buffalo mozzarella, upholding its unique quality and integrity. Full article