Biosensors, Volume 11, Issue 2 (February 2021) – 32 articles

Vital signs not only reflect essential functions of the human body but also symptoms of a more serious problem within the anatomy; they are well used for physical monitoring, caloric expenditure, and performance before a possible symptom of a massive failure—a great variety of possibilities that together form a first line of basic diagnosis and follow-up on the health and general condition of a person. This review includes a brief theory about fiber optic sensors’ operation and summarizes many research works carried out with them in which their operation and effectiveness are promoted to register some vital sign(s) as a possibility for their use in the medical, health care, and life support fields. The review presents methods and techniques to improve sensitivity in monitoring vital signs, such as the use of doping agents or coatings for optical fiber (OF) that provide stability and resistance to the external factors from which they must be protected in in vivo situations. It has been observed that most of these sensors work with single-mode optical fibers (SMF) in a spectral range of 1550 nm, while only some work in the visible spectrum (Vis); the vast majority, operate through fiber Bragg gratings (FBG), long-period fiber gratings (LPFG), and interferometers. These sensors have brought great advances to the measurement of vital signs, especially with regard to respiratory rate; however, many express the possibility of monitoring other vital signs through mathematical calculations, algorithms, or auxiliary devices. Their advantages due to miniaturization, immunity to electromagnetic interference, and the absence of a power source makes them truly desirable for everyday use at all times. Full article

Paper substrates are promising for development of cost-effective and efficient point-of-care biosensors, essential for public healthcare and environmental diagnostics in emergency situations. Most paper-based biosensors rely on the natural capillarity of paper to perform qualitative or semi-quantitative colorimetric detections. To achieve quantification and better sensitivity, technologies combining paper-based substrates and electrical detection are being developed. In this work, we demonstrate the potential of electrical measurements by means of a simple, parallel-plate electrode setup towards the detection of whole-cell bacteria captured in nitrocellulose (NC) membranes. Unlike current electrical sensors, which are mostly integrated, this plug and play system has reusable electrodes and enables simple and fast bacterial detection through impedance measurements. The characterized NC membrane was subjected to (i) a biofunctionalization, (ii) different saline solutions modelling real water samples, and (iii) bacterial suspensions of different concentrations. Bacterial detection was achieved in low conductivity buffers through both resistive and capacitive changes in the sensed medium. To capture Bacillus thuringiensis, the model microorganism used in this work, the endolysin cell-wall binding domain (CBD) of Deep-Blue, a bacteriophage targeting this bacterium, was integrated into the membranes as a recognition bio-interface. This experimental proof-of-concept illustrates the electrical detection of 10⁷ colony-forming units (CFU) mL⁻¹ bacteria in low-salinity buffers within 5 min, using a very simple setup. This offers perspectives for affordable pathogen sensors that can easily be reconfigured for different bacteria. Water quality testing is a particularly interesting application since it requires frequent testing, especially in emergency situations. Full article

Microbial reactor sensors (based on freshly harvested intact microbial cells) or microbial membrane sensors (based on immobilized microbial cells) can be used as convenient instruments for studying processes that cause the response of a biosensor, such as the properties of enzymes or the characteristics of metabolism. However, the mechanisms of the formation of biosensors responses have not yet been fully understood to study only one of these processes. In this work, the results of studies on the formation of a response to juglone for intact and immobilized bacterial cells used as receptors are presented. It was shown that the contribution of reactive oxygen species (ROS) to the formation of the biosensor response depends on the culture receptor and the form of juglone, quinone, or phenolate used. The response to the quinone form of juglone both for intact and immobilized cells of catalase-positive actinobacterium is formed regardless of the presence of ROS. The response of freshly harvested intact actinobacterial cells was caused by the rate of the enzymatic conversion of juglone. The rate of the response of immobilized actinobacterial cells was influenced by the activity of transport systems and metabolism. The response of immobilized pseudomonad cells was caused by the transport of juglone into cells, the inhibitory effect of juglone-induced ROS, and juglone metabolism. Full article

