A novel biomimetic electrochemical taste-biosensor based on conformational changes of the taste receptor.

Taste sensor, a useful tool which could detect and identify thousands of different chemical substances in liquid environments, has attracted continuous concern from beverage and foodstuff industry and its consumers. Although many taste sensing methods have been extensively developed, the assessment of tastant content remains challenging due to the limitations of sensor selectivity and sensitivity. Here we present a novel biomimetic electrochemical taste-biosensor based on bioactive sensing elements and immune amplification with nanomaterials carrier to address above concerns, while taking sweet taste perception as a model. The proposed biosensor based on ligand binding domain (T1R2 VFT) of human sweet taste receptor protein showed human mimicking character and initiated the application of immune recognition in gustation biosensor, which can precisely and sensitively distinguish sweet substances against other related gustation substances with detection limit of 5.1 pM, far less than that of taste sensors without immune amplification whose detection limit was 0.48 nM. The performance test demonstrated the biosensor has the capacity of monitoring the response of sweet substances in real food environments, which is crucial in practical. This biomimetic electrochemical taste-biosensor can work as a new screening platform for newly developed tastants and disclose sweet perception mechanism.

Rationally Designed Dual Base Pair Mismatch Enables Toehold-Mediated Strand Displacement to Efficiently Recognize Single-Nucleotide Polymorphism without Enzymes.

The efficiency of the enzyme-free toehold-mediated strand displacement (TMSD) technique is often insufficient to detect single-nucleotide polymorphism (SNP) that possesses only single base pair mismatch discrimination. Here, we report a novel dual base pair mismatch strategy enabling TMSD biosensing for SNP detection under enzyme-free conditions when coupled with catalytic hairpin assembly (CHA) and fluorescence resonance energy transfer (FRET). The strategy is based on a competitive strand displacement reaction mechanism, affected by the thermodynamic stability originating from rationally designed dual base pair mismatch, for the specific recognition of mutant-type DNA. In particular, enzyme-free nucleic acid circuits, such as CHA, emerge as a powerful method for signal amplification. Eventually, the signal transduction of this proposed biosensor was determined by FRET between streptavidin-coated 605 nm emission quantum dots (605QDs, donor) and Cy5/biotin hybridization (acceptor, from CHA) when incubated with each other. The proposed biosensor displayed high sensitivity to the mutant target (MT) with a detection concentration down to 4.3 fM and led to high discrimination factors for all types of mismatches in multiple sequence contexts. As such, the application of this proposed biosensor to investigate mechanisms of the competitive strand displacement reaction further illustrates the versatility of our dual base pair mismatch strategy, which can be utilized for the creation of a new class of biosensors.

A biosensing system employing nonlinear dynamic analysis-assisted neural network for drug-induced cardiotoxicity assessment.

Preclinical investigation of drug-induced cardiotoxicity is of importance for drug development. To evaluate such cardiotoxicity, in vitro high-throughput interdigitated electrode-based recording of cardiomyocytes mechanical beating is widely used. To automatically analyze the features from the beating signals for drug-induced cardiotoxicity assessment, artificial neural network analysis is conventionally employed and signals are segmented into cycles and feature points are located in the cycles. However, signal segmentation and location of feature points for different signal shapes require design of specific algorithms. Consequently, this may lower the efficiency of research and the applications of such algorithms in signals with different morphologies are limited. Here, we present a biosensing system that employs nonlinear dynamic analysis-assisted neural network (NDANN) to avoid the signal segmentation process and directly extract features from beating signal time series. By processing beating time series with fixed time duration to avoid the signal segmentation process, this NDANN-based biosensing system can identify drug-induced cardiotoxicity with accuracy over 0.99. The individual drugs were classified with high accuracies over 0.94 and drug-induced cardiotoxicity levels were accurately predicted. We also evaluated the generalization performance of the NDANN-based biosensing system in assessing drug-induced cardiotoxicity through an independent dataset. This system achieved accuracy of 0.85-0.95 for different drug concentrations in identification of drug-induced cardiotoxicity. This result demonstrates that our NDANN-based biosensing system has the capacity of screening newly developed drugs, which is crucial in practical applications. This NDANN-based biosensing system can work as a new screening platform for drug-induced cardiotoxicity and improve the efficiency of bio-signal processing.

