Highly Light-Harvesting MOF-on-MOF Heterostructure: Cascading Functionality to Flexible Photogating of Organic Photoelectrochemical Transistor and Bienzyme Cascade Detection.

Recently, organic photoelectrochemical transistor (OPECT) bioanalysis has become a prominent technique for the high-performance detection of biomolecules. However, as a sensitive index of the OPECT, the dynamic regulation transconductance (gm) is still severely deficient. Herein, this work reports a new photosensitive metal-organic framework (MOF-on-MOF) heterostructure for the effective modulation of maximum gm and natural bienzyme interfacing toward choline detection. Specifically, the bidentate ligand MOF (b-MOF) was assembled onto the UiO-66 MOF (u-MOF) by a modular assembly method, which could facilitate the charge separation and generate enhanced photocurrents and offer a biophilic environment for the immobilization of choline oxidase (ChOx) and horseradish peroxidase (HRP) through hydrogen-bonded bridges. The transconductance of the OPECT could be flexibly altered by increased light intensity to maximal value at zero gate bias, and sensitive choline detection was achieved with a detection limit of 0.2 µM. This work reveals the potential of MOF-on-MOF heterostructures for futuristic optobioelectronics.

Electrochemical Single-Cell Protein Therapeutics Using a Double-Barrel Nanopipette.

Single-cell protein therapeutics is expected to promote our in-depth understanding of how a specific protein with a therapeutic dosage treats the cell without population averaging. However, it has not yet been tackled by current single-cell nanotools. We address this challenge by the use of a double-barrel nanopipette, in which one lumen was used for electroosmotic cytosolic protein delivery and the other was customized for ionic evaluation of the consequence. Upon injection of protein DJ-1 through the delivery lumen, upregulation of the antioxidant protein could protect neural PC-12 cells against oxidative stress from phorbol myristate acetate exposure, as deduced by targeting of the cytosolic hydrogen peroxide by the detecting lumen. The nanotool developed in this study for single-cell protein therapeutics provides a perspective for future single-cell therapeutics involving different therapeutic modalities, such as peptides, enzymes and nucleic acids.

An Integrated Dual-Functional Nanotool Capable of Studying Single-Cell Epigenetics and Programmable Gene Regulation.

Single-cell epigenetics is envisioned to decipher manifold epigenetic phenomena and to contribute to our accurate knowledge about basic epigenetic mechanisms. Engineered nanopipette technology has gained momentum in single-cell studies; however, solutions to epigenetic questions remain unachieved. This study addresses the challenge by exploring N6-methyladenine (m6 A)-bearing deoxyribozyme (DNAzyme) confined within a nanopipette for profiling a representative m6 A-modifying enzyme, fat mass and obesity-associated protein (FTO). Electroosmotic intracellular extraction of FTO could remove the m6 A and cause DNAzyme cleavage, leading to the altered ionic current signal. Because the cleavage can release a DNA sequence, we simultaneously program it as an antisense strand against FTO-mRNA, intracellular injection of which has been shown to induce early stage apoptosis. This nanotool thus features the dual functions of studying single-cell epigenetics and programmable gene regulation.

A Photoelectrochemical Nanoreactor for Single-Cell Sampling and Near Zero-Background Faradaic Detection of Intracellular microRNA.

Rational utilization of the rich light-bio-matter interplay taking place in single-cell analysis represents a new technological direction in the field. The light-fueled operation is expected to achieve advanced photoelectrochemical (PEC) single-cell analysis with unknown possibilities. Here, a PEC nanoreactor capable of single-cell sampling and near zero-background Faradaic detection of intracellular microRNA (miR) is devised by the construction of a small reaction chamber accommodating the target-triggered hybridization chain reaction for binding the metallointercalator of [Ru(bpy)2 (dppz)]2+ as the signal reporter. Light stimulation of the dsDNA/metallointercalator adduct will induce the generation of photocurrents, underpinning a zero-biased and near zero-background PEC method toward Faradaic detection of non-electrogenic miR at the single-cell level. Using this nanotool, lower miR concentration in the near-nucleus region than that in the main cytosol was revealed.

Peptide-Based Photocathodic Biosensors: Integrating a Recognition Peptide with an Antifouling Peptide.