Osteosarcoma (OSA) is a type of bone cancer that begins in the cells that form bones. OSA is a rare mesenchymal bone neoplasm derived from mesenchymal stem cells. Genome disorganization, chromosomal modifications, deregulation of tumor suppressor genes, and DNA repair defects are the factors most responsible for OSA development. Despite significant advances in the diagnosing and treatment of OSA, patients’ overall survival has not improved within the last twenty years. Lately, advances in modern nanotechnology have spurred development in OSA management and offered several advantages to overcome the drawbacks of conventional therapies. This technology has allowed the practical design of nanoscale devices combined with numerous functional molecules, including tumor-specific ligands, antibodies, anti-cancer drugs, and imaging probes. Thanks to their small sizes, desirable drug encapsulation efficiency, and good bioavailability, functionalized nanomaterials have found wide-spread applications for combating OSA progression. This review invokes the possible utility of engineered nanomaterials in OSA diagnosis and treatment, motivating the researchers to seek new strategies for tackling the challenges associated with it. Full article

In this report, we summarized our development of biosensors for Rhodamine 6G and in vitro glucose detection based on surface-enhanced Raman scattering technology. For the detection of both Rhodamine 6G and in vitro glucose, a nature-patterned substrate with gold films over nanostructures (NPS-AuFON) was used as the surface-enhanced Raman scattering sensor platform. The enhancement factor was calculated at 9 × 10⁷. In the processing of the substrate, cyclic voltammetry was used to form nano-gold particles under different conditions. The Rhodamine 6G and glucose detection were then achieved on this substrate. Furthermore, we combined the potentiostatic technique and electrochemical adsorption to best detect glucose in low concentrations. The glucose oxidation potential (100 mV) was used to capture glucose close to the surface of the NPS-AuFON. The quantitative detection of glucose in solution and in situ inspection were confirmed. Further, we determined that this surface modification technology can reach the goal of experiments set by the World Health Organization to judge whether or not a patient is a diabetic by detecting a glucose concentration of 11.1 mmol/L (mg/dL) at a minimum. Full article

This research reports a portable immunochip, based on quartz crystal microbalance (QCM) for label-free, low-cost qualitative detection of zearalenone (ZEN) in food samples. The experimental parameters in the functionalization and working process were evaluated in detail, in order to achieve a high accuracy and sensitivity. Under optimal conditions, the ZEN concentration at an inhibition ratio of 50% and 15% of the proposed QCM immunochip achieved 3.41 µg L⁻¹ and 0.37 µg L⁻¹, respectively. This portable QCM immunochip also exhibited high specificity, no obvious cross-reaction to five structural analogs of ZEN, and showed other mycotoxins. It could finish the whole qualitative measurement within 30 min, showed good stability during the processes of preparation (SD < 5%, n = 9), storage (frequency response >90%, in PBS at 4 °C for 15 days), and application (frequency response >90% after being reused 6 times). The developed QCM immunochip obtained accurate and repeatable recovery results in ZEN analysis in the chosen food samples (corn, wheat flour, soy sauce, and milk), which had a high correlation (R² = 0.9844) with that achieved by the HPLC–MS/MS method. In short, this work developed a portable, stable, and reproducible QCM immunochip that could be used for rapid, low-cost, and sensitively measurement of ZEN content in real food samples. Full article

In this study, we combined quantitative ultrashort echo time (UTE) magnetic resonance (MR) imaging and an investigation by a probing device with tri-axial force sensor to seek correlations with mechanical properties of human patellar cartilage for in situ evaluation of biomechanical properties. Cartilage blocks (15 × 20 × 5 mm³) were dissected from the patella of six donors; 5 mm square regions of interest from the cartilage blocks were imaged using UTE-MR imaging sequences (T2* and magnetization transfer ratio (MTR)), and mechanical properties were measured using a micro indentation device. Then, the vertical reaction force on the cartilage surface was measured while push-probing forward 3 mm with the probing device at a 30° tilt to the horizontal plane. The results showed a positive correlation between stiffness/elastic modulus and each predictor variable (UTE-T2* (r = 0.240 and 0.255, respectively, UTE-MTR (r = 0.378 and 0.379, respectively), and probing device force (r = 0.426 and 0.423, respectively). Furthermore, multiple linear regression analysis showed the combination of the three predictors had stronger correlation (adjusted r² = 0.314 (stiffness), 0.323 (elastic), respectively). Our results demonstrate the potential for these non- and less- invasive methods for in situ evaluation of the mechanical properties of cartilage tissue. Full article