A smart tablet-phone-based system using dynamic light modulation for highly sensitive colorimetric biosensing.

Facile, efficient, and inexpensive biosensing systems are in high demand for biomedical test. In recent years, numerous smartphone-based biosensing systems have been developed to match demand for biomedical test in source-limited environment. However, application of these smartphone-based biosensing systems was limited because of performance gap between the smartphone-based systems and commercial plate readers. In this study, we have developed a smart tablet-phone-based colorimetric plate reader (STPCPR) with intelligent and dynamic light modulation for broad-range colorimetric assays. The STPCPR allows controllable modulation of exciting light in three different color channels that is lack in conventional smartphone-based system. Using optimized exciting modulation, the STPCPR shows higher sensitivities, lower detection limits, and broader detection ranges in test of pigments, proteins, and cells when compared to conventional plate readers and smartphone-based system. Therefore, the developed STPCPR can serve as an ideal next-generation smartphone-based biosensing system for point-of-care colorimetric test in diverse biomedical applications in source-limited environment.

Recent Advances of Organ-on-a-Chip in Cancer Modeling Research.

Although many studies have focused on oncology and therapeutics in cancer, cancer remains one of the leading causes of death worldwide. Due to the unclear molecular mechanism and complex in vivo microenvironment of tumors, it is challenging to reveal the nature of cancer and develop effective therapeutics. Therefore, the development of new methods to explore the role of heterogeneous TME in individual patients' cancer drug response is urgently needed and critical for the effective therapeutic management of cancer. The organ-on-chip (OoC) platform, which integrates the technology of 3D cell culture, tissue engineering, and microfluidics, is emerging as a new method to simulate the critical structures of the in vivo tumor microenvironment and functional characteristics. It overcomes the failure of traditional 2D/3D cell culture models and preclinical animal models to completely replicate the complex TME of human tumors. As a brand-new technology, OoC is of great significance for the realization of personalized treatment and the development of new drugs. This review discusses the recent advances of OoC in cancer biology studies. It focuses on the design principles of OoC devices and associated applications in cancer modeling. The challenges for the future development of this field are also summarized in this review. This review displays the broad applications of OoC technique and has reference value for oncology development.

A Portable Smartphone-Based System for the Detection of Blood Calcium Using Ratiometric Fluorescent Probes.

Hypocalcemia is a disease that adversely affects the production and reproduction of dairy cows. A portable device for rapid bovine blood calcium sensing has been growing in demand. Herein, we report a smartphone-based ratiometric fluorescence probe (SRFP) platform as a new way to detect and quantify calcium ions (Ca2+) in blood serum. Specifically, we employed a cost-effective and portable smartphone-based platform coupled with customized software that evaluates the response of Ca2+ ions to ratiometric fluorescence probe in bovine serum. The platform consists of a three-dimensional (3D) printed housing and low-cost optical components that excite fluorescent probe and selectively transmit fluorescence emissions to smartphones. The customized software is equipped with a calibration model to quantify the acquired fluorescence images and quantify the concentration of Ca2+ ions. The ratio of the green channel to the red channel bears a highly reproducible relationship with Ca2+ ions concentration from 10 µM to 40 µM in bovine serum. Our detection system has a limit of detection (LOD) of 1.8 µM in bovine serum samples and the recoveries of real samples ranged from 92.8% to 110.1%, with relative standard deviation (RSD) ranging from 1.72% to 4.89%. The low-cost SRFP platform has the potential to enable campesino to rapidly detect Ca2+ ions content in bovine serum on-demand in any environmental setting.

A biosensing system employing nanowell microelectrode arrays to record the intracellular potential of a single cardiomyocyte.