Accurate and sensitive detection of targets in practical biological matrixes such as blood, plasma, serum, or tissue fluid is a frontier issue for most biosensors since the coexistence of both potential reducing agents and protein molecules has the possibility of causing signal interference. Herein, aiming at detection in a complex environment, an advanced and robust peptide-based photocathodic biosensor, which integrated a recognition peptide with an antifouling peptide in one probe electrode, was first proposed. Selecting human chorionic gonadotropin (hCG) as a model target, the recognition peptide with the sequence PPLRINRHILTR was first anchored on the CuBi2O4/Au (CBO/Au) photocathode and then the antifouling peptide with the sequence EKEKEKEPPPPC was further anchored to generate an antifouling biointerface. The peptide-based photocathodic biosensor demonstrated excellent anti-interference to both nonspecific proteins and reducing agents because of the capability of the antifouling peptide. It also exhibited good sensitivity owing to the utilization of the recognition peptide rather than an antibody probe. This peptide-integrated method offers a new perspective for practical applications of photocathodic biosensors.

Target-Triggered Assembly in a Nanopipette for Electrochemical Single-Cell Analysis.

Engineered nanopipette tools have recently emerged as a powerful approach for electrochemical nanosensing, which has major implications in both fundamental biological research and biomedical applications. Herein, we describe a generic method of target-triggered assembly of aptamers in a nanopipette for nanosensing, which is exemplified by sensitive and rapid electrochemical single-cell analysis of adenosine triphosphate (ATP), a ubiquitous energy source in life and important signaling molecules in many physiological processes. Specifically, a layer of thiolated aptamers is immobilized onto a Au-coated interior wall of a nanopipette tip. With backfilled pairing aptamers, the engineered nanopipette is then used for probing intracellular ATP via the ATP-dependent linkage of the split aptamers. Due to the higher surface charge density from the aptamer assembly, the nanosensor would exhibit an enhanced rectification signal. Besides, this ATP-responsive nanopipette tool possesses excellent selectivity and stability as well as high recyclability. This work provides a practical single-cell nanosensor capable of intracellular ATP analysis. More generally, integrated with other split recognition elements, the proposed mechanism could serve as a viable basis for addressing many other important biological species.

An Integrated Electrochemical Nanodevice for Intracellular RNA Collection and Detection in Single Living Cell.

New tools for single-cell interrogation enable deeper understanding of cellular heterogeneity and associated cellular behaviors and functions. Information of RNA expression in single cell could contribute to our knowledge of the genetic regulatory circuits and molecular mechanism of disease development. Although significant progresses have been made for intracellular RNA analysis, existing methods have a trade-off between operational complexity and practical feasibility. We address this challenge by combining the ionic current rectification property of nanopipette reactor with duplex-specific nuclease-assisted hybridization chain reaction for signal amplification to realize a simple and practical intracellular nanosensor with minimal invasiveness, which enables single-cell collection and electrochemical detection of intracellular RNA with cell-context preservation. Systematic studies on differentiation of oncogenic miR-10b expression levels in non-malignant breast cells, metastatic breast cancer cells as well as non-metastatic breast cancer cells were then realized by this nanotool accompanied by assessment of different drugs effects. This work has unveiled the ability of electrochemistry to probe intracellular RNA and opened new opportunities to study the gene expression and heterogeneous complexity under physiological conditions down to single-cell level.

Gold Nanoparticle-Induced Photocurrent Quenching and Recovery of Polymer Dots: Toward Signal-On Energy-Transfer-Based Photocathodic Bioanalysis of Telomerase Activity in Cell Extracts.

Energy transfer (ET) in photoelectrochemical (PEC) bioanalysis is usually generated between noble metal nanoparticles (NPs) and traditional inorganic quantum dots (QDs). Using the innovative polymer dot (Pdot)-involved ET, this work reports the first signal-on and cathodic PEC bioanalysis toward telomerase (TE) activity in cell extracts. Specifically, the sequential binding of capture DNA (cDNA), telomerase primer sequence (TS), and Au NP-labeled probe DNA (Au NP-pDNA) on the electrode would place the Au NPs in close proximity of the Pdots, leading to obvious quenching of the cathodic photocurrent. The subsequent extension of the TS by TE in the presence of deoxyribonucleoside triphosphates (dNTPs) would then release the Ag NP-pDNA from the electrode, leading to the recovery of the photocurrent. On the basis of the Au NP-induced photocurrent quenching and the recovery of Pdots, a sensitive biosensor could thus be developed by tracking the photocurrents to probe the TE activity. This strategy allows for signal-on and cathodic PEC bioanalysis of TE, which can be easily extended for numerous other targets of interest. We believe this work could offer a new perspective for the rational implementation of Pdot-involved ET for advanced PEC bioanalysis.

Gold Nanoparticle Couples with Entropy-Driven Toehold-Mediated DNA Strand Displacement Reaction on Magnetic Beads: Toward Ultrasensitive Energy-Transfer-Based Photoelectrochemical Detection of miRNA-141 in Real Blood Sample.