Screen-printed technology has helped considerably to the development of portable electrochemical sensors since it provides miniaturized but robust and user-friendly electrodes. Moreover, this technology allows to obtain very versatile transducers, not only regarding their design, but also their ease of modification. Therefore, in the last decades, the use of screen-printed electrodes (SPEs) has exponentially increased, with ceramic as the main substrate. However, with the growing interest in the use of cheap and widely available materials as the basis of analytical devices, paper or other low-cost flat materials have become common substrates for SPEs. Thus, in this revision, a comprehensive overview on paper-based SPEs used for analytical proposes is provided. A great variety of designs is reported, together with several examples to illustrate the main applications. Full article

Tendons are collagenous musculoskeletal tissues that connect muscles to bones and transfer the forces necessary for movement. Tendons are susceptible to injury and heal poorly, with long-term loss of function. Mesenchymal stem cell (MSC)-based therapies are a promising approach for treating tendon injuries but are challenged by the difficulties of controlling stem cell fate and of generating homogenous populations of stem cells optimized for tenogenesis (differentiation toward tendon). To address this issue, we aim to explore methods that can be used to identify and ultimately separate tenogenically differentiated MSCs from non-tenogenically differentiated MSCs. In this study, baseline and tenogenically differentiating murine MSCs were characterized for dielectric properties (conductivity and permittivity) of their outer membrane and cytoplasm using a dielectrophoretic (DEP) crossover technique. Experimental results showed that unique dielectric properties distinguished tenogenically differentiating MSCs from controls after three days of tenogenic induction. A single shell model was used to quantify the dielectric properties and determine membrane and cytoplasm conductivity and permittivity. Together, cell responses at the crossover frequency, cell morphology, and shell models showed that changes potentially indicative of early tenogenesis could be detected in the dielectric properties of MSCs as early as three days into differentiation. Differences in dielectric properties with tenogenesis indicate that the DEP-based label-free separation of tenogenically differentiating cells is possible and avoids the complications of current label-dependent flow cytometry-based separation techniques. Overall, this work illustrates the potential of DEP to generate homogeneous populations of differentiated stem cells for applications in tissue engineering and regenerative medicine. Full article

This paper presents a lateral flow assay (LFA) for the quantitative, fluorescence-based detection of the cardiac biomarker troponin I (cTnI) that features an analytical strip made of cellulose filter paper. The results show that the wicking and test time are comparable to those obtained with conventional nitrocellulose (NC)-based LFAs. Further, the cellulose paper provides an excellent background with no auto-fluorescence that is very adequate in detecting fluorescent lines. While fluorescence that was generated with cellulose strips was lower when compared to that generated in NC strips, signals could be improved by layering carbon nanofibers (CNF) on the cellulose. A nonlinear behavior of the concentration–response relationship was observed for the LFA architectures with NC, cellulose, and cellulose-CNF in the 0 to 200 ng/mL cTnI concentration range. The measurements were consistent and characterized by coefficients of variation lower than 2.5%. Detection and quantitation limits that were in the range 1.28–1.40 ng/mL and 2.10–2.75 ng/mL were obtained for LFA with cellulose and cellulose CNF strips that are equivalent to the limits obtained with the standard NC LFA. Overall, we showed that commercially available filter paper can be used in the analytical strip of LFA. Full article

Halophilic bacteria are remarkable organisms that have evolved strategies to survive in high saline concentrations. These bacteria offer many advances for microbial-based biotechnologies and are commonly used for industrial processes such as compatible solute synthesis, biofuel production, and other microbial processes that occur in high saline environments. Using halophilic bacteria in electrochemical systems offers enhanced stability and applications in extreme environments where common electroactive microorganisms would not survive. Incorporating halophilic bacteria into microbial fuel cells has become of particular interest for renewable energy generation and self-powered biosensing since many wastewaters can contain fluctuating and high saline concentrations. In this perspective, we highlight the evolutionary mechanisms of halophilic microorganisms, review their application in microbial electrochemical sensing, and offer future perspectives and directions in using halophilic electroactive microorganisms for high saline biosensing. Full article