Electrophysiological recording is a widely used method to investigate cardiovascular pathology, pharmacology and developmental biology. Microelectrode arrays record the electrical potential of cells in a minimally invasive and high-throughput way. However, commonly used microelectrode arrays primarily employ planar microelectrodes and cannot work in applications that require a recording of the intracellular action potential of a single cell. In this study, we proposed a novel measuring method that is able to record the intracellular action potential of a single cardiomyocyte by using a nanowell patterned microelectrode array (NWMEA). The NWMEA consists of five nanoscale wells at the center of each circular planar microelectrode. Biphasic pulse electroporation was applied to the NWMEA to penetrate the cardiomyocyte membrane, and the intracellular action potential was continuously recorded. The intracellular potential recording of cardiomyocytes by the NWMEA measured a potential signal with a higher quality (213.76 ± 25.85%), reduced noise root-mean-square (~33%), and higher signal-to-noise ratio (254.36 ± 12.61%) when compared to those of the extracellular recording. Compared to previously reported nanopillar microelectrodes, the NWMEA could ensure single cell electroporation and acquire high-quality action potential of cardiomyocytes with reduced fabrication processes. This NWMEA-based biosensing system is a promising tool to record the intracellular action potential of a single cell to broaden the usage of microelectrode arrays in electrophysiological investigation.

A biosensing system using a multiparameter nonlinear dynamic analysis of cardiomyocyte beating for drug-induced arrhythmia recognition.

Cardiovascular disease is the number one cause of death in humans. Therefore, cardiotoxicity is one of the most important adverse effects assessed by arrhythmia recognition in drug development. Recently, cell-based techniques developed for arrhythmia recognition primarily employ linear methods such as time-domain analysis that detect and compare individual waveforms and thus fall short in some applications that require automated and efficient arrhythmia recognition from large datasets. We carried out the first report to develop a biosensing system that integrated impedance measurement and multiparameter nonlinear dynamic algorithm (MNDA) analysis for drug-induced arrhythmia recognition and classification. The biosensing system cultured cardiomyocytes as physiologically relevant models, used interdigitated electrodes to detect the mechanical beating of the cardiomyocytes, and employed MNDA analysis to recognize drug-induced arrhythmia from the cardiomyocyte beating recording. The best performing MNDA parameter, approximate entropy, enabled the system to recognize the appearance of sertindole- and norepinephrine-induced arrhythmia in the recording. The MNDA reconstruction in phase space enabled the system to classify the different arrhythmias and quantify the severity of arrhythmia. This new biosensing system utilizing MNDA provides a promising and alternative method for drug-induced arrhythmia recognition and classification in cardiological and pharmaceutical applications.

An Improved Automated High-Throughput Efficient Microplate Reader for Rapid Colorimetric Biosensing.

A high-throughput instrument to measure the full spectral properties of biochemical agents is necessary for fast screening in fields such as medical tests, environmental monitoring, and food analysis. However, this need has currently not been fully met by the commercial microplate reader (CMR). In this study, we have developed an automated high-throughput efficient microplate reader (AHTEMR) platform by combining a spectrometer and high-precision ball screw two-dimensional motion slide together, for high-throughput and full-spectrum-required biochemical assays. A two-dimensional slide working on a ball screw was driven by a stepper motor with a custom-designed master control circuit and used as a motion system of the AHTEMR platform to achieve precise positioning and fast movement of the microplate during measurements. A compact spectrometer was coupled with an in-house designed optical pathway system and used to achieve rapid capture of the full spectral properties of biochemical agents. In a performance test, the AHTEMR platform successfully measured the full spectral absorbance of bovine serum albumin (BSA) and glucose solution in multiple wells of the microplate within several minutes and presented the real-time full spectral absorbance of BSA and glucose solution. Compared with the CMR, the AHTEMR is 79 times faster in full-spectrum measurements and 2.38 times more sensitive at the optimal wavelength of 562 nm. The rapid measurement also demonstrated the great capacity of the AHTEMR platform for screening out the best colorimetric wavelengths for tests of BSA and glucose development, which will provide a promising approach to achieving high-throughput and full-spectrum-required biochemical assays.

Electromechanical integrated recording of single cardiomyocyte in situ by multimodal microelectrode biosensing system.