Highly stable circulating microRNAs (miRNAs) are currently recognized as a novel potential biomarker for clinical cancer diagnosis in the early stage. However, limited by its low concentration, high sequence similarity, as well as the numerous interferences in body fluids, detection of miRNA in whole blood with sufficient selectivity and sensitivity is still challenging. Herein, we reported the integration of entropy-driven toehold-mediated DNA strand displacement (ETSD) reaction with magnetic beads (MB) toward the energy-transfer-based photoelectrochemical (PEC) detection of the prostate carcinoma (PCa) biomarker miRNA-141 in a real blood sample. In this protocol, the ETSD reaction was divided into two steps, and cooperated with magnetic separation, target extraction and amplification could be realized in a single test and ultrasensitive detection of miRNA-141 could be achieved in undiluted whole blood sample. This work proposed a new solution for sensitive biomolecular detection in a complex biological milieu and exhibited great promise for future clinical cancer diagnosis.

Energy Transfer between Semiconducting Polymer Dots and Gold Nanoparticles in a Photoelectrochemical System: A Case Application for Cathodic Bioanalysis.

We report herein the energy transfer (ET) between semiconducting polymer dots (Pdots) and gold nanoparticles (Au NPs) in a photoelectrochemical (PEC) system and its feasibility for cathodic bioanalysis application. Specifically, COOH-capped Pdots were first fabricated and then assembled onto the indium-tin oxide (ITO) surface, followed by the modification of single-strand (ss) DNA probe (pDNA). After the DNA hybridization with the Au NP-tethered complementary ssDNA (Au NP-tDNA), the Au NPs were brought into the close proximity of Pdots. Upon light stimulation, photoluminescence (PL) was annihilated, fluorescence was attenuated, and the photocurrent intensity was evidently decreased. This ET-based PEC DNA sensor exhibited a linear range from 1 fM to 10 pM with a detection limit of 0.97 fM at a signal-to-noise ratio of 3. The present work first exploited the ET between Pdots and Au NPs, and we believe this phenomenon will spark new interest in the study of various Pdots-based ET-influenced PEC systems and thus catalyze increasing studies for specific bioanalytical purposes.

A Polymer Dots-Based Photoelectrochemical pH Sensor: Simplicity, High Sensitivity, and Broad-Range pH Measurement.

This work reported the photoelectrochemical (PEC) pH sensor for sensitive and broad-range pH measurement on the basis of semiconducting polymer dots (Pdots). The sensor was fabricated by immobilizing Pdots onto the surface of indium tin oxide (ITO). Experimental results revealed that the carboxylic acid groups of Pdots were sensitive to pH variation, which could result in conformational changes and further diffusion of carriers. Besides, different pH value could change the redox properties of the Pdots, and the photocurrent response was hence altered by the carriers produced on the Pdots. Further results demonstrated that the developed sensor exhibited variable photocurrent sensitively by responding to different pH values. This pH sensor is of high sensitivity, stability, and reversibility, which provides a bright prospect for future pH measurements in the bioanalytical field.

3D Semiconducting Polymer/Graphene Networks: Toward Sensitive Photocathodic Enzymatic Bioanalysis.

This work reports the development of three-dimensional (3D) semiconducting polymer/graphene (SP/G) networks toward sensitive photocathodic enzymatic bioanalysis. Specifically, the porous 3D graphene was first synthesized via the hydrothermal and freeze-dry processes and then mixed with semiconducting polymer to obtain the designed hierarchical structure with unique porosity and large surface area. Afterward, the as-prepared hybrid was immobilized onto the indium tin oxide (ITO) for further characterizations. Exemplified by sarcosine oxidase (SOx) as a model biocatalyst, an innovative 3D SP/G-based photocathodic bioanalysis capable of sensitive and specific sarcosine detection was achieved. The suppression of cathodic photocurrent was observed in the as-developed photocathodic enzymatic biosystem due to the competition of oxygen consumption between the enzyme-biocatalyst process and O2-dependent photocathodic electrode. This work not only presented a unique protocol for 3D SP/G-based photocathodic enzymatic bioanalysis but also provided a new horizon for the design, development, and utilization of numerous 3D platforms in the broad field of general photoelectrochemical (PEC) bioanalysis.

Optimization of Graphene Conductive Ink with 73 wt% Graphene Contents.

