Label-free approaches for extracellular vesicle detection

Extracellular vesicles (EVs) represent pivotal mediators in cell-to-cell communication. They are lipid-membranous carriers of several biomolecules, which can be produced by almost all cells. In the current Era of precision medicine, EVs gained growing attention thanks to their potential in both biomarker discovery and nanotherapeutics applications. However, current technical limitations in isolating and/or detecting EVs restrain their standard use in clinics. This review explores all the state-of-the-art analytical technologies which are currently overcoming these issues. On one end, several innovative optical-, electrical- and spectroscopy-based detection methods represent advantageous label-free methodologies for faster EV detection. On the other end, microfluidics-based lab-on-a-chip tools support EV purification from low-concentrated samples. Altogether, these technologies will strengthen the routine application of EVs in clinics

Gas sensing technologies -- status, trends, perspectives and novel applications

The strong, continuous progresses in gas sensors and electronic noses resulted in improved performance and enabled an increasing range of applications with large impact on modern societies, such as environmental monitoring, food quality control and diagnostics by breath analysis. Here we review this field with special attention to established and emerging approaches as well as the most recent breakthroughs, challenges and perspectives. In particular, we focus on (1) the transduction principles employed in different architectures of gas sensors, analysing their advantages and limitations; (2) the sensing layers including recent trends toward nanostructured, low-dimensional and composite materials; (3) advances in signal processing methodologies, including the recent advent of artificial neural networks. Finally, we conclude with a summary on the latest achievements and trends in terms of applications.Comment: arXiv admin comment: This version has been removed by arXiv administrators as the submitter did not have the rights to agree to the license at the time of submissio

Improving the early diagnostic of prostate cancer by multiple biomarker detection with new biosensing devices

Prostate cancer (PCa) is the most common form of cancer in men, in Europe (World Health Organization data). The most recent statistics, in Portuguese territory, confirm this scenario, which states that about 50% of Portuguese men may suffer from prostate cancer and 15% of these will die from this condition. Its early detection is therefore fundamental. This is currently being done by Prostate Specific Antigen (PSA) screening in urine but false positive and negative results are quite often obtained and many patients are sent to unnecessary biopsy procedures. This early detection protocol may be improved, by the development of point-of-care cancer detection devices, not only to PSA but also to other biomarkers recently identified. Thus, the present work aims to screen several biomarkers in cultured human prostate cell lines, serum and urine samples, developing low cost sensors based on new synthetic biomaterials. Biomarkers considered in this study are the following: prostate specific antigen (PSA), annexin A3 (ANXA3), microseminoprotein-beta (MSMB) and sarcosine (SAR). The biomarker recognition may occurs by means of molecularly imprinted polymers (MIP), which are a kind of plastic antibodies, and enzymatic approaches. The growth of a rigid polymer, chemically stable, using the biomarker as a template allows the synthesis of the plastic antibody. MIPs show high sensitivity/selectivity and present much longer stability and much lower price than natural antibodies. This nanostructured material was prepared on a carbon solid. The interaction between the biomarker and the sensing-material produces electrical signals generating quantitative or semi-quantitative data. These devices allow inexpensive and portable detection in point-of-care testing

Electronic nose implementation for biomedical applications

The growing rate of diabetes and undiagnosed diabetes related diseases is becoming a worldwide major health concern. The motivation of this thesis was to make use of a technology called the ‘electronic nose’ (eNose) for diagnosing diseases. It presents a comprehensive study on metabolic and gastro-intestinal disorders, choosing diabetes as a target disease. Using eNose technology with urinary volatile organic compounds (VOCs) is attractive as it allows non-invasive monitoring of various molecular constituents in urine. Trace gases in urine are linked to metabolic reactions and diseases. Therefore, urinary volatile compounds were used for diagnosis purposes in this thesis. The literature on existing eNose technologies, their pros and cons and applications in biomedical field was thoroughly reviewed, especially in detecting headspace of urine. Since the thesis investigates urinary VOCs, it is important to discover the stability of urine samples and their VOCs in time. It was discovered that urine samples lose their stability and VOCs emission after 9 months. A comprehensive study with 137 diabetic and healthy control urine samples was done to access the capability of commercially available eNose instruments for discrimination between these two groups. Metal oxide gas sensor based commercial eNose (Fox 4000, AlphaMOS Ltd) and field asymmetric ion mobility spectrometer (Lonestar, Owlstone Ltd) were used to analyse volatiles in urinary headspace. Both technologies were able to distinguish both groups with sensitivity and specificity of more than 90%. Then the project moved onto developing a Non-dispersive infrared (NDIR) sensor system that is non-invasive, low-cost, precise, rapid, simple and patient friendly, and can be used at both hospitals and homes. NDIR gas sensing is one of the most widely used optical gas detection techniques. NDIR system was used for diagnosing diabetes and gastro related diseases from patient’s wastes. To the best of the authors’ knowledge, this is the first and only developed tuneable NDIR eNose system. The developed optical eNose is able to scan the whole infrared range between 3.1μm and 10.5 μm with step size of 20 nm. To simulate the effect of background humidity and temperature on the sensor response, a gas test rig system that includes gas mixture, VOC generator, humidity generator and gas analyser was designed to enable the user to have control of gas flow, humidity and temperature. This also helps to find out system’s sensitivity and selectivity. Finally, after evaluating the sensitivity and selectivity of optical eNose, it was tested on simple and complex odours. The results were promising in discriminating the odours. Due to insufficient sample batches received from the hospital, synthetic urine samples were purchased, and diabetic samples were artificially made. The optical eNose was able to successfully separate artificial diabetic samples from non-diabetic ones

