This article explores the development and application of innovative piezoelectric sensors in point-of-care diagnostics. It highlights the significance of bedside tests, such as lateral flow and electrochemical tests, in providing rapid and accurate results directly at the patient's location. This paper delves into the principles of piezoelectric assays, emphasizing their ability to detect disease-related biomarkers through mechanical stress-induced electrical signals. Various applications of piezoelectric chemosensors and biosensors are discussed, including their use in the detection of cancer biomarkers, pathogens, and other health-related analytes. This article also addresses the integration of piezoelectric materials with advanced sensing technologies to improve diagnostic accuracy and efficiency, offering a comprehensive overview of current advances and future directions in medical diagnostics.

• MeSH
• biologické markery analýza   MeSH
• biosenzitivní techniky * MeSH
• elektrochemické techniky MeSH
• lidé MeSH
• vyšetření u lůžka MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• přehledy MeSH

OBJECTIVES: Risk-based stratification approaches using measurable residual disease (MRD) successfully help to identify T-acute lymphoblastic leukemia (T-ALL) patients at risk of relapse, whose treatment outcomes are very poor. Because of T-ALL heterogeneity and rarity, a reliable and standardized approach for flow cytometry (FC)-based MRD measurement and analysis is often missing. METHODS: Within the international AIEOP-BFM-ALL-FLOW study group we made a consensus on markers and a standard operating procedure for common 8- and 12-color T-ALL MRD panels. Custom manufactured tubes with dried backbone antibodies were tested in parallel to local FC standards. RESULTS: Altogether, 66 diagnostic and 67 day 15 samples were analyzed. We designed two guided MRD gating strategies to identify blast cells in parallel to expert-based evaluation. We proved that the optimized tubes allowed the correct identification of blast cells in all diagnostic samples. Both, expert and guided analysis of day 15 samples correlated to local standard (Spearman R=0.98 and R=0.94, respectively). Only in 2 (3 %) and 4 (6 %) patients expert gating and guided analysis results were substantially discordant from local standard, respectively. The cases that require an individualized approach may be partially identified at diagnosis through a rare immunophenotype or mixed phenotype acute leukemia status. CONCLUSIONS: Our work shows that standardized operating procedures together with guided analysis are applicable in a great majority of T-ALL cases. Further improvement of MRD detection is needed, as in some cases an individualized analytical approach is still required due to the challenging nature of the T-ALL phenotype.

• MeSH
• dítě MeSH
• kojenec MeSH
• konsensus MeSH
• lidé MeSH
• lymfoblastická leukemie-lymfom z prekurzorových T-buněk * diagnóza   patologie   MeSH
• mladiství MeSH
• předškolní dítě MeSH
• průtoková cytometrie * normy   metody   MeSH
• reziduální nádor * diagnóza   MeSH
• Check Tag
• dítě MeSH
• kojenec MeSH
• lidé MeSH
• mladiství MeSH
• mužské pohlaví MeSH
• předškolní dítě MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• multicentrická studie MeSH

The review focuses on the design of detection cells, the use of microcontrollers for processing and evaluation of the detection signal, and the development of multi-detection systems for electromigration, liquid chromatography, flow-through and microfluidic techniques. A separate section is the introduction of modern 3D printing techniques and the use of new printing materials for the design of multidetection systems. In addition to traditional utilisation in separation techniques, new versions of contactless conductivity detectors are finding applications in FIA, SIA, portable and paper based analytical systems or as independent sensors. Applicationwise, C4Ds find new use in gas detection, segmented flow monitoring, as part of point of care devices, and in many other biomedical, environmental, agricultural and industrial applications.

