Development of Essential Oil Delivery Systems by 'Click Chemistry' Methods: Possible Ways to Manage Duchenne Muscular Dystrophy.

Recently, rare diseases have received attention due to the need for improvement in diagnosed patients' and their families' lives. Duchenne muscular dystrophy (DMD) is a rare, severe, progressive, muscle-wasting disease. Today, the therapeutic standard for treating DMD is corticosteroids, which cause serious adverse side effects. Nutraceuticals, e.g., herbal extracts or essential oils (EOs), are possible active substances to develop new drug delivery systems to improve DMD patients' lives. New drug delivery systems lead to new drug effects, improved safety and accuracy, and new therapies for rare diseases. Herbal extracts and EOs combined with click chemistry can lead to the development of safer treatments for DMD. In this review, we focus on the need for novel drug delivery systems using EOs as the therapy for DMD and the potential use of click chemistry for drug delivery systems. New EO complex drug delivery systems may offer a new approach for improving muscle conditions and mental health issues associated with DMD. However, further research should identify the potential of these systems in the context of DMD. In this review, we discuss possibilities for applying EOs to DMD before implementing expensive research in a theoretical way.

Molecularly imprinted polymers for the recognition of biomarkers of certain neurodegenerative diseases.

The appearance of the biomarkers in body fluids like blood, urine, saliva, tears, etc. can be used for the identification of many diseases. This article aimed to summarize the studies about electrochemical biosensors with molecularly imprinted polymers as sensitive and selective layers on the electrode to detect protein-based biomarkers of such neurodegenerative diseases as Alzheimer's disease, Parkinson's disease, and stress. The main attention in this article is focused on the detection methods of amyloid-ß oligomers and p-Tau which are representative biomarkers for Alzheimer's disease, α-synuclein as the biomarker of Parkinson's disease, and α-amylase and lysozyme as the biomarkers of stress using molecular imprinting technology. The research methods, the application of different electrodes, the influence of the polymers, and the established detection limits are reviewed and compared.

Electrochemical Determination of Interaction between SARS-CoV-2 Spike Protein and Specific Antibodies.

The serologic diagnosis of coronavirus disease 2019 (COVID-19) and the evaluation of vaccination effectiveness are identified by the presence of antibodies specific to severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). In this paper, we present the electrochemical-based biosensing technique for the detection of antibodies specific to the SARS-CoV-2 proteins. Recombinant SARS-CoV-2 spike proteins (rSpike) were immobilised on the surface of a gold electrode modified by a self-assembled monolayer (SAM). This modified electrode was used as a sensitive element for the detection of polyclonal mouse antibodies against the rSpike (anti-rSpike). Electrochemical impedance spectroscopy (EIS) was used to observe the formation of immunocomplexes while cyclic voltammetry (CV) was used for additional analysis of the surface modifications. It was revealed that the impedimetric method and the elaborate experimental conditions are appropriate for the further development of electrochemical biosensors for the serological diagnosis of COVID-19 and/or the confirmation of successful vaccination against SARS-CoV-2.

Electrochemical molecularly imprinted polymer based sensors for pharmaceutical and biomedical applications (review).

Recent challenges in the pharmaceutical and biomedical fields require the development of new analytical methods. Therefore, the development of new sensors is a very important task. In this paper, we are outlining the development of molecularly imprinted polymer (MIP) based sensors, which belongs to important branch of affinity sensors. In this review, recent advances in the design of MIP-based sensors are overviewed. MIPs-based sensing structures can replace expensive natural affinity compounds such as receptors or antibodies. Among many different polymers, conducting polymers show the most versatile properties, which are suitable for sensor application. Therefore, significant attention is paid towards MIPs based on conducting polymers, namely polypyrrole, polythiophene, poly(3,4-ethylenedioxythiophene), polyaniline and ortho-phenylenediamine. Moreover, many other materials, which could be imprinted analyte molecules, are overviewed. Among many conducting polymers, polypyrrole is highlighted as one of the most suitable for molecular imprinting. Some attention is dedicated to overview polymerization methods applied for the design of sensing structures used in various affinity sensors. The transduction of analytical signal is an important issue, therefore, physicochemical methods suitable for analytical signal transduction are also outlined. Advances, trends and perspectives in MIP application are discussed.

