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This letter introduces the pre-steady-state kinetic approach, which is traditional for evaluation of elementary constants in molecular (enzyme) catalysis, for nanozymes. Apparently, the most active peroxidase-mimicking nanozyme based on catalytically synthesized Prussian Blue nanoparticles has been chosen. The elementary constants ( k 1 ) linearly decays with an increase of the substrate redox potential (cotangent value ≈125 mV). On the contrary, for the largest nanozymes with a diameter above 150 nm, k 1 ) linearly decays with an increase of the substrate redox potential (cotangent value ≈125 mV). On the contrary, for the largest nanozymes with a diameter above 150 nm, k 1 is almost independent of it. Moreover, for the substrate with the lowest redox potential (K 4 [Fe(CN) 6 ]), the rate constant under discussion ( k 1 ) is almost independent of the nanozymes' size. Perhaps, the rate of the intrananozyme electron transfer causing bleaching becomes comparative or even lower than that of the nanoparticle interaction with the fastest substrate. Anyway, the elementary constant of nanozyme reduction with potassium ferrocyanide ( k 1 ) reaches the value of 1 × 10 10 M -1 s -1 , which is 3-4 orders of magnitude faster than for enzymes peroxidases. The obtained results obviously demonstrate that the pre-steady-state kinetic approach is able to discover novel advantages of nanozymes from both fundamental and practical points of view.

Many physical and chemical properties of solids, such as strength, plasticity, dispersibility, solubility and dissolution are determined by defects in the crystal structure. The aim of this work is to study in situ dynamic, dispersion, chemical, biological and surface properties of lacosamide powder after a complete cycle of mechanical loading by laser scattering, electron microscopy, FR-IR and biopharmaceutical approaches. The SLS method demonstrated the spontaneous tendency toward surface-energy reduction due to aggregation during micronisation. DLS analysis showed conformational changes of colloidal particles as supramolecular complexes depending on the loading time on the solid. SEM analysis demonstrated the conglomeration of needle-like lacosamide particles after 60 min of milling time and the transition to a glassy state with isotropy of properties by the end of the tribochemistry cycle. The following dynamic properties of lacosamide were established: elastic and plastic deformation boundaries, region of inhomogeneous deformation and fracture point. The ratio of dissolution-rate constants in water of samples before and after a full cycle of loading was 2.4. The lacosamide sample, which underwent a full cycle of mechanical loading, showed improved kinetics of API release via analysis of dissolution profiles in 0.1 M HCl medium. The observed activation-energy values of the cell-death biosensor process in aqueous solutions of the lacosamide samples before and after the complete tribochemical cycle were 207 kJmol -1 and 145 kJmol -1 , respectively. The equilibrium time of dissolution and activation of cell-biosensor death corresponding to 20 min of mechanical loading on a solid was determined. The current study may have important practical significance for the transformation and management of the properties of drug substances in solid form and in solutions and for increasing the strength of drug matrices by pre-strain hardening via structural rearrangements during mechanical loading.

We report on the drop-cast production of glucose biosensors based on the most efficient bioelectrocatalysis by pyrroloquinoline quinone dependent glucose dehydrogenase (PQQ GDH). To orient the enzyme upon immobilization we suggest using poly(Methylene Green) (p(MG)) nanoparticles acting as anchors. Synthesis of polymeric anchors has been carried out in course of Methylene Green electropolymerization, which allowed to tune polymer-to-monomer ratio in the drop-cast mixtures varying the monomer concentration and applying different number of potential sweep cycles. Except for elimination of electrochemical step for the electrode modification, opening the prospects for mass-production, the use of drop-cast enzyme anchors provides advantageous characteristics of the resulting biosensors. Catalytic current of PQQ GDH immobilized over p(MG) nanoparticles obtained in optimal conditions is increased only 2-2.5 times after addition of the freely diffusing mediator. Obviously, the lowest ratio of mediated-to-reagentless current points to the most efficient bioelectrocatalysis. The obtained ratio is 2.5 times lower than that for biosensors based on electropolymerized p(MG) films and practically an order of magnitude lower than that for the best reagentless sensors based on PQQ GDH immobilized over conductive nanomaterials. The achieved most efficient bioelectrocatalysis provides high sensitivity of the elaborated biosensors even at 0.0 V potential, which allows to operate them in power generation mode and control relative sweat glucose variation as a tool for non-invasive diabetes monitoring., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier B.V. All rights reserved.)

