Magnetic nanoarrays on flexible substrates.

Abstract: In this work, we used nanosphere lithography to fabricate large area 2-D magnetic nanoparticle (MNP) arrays on a flexible polyimide substrate (Kapton). Samples were fabricated by assembling polystyrene (PS) spheres on thin films of Co capped with Au. Etched PS spheres were used to mask Co-Au particle arrays. The MNP arrays were subjected to superconducting quantum interference device measurements; flat samples (10 nm Co coated with 10 nm Au) exhibited an M s of 117.3 emu g-1, which was lower than the reported literature value for bulk Co (162.7 emu g-1). When compared to the flat film, coercivity, H c, increased in a linear fashion with respect to particle size. These preliminary results reveal that future investigations of the magnetic properties on flexible substrates should account for residual Co remaining in the polymeric material, the unique MNP shape, the effect of order (or lack or order) of the 2D array, and positioning with respect to the direction of the magnetic field. Supplementary Information: The online version contains supplementary material available at 10.1557/s43580-021-00193-z.

Gold nanostar-based voltammetric sensor for chromium(VI).

This paper presents an electrochemical sensor for Cr(VI) (chromate ion) in water. A disposable screen-printed electrode was modified with gold nanostars (AuNSs) that were synthesized by Good's buffer method. Linear sweep voltammetry (LSV) was employed for the detection of Cr(VI) in 0.1 M sulfuric acid solution. The AuNSs are shown to provide higher current response to Cr(VI) than spherically shaped gold nanoparticles. The sensor gives the strongest response at a scan rate of 0.05 V (vs Ag/AgCl) and exhibits minimal interference from other electroactive species. The linear range extends from 10 to 75,000 ppb, and the limit of detection is 3.5 ppb. This is well below the provisional guideline value given by the World Health Organization. Excellent recoveries (ranging between 95 and 97%) were found when analyzing contaminated ground water samples obtained from a site situated in Wellesley, MA. Graphical abstract Schematic presentation of preparation of gold nanostars (AuNS) on carbon paste screen printed electrode (CPSPE) by drop casting and electrochemical detection of chromium (VI) using linear sweep voltammetry (LSV).

Potential of using cerium oxide nanoparticles for protecting healthy tissue during accelerated partial breast irradiation (APBI).

The purpose of this study is to investigate the feasibility of using cerium oxide nanoparticles (CONPs) as radical scavengers during accelerated partial breast irradiation (APBI) to protect normal tissue. We hypothesize that CONPs can be slowly released from the routinely used APBI balloon applicators-via a degradable coating-and protect the normal tissue on the border of the lumpectomy cavity over the duration of APBI. To assess the feasibility of this approach, we analytically calculated the initial concentration of CONPs required to protect normal breast tissue from reactive oxygen species (ROS) and the time required for the particles to diffuse to various distances from the lumpectomy wall. Given that cerium has a high atomic number, we took into account the possible inadvertent dose enhancement that could occur due to the photoelectric interactions with radiotherapy photons. To protect against a typical MammoSite treatment fraction of 3.4Gy, 5ng·g(-1) of CONPs is required to scavenge hydroxyl radicals and hydrogen peroxide. Using 2nm sized NPs, with an initial concentration of 1mg·g(-1), we found that 2-10days of diffusion is required to obtain desired concentrations of CONPs in regions 1-2cm away from the lumpectomy wall. The resultant dose enhancement factor (DEF) is less than 1.01 under such conditions. Our results predict that CONPs can be employed for radioprotection during APBI using a new design in which balloon applicators are coated with the NPs for sustained/controlled in-situ release from within the lumpectomy cavity.

Modification of carbon nanotube electrodes with 1-pyrenebutanoic acid, succinimidyl ester for enhanced bioelectrocatalysis.

Conductive materials functionalized with redox enzymes provide bioelectronic architectures with application to biological fuel cells and biosensors. Effective electron transfer between the enzyme (biocatalyst) and the conductive materials is imperative for function. Various nanostructured carbon materials are common electrode choices for these applications as both the materials' inherent conductivity and physical integrity aids optimal performance. The following chapter presents a method for the use of carbon nanotube buckypaper as a conductive architecture suitable for biocatalyst functionalization. In order to securely attach the biocatalyst to the carbon nanotube surface, the conductive buckypaper is modified with the heterobifunctional cross-linker, 1-pyrenebutanoic acid, succinimidyl ester. The technique effectively tethers the enzyme to the carbon nanotube which enhances bioelectrocatalysis, preserves the conductive nature of the carbon surface, and facilities direct electron transfer between the catalyst and material interface. The approach is demonstrated using phenol oxidase (laccase) and pyrroloquinoline quinone-dependent glucose dehydrogenase PQQ-GDH, as representative biocatalysts.

Power generation from a hybrid biological fuel cell in seawater.