Microbial Fuel Cell Based on Nitrogen-Fixing Rhizobium anhuiense Bacteria.

In this study, the nitrogen-fixing, Gram-negative soil bacteria Rhizobium anhuiense was successfully utilized as the main biocatalyst in a bacteria-based microbial fuel cell (MFC) device. This research investigates the double-chambered, H-type R. anhuiense-based MFC that was operated in modified Norris medium (pH = 7) under ambient conditions using potassium ferricyanide as an electron acceptor in the cathodic compartment. The designed MFC exhibited an open-circuit voltage (OCV) of 635 mV and a power output of 1.07 mW m-2 with its maximum power registered at 245 mV. These values were further enhanced by re-feeding the anode bath with 25 mM glucose, which has been utilized herein as the main carbon source. This substrate addition led to better performance of the constructed MFC with a power output of 2.59 mW m-2 estimated at an operating voltage of 281 mV. The R. anhuiense-based MFC was further developed by improving the charge transfer through the bacterial cell membrane by applying 2-methyl-1,4-naphthoquinone (menadione, MD) as a soluble redox mediator. The MD-mediated MFC device showed better performance, resulting in a slightly higher OCV value of 683 mV and an almost five-fold increase in power density to 4.93 mW cm-2. The influence of different concentrations of MD on the viability of R. anhuiense bacteria was investigated by estimating the optical density at 600 nm (OD600) and comparing the obtained results with the control aliquot. The results show that lower concentrations of MD, ranging from 1 to 10 µM, can be successfully used in an anode compartment in which R. anhuiense bacteria cells remain viable and act as a main biocatalyst for MFC applications.

Electrochemically Deposited Molecularly Imprinted Polymer-Based Sensors.

This review is dedicated to the development of molecularly imprinted polymers (MIPs) and the application of MIPs in sensor design. MIP-based biological recognition parts can replace receptors or antibodies, which are rather expensive. Conducting polymers show unique properties that are applicable in sensor design. Therefore, MIP-based conducting polymers, including polypyrrole, polythiophene, poly(3,4-ethylenedioxythiophene), polyaniline and ortho-phenylenediamine are frequently applied in sensor design. Some other materials that can be molecularly imprinted are also overviewed in this review. Among many imprintable materials conducting polymer, polypyrrole is one of the most suitable for molecular imprinting of various targets ranging from small organics up to rather large proteins. Some attention in this review is dedicated to overview methods applied to design MIP-based sensing structures. Some attention is dedicated to the physicochemical methods applied for the transduction of analytical signals. Expected new trends and horizons in the application of MIP-based structures are also discussed.

Biosensors for the Determination of SARS-CoV-2 Virus and Diagnosis of COVID-19 Infection.

Monitoring and tracking infection is required in order to reduce the spread of the coronavirus disease 2019 (COVID-19), induced by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). To achieve this goal, the development and deployment of quick, accurate, and sensitive diagnostic methods are necessary. The determination of the SARS-CoV-2 virus is performed by biosensing devices, which vary according to detection methods and the biomarkers which are inducing/providing an analytical signal. RNA hybridisation, antigen-antibody affinity interaction, and a variety of other biological reactions are commonly used to generate analytical signals that can be precisely detected using electrochemical, electrochemiluminescence, optical, and other methodologies and transducers. Electrochemical biosensors, in particular, correspond to the current trend of bioanalytical process acceleration and simplification. Immunosensors are based on the determination of antigen-antibody interaction, which on some occasions can be determined in a label-free mode with sufficient sensitivity.

Towards application of CRISPR-Cas12a in the design of modern viral DNA detection tools (Review).