Influenza virus poses a threat to global health by causing seasonal outbreaks as well as three pandemics in the 20th century. In humans, disease is primarily caused by influenza A and B viruses, while influenza C virus causes mild disease mostly in children. Influenza D is an emerging virus found in cattle and pigs. To mitigate the morbidity and mortality associated with influenza, rapid and accurate diagnostic tests need to be deployed. However, the high genetic diversity displayed by influenza viruses presents a challenge to the development of a robust diagnostic test. Nucleic acid-based tests are more accurate than rapid antigen tests for influenza and are therefore better candidates to be used in both diagnostic and surveillance applications. Here, we review various nucleic acid-based techniques that have been applied towards the detection of influenza viruses in order to evaluate their utility as both diagnostic and surveillance tools. We discuss both traditional as well as novel methods to detect influenza viruses by covering techniques that require nucleic acid amplification or direct detection of viral RNA as well as comparing advantages and limitations for each method. There has been substantial progress in the development of nucleic acid-based sensing techniques for the detection of influenza virus. However, there is still an urgent need for a rapid and reliable influenza diagnostic test that can be used at point-of-care in order to enhance responsiveness to both seasonal and pandemic influenza outbreaks. Full article

Foodborne illnesses represent high costs worldwide in terms of medical care and productivity. To ensure safety along the food chain, technologies that help to monitor and improve food preservation have emerged in a multidisciplinary context. These technologies focus on the detection and/or removal of either biological (e.g., bacteria, virus, etc.) or chemical (e.g., drugs and pesticides) safety hazards. Imprinted polymers are synthetic receptors able of recognizing both chemical and biological contaminants. While numerous reviews have focused on the use of these robust materials in extraction and separation applications, little bibliography summarizes the research that has been performed on their coupling to sensing platforms for food safety. The aim of this work is therefore to fill this gap and highlight the multidisciplinary aspects involved in the application of imprinting technology in the whole value chain ranging from IP preparation to integrated sensor systems for the specific recognition and quantification of chemical and microbiological contaminants in food samples. Full article

This paper reports the analytical detection and energetic properties of a glucose-fed Direct Catalytic Fuel Cell (DCFC) operated in association with yeast cells (Saccharomyces Cerevisiae). The cell was tested in a potentiostatic mode, and the operating conditions were optimized to maximize the current produced by a given concentration of glucose. Results indicate that the DCFC is characterized by a glucose detection limit of the order to 21 mmol L⁻¹. The cell was used to estimate the “pool” of carbohydrate content in commercial soft drinks. Furthermore, the use of different carbohydrates, such as fructose and sucrose, has been shown to result in a good current yield. Full article

An origami three-dimensional design of a paper-based potentiometric sensor is described. In its simplest form, this electrochemical paper-based analytical device (ePAD) is made from three small parts of the paper. Paper layers are folded on each other for the integration of a solid contact ion selective electrode (here a carbon-paste composite electrode) and a solid-state pseudo-reference electrode (here writing pencil 6B on the paper), which are in contact with a hydrophilic channel fabricated on the middle part (third part) of the paper. In this case, the pseudo-reference and working electrodes are connected to the two sides of the hydrophilic channel and hence the distance between them is as low as the width of paper. The unmodified carbon paste electrode (UCPE) and modification with the crown ether benzo15-crown-5 (B15C5) represented a very high sensitivity to Cu (II) and Cd²⁺ ions, respectively. The sensor responded to H₂O₂ using MnO₂-doped carbon paste electrode (CPE). Furthermore, a biosensor was achieved by the addition of glucose oxidase to the MnO₂-doped CPE and hence made it selective to glucose with ultra-sensitivity. In addition to very high sensitivity, our device benefits from consuming a very low volume of sample (10.0 µL) and automatic sampling without need for sampling devices. Full article

