CRISPR/Cas12a-Assisted Visual Logic-Gate Detection of Pathogenic Microorganisms Based on Water-Soluble DNA-Binding AIEgens

Here, we developed a rapid, visual and double-checked Logic Gate detection platform for detection of pathogenic microorganisms by aggregation-induced emission luminogens (AIEgens) in combination with Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)/CRISPR associated (Cas). DNA light-up AIEgens (1,1,2,2-tetrakis[4-(2-bromo-ethoxy) phenyl]ethene, TTAPE) was non-emissive but the emission was turned on in the presence of large amount of DNA produced by recombinase polymerase amplification (RPA). When CRISPR/Cas12a was added, all long-stranded DNA were cut leading to the emission quenched. Thus, a method that can directly observe the emission changes with the naked eye has been successfully constructed. The detection is speedy within only 20 min, and has strong specificity to the target. The result can be judged by Logic Gate. Only when the output signal is (1,0), does it represent the presence of pathogenic microorganisms in the test object. Finally, the method was applied to the detect pathogenic microorganisms in environmental water samples, which proved that this method has high selectivity, specificity and applicability for the detection of pathogenic microorganisms in environmental water samples.

Towards Multi-Analyte Detection with Field-Effect Capacitors Modified with Tobacco Mosaic Virus Bioparticles as Enzyme Nanocarriers

Utilizing an appropriate enzyme immobilization strategy is crucial for designing enzyme-based biosensors. Plant virus-like particles represent ideal nanoscaffolds for an extremely dense and precise immobilization of enzymes, due to their regular shape, high surface-to-volume ratio and high density of surface binding sites. In the present work, tobacco mosaic virus (TMV) particles were applied for the co-immobilization of penicillinase and urease onto the gate surface of a field-effect electrolyte-insulator-semiconductor capacitor (EISCAP) with a p-Si-SiO2-Ta2O5 layer structure for the sequential detection of penicillin and urea. The TMV-assisted bi-enzyme EISCAP biosensor exhibited a high urea and penicillin sensitivity of 54 and 85 mV/dec, respectively, in the concentration range of 0.1–3 mM. For comparison, the characteristics of single-enzyme EISCAP biosensors modified with TMV particles immobilized with either penicillinase or urease were also investigated. The surface morphology of the TMV-modified Ta2O5-gate was analyzed by scanning electron microscopy. Additionally, the bi-enzyme EISCAP was applied to mimic an XOR (Exclusive OR) enzyme logic gate.

Double-Gate Operation of Metal Nanodot-Array-Based Single-Electron Device

Multidot single-electron devices (SEDs) can realize new types of computing technologies, such as reconfigurable and reservoir computing. The self-assembled metal nanodot-array film attached with multiple gates is a candidate for use in such SEDs to achieve high functionality. However, the single-electron properties of such a film have not yet been investigated in use with optimally controlled multiple gates because of structural complexity having many nanodots. In this study, Fe nanodot-array-based double-gate SEDs were fabricated and their single-electron properties modulated by the top- and bottom-gate voltages (VT and VB, respectively) were investigated. As reported in our previous study, the drain current (ID) exhibited clear oscillations against VB (i.e., Coulomb blockade oscillation) in a part of the devices, originating from a single dot among several dots. The phase of the Coulomb blockade oscillation systematically shifted with VT, indicating that the charge state of the single dot was clearly controlled by both the gate voltages despite the multidot structure and the metal multidot SED has potential for logic-gate operation. The top and bottom gates affected the electronic state of the dot unevenly owing to the geometrical effect caused by the dot shape and size of the surrounding dots.

Terminal Deoxynucleotidyl Transferase Extension-Dominated In Situ Signal Attenuation-Free Electrochemical Platform and Its Logic Gate Manipulation