Early detection of viral pathogens by DNA-sensors in clinical samples, contaminated foods, soil or water can dramatically improve clinical outcomes and reduce the socioeconomic impact of diseases such as COVID-19. Clustered regularly interspaced short palindromic repeat (CRISPR) and its associated protein Cas12a (previously known as CRISPR-Cpf1) technology is an innovative new-generation genomic engineering tool, also known as 'genetic scissors', that has demonstrated the accuracy and has recently been effectively applied as appropriate (E-CRISPR) DNA-sensor to detect the nucleic acid of interest. The CRISPR-Cas12a from Prevotella and Francisella 1 are guided by a short CRISPR RNA (gRNA). The unique simultaneous cis- and trans- DNA cleavage after target sequence recognition at the PAM site, sticky-end (5-7 bp) employment, and ssDNA/dsDNA hybrid cleavage strategies to manipulate the attractive nature of CRISPR-Cas12a are reviewed. DNA-sensors based on the CRISPR-Cas12a technology for rapid, robust, sensitive, inexpensive, and selective detection of virus DNA without additional sample purification, amplification, fluorescent-agent- and/or quencher-labeling are relevant and becoming increasingly important in industrial and medical applications. In addition, CRISPR-Cas12a system shows great potential in the field of E-CRISPR-based bioassay research technologies. Therefore, we are highlighting insights in this research direction.

Assessment of Rhizobium anhuiense Bacteria as a Potential Biocatalyst for Microbial Biofuel Cell Design.

The development of microbial fuel cells based on electro-catalytic processes is among the novel topics, which are recently emerging in the sustainable development of energetic systems. Microbial fuel cells have emerged as unique biocatalytic systems, which transform the chemical energy accumulated in renewable organic fuels and at the same time reduce pollution from hazardous organic compounds. However, not all microorganisms involved in metabolic/catalytic processes generate sufficient redox potential. In this research, we have assessed the applicability of the microorganism Rhizobium anhuiense as a catalyst suitable for the design of microbial fuel cells. To improve the charge transfer, several redox mediators were tested, namely menadione, riboflavin, and 9,10-phenanthrenequinone (PQ). The best performance was determined for a Rhizobium anhuiense-based bio-anode mediated by menadione with a 0.385 mV open circuit potential and 5.5 µW/cm2 maximal power density at 0.35 mV, which generated 50 µA/cm2 anode current at the same potential.

Advances and insights in the diagnosis of viral infections.

Viral infections are the most common among diseases that globally require around 60 percent of medical care. However, in the heat of the pandemic, there was a lack of medical equipment and inpatient facilities to provide all patients with viral infections. The detection of viral infections is possible in three general ways such as (i) direct virus detection, which is performed immediately 1-3 days after the infection, (ii) determination of antibodies against some virus proteins mainly observed during/after virus incubation period, (iii) detection of virus-induced disease when specific tissue changes in the organism. This review surveys some global pandemics from 1889 to 2020, virus types, which induced these pandemics, and symptoms of some viral diseases. Non-analytical methods such as radiology and microscopy also are overviewed. This review overlooks molecular analysis methods such as nucleic acid amplification, antibody-antigen complex determination, CRISPR-Cas system-based viral genome determination methods. Methods widely used in the certificated diagnostic laboratory for SARS-CoV-2, Influenza A, B, C, HIV, and other viruses during a viral pandemic are outlined. A comprehensive overview of molecular analytical methods has shown that the assay's sensitivity, accuracy, and suitability for virus detection depends on the choice of the number of regions in the viral open reading frame (ORF) genome sequence and the validity of the selected analytical method.

Molecular Imprinting Technology for Determination of Uric Acid.

The review focuses on the overview of electrochemical sensors based on molecularly imprinted polymers (MIPs) for the determination of uric acid. The importance of robust and precise determination of uric acid is highlighted, a short description of the principles of molecular imprinting technology is presented, and advantages over the others affinity-based analytical methods are discussed. The review is mainly concerned with the electro-analytical methods like cyclic voltammetry, electrochemical impedance spectroscopy, amperometry, etc. Moreover, there are some scattered notes to the other electrochemistry-related analytical methods, which are capable of providing additional information and to solve some challenges that are not achievable using standard electrochemical methods. The significance of these overviewed methods is highlighted. The overview of the research that is employing MIPs imprinted with uric acid is mainly targeted to address these topics: (i) type of polymers, which are used to design uric acid imprint structures; (ii) types of working electrodes and/or other parts of signal transducing systems applied for the registration of analytical signal; (iii) the description of the uric acid extraction procedures applied for the design of final MIP-structure; (iv) advantages and disadvantages of electrochemical methods and other signal transducing methods used for the registration of the analytical signal; (vi) overview of types of interfering molecules, which were analyzed to evaluate the selectivity; (vi) comparison of analytical characteristics such as linear range, limits of detection and quantification, reusability, reproducibility, repeatability, and stability. Some insights in future development of uric acid sensors are discussed in this review.