Chlorophene is an important antimicrobial agent present in disinfectant products which has been related to health and environmental effects, and its detection has been limited to chromatographic techniques. Thus, there is a lack of research that attempts to develop new analytical tools, such as biosensors, that address the detection of this emerging pollutant. Therefore, a new biosensor for the direct detection of chlorophene in real water is presented, based on surface plasmon resonance (SPR) and using a laccase enzyme as a recognition element. The biosensor chip was obtained by covalent immobilization of the laccase on a gold-coated surface through carbodiimide esters. The analytical parameters accomplished resulted in a limit of detection and quantification of 0.33 mg/L and 1.10 mg/L, respectively, fulfilling the concentrations that have already been detected in environmental samples. During the natural river’s measurements, no significant matrix effects were observed, obtaining a recovery percentage of 109.21% ± 7.08, which suggested that the method was suitable for the fast and straightforward analysis of this contaminant. Finally, the SPR measurements were validated with an HPLC method, which demonstrated no significant difference in terms of precision and accuracy, leading to the conclusion that the biosensor reflects its potential as an alternative analytical tool for the monitoring of chlorophene in aquatic environments. Full article

Circulating tumour DNA (ctDNA) is widely used in liquid biopsies due to having a presence in the blood that is typically in proportion to the stage of the cancer and because it may present a quick and practical method of capturing tumour heterogeneity. This paper outlines a simple electrochemical technique adapted towards point-of-care cancer detection and treatment monitoring from biofluids using a label-free detection strategy. The mutations used for analysis were the KRAS G12D and G13D mutations, which are both important in the initiation, progression and drug resistance of many human cancers, leading to a high mortality rate. A low-cost DNA sensor was developed to specifically investigate these common circulating tumour markers. Initially, we report on some developments made in carbon surface pre-treatment and the electrochemical detection scheme which ensure the most sensitive measurement technique is employed. Following pre-treatment of the sensor to ensure homogeneity, DNA probes developed specifically for detection of the KRAS G12D and G13D mutations were immobilized onto low-cost screen printed carbon electrodes using diazonium chemistry and 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride/N-hydroxysuccinimide coupling. Prior to electrochemical detection, the sensor was functionalised with target DNA amplified by standard and specialist PCR methodologies (6.3% increase). Assay development steps and DNA detection experiments were performed using standard voltammetry techniques. Sensitivity (as low as 0.58 ng/μL) and specificity (>300%) was achieved by detecting mutant KRAS G13D PCR amplicons against a background of wild-type KRAS DNA from the representative cancer sample and our findings give rise to the basis of a simple and very low-cost system for measuring ctDNA biomarkers in patient samples. The current time to receive results from the system was 3.5 h with appreciable scope for optimisation, thus far comparing favourably to the UK National Health Service biopsy service where patients can wait for weeks for biopsy results. Full article

Biofuel cells use chemical reactions and biological catalysts (enzymes or microorganisms) to produce electrical energy, providing clean and renewable energy. Enzymatic biofuel cells (EBFCs) have promising characteristics and potential applications as an alternative energy source for low-power electronic devices. Over the last decade, researchers have focused on enhancing the electrocatalytic activity of biosystems and on increasing energy generation and electronic conductivity. Self-powered biosensors can use EBFCs while eliminating the need for an external power source. This review details improvements in EBFC and catalyst arrangements that will help to achieve complete substrate oxidation and to increase the number of collected electrons. It also describes how analytical techniques can be employed to follow the intermediates between the enzymes within the enzymatic cascade. We aim to demonstrate how a high-performance self-powered sensor design based on EBFCs developed for ethanol detection can be adapted and implemented in power devices for biosensing applications. Full article