• Publikační typ
• časopisecké články MeSH
• přehledy MeSH

Early-stage diagnosis of prostatic carcinoma is essential for successful treatment and, thus, significant prognosis improvement. In laboratory practice, the standard non-invasive diagnostic approach is the immunochemical detection of the associated biomarker, prostate-specific antigen (PSA). Ultrasensitive detection of PSA is essential for both diagnostic and recurrence monitoring purposes. To achieve exceptional sensitivity, we have developed a microfluidic device with a flow-through cell for single-molecule analysis using photon-upconversion nanoparticles (UCNPs) as a detection label. For this purpose, magnetic microparticles (MBs) were first optimized for the capture and preconcentration of PSA and then used to implement a bead-based upconversion-linked immunoassay (ULISA) in the microfluidic device. The digital readout based on counting single nanoparticle-labeled PSA molecules on MBs enabled a detection limit of 1.04 pg mL-1 (36 fM) in 50% fetal bovine serum, which is an 11-fold improvement over the respective analog MB-based ULISA. The microfluidic technique conferred several other advantages, such as easy implementation and the potential for achieving high-throughput analysis. Finally, it was proven that the microfluidic setup is suitable for clinical sample analysis, showing a good correlation with a reference electrochemiluminescence assay (recovery rates between 97% and 105%).

• MeSH
• imunoanalýza přístrojové vybavení   metody   MeSH
• lidé MeSH
• limita detekce MeSH
• mikrofluidní analytické techniky přístrojové vybavení   MeSH
• nádory prostaty diagnóza   krev   MeSH
• nanočástice chemie   MeSH
• prostatický specifický antigen * analýza   krev   MeSH
• Check Tag
• lidé MeSH
• mužské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Advances in single-cell level analytical techniques, especially cytometric approaches, have led to profound innovation in biomedical research, particularly in the field of clinical immunology. This has resulted in an expansion of high-dimensional data, posing great challenges for comprehensive and unbiased analysis. Conventional manual analysis is thus becoming untenable to handle these challenges. Furthermore, most newly developed computational methods lack flexibility and interoperability, hampering their accessibility and usability. Here, we adapted Seurat, an R package originally developed for single-cell RNA sequencing (scRNA-seq) analysis, for high-dimensional flow cytometric data analysis. Based on a 20-marker antibody panel and analyses of T-cell profiles in both adult blood and cord blood (CB), we showcased the robust capacity of Seurat in flow cytometric data analysis, which was further validated by Spectre, another high-dimensional cytometric data analysis package, and conventional manual analysis. Importantly, we identified a unique CD8+ T-cell population defined as CD8+CD45RA+CD27+CD161+ T cell that was predominantly present in CB. We characterised its IFN-γ-producing and potential cytotoxic properties using flow cytometry experiments and scRNA-seq analysis from a published dataset. Collectively, we identified a unique human CB CD8+CD45RA+CD27+CD161+ T-cell subset and demonstrated that Seurat, a widely used package for scRNA-seq analysis, possesses great potential to be repurposed for cytometric data analysis. This facilitates an unbiased and thorough interpretation of complicated high-dimensional data using a single analytical pipeline and opens a novel avenue for data-driven investigation in clinical immunology.

• MeSH
• analýza jednotlivých buněk metody   MeSH
• antigeny CD27 metabolismus   imunologie   MeSH
• antigeny CD45 * metabolismus   imunologie   MeSH
• CD8-pozitivní T-lymfocyty * imunologie   MeSH
• dospělí MeSH
• fetální krev * imunologie   cytologie   MeSH
• imunofenotypizace metody   MeSH
• lektinové receptory NK-buněk - podrodina B imunologie   metabolismus   MeSH
• lidé MeSH
• průtoková cytometrie * metody   MeSH
• software MeSH
• T-lymfocyty - podskupiny imunologie   metabolismus   MeSH
• Check Tag
• dospělí MeSH
• lidé MeSH
• Publikační typ
• časopisecké články MeSH