With the pace of development accelerating in printed electronics, the fabrication and application of conductive ink have been brought into sharp focus in recent years. The discovery of graphene also unfolded a vigorous research campaign. In this paper, we prepared graphene conductive ink and explored the feasibility of applying the ink to flexible paper-based circuit. Since experimental study concentrating upon ink formulation was insufficient, orthogonal test design was used in the optimization of preparation formula of conductive ink for the first time. The purpose of this study was to determine the effect of constituent dosage on conductivity of graphene conductive ink, so as to obtain the optimized formula and prepare graphene conductive ink with good conductivity. Characterization of optimized graphene conductive ink we fabricated showed good adhesion to substrate and good resistance to acid and water. The graphene concentration of the optimized ink reached 73.17 wt% solid content. Particle size distribution of graphene conductive ink was uniform, which was about 1940 nm. Static surface tension was 28.9 mN/m and equilibrium contact angle was 23°, demonstrating that conductive ink had good wettability. Scanning Electron Microscope (SEM) analysis was also investigated, moreover, the feasibility of lightening a light-emitting diode (LED) light was verified. The graphene conductive ink with optimized formula can be stored for almost eight months, which had potential applications in flexible paper-based circuit in the future.

Cathode Photoelectrochemical Immunosensing Platform Integrating Photocathode with Photoanode.

Generally, photoanode-based photoelectrochemical immunoassay possesses obvious photocurrent response and lower detection limit for ideal sample detection, but it has the inherent imperfection of poor anti-interference capability for real sample detection. Photocathode-based immunoassay can well avoid the intrinsic drawback of photoanode-based immunoassay, but it has low photocurrent response resulting in less good sensitivity. Herein, a promising new cathode photoelectrochemical immunosensing platform integrating photocathode with photoanode was reported for accurate and sensitive detection of biomarkers. In this proposal, prostate-specific antigen (PSA, Ag) was chosen as a model of target analyte to exhibit the analytical performances of this platform. TiO2/CdS:Mn hybrid structure modified indium-tin oxide (ITO) electrode served as photoanode, whereas CuInS2 microflowers modified ITO electrode was selected as photocathode. The transducer elements of PSA antibody (Ab) were modified on photocathode to fabricate a label-free cathode immunosensing electrode. The proposed immunosensing platform possesses two distinct advantages simultaneously. First, it has good anti-interference capability for the detection of real biological samples, since the biorecognition events occurred on photocathode. Second, the photoelectrochemical system owns evident photocurrent response and low detection limit for target Ag detection thanks to the introduction of the photoanode. Moreover, the proposed immunosensing platform also exhibits good specificity, reproducibility, and stability, and meanwhile it opens up a new horizon to construct other kinds of photoelectrochemical biosensors.

[Investigation of ultrasound markers in screening fetal trisomy 21].

OBJECTIVE: To investigate the clinical value of ultrasound markers in screening fetal trisomy 21. METHODS: From Jan. 2001 to Dec. 2011, a retrospective study about sonographic information of 138 fetuses diagnosed as trisomy 21 was taken in the First Affiliated Hospital of Sun Yat-sen University. All fetuses were divided into 3 groups: isolated ultrasound markers, non-isolated ultrasound markers, and isolated structural malformations or other abnormalities. The relationship between trisomy 21 and ultrasound markers as well as structural anomalies or other abnormalities was analyzed. RESULTS: Sonographic anomalies were detected in 132 fetuses (95.7%, 132/138), including ultrasound markers and structural malformations or other abnormalities. One hundred and twenty cases (87.0%, 120/138) had ultrasound markers, 38 (31.7%, 38/120) had one marker and 82 (68.3%, 82/120) had more than one marker (P < 0.01). Fifty-one fetuses (37.0%, 51/138) had isolated ultrasound markers and non-isolated markers were found in 69 fetuses (50.0%, 69/138). Only 12 fetuses (8.7%, 12/138) had isolated structural malformations or other abnormalities. In 20 fetuses on whom the first-trimester ultrasound screening were performed, all had ultrasound markers, 95% (19/20) had thickened nuchal translucency and 55% (11/20) had nasal bone hypoplasia. The most common ultrasound markers on the second-trimester screening were nasal bone hypoplasia, which accounted for 41.9% (52/124) cases, followed by thickened nuchal fold (25.0%, 31/124), short femurs and humerus (24.2%, 30/124), echogenic intracardiac focus (16.1%, 20/124), mild ventriculomegaly (15.3%, 19/124), hyperechoic bowel (12.9%, 16/124), mild renal pyelectasis (12.1%, 15/124). Furthermore, the common structural malformations or other abnormalities were as follows: cardiac defects (33.1%, 41/124), digestive system (26.6%, 33/124). CONCLUSIONS: Ultrasound markers are valuable for screening fetal trisomy 21. The fetuses of trisomy 21 usually had more than one ultrasound markers or associated with other abnormalities. Combinations of ultrasound markers with the results of serum screening and maternal age are necessary for evaluation.