Major advances in cancer control can be greatly aided by early diagnosis and effective treatment in its pre-invasive state. Lung cancer (small cell and non-small cell) is a leading cause of cancer-related deaths among both men and women around the world. A lot of research attention has been directed toward diagnosing and treating lung cancer. A common method of lung cancer treatment is based on COX-2 (cyclooxygenase-2) inhibitors. This is because COX-2 is commonly overexpressed in lung cancer and also the abundance of its enzymatic product prostaglandin E2 (PGE₂). Instead of using traditional COX-2 inhibitors to treat lung cancer, here, we introduce a new anti-cancer strategy recently developed for lung cancer treatment. It adopts more abundant omega-6 (ω-6) fatty acids such as dihomo-γ-linolenic acid (DGLA) in the daily diet and the commonly high levels of COX-2 expressed in lung cancer to promote the formation of 8-hydroxyoctanoic acid (8-HOA) through a new delta-5-desaturase (D5Di) inhibitor. The D5Di does not only limit the metabolic product, PGE_2, but also promote the COX-2 catalyzed DGLA peroxidation to form 8-HOA, a novel anti-cancer free radical byproduct. Therefore, the measurement of the PGE₂ and 8-HOA levels in cancer cells can be an effective method to treat lung cancer by providing in-time guidance. In this paper, we mainly report on a novel sensor, which is based on a newly developed functionalized nanomaterial, 2-dimensional nanosheets, or Ti₃C₂ MXene. The preliminary results have proven to sensitively, selectively, precisely, and effectively detect PGE₂ and 8-HOA in A549 lung cancer cells. The capability of the sensor to detect trace level 8-HOA in A549 has been verified in comparison with the traditional gas chromatography–mass spectrometry (GC–MS) method. The sensing principle could be due to the unique structure and material property of Ti₃C₂ MXene: a multilayered structure and extremely large surface area, metallic conductivity, and ease and versatility in surface modification. All these make the Ti₃C₂ MXene-based sensor selectively adsorb 8-HOA molecules through effective charge transfer and lead to a measurable change in the conductivity of the material with a high signal-to-noise ratio and excellent sensitivity. Full article

Cyclic adenosine monophosphate (cAMP) plays a key role in signal transduction pathways as a second messenger. Studies on the cAMP dynamics provided useful scientific insights for drug development and treatment of cAMP-related diseases such as some cancers and prefrontal cortex disorders. For example, modulation of cAMP-mediated intracellular signaling pathways by anti-tumor drugs could reduce tumor growth. However, most early stage tools used for measuring the cAMP level in living organisms require cell disruption, which is not appropriate for live cell imaging or animal imaging. Thus, in the last decades, tools were developed for real-time monitoring of cAMP distribution or signaling dynamics in a non-invasive manner. Genetically-encoded sensors based on fluorescent proteins and luciferases could be powerful tools to overcome these drawbacks. In this review, we discuss the recent genetically-encoded cAMP sensors advances, based on single fluorescent protein (FP), Föster resonance energy transfer (FRET), single luciferase, and bioluminescence resonance energy transfer (BRET) for real-time non-invasive imaging. Full article

In light of future missions beyond low Earth orbit (LEO) and the potential establishment of bases on the Moon and Mars, the effects of the deep space environment on biology need to be examined in order to develop protective countermeasures. Although many biological experiments have been performed in space since the 1960s, most have occurred in LEO and for only short periods of time. These LEO missions have studied many biological phenomena in a variety of model organisms, and have utilized a broad range of technologies. However, given the constraints of the deep space environment, upcoming deep space biological missions will be largely limited to microbial organisms and plant seeds using miniaturized technologies. Small satellites such as CubeSats are capable of querying relevant space environments using novel, miniaturized instruments and biosensors. CubeSats also provide a low-cost alternative to larger, more complex missions, and require minimal crew support, if any. Several have been deployed in LEO, but the next iterations of biological CubeSats will travel beyond LEO. They will utilize biosensors that can better elucidate the effects of the space environment on biology, allowing humanity to return safely to deep space, venturing farther than ever before. Full article

Surface-enhanced Raman scattering (SERS) nanoprobes have shown tremendous potential in in vivo imaging. The development of single oligomer resolution in the SERS promotes experiments on DNA and protein identification using SERS as a nanobiosensor. As Raman scanners rely on a multiple spectrum acquisition, faster imaging in real-time is required. SERS weak signal requires averaging of the acquired spectra that erases information on conformation and interaction. To build spectral libraries, the simulation of measurement conditions and conformational variations for the nucleotides relative to enhancer nanostructures would be desirable. In the molecular dynamic (MD) model of a sensing system, we simulate vibrational spectra of the cytosine nucleotide in FF2/FF3 potential in the dynamic interaction with the Au20 nanoparticles (NP) (EAM potential). Fourier transfer of the density of states (DOS) was performed to obtain the spectra of bonds in reaction coordinates for nucleotides at a resolution of 20 to 40 cm⁻¹. The Au₂₀ was optimized by ab initio density functional theory with generalized gradient approximation (DFT GGA) and relaxed by MD. The optimal localization of nucleotide vs. NP was defined and the spectral modes of both components vs. interaction studied. Bond-dependent spectral maps of nucleotide and NP have shown response to interaction. The marker frequencies of the Au₂₀—nucleotide interaction have been evaluated. Full article