BACKGROUND: The number of older people with cognitive impairment is increasing worldwide. Impaired lung function might be associated with cognitive decline in older age; however, results from large longitudinal studies are lacking. In this study, we examined the longitudinal associations between pulmonary function and the trajectories of cognitive decline using prospective population-based SHARE data from 14 countries. METHODS: The analytic sample included N = 32,049 older adults (Mean age at baseline = 64.76 years). The dependent variable was cognitive performance, measured repeatedly across six waves in three domains: verbal fluency, memory, and numeracy. The main predictor of interest was peak expiratory flow (PEF). The data were analyzed in a multilevel accelerated longitudinal design, with models adjusted for a variety of covariates. RESULTS: A lower PEF score was associated with lower cognitive performance for each domain as well as a lower global cognitive score. These associations remained statistically significant after adjusting for all covariates Q4 vs Q1 verbal fluency: unstandardized coefficient B = -3.15; numeracy: B = -0.52; memory: B = -0.64; global cognitive score B = -2.65, all p < .001). However, the PEF score was not found to be associated with the rate of decline for either of the cognitive outcomes. CONCLUSIONS: In this large multi-national longitudinal study, the PEF score was independently associated with lower levels of cognitive functions, but it did not predict a future decline. The results suggest that pre-existing differences in lung functions are responsible for variability in cognitive functions and that these differences remained stable across aging.

• MeSH
• kognice MeSH
• kognitivní dysfunkce * epidemiologie   MeSH
• lidé MeSH
• longitudinální studie MeSH
• plíce MeSH
• prospektivní studie MeSH
• senioři MeSH
• stárnutí * psychologie   MeSH
• Check Tag
• lidé MeSH
• senioři MeSH
• Publikační typ
• časopisecké články MeSH

An instrument integrating thermal desorption (TD) to selected ion flow tube mass spectrometry (SIFT-MS) is presented, and its application to analyze volatile organic compounds (VOCs) in human breath is demonstrated for the first time. The rationale behind this development is the need to analyze breath samples in large-scale multicenter clinical projects involving thousands of patients recruited in different hospitals. Following adapted guidelines for validating analytical techniques, we developed and validated a targeted analytical method for 21 compounds of diverse chemical class, chosen for their clinical and biological relevance. Validation has been carried out by two independent laboratories, using calibration standards and real breath samples from healthy volunteers. The merging of SIFT-MS and TD integrates the rapid analytical capabilities of SIFT-MS with the capacity to collect breath samples across multiple hospitals. Thanks to these features, the novel instrument has the potential to be easily employed in clinical practice.

• MeSH
• dechové testy metody   MeSH
• hmotnostní spektrometrie metody   MeSH
• lidé MeSH
• těkavé organické sloučeniny * analýza   MeSH
• tělesné tekutiny * chemie   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• multicentrická studie MeSH
• práce podpořená grantem MeSH

Byla zavedena a validována jednoduchá metoda s použitím kapalinové chromatografie a tandemové hmotnostní spektrometrie (LC-MS/MS) pro stanovení kolistinu A a kolistinu B v plazmě. Extrakce proteinů z plazmy byla provedena pomocí 1 ml kazet Oasis HLB a chromatografická separace byla provedena na koloně Arion® Polar C18 (250 × 4,6 mm; 5 mm) při 35 °C. Mobilní fáze se skládala z vody obsahující 0,1 % kyseliny mravenčí a methanolu obsahujícího 0,1 % kyseliny mravenčí v poměru 40 : 60 (v/v), při průtoku 0,8 ml/minutu. Eluent byl detekován v režimu pozitivních iontů pomocí ionizace elektrosprejem s následujícími iontovými přechody m/z: kolistin A 585,55 → 101,05; kolistin B 578,5 → 101,15; a IS 602,4 → 101,1; 120,15; 86,15. Byly provedeny testy krátkodobé stability kolistinu a CMS, a to při pokojové teplotě a 37 °C, kdy se stabilita obou složek se zvyšující se teplotou snižuje. Předložený příspěvek je součástí studie Pharmacokinetics of Colistin in Critically Ill Patients With Extracorporeal Membrane Oxygenation (COL-ECMO2022), v níž budou prezentovány další výsledky.