Recent developments in biosensing methods for extracellular vesicle protein characterization

Research into extracellular vesicles (EVs) has grown significantly over the last few decades with EVs being widely regarded as a source of biomarkers for human health and disease with massive clinical potential. Secreted by every cell type in the body, EVs report on the internal cellular conditions across all tissue types. Their presence in readily accessible biofluids makes the potential of EV biosensing highly attractive as a noninvasive diagnostic platform via liquid biopsies. However, their small size (50-250 nm), inherent heterogeneity, and the complexity of the native biofluids introduce challenges for effective characterization, thus, limiting their clinical utility. This has led to a surge in the development of various novel EV biosensing techniques, with capabilities beyond those of conventional methods that have been directly transferred from cell biology. In this review, key detection principles used for EV biosensing are summarized, with a focus on some of the most recent and fundamental developments in the field over the last 5 years. This article is categorized under: Diagnostic Tools > Biosensing Diagnostic Tools > In Vitro Nanoparticle-Based Sensing

Biomarkers Used for the Diagnosis of Diseases

The detection and quantification of with high precision nucleic acid biomarkers and protein biomarkers in resource-limited settings is key to the early diagnosis of diseases and for monitoring the effects of treatments. As there is an enormous demand for high-quality biomarker detection platforms that are robust and highly applicable in resource-limited settings, this book is devoted to exploring methods for detection and quantification of biomarkers, focusing on the recent advances in this field

Advances in Chemical Analysis Procedures (Part I)

The availability (and the development) of innovative approaches to quantitative analyses and the data processing are often mandatory to deeply characterize a sample and to correctly highlight the analytical target. These objectives are carried out either by simply improving a single aspect of the analytical protocol or by developing a synergy of steps (from extraction to instrumental configuration to chemometric approaches) to obtain the maximum analytical information sought. Examples are innovative extraction protocols (also following the recent guidelines on green analytical chemistry) or new materials for the selective extraction of target compounds, multi-analytes screening methods, and "untargeted" approaches for food applications. In this text, the various articles are attributable to these elements, in particular, we start with a multi-analyte method for the determination of 10 different cannabinoids from Cannabis sativa L. by means of conventional techniques (Mandrioli and coworkers), to then see the application of techniques hyphenated "ultra-fast" by UPLC-MS for the authentication of food products (Xue and coworkers). The work of Song and coworkers on these applications in food products is also interesting, as it highlights how the collection process (and the timing of this passage) can affect the chemical profile and, consequently, the biological activity of Panax ginseng. Mocan and coworkers, applying an innovative extraction technique based on microwaves and applying well-known, robust, and easy-to-use instrumentation, have demonstrated how it is possible to discriminate between various species of Galium and how the chemical profiles obtained can support the biological activities observed. Similarly, but with the aim of developing new sample pretreatment procedures, Maggira and collaborators have developed graphene oxide-based materials for the selective extraction of sulfonamides in milk. Shen and coworkers apply a different type of approach, the "untargeted" one, for the geographical characterization of the Gentian Rigescens for which they combine chemometric techniques for the processing of raw chemical profile data. Wang and coworkers report a multiclass screening of drugs with high-resolution mass spectrometry through which they manage to obtain a high-scale, fast screening method for pesticides in fishery drugs based on ultrahigh-performance liquid chromatography tandem quadrupole-orbitrap high-resolution mass spectrometer

Review—Non-Invasive Monitoring of Human Health by Exhaled Breath Analysis: A Comprehensive Review

Exhaled human breath analysis is a very promisingfield of research work having great potential for diagnosis of diseases in non-invasive way. Breath analysis has attracted huge attention in thefield of medical diagnosis and disease monitoring in the last twodecades. VOCs/gases (Volatile Organic Compounds) in exhaled breath bear thefinger-prints of metabolic and biophysicalprocesses going on in human body. It’s a non-invasive, fast, non-hazardous, cost effective, and point of care process for diseasestate monitoring and environmental exposure assessment in human beings. Some VOCs/gases in exhaled breath are bio-markers ofdifferent diseases and their presence in excess amount is indicative of un-healthiness. Breath analysis has the potential for earlydetection of diseases. However, it is still underused and commercial device is yet not available owing to multiferrious challenges.This review is intended to provide an overview of major biomarkers (VOCs/gases) present in exhaled breath, importance of theiranalysis towards disease monitoring, analytical techniques involved, promising materials for breath analysis etc. Finally, relatedchallenges and limitations along with future scope will be touched upon.will be touched upon