The paper describes a comparative study of swelling processes in free-standing graphene oxide (GO) membranes and GO laminates encapsulated with epoxy glue. For free-standing graphene oxide membranes, a huge variation in d -spacing in the range of 8-12 Å depending on the ambient humidity and from 12 to >30 Å depending on the electrolyte type and its concentration was revealed using direct in situ and in operando XRD studies. Limited swelling at various humidity levels as well as in electrolyte solution with low constriction/expansion of epoxy-encapsulated GO is counterposed to that of free-standing graphene oxides. The swelling suppression was explained by both physical constriction and the intercalation of amines into GO laminates, which was proved by local EDX studies. This results in ion diffusivity variation for over 2 orders of magnitude in free-standing and constrained graphene oxide membranes and provides factual evidence for tunable sieving of ions with confined graphene oxides.

We propose pulse power generation (PPG) amperometry as an advanced readout realized for Prussian blue (PB)-based (bio)sensors. In contrast to the conventional power generation mode, when the current response is generated upon continuous short-circuiting, the suggested pulse regime is fulfilled by periodic opening and shorting of the circuit. Despite PB being electroactive, the pulse readout is advantageous over conventional steady-state power generation, providing up to a 15-fold increased signal-to-background ratio as well as dramatically improved sensitivity exceeding 10 A·M -1 ·cm -2 for H 2 O 2 sensors and 3.9 A·M -1 ·cm -2 for glucose biosensors. Such analytical performance characteristics are, most probably, achieved due to the enrichment of the diffusion layer by analyte mass transfer from the bulk upon opening of the circuit. Due to an improved sensitivity-to-background ratio, reduced flow-rate dependence, and enhanced operational stability, the regime allows reliable monitoring of blood glucose variations through sweat analysis with the on-skin device.

We propose catalytically synthesized nanozymes based on Prussian Blue (PB) and azidomethyl-substituted poly (3,4-ethylenedioxythiophene) (azidomethyl-PEDOT) as novel electrocatalytic labels for DNA/RNA sensors. Catalytic approach allowed to synthesize highly redox and electrocatalytically active Prussian Blue nanoparticles functionalized with azide groups that enable 'click' conjugation with alkyne-modified oligonucleotides. Both competitive and sandwich-type schemes were realized. As the sensor response the direct (mediator-free) electrocatalytic current of H 2 O 2 reduction can be measured, which is proportional to the concentration of the hybridized labeled sequences. The current of H 2 O 2 electrocatalytic reduction is only 3-8 times increased in the presence of the freely diffusing mediator catechol, which indicates high efficiency of direct electrocatalysis with the elaborated labels. Electrocatalytic amplification of the signal allows robust detection of (63-70)-base target sequences with concentrations below 0.2 nM in blood serum within an hour. We believe, the use of advanced Prussian Blue based electrocatalytic labels sets new avenues for point-of-care DNA/RNA sensing., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier B.V. All rights reserved.)

Despite non-invasive instant monitoring of sweat metabolites is becoming a general trend in early diagnostics and screening, the reliability and accuracy of the on-skin electrochemical biosensors in real-life scenarios still remain questionable. As a rule, mass transport effects in scantily excreted liquids are ignored, when considering the design of such wearable setups. Here we provide a comprehensive investigation of the disruption factors for commonly used Prussian Blue based (bio)sensors under different hydrodynamic conditions (2 × 10 -5 - 5 × 10 0  mm s -1 electrolyte velocity). A huge effect of flow on the (bio)sensors response has been revealed and explained with transport limitations for both analyte influx and reaction product outflux. It suggests no need for improving the sensor sensitivity, while minimizing analyte consumption and enhancing product withdrawal. Some strategies concerning measurement schemes and sensor design ensuring reliable sweat analysis have been discussed and illustrated for lactate and glucose on-skin monitoring., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier B.V. All rights reserved.)