A simple liquid chromatography tandem mass spectrometry (LC-MS/MS) assay for the determination of colistin A and colistin B in human plasma was developed and validated. Plasma extraction was performed using Oasis HLB 1 ml cartridges, analysis was performed using Arion® Polar C18 (250×4,6mm; 5mm) column at 35°C. Mobile phases consisted of water containing 0,1% formic acid and methanol containing 0,1% formic acid (40:60, v/v) delivered at a flow rate of 0,8 ml/minute. Eluent was detected in the positive ion mode using electrospray ionization at the following transitions of mass to charge (m/z): colistin A 585,55 → 101,05; colistin B 578,5 → 101,15; and IS 602,4 → 101,1; 120,15; 86,15. Short-term stability tests of colistin and CMS were performed, at room temperature and 37°C, where the stability of both components decreases with increasing temperature. The presented paper is part of the Pharmacokinetics of Colistin in Critically Ill Patients With Extracorporeal Membrane Oxygenation (COL-ECMO2022) study in which further results will be presented.

• MeSH
• chemické techniky analytické * metody   přístrojové vybavení   MeSH
• chromatografie kapalinová metody   přístrojové vybavení   MeSH
• hydrolýza MeSH
• kolistin * krev   MeSH
• lidé MeSH
• methansulfonáty krev   MeSH
• stabilita proteinů MeSH
• statistika jako téma MeSH
• tandemová hmotnostní spektrometrie MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• klinická studie MeSH
• práce podpořená grantem MeSH

Short-chain fatty acids (SCFAs) are the main metabolites produced by bacterial fermentation of non-digestible carbohydrates in the gastrointestinal tract. They can be seen as the major flow of carbon from the diet, through the microbiome to the host. SCFAs have been reported as important molecules responsible for the regulation of intestinal homeostasis. Moreover, these molecules have a significant impact on the immune system and are able to affect inflammation, cardiovascular diseases, diabetes type II, or oncological diseases. For this purpose, SCFAs could be used as putative biomarkers of various diseases, including cancer. A potential diagnostic value may be offered by analyzing SCFAs with the use of advanced analytical approaches such as gas chromatography (GC), liquid chromatography (LC), or capillary electrophoresis (CE) coupled with mass spectrometry (MS). The presented review summarizes the importance of analyzing SCFAs from clinical and analytical perspective. Current advances in the analysis of SCFAs focused on sample pretreatment, separation strategy, and detection methods are highlighted. Additionally, it also shows potential areas for the development of future diagnostic tools in oncology and other varieties of diseases based on targeted metabolite profiling.

• Publikační typ
• časopisecké články MeSH
• přehledy MeSH

OBJECTIVES: The analysis of organic acids in urine is an important part of the diagnosis of inherited metabolic disorders (IMDs), for which gas chromatography coupled with mass spectrometry is still predominantly used. METHODS: Ultra-performance liquid chromatography-tandem mass spectrometry (LC-MS/MS) assay for urinary organic acids, acylcarnitines and acylglycines was developed and validated. Sample preparation consists only of dilution and the addition of internal standards. Raw data processing is quick and easy using selective scheduled multiple reaction monitoring mode. A robust standardised value calculation as a data transformation together with advanced automatic visualisation tools are applied for easy evaluation of complex data. RESULTS: The developed method covers 146 biomarkers consisting of organic acids (n=99), acylglycines (n=15) and acylcarnitines (n=32) including all clinically important isomeric compounds present. Linearity with r2>0.98 for 118 analytes, inter-day accuracy between 80 and 120 % and imprecision under 15 % for 120 analytes were achieved. Over 2 years, more than 800 urine samples from children tested for IMDs were analysed. The workflow was evaluated on 93 patient samples and ERNDIM External Quality Assurance samples involving a total of 34 different IMDs. CONCLUSIONS: The established LC-MS/MS workflow offers a comprehensive analysis of a wide range of organic acids, acylcarnitines and acylglycines in urine to perform effective, rapid and sensitive semi-automated diagnosis of more than 80 IMDs.

• MeSH
• chromatografie kapalinová metody   MeSH
• dítě MeSH
• lidé MeSH
• metabolické nemoci * MeSH
• organické látky MeSH
• průběh práce MeSH
• tandemová hmotnostní spektrometrie * metody   MeSH
• Check Tag
• dítě MeSH
• lidé MeSH
• Publikační typ
• časopisecké články MeSH