A graphene sample (EGr) was prepared by electrochemical exfoliation of graphite rods in solution containing 0.05 M (NH₄)₂SO₄ + 0.1 M H₃BO₃ + 0.05 M NaCl. The exfoliation was performed by applying a constant voltage (12 V) between the graphite rods, while the temperature was kept constant (18 °C) with a temperature-controlled cryostat. The structural investigation of the graphene sample, performed by X-ray powder diffraction (XRD), revealed that the sample consists of a mixture of few-layer (69%), multi-layer graphene (14%) and graphene oxide (17%). In addition, XPS analysis proved that the sample was triple-doped with heteroatoms such as nitrogen (1.7 at%), sulfur (2.5 at%), and boron (3 at%). The sample was deposited onto the surface of a clean, glassy carbon electrode (GC) and investigated for the non-enzymatic electrochemical detection of L-tryptophan (TRP). The electrocatalytic properties of the EGr/GC electrode led to a considerable decrease in the oxidation potential from +0.9 V (bare GC) to +0.72 V. In addition, the EGr/GC electrode has higher sensitivity (two times) and a lower detection limit (ten times) in comparison with the bare GC electrode. Full article

Recently, a smart-device-based chest compression depth (CCD) feedback system that helps ensure that chest compressions have adequate depth during cardiopulmonary resuscitation (CPR) was developed. However, no CCD feedback device has been developed for infants, and many feedback systems are inconvenient to use. In this paper, we report the development of a smart-ring-based CCD feedback device for CPR based on an inertial measurement unit, and propose a high-quality chest compression depth estimation algorithm that considers the orientation of the device. The performance of the proposed feedback system was evaluated by comparing it with a linear variable differential transformer in three CPR situations. The experimental results showed compression depth errors of 2.0 ± 1.1, 2.2 ± 0.9, and 1.4 ± 1.1 mm in the three situations. In addition, we conducted a pilot test with an adult/infant mannequin. The results of the experiments show that the proposed smart-ring-based CCD feedback system is applicable to various chest compression methods based on real CPR situations. Full article

The detection of water contamination with Legionella pneumophila is of critical importance to manufacturers of water processing equipment and public health entities dealing with water networks and distribution systems. Detection methods based on polymerase chain reaction or biosensor technologies require preconcentration steps to achieve attractive sensitivity levels. Preconcentration must also be included in protocols of automated collection of water samples by systems designed for quasi-continuous monitoring of remotely located water reservoirs for the presence of L. pneumophila. We designed and characterized a water sampling module for filtration and backwashing intended for analysis of low-to-medium contaminated water, typically with L. pneumophila bacteria not exceeding 50 colony-forming units per milliliter. The concentration factors of 10× and 21× were achieved with 0.22 and 0.45 µm filters, respectively, for samples of bacteria prepared in clean saline solutions. However, a 5× concentration factor was achieved with 0.45 µm filters for a heavily contaminated or turbid water typical of some industrial water samples. Full article

Worldwide infection disease due to SARS-CoV-2 is tremendously affecting our daily lives. High-throughput detection methods for nucleic acids are emergently desired. Here, we show high-sensitivity and high-throughput metasurface fluorescence biosensors that are applicable for nucleic acid targets. The all-dielectric metasurface biosensors comprise silicon-on-insulator nanorod array and have prominent electromagnetic resonances enhancing fluorescence emission. For proof-of-concept experiment on the metasurface biosensors, we have conducted fluorescence detection of single-strand oligoDNAs, which model the partial sequences of SARS-CoV-2 RNA indicated by national infection institutes, and succeeded in the high-throughput detection at low concentrations on the order of 100 amol/mL without any amplification technique. As a direct detection method, the metasurface fluorescence biosensors exhibit high performance. Full article