The pathway from the advanced electrocatalyst to nanozymes defeating natural enzyme is reviewed. Prussian blue, being the most advantageous electrocatalyst for hydrogen peroxide reduction, is obviously the best candidate for mimicking peroxidase activity. Indeed, catalytically synthesized Prussian blue nanoparticles are characterized by the catalytic rate constants, which are significantly (up to 4 orders of magnitude) higher than for enzyme peroxidase. Displaying in addition the enzymatic specificity in terms of an absence of oxidase-like activity, catalytically synthesized Prussian blue nanoparticles can be referred to as nanozymes. The latter provide the most versatile method for surface covering with the electrocatalyst, allowing to modify non-traditional materials like boron-doped diamond. For stabilization, Prussian blue core can be covered with nickel hexacyanoferrate shell; the resulting core-shell nanozymes still defeat natural enzyme in terms of activity. Discovering the catalytic pathway of nanozymes "artificial peroxidase" action, we have found the novel advantage of nanozymes over the corresponding biological catalysts: their dramatically (100 times) improved bimolecular rate constants., (© 2022. The Author(s), under exclusive licence to Springer-Verlag GmbH Austria, part of Springer Nature.)

We report on the simultaneous monitoring of sweat lactate concentration and sweat secretion rate. For this aim lactate oxidase-Prussian Blue enzyme-nanozyme type lactate biosensors were elaborated. The use of siloxane-perfluorosulfonated ionomer composite membrane for enzyme-nanozyme immobilization results in the biosensor displaying flux independence in the whole range of physiological sweat secretion rates (0.025-2 μl cm -2 min -1 ). On the contrary, current response of the biosensor based on solely siloxane membranes becomes saturated at physiological sweat lactate concentration, depending mostly on the flow rate. Accordingly, for simultaneous monitoring of sweat lactate concentration and its secretion rate both flow-through biosensors were integrated with high-accuracy wearable electronic devices allowing real-time remote monitoring. As found, during exhaustive physical exercise sweat secretion rate and lactate content are independent of each other, thus, confirming that this excretory liquid is suitable for non-invasive diagnostics., (Copyright © 2022 Elsevier B.V. All rights reserved.)

We report on fully electrochemical flow-through synthesis of Prussian Blue based nanozymes defeating peroxidase in terms of more than 200 times higher catalytic rate constant (k = 6 × 10 4 s -1 ). Being reagentless, reproducible, simple and scalable, the proposed approach blazes new trails for the electrosynthesis of functional conductive and electroactive nanomaterials.

Catalytic current of pyrroloquinoline quinone (PQQ)-glucose dehydrogenase (PQQ-GDH) immobilized over electropolymerized methylene green (MG) is increased only five times after the addition of the freely diffusing mediator. This value, being an efficiency criterion for bioelectrocatalysis, is several (three to six) times lower than that for the best reagentless glucose electrodes reported for this enzyme. Thermodynamics of the polyMG|PQQ-GDH electrode is determined by the enzyme-catalyzed reaction pointing to the direct bioelectrocatalysis. PQQ-GDH immobilized over polyMG displays the current plateau region from 0.0 to 0.2 V in the presence of glucose; at 0.00 V, being the optimal potential for biosensing applications, the catalytic current of the polyMG|PQQ-GDH electrode is 700-fold higher than that for the enzyme on a blank electrode. Successful glucose detection in human sweat by means of the corresponding enzyme electrode confirms that the reported bioelectrocatalytic system is attractive for advanced biosensors, as well as for biofuel cells.

We report on the nanoparticles composed of the catalytically synthesized Prussian Blue (PB) core stabilized with the nickel hexacyanoferrate (NiHCF) shell. Catalyzing hydrogen peroxide reduction, the resulting nanozymes (ø = 66 nm) display catalytic rate constants, which for pyrogallol or ferrocyanide are, respectively, 25 and 35 times higher than those for peroxidase enzyme. After more than half a year of storage at a room temperature, the core-shell PB-NiHCF nanozymes retain both their size and physicochemical properties; such stability is unreachable for the enzymes. Being immobilized, core-shell PB-NiHCF nanozymes (ø = 45 nm) result in a hydrogen peroxide sensor with a sensitivity similar to that of the sensor based on sole PB nanoparticles. However, whereas the latter response in hard inactivating conditions (25 min in 1 mM H 2 O 2 ) drops down to 7.5%, the PB-NiHCF nanozymes-based sensor retains >75% of initial sensitivity. Application of the core-shell PB-NiHCF nanozymes "artificial peroxidase" would obviously open new horizons in elaboration of anti-inflammatory drugs and (bio)sensors.