The development of new drugs is a lengthy process, while the observation of serious side effects, such as cardiotoxicity, can result in the drug to be withdrawn even after development, leading to heavy burden on human health and social economy. To assess the drug cardiotoxicity, the electrical and mechanical properties of cardiomyocytes are increasingly being used to investigate the mechanisms and potential toxicity of drugs. Conventional non-invasive and label-free recording strategies are not well suited to record the integrated electromechanical signals from the single cell in a high-throughput manner, whereas label-based recordings strategies suffer from phototoxicity and drug side effects, precluding their long-time detection. In this study, we established a new multimodal microelectrode biosensing system to achieve the simultaneous and dynamic interrogation of electromechanical signals from multisite single cardiomyocytes. This multimodal device can detect subtle changes in the electromechanical signals induced by ion channel drugs during the excitation-contraction coupling of cardiomyocytes. The use of electromechanical integrated single cell signals for drug assessment was compared with commercial drug assays, and our multimodal microelectrode biosensing system can afford record electromechanical integrated signals as well as efficiently identify the effects of ion channel-blocking drugs on the electrical and mechanical properties of cardiomyocytes. Our multimodal microelectrode biosensing system is a potential valuable platform in the fields of cardiology and pharmacology.

Multi-labeled neural network model for automatically processing cardiomyocyte mechanical beating signals in drug assessment.

High-throughput cardiotoxicity assessment is important for large-scale preclinical screening in novel drug development. To improve the efficiency of drug development and avoid drug-induced cardiotoxicity, there is a huge demand to explore the automatic and intelligent drug assessment platforms for preclinical cardiotoxicity investigations. In this work, we proposed an automatic and intelligent strategy that combined automatic feature extraction and multi-labeled neural network (MLNN) to process cardiomyocytes mechanical beating signals detected by an interdigital electrode biosensor for the assessment of drug-induced cardiotoxicity. Taking advantages of artificial neural network, our work not only classified different drugs inducing different cardiotoxicities but also predicted drug concentrations representing severity of cardiotoxicity. This has not been achieved by conventional strategies like principal component analysis and visualized heatmap. MLNN analysis showed high accuracy (up to 96%) and large AUC (more than 98%) for classification of different drug-induced cardiotoxicities. There was a high correlation (over 0.90) between concentrations reported by MLNN and experimentally treated concentrations of various drugs, demonstrating great capacity of our intelligent strategy to predict the severity of drug-induced cardiotoxicity. This new intelligent bio-signal processing algorithm is a promising method for identification and classification of drug-induced cardiotoxicity in cardiological and pharmaceutical applications.

Electrochemistry Coupling Localized Surface Plasmon Resonance for Biochemical Detection.

Localized surface plasmon resonance (LSPR) associated with metal nanostructures has developed into highly useful sensor techniques. LSPR spectroscopy often shows absorption peaks which could be used for biomedical detection. Here we report nanoplasmonic sensors using LSPR on nanostructures such as nanoparticles, nanocups, and nanocones to recognize biomolecular. These sensors can be modified for quantitative detection of explosives and evaluation of enzymatic activity. Electrochemical LSPR sensors can also be designed by coupling electrochemistry and LSPR spectroscopy measurements for biochemical detection. Multiple sensing information can be obtained and electrochemical LSPR property can be investigated for biosensors. In some applications, the electrochemical LSPR biosensor can be used to quantify heavy metal ions, neurotransmitters, and sialic acid. The biosensors exhibit better performance than those of conventional optical LSPR measurements. With multitransducers, the nanoplasmonic biosensor can provide a promising approach for biochemical detection in environmental monitoring, healthcare diagnostics, and food quality control.

Smartphone-Based Electrochemical System for Biosensors and Biodetection.

With the advantages of high popularity, convenient operation, open-source operation systems, high resolution imaging, and excellent computing capabilities, smartphones have been widely used as the core of detection system for calculation, control, and real-time display. Hence, smartphones play an important role in electrochemical detection and optical detection. Smartphone-based electrochemical systems were combined with screen-printed electrode and interdigital electrodes for in situ detection. The electrodes were modified with biomaterials, chemical materials, and nanomaterials for biosensors and biodetection, such as 3-amino phenylboronic acid nanocomposites, graphene, gold nanoparticles, zinc oxide nanoparticles, carbon nanotubes, proteins, peptides, and antibodies. With the modified electrodes, the smartphone-based impedance system was used to detect acetone, bovine serum albumin, human serum albumin, and trinitrotoluene, while smartphone-based amperometric system was employed to monitor glucose, ascorbic acid, dopamine, uric acid, and levodopa. The smartphone-based electrochemical system for biosensors and biodetection has provided miniaturized and portable alternative for diagnosis, which is promising to find application in point-of-care testing (POCT).