Evaluation of Electrochromic Properties of Polypyrrole/Poly(Methylene Blue) Layer Doped by Polysaccharides.

Polypyrrole (Ppy) and poly(methylene blue) (PMB) heterostructure (Ppy-PMB) was electrochemically formed on the indium tin oxide (ITO) coated glass slides, which served as working electrodes. For electropolymerization, a solution containing pyrrole, methylene blue, and a saccharide (lactose, sucrose, or heparin) that served as dopant was used. The aim of this study was to compare the effect of the saccharides (lactose, sucrose, and heparin) on the electrochromic properties of the Ppy-PMB layer. AFM and SEM have been used for the analysis of the surface dominant features of the Ppy-PMB layers. From these images, it was concluded that the saccharides used in this study have a moderate effect on the surface morphology. Electrochromic properties were analyzed with respect to the changes of absorbance of the layer at two wavelengths (668 nm and 750 nm) by changing the pH of the surrounding solution and the potential between +0.8 V and -0.8 V. It was demonstrated that the highest absorbance changes are characteristic for all layers in the acidic media. Meanwhile, the absorbance changes of the layers were decreased in the more alkaline media. It was determined that the Ppy-PMB layers with heparin as a dopant were more mechanically stable in comparison to the layers doped with lactose and sucrose. Therefore, the Ppy-PMB layer doped with heparin was selected for the further experiment and it was applied in the design of electrochromic sensors for the determination of three xanthine derivatives: caffeine, theobromine, and theophylline. A linear relationship of ΔA (∆A = A+0.8V - A-0.8V) vs. concentration was determined for all three xanthine derivatives studied. The largest change in optical absorption was observed in the case of theophylline determination.

Polyethylene-Carbon Composite (Velostat®) Based Tactile Sensor.

The progress observed in 'soft robotics' brought some promising research in flexible tactile, pressure and force sensors, which can be based on polymeric composite materials. Therefore, in this paper, we intend to evaluate the characteristics of a force-sensitive material-polyethylene-carbon composite (Velostat®) by implementing this material into the design of the flexible tactile sensor. We have explored several possibilities to measure the electrical signal and assessed the mechanical and time-dependent properties of this tactile sensor. The response of the sensor was evaluated by performing tests in static, long-term load and cyclic modes. Experimental results of loading cycle measurements revealed the hysteresis and nonlinear properties of the sensor. The transverse resolution of the sensor was defined by measuring the response of the sensor at different distances from the loaded point. Obtained dependencies of the sensor's sensitivity, hysteresis, response time, transversal resolution and deformation on applied compressive force promise a practical possibility to use the polyethylene-carbon composite as a sensitive material for sensors with a single electrode pair or its matrix. The results received from experimental research have defined the area of the possible implementation of the sensor based on a composite material-Velostat®.

Evaluation of electrochemical quartz crystal microbalance based sensor modified by uric acid-imprinted polypyrrole.