We report on the kinetic mechanism of the catalytically synthesized Prussian Blue nanoparticles denoted as "artificial peroxidase". In contrast to the enzyme, whose active site first interacts with hydrogen peroxide forming the so-called Compound I, in the case of the nanozymes, H 2 O 2 oxidizes their complex with reducing substrate. Slow release of the product (oxidized form of the latter) from the nanozymes has been registered. The interaction of substrates with the nanozymes is 100 times faster than with enzyme peroxidases, and the rate-limiting constant for the nanozymes is also 2 orders of magnitude greater: for pyrogallol k 2 = 1.3 ± 0.1 × 10 8 M -1 s -1 and for ferrocyanide k 2 = 1.9 ± 0.1 × 10 8 M -1 s -1 . Thus, the discovered novel advantage of nanozymes over the corresponding enzymes is the 100-fold greater bimolecular rate constants, resulting, most probably, from their uniformly accessible surface, avoiding the effect of rotation on the diffusion-controlled rate.

In contrast to bienzyme biosensors, we propose the nanozyme-enzyme based ones substituting the enzyme peroxidase with the more active and stable nanoparticles "artificial peroxidase". The use of catalytically synthesized Prussian Blue based nanozymes simplifies assembling of hydrogen peroxide transducer providing its higher sensitivity. For immobilization of lactate oxidase the composite alkoxysilane - perfluorosulfonated ionomer (PFSI) membranes are proposed achieving the significantly improved operating stability. The resulting nanozyme-enzyme lactate biosensor displays twice higher sensitivity (>0.2 A M -1  cm -2 ) compared to the Prussian Blue film based one. Nanozymes "artificial peroxidase" are expected to find wide use in elaboration of oxidase based biosensors., (Copyright © 2019 Elsevier B.V. All rights reserved.)

We synthesized Prussian Blue (PB) nanoparticles through catalytic reaction involving hydrogen peroxide (H 2 O 2 ) activation. The resulting nanoparticles display the size-dependent catalytic rate constants in H 2 O 2 reduction, which are significantly improved compared to natural enzyme peroxidase: for PB nanoparticles 200 nm in diameter, the turnover number is 300 times higher; for 570 nm diameter nanoparticles, it is 4 orders of magnitude higher. Comparing to the known peroxidase-like nanozymes, the advantages of the reported PB nanoparticles are their true enzymatic properties: (1) enzymatic specificity (an absence of oxidase-like activity) and (2) an ability to operate in physiological solutions. The ultrahigh activity and enzymatic specificity of the catalytically synthesized PB nanoparticles together with high stability and low cost, obviously peculiar to noble metal free inorganic materials, would allow the substitution of natural and recombinant peroxidases in biotechnology and analytical sciences.

In contrast to "self-powered" (bio)sensors aiming to generate maximum energy output, we propose the systems with the lowest potential difference between the working and the counter electrodes, which in galvanic mode would provide achievement of the best analytical performance characteristics. Prussian Blue based (bio)sensors known to operate at 0.00 V versus Ag|AgCl reference, in the short-circuit regime generate the current proportional to analyte concentration. Sensitivity and dynamic range of Prussian Blue based (bio)sensors in power generation mode are, respectively, even slightly higher and wider compared to the same (bio)sensors operated in the conventional three-electrode regime powered by a potentiostat. Selectivity of the (bio)sensors in power generation mode is similarly high relative to both oxygen, allowing H 2 O 2 detection by its reduction, and reductants. Among the most important advantages of the proposed power generation mode is an order of magnitude decreased noise compared to performance in a conventional three-electrode setup powered by a potentiostat. Noiseless performances of Prussian Blue based (bio)sensors would open new horizons for electrochemical analysis.