Photoanode-supported cathodic immunosensor for sensitive and specific detection of human chorionic gonadotropin.

Target biomarker detection with high accuracy in biological sample is necessary for the constructed immunoassays. Herein, a novel and enhanced cathodic immunosensor supported by photoanode was designed for sensitive and specific detection of human chorionic gonadotropin (HCG). Specifically, the electrode of TiO2 nanotube with N doping (TiO2:N) was fabricated and assembled with AgInS2 quantum dots (QDs) to acquire the TiO2:N/AgInS2 photoanode. For the sensing cathode, Pt nanoparticles (NPs) were decorated on carbon nanotubes (CNTs) to prepare the CNT/Pt cathodic matrix and was used to modify capture HCG antibody (Ab). In this photoelectrochemical (PEC) sensing system, the TiO2:N/AgInS2 photoanode served as the signal-converting element to produce prominent current signal, while the immune recognition events occurred on the sensing cathode to evidently change the initial current signal from steric hindrance effect. Profiting by excellent photoelectric property and good anti-interference ability of this featured PEC system, the developed cathodic immunosensor demonstrated high sensitivity and specificity for the detection of target HCG antigen (Ag). This photoanode-supported cathodic sensing strategy provided a potential path forward to exploit other enhanced PEC immunosensors in the application of biological samples.

RSK2 promotes melanoma cell proliferation and vemurafenib resistance via upregulating cyclin D1.

BRAF inhibitors are commonly used in targeted therapies for melanoma patients harboring BRAFV600E mutant. Despite the benefit of vemurafenib therapy, acquired resistance during or after treatment remains a major obstacle in BRAFV600E mutant melanoma. Here we found that RSK2 is overexpressed in melanoma cells and the high expression of RSK2 indicates poor overall survival (OS) in melanoma patients. Overexpression of RSK2 leads to vemurafenib resistance, and the deletion of RSK2 inhibits cell proliferation and sensitizes melanoma cells to vemurafenib. Mechanistically, RSK2 enhances the phosphorylation of FOXO1 by interacting with FOXO1 and promoting its subsequent degradation, leading to upregulation of cyclin D1 in melanoma cells. These results not only reveal the presence of a RSK2-FOXO1-cyclin D1 signaling pathway in melanoma, but also provide a potential therapeutic strategy to enhance the efficacy of vemurafenib against cancer.

Functional nucleic acid engineered double-barreled nanopores for measuring sodium to potassium ratio at single-cell level.

The use of double-barreled nanopipette (θ-nanopipette) to electrically sample, manipulate, or detect biomaterials has recently seen strong growth in single-cell studies, driven by the potential of the nanodevices and applications that they may enable. Considering the pivotal roles of Na/K ratio (RNa/K) at cellular level, herein we describe an engineered θ-nanopipette for measuring single-cell RNa/K. The two independently addressable nanopores, located within one nanotip, allow respective customization of functional nucleic acids but simultaneous deciphering of Na and K levels inside a single cell of a non-Faradic manner. Two ionic current rectification signals, corresponding to the Na- and K-specific smart DNA responses, could be easily used to derive the RNa/K. The applicability of this nanotool is validated by practical probing intracellular RNa/K during the drug-induced primary stage of apoptotic volume decrease. Especially, the RNa/K has been shown by our nanotool to be different in cell lines with different metastatic potential. This work is expected to contribute to futuristic study of single-cell RNa/K in various physiological and pathological processes.

[Spectroscopy study of binding mechanisms and molecular recognition of class-specific molecularly imprinted polymer beads].

A novel class-specific molecularly imprinted polymer (MIP) beads for sulfonylurea herbicides was synthesized by precipitation polymerization using chlorsulfuron as the template molecule, 2-(diethylamino) ethyl methacrylate (DEAEMA) as the functional monomer, and trimethylolpropane trimethacrylate (TRIM) as the cross-linker. The mechanisms of recognition of MIP beads to the template molecule were evaluated by UV-spectrum and FTIR in the choosing and optimizing polymerization system experiment. The results showed that MIP beads contained the group which could interact with template molecule and its analogue by the hydrogen bonding specifically.

CdS Quantum Dots Modified Photoelectrochemical Biosensor for TATA-Binding Protein Probing.

The photoelectrochemical (PEC) biosensor, in which light is utilized to excite the photoactive species and current is employed as the detection signal, is a newly appeared yet dynamically developing technique for biological analysis. Based on the assay of DNA binding proteins upon visible light irradiation, a PEC biosensor is constructed for successfully probing a DNA-protein interaction.