Uric acid-imprinted polypyrrole-based (MIP(UA)-Ppy) electrochemical quartz crystal microbalance sensor (EQCM) was developed. Experiments and theoretical calculations were focused on molecular interactions between uric acid molecule and: i) polypyrrole imprinted by uric acid (MIP(UA)-Ppy) ii) polypyrrole film without any molecular imprints (NIP-Ppy). Resonant frequency differences during electrochemical deposition of MIP(UA)-Ppy and NIP-Ppy films were observed and were attributed to the phenomenon of molecule capture within formed Ppy matrix. EQCM-resonators modified by MIP-Ppy showed the following advantages: selectivity, qualitative response, cost-effectiveness, and simple procedure. The selectivity of MIP(UA)-Ppy was tested by the replacement of uric acid in the PBS solution with several different concentrations of caffeine and glucose. Langmuir isotherm based molecular adsorption model was applied to evaluate the interaction of MIP(UA)-Ppy with uric acid. From experimental results calculated the standard Gibbs free energy of association (ΔGa) of uric acid with MIP(UA)-Ppy is -16.4 ± 2.05 kJ/mol and with NIP-Ppy is -13.3 ± 8.56 kJ/mol ΔG values illustrate that the formation of uric acid complex with MIP(UA)-Ppy is thermodynamically more favourable than that for complexation with NIP-Ppy.

Time-resolved fluorescence spectroscopy based evaluation of stability of glucose oxidase.

Glucose oxidase (GOx) is one of the most frequently used enzymes in a design of enzymatic biosensors and biofuel cells, which are novel electrical energy generation systems. Therefore, a better understanding of the mode of action of this enzyme is very important for further development of GOx-based sensors. In this research fluorescence properties of GOx in different acidic media have been estimated by the evaluation of redox states of active center that is flavine adenine dinucleotide (FAD). Steady-state fluorescence spectroscopy was applied to monitor the activity of GOx. A variation of pH has been invoked to gain a better understanding in the variations of GOx activity. The tendency of GOx activity to decrease over the time was determined, while increased intensity of the fluorescence band of GOx at 530 nm was associated with a decreased activity of the enzyme. The changes in fluorescence intensity of this band are caused by the dissociation of FAD from the enzyme. This process is not reversible, therefore, the decrease in the fluorescence intensity can be also associated with structural changes of the FAD during its reduction.

Tuning the Photo-Luminescence Properties of WO3 Layers by the Adjustment of Layer Formation Conditions.

In this research we have applied sol-gel synthesis for the deposition of tungsten (VI) oxide (WO3) layers using two different reductants (ethanol and propanol) and applying different dipping times. WO3 samples were characterized by X-ray diffraction (XRD), scanning electron microscopy (SEM), Fourier Transform Infrared spectroscopy (FTIR), photoluminescence (PL) and time-resolved photoluminescence decay methods. Photoelectrochemical (PEC) behaviour of synthesized coatings was investigated using cyclic voltammetry in the dark and under illumination. Formation of different structures in differently prepared samples was revealed and significant differences in the PL spectra and PEC performance of the samples were observed. The results showed that reductant used in the synthesis and dipping time strongly influenced photo-electrochemical properties of the coatings. Correlation between the morphology, PL and PEC behaviour has been explained.

Single-walled carbon nanotube based coating modified with reduced graphene oxide for the design of amperometric biosensors.

In this study a polycarbonate filter membrane (PcFM) with 400 nm diameter holes was covered/protruded by single walled carbon nanotubes (SWCNT) and then formed PcFM/SWCNT structure was covered by thin layer of graphene oxide (GO) or reduced graphene oxide (rGO) in order to get the multilayered PcFM/SWCNT/GO and PcFM/SWCNT/rGO coatings, respectively. It was determined that the SWCNTs filaments were able to form a layer on the polycarbonate membrane having a number of carbon nanotube arranged in different orientations. A fraction of SWCNT filaments protruded through the holes of polycarbonate membrane and in such way significantly enhanced the adhesion of SWCNT-based layer and provided electrical conductivity across the PcFM. Atomic force microscopy (AFM), scanning electron microscopy (SEM) images and Raman spectroscopy-based evaluation revealed the characteristic morphology features: wide distribution of height profile, separate GO/rGO flakes on the top of PcFM/SWCNT/GO structure and close attachment of rGO flakes on the top of multilayered PcFM/SWCNT/rGO coating. Performed contact angle measurement (CAM) enabled to determine the surface energy components and wettability data of prepared coatings. Both PcFM/SWCNT/GO and PcFM/SWCNT/rGO coatings were modified with glucose oxidase (GOx). Amperometric measurements revealed that multilayered PcFM/SWCNT/rGO/GOx coating is the most suitable structure for glucose biosensor design.