We report on the novel reagentless and label-free detection principle based on electroactive (conducting) polymers considering sensors for polyols, particularly, saccharides and hydroxy acids. Unlike the majority of impedimetric and conductometric (bio)sensors, which specific and unspecific signals are directed in the same way (resistance increase), making doubtful their real applications, the response of the reported system results in resistance decrease, which is directed oppositely to the background. The mechanism of the resistance decrease is the polyaniline self-doping, i.e., as an alternative to proton doping, an appearance of the negatively charged aromatic ring substituents in polymer chain. Negative charge "freezing" at the boron atom is indeed a result of complex formation with di- and polyols, specific binding. Changes in Raman spectra of boronate-substituted polyaniline after addition of glucose are similar to those caused by proton doping of the polymer. Thermodynamic data on interaction of the electropolymerized 3-aminophenylboronic acid with saccharides and hydroxy acids also confirm that the observed resistance decrease is due to polymer interaction with polyols. The first reported conductivity increase as a specific signal opens new horizons for reagentless affinity sensors, allowing the discrimination of specific affinity bindings from nonspecific interactions.

The ability of Prussian Blue, ferric hexacyanoferrate (FeHCF), to sensitively and selectively detect hydrogen peroxide by its reduction in the presence of oxygen is of high importance for analytical chemistry. Success with Prussian Blue (PB) provided an appearance of contradictory reports concerning electrocatalysis of the other transition metal hexacyanoferrates (HCFs) in H2O2 reduction. Investigating thermodynamics of the catalyzed reactions as well as electrochemical properties of the hexacyanoferrates, we are able to conclude that the noniron hexacyanoferrates themselves are completely electrocatalytically inactive, except for a minor electrocatalysis in the opposite reaction, hydrogen peroxide oxidation, registered for NiHCF. Concerning the most important reaction, H2O2 reduction, the observed electrocatalytic activity (by the way, 100 times decreased compared to PB) is due to the presence of FeHCF (Prussian Blue) as defects in the structure of noniron hexacyanoferrates. This finding, considering other unique properties of transition metal HCFs, will provide a systematic search for sensing materials with improved analytical performance characteristics.

We report here a way for improving the stability of ultramicroelectrodes (UME) based on hexacyanoferrate-modified metals for the detection of hydrogen peroxide. The most stable sensors were obtained by electrochemical deposition of six layers of hexacyanoferrates (HCF), more specifically, an alternating pattern of three layers of Prussian Blue and three layers of Ni-HCF. The microelectrodes modified with mixed layers were continuously monitored in 1 mM hydrogen peroxide and proved to be stable for more than 5 h under these conditions. The mixed layer microelectrodes exhibited a stability which is five times as high as the stability of conventional Prussian Blue-modified UMEs. The sensitivity of the mixed layer sensor was 0.32 A·M(-1)·cm(-2), and the detection limit was 10 µM. The mixed layer-based UMEs were used as sensors in scanning electrochemical microscopy (SECM) experiments for imaging of hydrogen peroxide evolution.

We report on a superstable hydrogen peroxide (H(2)O(2)) transducer made by sequential deposition of the iron- and nickel-hexacyanoferrate (NiHCF) layers. Both chemical and mechanical stability of the latter, as well as similarity of its structure to Prussian Blue (PB) provide a substantial stabilization of the most advantageous H(2)O(2) transducer. The electrochemically deposited five bilayers of PB-NiHCF exhibit a complete stability under the continuous wall-jet flow of 1 mM of H(2)O(2) during more than 2 h, maintaining current at a level of 0.2 mA cm(-2), whereas common Prussian Blue loses half of its response within the first 20-25 min. Even being deposited in the open circuit regime on screen-printed electrodes, PB-NiHCF bilayers dramatically improve tolerance of the resulting transducer to alkaline solutions and iron ligands. Despite their 2-2.5 times decreased sensitivity (compared to common Prussian Blue), the sequentially deposited bilayers of PB-NiHCF provide a similar dynamic range of the transducer due to the decreased noise level.