Rational engineering of ratiometric calcium sensors with bright green and red fluorescent proteins.

Ratiometric genetically encoded calcium indicators (GECIs) record neural activity with high brightness while mitigating motion-induced artifacts. Recently developed ratiometric GECIs primarily employ cyan and yellow-fluorescent fluorescence resonance energy transfer pairs, and thus fall short in some applications that require deep tissue penetration and resistance to photobleaching. We engineered a set of green-red ratiometric calcium sensors that fused two fluorescent proteins and calcium sensing domain within an alternate configuration. The best performing elements of this palette of sensors, Twitch-GR and Twitch-NR, inherited the superior photophysical properties of their constituent fluorescent proteins. These properties enabled our sensors to outperform existing ratiometric calcium sensors in brightness and photobleaching metrics. In turn, the shot-noise limited signal fidelity of our sensors when reporting action potentials in cultured neurons and in the awake behaving mice was higher than the fidelity of existing sensors. Our sensor enabled a regime of imaging that simultaneously captured neural structure and function down to the deep layers of the mouse cortex.

The Early Diagnostic Value of Procalcitonin in Pneumonia After Off-Pump Coronary Artery Bypass Grafting Surgery.

BACKGROUND The incidence of early postoperative pneumonia (EPOP) after off-pump coronary artery bypass grafting surgery (CABG) is relatively high, but its diagnosis by traditional methods remains difficult, which could be deleterious to the prognosis. Moreover, few data exist regarding procalcitonin (PCT) in early diagnosis of pneumonia after off-pump CABG. Thus, this study was performed to evaluate the value of PCT in diagnosing EPOP after off-pump CABG. MATERIAL AND METHODS A total of 402 consecutive patients undergoing off-pump CABG were retrospectively enrolled. Forty-four patients were diagnosed with EPOP and 112 patients were diagnosed with systemic inflammatory response syndrome (SIRS). Chest roentgenogram, serum PCT, white blood cells, neutral granulocyte ratio, and daily maximum body temperature were recorded. The ability of PCT to diagnose EPOP was evaluated by receiver operating characteristic (ROC) analyses in comparison with traditional methods. Clinical net benefits were estimated via decision curve analysis (DCA). RESULTS PCT presented satisfying accuracy in diagnosing EPOP with a cutoff value of 1.585 ng/mL (area under the curve [AUC] 0.808, 95% confidence interval [CI] 0.724-0.891, sensitivity 73%, specificity 86%). PCT performed better in diagnosing EPOP among SIRS patients (AUC 0.868, 95% CI 0.748-0.988, sensitivity 85%, specificity 89%). DCA showed valuable clinical net benefits of PCT in diagnosing EPOP after off-pump CABG regardless of threshold selected. CONCLUSIONS PCT could be a diagnostic marker for EPOP after off-pump CABG. The optimal cutoff value for diagnosing EPOP was 1.585 ng/mL. The application of PCT in diagnosing EPOP in SIRS patients was also satisfying with a cutoff value of 1.775 ng/mL.

Enhanced genetically encoded voltage indicators advance their applications in neuroscience.

Genetically encoded voltage indicators report membrane voltage with high spatiotemporal resolution. Extensive recent efforts to improve the GEVIs' brightness, sensitivity, and kinetics have greatly increased the GEVIs' signal-to-noise performance over ten-fold and lowered their response time to the sub-millisecond regime. Such capabilities have broadened the GEVIs' ability to measure membrane voltage of neural populations at cellular resolution in vitro and in vivo, all at high speeds. The GEVIs' high voltage fidelity and fast response have revealed novel physiological phenomena in multiple neuroscientific applications. Such applications portend future targeted studies of voltage activity that take advantage of the GEVIs' ability to report rapid dynamics from genetically-targeted neural populations.

Nanoplasmonic Biosensor Using Localized Surface Plasmon Resonance Spectroscopy for Biochemical Detection.