A hybrid biological fuel cell (HBFC) comprised of a microbial anode for lactate oxidation and an enzymatic cathode for oxygen reduction was constructed and then tested in a marine environment. Shewanella oneidensis DSP-10 was cultivated in laboratory medium and then fixed on a carbon felt electrode via a silica sol-gel process in order to catalyze anodic fuel cell processes. The cathode electrocatalyst was composed of bilirubin oxidase, fixed to a carbon nanotube electrode using a heterobifunctional cross linker, and then stabilized with a silica sol-gel coating. The anode and cathode half-cells provided operating potentials of -0.44 and 0.48 V, respectively (vs. Ag/AgCl). The HBFC maintained a reproducible open circuit voltage >0.7 V for 9 d in laboratory settings and sustained electrocatalytic activity for >24h in open environment tests.

Realization and properties of biochemical-computing biocatalytic XOR gate based on enzyme inhibition by a substrate.

We consider a realization of the XOR logic gate in a process biocatalyzed by an enzyme which can be inhibited by a substrate when the latter is inputted at large enough concentrations. A model is developed for describing such systems in an approach suitable for evaluation of the analog noise amplification properties of the gate. The obtained data are fitted for gate quality evaluation within the developed model, and we discuss aspects of devising XOR gates for functioning in "biocomputing" systems utilizing biomolecules for information processing.

Alert-type biological dosimeter based on enzyme logic system.

A cooperative effect of two biomarkers, α-amylase and lactate dehydrogenase, was used to analyze radiation-caused tissue damage in vitro in model solutions of human serum. The analytical system was based on the recently emerged biocomputing concept applying biocatalytic cascades for logic processing of biochemical input signals. The studied system resembled a Boolean NAND logic gate in which the change of the optical output signal from a high level (logic value 1) to a low level (logic value 0) confirmed the presence of both biomarkers at pathological concentrations (1,1 input signals), thus yielding the conclusion about radiation tissue damage. The system operates in a digital YES/NO format as an alert-type biosensor with a built-in Boolean logic.

Bioelectrocatalytic generation of directly readable code: harnessing cathodic current for long-term information relay.

Here we present an exceptionally stable bioelectrocatalytic architecture for electrocatalytic oxygen reduction using a carbon nanotube electrode as the electron donor and a fungal enzyme as electrocatalyst. Controlling oxygen content in the electrolyte enables generation of a directly readable barcode from monitoring the enzyme response.

Multi-enzyme logic network architectures for assessing injuries: digital processing of biomarkers.

A multi-enzyme biocatalytic cascade processing simultaneously five biomarkers characteristic of traumatic brain injury (TBI) and soft tissue injury (STI) was developed. The system operates as a digital biosensor based on concerted function of 8 Boolean AND logic gates, resulting in the decision about the physiological conditions based on the logic analysis of complex patterns of the biomarkers. The system represents the first example of a multi-step/multi-enzyme biosensor with the built-in logic for the analysis of complex combinations of biochemical inputs. The approach is based on recent advances in enzyme-based biocomputing systems and the present paper demonstrates the potential applicability of biocomputing for developing novel digital biosensor networks.

Enzymatic AND logic gates operated under conditions characteristic of biomedical applications.

Experimental and theoretical analyses of the lactate dehydrogenase and glutathione reductase based enzymatic AND logic gates in which the enzymes and their substrates serve as logic inputs are performed. These two systems are examples of the novel, previously unexplored class of biochemical logic gates that illustrate potential biomedical applications of biochemical logic. They are characterized by input concentrations at logic 0 and 1 states corresponding to normal and pathophysiological conditions. Our analysis shows that the logic gates under investigation have similar noise characteristics. Both significantly amplify random noise present in inputs; however, we establish that for realistic widths of the input noise distributions, it is still possible to differentiate between the logic 0 and 1 states of the output. This indicates that reliable detection of pathophysiological conditions is indeed possible with such enzyme logic systems.

Multiplexing of injury codes for the parallel operation of enzyme logic gates.

The development of a highly parallel enzyme logic sensing concept employing a novel encoding scheme for the determination of multiple pathophysiological conditions is reported. The new concept multiplexes a contingent of enzyme-based logic gates to yield a distinct 'injury code' corresponding to a unique pathophysiological state as prescribed by a truth table. The new concept is illustrated using an array of NAND and AND gates to assess the biomedical significance of numerous biomarker inputs including creatine kinase, lactate dehydrogenase, norepinephrine, glutamate, alanine transaminase, lactate, glucose, glutathione disulfide, and glutathione reductase to assess soft-tissue injury, traumatic brain injury, liver injury, abdominal trauma, hemorrhagic shock, and oxidative stress. Under the optimal conditions, physiological and pathological levels of these biomarkers were detected through either optical or electrochemical techniques by monitoring the level of the outputs generated by each of the six logic gates. By establishing a pathologically meaningful threshold for each logic gate, the absorbance and amperometric assays tendered the diagnosis in a digitally encoded 6-bit word, defined as an 'injury code'. This binary 'injury code' enabled the effective discrimination of 64 unique pathological conditions to offer a comprehensive high-fidelity diagnosis of multiple injury conditions. Such processing of relevant biomarker inputs and the subsequent multiplexing of the logic gate outputs to yield a comprehensive 'injury code' offer significant potential for the rapid and reliable assessment of varied and complex forms of injury in circumstances where access to a clinical laboratory is not viable. While the new concept of parallel and multiplexed enzyme logic gates is illustrated here in connection to multi-injury diagnosis, it could be readily extended to a wide range of practical medical, industrial, security and environmental applications.