The binding of integrin proteins to peptide sequences such as arginine-glycine-aspartic acid (RGD) is a crucial step in the adhesion process of mammalian cells. While these bonds can be examined between purified proteins and their ligands, live-cell assays are better suited to gain biologically relevant information. Here we apply a computer-controlled micropipette (CCMP) to measure the dissociation constant (K_d) of integrin-RGD-binding. Surface coatings with varying RGD densities were prepared, and the detachment of single cells from these surfaces was measured by applying a local flow inducing hydrodynamic lifting force on the targeted cells in discrete steps. The average behavior of the populations was then fit according to the chemical law of mass action. To verify the resulting value of K_d^2d = (4503 ± 1673) 1/µm², a resonant waveguide grating based biosensor was used, characterizing and fitting the adhesion kinetics of the cell populations. Both methods yielded a K_d within the same range. Furthermore, an analysis of subpopulations was presented, confirming the ability of CCMP to characterize cell adhesion both on single cell and whole population levels. The introduced methodologies offer convenient and automated routes to quantify the adhesivity of living cells before their further processing. Full article

This systematic review aimed to assess the advantages of biosensors in detecting biomarkers for the early diagnosis of osteoarthritis (OA). OA is the most prevalent musculoskeletal disease and is a leading cause of disability and pain worldwide. The diagnosis of OA could be performed through clinical examinations and imaging only during the late stages of the disease. Biomarkers could be used for the diagnosis of the disease in the very early stages. Biosensors could detect biomarkers with high accuracy and low costs. This paper focuses on the biosensors mainly adopted to detect OA markers (electrochemical, optical, Quartz crystal microbalance, molecular and wearable biosensors). A comprehensive search on PubMed, Cochrane, CINAHL and Embase databases was conducted from the inception to November 2020. The Preferred Reporting Items for Systematic reviews and Meta-Analyses (PRISMA) guidelines were used to improve the reporting of the review. The Methodological Index for Non-Randomized Studies (MINORS) was used for quality assessment. From a total amount of 1086 studies identified, only 19 articles were eligible for this study. The main advantages of the biosensors reported were accuracy, limited cost and ease of use, compared to traditional methods (ELISA). Otherwise, due to the lack of data and the low level of evidence of the papers included, it was impossible to find significant results. Therefore, further high-quality studies are required. Full article

Reactive oxygen species (ROS) have been found in plants, mammals, and natural environmental processes. The presence of ROS in mammals has been linked to the development of severe diseases, such as diabetes, cancer, tumors, and several neurodegenerative conditions. The most common ROS involved in human health are superoxide ${(\mathrm{O}}_2^{•-})$, hydrogen peroxide (H₂O₂), and hydroxyl radicals (•OH). Organic and inorganic molecules have been integrated with various methods to detect and monitor ROS for understanding the effect of their presence and concentration on diseases caused by oxidative stress. Among several techniques, fluorescence and electrochemical methods have been recently developed and employed for the detection of ROS. This literature review intends to critically discuss the development of these techniques to date, as well as their application for in vitro and in vivo ROS detection regarding free-radical-related diseases. Moreover, important insights into and further steps for using fluorescence and electrochemical methods in the detection of ROS are presented. Full article

Antibiotics are classes of antimicrobial substances that are administered widely in the field of veterinary science to promote animal health and feed efficiency. Cattle-administered antibiotics hold a risk of passing active residues to milk, during the milking process. This becomes a public health concern as these residues can cause severe allergic reactions to sensitive groups and considerable economic losses to the farmer. Hence, to ensure that the produced milk is safe to consume and adheres to permissible limits, an on-farm quick and reliable test is essential. This study illustrates the design and development of a microfluidic paper biosensor as a proof-of-concept detection system for gentamicin in milk. Localized surface plasmon resonance (LSPR) properties of gold nanoparticles have been explored to provide the user a visual feedback on the test, which was also corroborated by RGB analysis performed using Image J. The assay involves the use of a short stretch of single stranded DNA, called aptamer, which is very specific to the gentamicin present in the milk sample. The camera-based LOD for the fabricated paper device for milk samples spiked with gentamicin was calculated to be 300 nM, with a reaction time of 2 min. Full article