Localized surface plasmon resonance (LSPR) associated with metal nanostructures has developed into a highly useful sensor technique. Optical LSPR spectroscopy of nanostructures often shows sharp absorption and scattering peaks, which can be used to probe several bio-molecular interactions. Here, we report nanoplasmonic biosensors using LSPR on nanocup arrays (nanoCA) to recognize bio-molecular binding for biochemical detection. These sensors can be modified to quantify binding of small molecules to proteins for odorant and explosive detections. Electrochemical LSPR biosensors can also be designed by coupling electrochemistry and LSPR spectroscopy measurements. Multiple sensing information can be obtained and electrochemical LSPR property can be investigated for biosensors. In some applications, the electrochemical LSPR biosensor can be used to quantify immunoreactions and enzymatic activity. The biosensors exhibit better performance than those of conventional optical LSPR measurements. With multi-transducers, the nanoplasmonic biosensor can provide a promising approach for bio-detection in environmental monitoring, healthcare diagnostics, and food quality control.

Smartphone-based sensing system using ZnO and graphene modified electrodes for VOCs detection.

Volatile organic compounds (VOCs) detection is in high demand for clinic treatment, environment monitoring, and food quality control. Especially, VOCs from human exhaled breath can serve as significant biomarkers of some diseases, such as lung cancer and diabetes. In this study, a smartphone-based sensing system was developed for real-time VOCs monitoring using alternative current (AC) impedance measurement. The interdigital electrodes modified with zinc oxide (ZnO), graphene, and nitrocellulose were used as sensors to produce impedance responses to VOCs. The responses could be detected by a hand-held device, sent out to a smartphone by Bluetooth, and reported with concentration on an android program of the smartphone. The smartphone-based system was demonstrated to detect acetone at concentrations as low as 1.56ppm, while AC impedance spectroscopy was used to distinguish acetone from other VOCs. Finally, measurements of the exhalations from human being were carried out to obtain the concentration of acetone in exhaled breath before and after exercise. The results proved that the smartphone-based system could be applied on the detection of VOCs in real settings for healthcare diagnosis. Thus, the smartphone-based system for VOCs detection provided a convenient, portable and efficient approach to monitor VOCs in exhaled breath and possibly allowed for early diagnosis of some diseases.

Zinc Nanoparticles-equipped Bioelectronic Nose Using a Microelectrode Array for Odorant Detection.

Bioelectronic noses, such as olfactory cell- and receptor-based biosensors, have important applications for biomimetic odorant detection in various fields. Here, a nanoparticle-equipped biosensor was designed to record extracellular potentials from olfactory receptor cells effectively. In this research, a microelectrode array (MEA) was combined with olfactory epitheliums as the olfactory biosensor to record electrophysiological signals of receptor cells in the epitheliums. Zinc nanoparticles (NanoZn) were employed along with the biosensor for different kinds of odorant measurements, which improved the electrophysiological responses to odor molecules. The NanoZn-equipped biosensor showed greater performance, such as a higher sensitivity and a larger signal-to-noise ratio, than that without the nanoparticles. Thus, this approach provided a promising method to improve the detecting performance of biosensors based on olfactory cells and receptors, which would bring broad application prospects for bioelectronic noses in environmental monitoring, food analysis, and healthcare diagnosis.

Impedance spectroscopy analysis of human odorant binding proteins immobilized on nanopore arrays for biochemical detection.

Human odorant-binding proteins (hOBPs) not only can bind and transport odorants in the surrounding environment for sensing smells, but also play important roles in transmitting lots of biomolecules in different organs. Utilizing the properties of hOBPs, an electrochemical biosensor with nanopore array was developed to detect specific biomolecular ligands, such as aldehydes and fatty acids. The highly ordered nanopores of anodic aluminum oxide with diameter of 20-40 nm were fabricated with two-step oxidation. Through 2-carboxyethyl phosphonic acid, hOBPs were self-assembled on nanopores as the sensing membrane. With nanopore arrays, the impedance spectra showed quite different electron transfer processes in the frequency spectra, which could be characterized by the electron transfer resistance and electrical resistance of the porous membrane. Under stimulation of biomolecular ligands, series resistance of nanopores and hOBPs increased and showed a concentration-dependence feature, while the electron transfer resistance hardly changed. The nanopore based biosensor could sensitively detect biological ligands of benzaldehyde, docosahexaenoic acid, and lauric acid, which were closely related to or were potential biomarkers for cancers and other serious diseases. Equipped with hOBPs, the sensor exhibited promising potentials both in odorant and biomolecule detection for olfactory biosensing and in disease diagnosis and evaluation for biochemical detection.