Signal generation of traditional electrochemical biosensors suffers from the random diffusion of electroactive probes in a electrolyte solution, which is accompanied by poor reaction kinetics and low signal stability from complex biological systems. Herein, a novel circuit system with autonomous compensation solution ohmic drop (noted as “fast-scan cyclic voltammetry (FSCV)”) is developed to solve the above problems, and employed to achieve terminal deoxynucleotide transferase (TdT) and its small molecule inhibitor analysis. At first, a typical TdT-mediated catalytic polymerization in the conditions of original DNA, deoxythymine triphosphate (dTTP) and Hg2+ is applied for the electrode assembly. The novel electrochemical method can provide some unattenuated signals due to in-situ Hg redox reaction, thus improving reaction kinetics and signal stability. This approach is mainly dependent on TdT-mediated reaction, so it can be applied properly for TdT investigation, and a detection limit of 0.067 U/mL (S/N=3) is achieved successfully. More interesting, we also mimic the function of TdT-related signal communication in various logic gates such as YES, NOT, AND, N-IMPLY, and AND-AND-N-IMPLY cascade circuit. This study provides a new method for the detection of TdT biomarkers in many types of diseases and the construction of a signal attenuation-free logic gate.

Fuzzy Matrix Game: A Fast Approach using Artificial Hybrid Neural-Net Logic-Gate Switching Circuit

Introducing neuro -fuzzy concept in decision making problems, makes a new way in artificial intelligence and expert systems. Sometimes, neural networks are used to optimize certain performances. In general, knowledge acquisition becomes difficult when problem's variables, constraints, environment, decision maker's attitude and complex behavior are encountered with. A sense of fuzziness prevails in these situations; sometimes numerically and sometimes linguistically. Neural networks (or neural nets) help to overcome this problem. Neural networks are explicitly and implicitly hyped to draw out fuzzy rules from numerical information and linguistic information. Logic-gate and switching circuit mobilize the fuzzy data in crisp environment and can be used in artificial neural network, also. Game theory has a tremendous scope in decision making; and consequently decision makers' hesitant characters play an important role in it. In this paper, a game situation is clarified under artificial neural network through logic-gate switching circuit in hesitant fuzzy environment with a suitable example; and this concept can be applied in future for real-life situations.

An efficient 2-aminothiazolesalicylaldehyde Fluorescent Chemosensor for Fe2+ ion Detection and Potential Inhibitor of NUDT5 Signaling Hormone for Breast Cancer Cell and Molecular Keypad Lock Application

A novel thiazole phenol conjugate 2-aminothiazolesalicylaldehyde (receptor1) was designed and synthesized for the first time through a single step process via schiff base condensation reaction. The formation of receptor1 was confirmed by FTIR, 13C NMR and 1H NMR. The receptor1 complexing with various metal ions studied through fluorescence spectroscopy showed its selectivity towards Fe2+ ion following a reverse photo induced electron transfer (PET) process compared to all other potentially competing ions. Furthermore, the receptor1 showed excellent reversibility property on adding EDTA solution. The binding affinity between receptor1-Fe2+ ion was found to be Ka = 2.22x102 M-1 and the sensing affinity of receptor1 towards Fe2+ ion at nanolevel concentration was 33.7x10-9 M which is lower than the detection limit of existing Schiff base. The receptor1 was used to detect Fe2+ ion in different water samples. Using molecular docking the chelating function of ligand (receptor1) was assessed as a potential inhibitor of NUDT5, a silence hormone signaling for breast cancer. The excellent reversibility property is used to construct INHIBIT molecular logic gate.

Dual-Channel Probe of Carbon Dots Cooperating with Lanthanide Complex Employed for Simultaneously Distinguishing and Sequentially Detecting Tetracycline and Oxytetracycline

Tetracycline (TC) and oxytetracycline (OTC) are the most widely used broad-spectrum antimicrobial agents in tetracycline drugs, and their structures and properties are very similar, so it is a great challenge to distinguish and detect these two antibiotics with a single probe at the same time. Herein, a dual-channel fluorescence probe (SiCDs@mMIPs-cit-Eu) was developed by integrating two independent reaction sites with SiCDs-doped mesoporous silica molecular imprinting group and europium complex group into a nanomaterial. The synergistic influence of inner filter effect and “antenna effect” can be guaranteed to solve the distinction between TC and OTC. Moreover, this novel strategy can also sequentially detect TC and OTC in buffer solution and real samples with high sensitivity and selectivity. This method revealed good responses to TC and OTC ranging from 0 to 5.5 μM with a detection limit of 5 and 16 nM, respectively. Combined with the smartphone color-scanning application, the portable and cheap paper-based sensor was designed to realize the multi-color visual on-site detection of TC and OTC. In addition, the logic gate device was constructed according to the fluorescence color change of the probe for TC and OTC, which provided the application possibility for the intelligent detection of the probe.