Isothermal amplification-assisted diagnostics for COVID-19.

The scenery of molecular diagnostics for infectious diseases is rapidly evolving to respond to the COVID-19 epidemic. The sensitivity and specificity of diagnostics, along with speed and accuracy, are crucial requirements for effective analytical tools to address the disease spreading around the world. Emerging diagnostic devices combine the latest trends in isothermal amplification methods for nucleic acids with state-of-the-art biosensing systems, intending to bypass roadblocks encountered in the last 2 years of the pandemic. Isothermal nucleic acid amplification is a simple procedure that quickly and efficiently accumulates nucleic acid sequences at a constant temperature, without the need for sophisticated equipment. The integration of isothermal amplification into portable biosensing devices confers high sensitivity and improves screening at the point of need in low-resource settings. This review reports the latest trends reached in this field with the latest examples of isothermal amplification-powered biosensors for detecting SARS-CoV-2, with different configurations, as well as their intrinsic advantages and disadvantages.

Innovative Eco-Friendly Conductive Ink Based on Carbonized Lignin for the Production of Flexible and Stretchable Bio-Sensors.

In this study, we report a novel way to produce carbon-based conductive inks for electronic and sensor technology applications. Carbonized lignin, obtained from the waste products of the Eucalyptus globulus tree paper industry, was used to produce a stable conductive ink. To this end, liquid-phase compositions were tested with different amounts of carbonized lignin powder to obtain an ink with optimal conductivity and rheological properties for different possible uses. The combination that showed the best properties, both regarding electrochemical properties and green compatibility of the materials employed, was cyclohexanone/cellulose acetate/carbonized lignin 5% (w/w), which was used to produce screen-printed electrodes. The electrodes were characterized from a structural and electrochemical point of view, resulting in an electrochemically active area of 0.1813 cm2, compared to the electrochemically active area of 0.1420 cm2 obtained by employing geometrically similar petroleum-based screen-printed electrodes and, finally, their performance was demonstrated for the quantification of uric acid, with a limit of detection of 0.3 µM, and their biocompatibility was assessed by testing it with the laccase enzyme and achieving a limit of detection of 2.01 µM for catechol as the substrate. The results suggest that the developed ink could be of great use in both sensor and electronic industries, reducing the overall ecological impact of traditionally used petroleum-based inks.

Decision making based on hybrid modeling approach applied to cellulose acetate based historical films conservation.

Preserving culture heritage cellulose acetate-based historical films is a challenge due to the long-term instability of these complex materials and a lack of prediction models that can guide conservation strategies for each particular film. In this work, a cellulose acetate degradation model is proposed as the basis for the selection of appropriate strategies for storage and conservation for each specimen, considering its specific information. Due to the formulation complexity and diversity of cellulose acetate-based films produced over the decades, we hereby propose a hybrid modeling approach to describe the films degradation process. The problem is addressed by a hybrid model that uses as a backbone a first-principles based model to describe the degradation kinetics of the pure cellulose diacetate polymer. The mechanistic model was successfully adapted to fit experimental data from accelerated aging of plasticized films. The hybrid model considers then the specificity of each historical film via the development of two chemometric models. These models resource on gas release data, namely acetic acid, and descriptors of the films (manufacturing date, AD-strip value and film type) to assess the current polymer degradation state and estimate the increase in the degradation rate. These estimations are then conjugated with storage conditions (e.g., temperature and relative humidity, presence of adsorbent in the film's box) and used to feed the mechanistic model to provide the required time degradation simulations. The developed chemometric models provided predictions with accuracy more than 87%. We have found that the storage archive as well as the manufacturing company are not determining factors for conservation but rather the manufacturing date, off gas data as well as the film type. In summary, this hybrid modeling was able to develop a practical tool for conservators to assess films conservation state and to design storage and conservation policies that are best suited for each cultural heritage film.

Photoautotrophs-Bacteria Co-Cultures: Advances, Challenges and Applications.

Photosynthetic microorganisms are among the fundamental living organisms exploited for millennia in many industrial applications, including the food chain, thanks to their adaptable behavior and intrinsic proprieties. The great multipotency of these photoautotroph microorganisms has been described through their attitude to become biofarm for the production of value-added compounds to develop functional foods and personalized drugs. Furthermore, such biological systems demonstrated their potential for green energy production (e.g., biofuel and green nanomaterials). In particular, the exploitation of photoautotrophs represents a concrete biorefinery system toward sustainability, currently a highly sought-after concept at the industrial level and for the environmental protection. However, technical and economic issues have been highlighted in the literature, and in particular, challenges and limitations have been identified. In this context, a new perspective has been recently considered to offer solutions and advances for the biomanufacturing of photosynthetic materials: the co-culture of photoautotrophs and bacteria. The rational of this review is to describe the recently released information regarding this microbial consortium, analyzing the critical issues, the strengths and the next challenges to be faced for the intentions attainment.

Quantum dots functionalised artificial peptides bioinspired to the D1 protein from the Photosystem II of Chlamydomonas reinhardtii for endocrine disruptor optosensing.