Self-powered biomolecular keypad lock security system based on a biofuel cell.

The enzyme-based keypad lock was integrated with a biofuel cell yielding a self-powered biomolecular information security system. The correct "password" introduced into the keypad lock resulted in the activation of the biofuel cell, while all other "wrong" permutations of the enzyme inputs preserved the "OFF" state of the biofuel cell.

Set-reset flip-flop memory based on enzyme reactions: toward memory systems controlled by biochemical pathways.

The enzyme-based set-reset flip-flop memory system was designed with the core part composed of horseradish peroxidase and diaphorase biocatalyzing oxidation and reduction of redox species (2,6-dichloroindophenol or ferrocyanide). The biocatalytic redox reactions were activated by H(2)O(2) and NADH produced in situ by different enzymatic reactions allowing transformation of various biochemical signals (glucose, lactate, d-glucose-6-phosphate, ethanol) into reduced or oxidized states of the redox species. The current redox state of the system, controlled by the set and reset signals, was read out by optical and electrochemical means. The multiwell setup with the flip-flop units separately activated by various set/reset signals allowed encoding of complex information. For illustrative purposes, the words "Clarkson" and then "University" were encoded using ASCII character codes. The present flip-flop system will allow additional functions of enzyme-based biocomputing systems, thus enhancing the performance of multisignal biosensors and actuators controlled by logically processed biochemical signals. The integrated enzyme logic systems and flip-flop memories associated with signal-responsive chemical actuators are envisaged as basic elements of future implantable biomedical devices controlled by immediate physiological conditions.

Biofuel cell logically controlled by antigen-antibody recognition: towards immune-regulated bioelectronic devices.

A switchable biofuel cell logically controlled by immune signals was developed as a model prototype for future adaptive implantable bioelectronic devices regulated by immune reactions. The cell demonstrated NOR Boolean logic operation in situ controlled by antibody signals.

Logic networks based on immunorecognition processes.

The biochemical system logically processing biochemical signals using immune-specific and biocatalytic reactions was designed, and the generated output signals were analyzed by AFM and optical means. Different patterns of immune signals resulted in the formation of various interfacial structures followed by biocatalytic reactions activated by the next set of biochemical inputs. The developed approach to multisignal biosensing allows qualitative evaluation of the biochemical information in terms of YES-NO, providing the base for novel molecular-level logic analysis of complex patterns of biochemical signals. Application of AFM to read out the structures generated on the interface could potentially lead to substantial miniaturization of the immune logic systems.

Analog noise reduction in enzymatic logic gates.

In this work, we demonstrate both experimentally and theoretically that the analog noise generation by a single enzymatic logic gate can be dramatically reduced to yield gate operation with virtually no input noise amplification. We demonstrate that when a cosubstrate with a much smaller affinity than the primary substrate is used, a negligible increase in the noise output from the logic gate is obtained, as compared to the input noise level. Our general theoretical conclusions were confirmed by experimental realizations of the AND logic gate based on the enzyme horseradish peroxidase using hydrogen peroxide as the substrate, with 2,2'-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) or ferrocyanide as cosubstrates with vastly different rate constants.

Optimization of enzymatic biochemical logic for noise reduction and scalability: how many biocomputing gates can be interconnected in a circuit?

We report an experimental evaluation of the "input-output surface" for a biochemical AND gate. The obtained data are modeled within the rate-equation approach, with the aim to map out the gate function and cast it in the language of logic variables appropriate for analysis of Boolean logic for scalability. In order to minimize "analog" noise, we consider a theoretical approach for determining an optimal set for the process parameters to minimize "analog" noise amplification for gate concatenation. We establish that under optimized conditions, presently studied biochemical gates can be concatenated for up to order 10 processing steps. Beyond that, new paradigms for avoiding noise buildup will have to be developed. We offer a general discussion of the ideas and possible future challenges for both experimental and theoretical research for advancing scalable biochemical computing.

Boolean logic gates that use enzymes as input signals.

Biochemical systems that demonstrate the Boolean logic operations AND, OR, XOR, and InhibA were developed by using soluble compounds, which represent the chemical "devices", and the enzymes glucose oxidase (GOx), glucose dehydrogenase (GDH), alcohol dehydrogenase (AlcDH), and microperoxidase-11 (MP-11), which operated as the input signals that activated the logic gates. The enzymes were used as soluble materials and as immobilized biocatalysts. The studied systems are proposed to be a step towards the construction of "smart" signal-responsive materials with built-in Boolean logic.

Biocomputing security system: concatenated enzyme-based logic gates operating as a biomolecular keypad lock.

A biomolecular security system mimicking a keypad lock device was developed using enzyme-based concatenated AND logic gates resulting in the implication logic network.