Herein we describe the design and synthesis of novel artificial peptides mimicking the plastoquinone binding niche of the D1 protein from the green photosynthetic alga Chlamydomonas reinhardtii, also able to bind herbicides. In particular, molecular dynamics (MD) simulations were performed to model in silico the behaviour of three peptides, D1Pep70-H, D1Pep70-S264K and D1Pep70-S268C, as genetic variants with different affinity towards the photosynthetic herbicide atrazine. Then the photosynthetic peptides were functionalised with quantum dots for the development of a hybrid optosensor for the detection of atrazine, one of the most employed herbicides for weed control in agriculture as well as considered as a putative endocrine disruptor case study. The excellent agreement between computational and experimental results self consistently shows resistance or super-sensitivity toward the atrazine target, with detection limits in the µg/L concentration range, meeting the requirements of E.U. legislation.

A microbial sensor platform based on bacterial bioluminescence (luxAB) and green fluorescent protein (gfp) reporters for in situ monitoring of toxicity of wastewater nitrification process dynamics.

To avoid the upset of nitrification process in wastewater treatment plants, monitoring of influent toxic chemicals is essential for stable operation. Toxic chemical compounds can interfere with the biological nitrogen removal, thus affecting plant efficiency and effluent water quality. Here we report the development of fluorescence and bioluminescence bioassays, based on E. coli engineered to contain the promoter region of ammonia oxidation pathway (AmoA1) of Nitrosomonas europaea and a reporter gene (lux or gfp). The fluorescence or bioluminescence signal was measured with newly designed optical devices. The microbial sensors were tested and validated at different concentrations of nitrification-inhibiting compounds such as allylthiourea, phenol, and mercury. The signal decrease was immediate and proportional to inhibitor concentration. The developed bacterial bioassays could detect the inhibition of the nitrification process in wastewater for allylthiourea concentrations of 1 µg/L for E.coli pMosaico-Pamo-gfp and 0.5 µg/L for E.coli pMosaico-Pamo-luxAB. The results were confirmed using water from a wastewater plant, containing nitrification-inhibiting compounds.

High-Tech and Nature-Made Nanocomposites and Their Applications in the Field of Sensors and Biosensors for Gas Detection.

Gas sensors have been object of increasing attention by the scientific community in recent years. For the development of the sensing element, two major trends seem to have appeared. On one hand, the possibility of creating complex structures at the nanoscale level has given rise to ever more sensitive sensors based on metal oxides and metal-polymer combinations. On the other hand, gas biosensors have started to be developed, thanks to their intrinsic ability to be selective for the target analyte. In this review, we analyze the recent progress in both areas and underline their strength, current problems, and future perspectives.

Deep eutectic solvents (DES) as green extraction media for antioxidants electrochemical quantification in extra-virgin olive oils.

A new electroanalytical method has been developed for the determination of polar antioxidant compounds in extra virgin olive oils. This method is based on the extraction of polar antioxidant compounds from extra-virgin olive oils by means of a deep eutectic solvent and their determination by a modified screen-printed electrode platform. The platform sensitivity was increased by modifying the working electrode with MWCNT and TiO2 nanoparticles as modifiers and Nafion as a binder. The platform showed very good sensitivity in detecting polar antioxidant compounds in extra-virgin olive oils in a fairly wide range of concentrations. The measurements were performed by using square wave voltammetry. The extraction was performed without using organic solvents, making the method environmentally friendly. The proposed method has been compared with a common spectrophotometric one, the results appeared in good agreement. The method is sufficiently easy and quick to be used for screening analyses of polar antioxidant compounds in extra-virgin olive oils on the field.

ZnO/polyaniline composite based photoluminescence sensor for the determination of acetic acid vapor.

In this study, we report a novel ZnO/polyaniline (PANI) nanocomposite optical gas sensor for the determination of acetic acid at room temperatures. ZnO nanorods, synthesized in powder form were coated by PANI (ZnO/PANI) by chemical polymerization method. The obtained nanocomposites were deposited on glass substrate and dried overnight at room temperature. Structure and optical properties of ZnO/PANI nanocomposite have been studied by using X-ray diffraction, transmission electron microscopy, scanning electron microscopy, diffuse reflectance and photoluminescence spectroscopy. Tests towards acetic acids were performed in the range of concentrations 1-13 ppm. The adsorption of acetic acid on the sensor's surface resulted in the decrease of ZnO/PANI photoluminescence. The response and recovery time of the sensor were in the range of 30-50 s and 5 min, respectively. The developed sensors showed sensitivity towards acetic acid in a range of 1-10 ppm with the limit of detection of 1.2 ppm. Specially designed miniaturized sensing system based on integrated sensing layer, light emission diode as excitation source and optical fiber spectrometer was developed for the measurement of the sensor signal. The developed sensing system was applied for the investigation of some real sample assessment including the evaluation of storage conditions of ancient cellulose acetate films, which during the degradation are releasing acetic acid. The obtained results suggest that the developed novel optical ZnO/PANI nanocompsite based sensor shows great potential for acetic acid determination in various samples.

The effect of ionic strength and phosphate ions on the construction of redox polyelectrolyte-enzyme self-assemblies.

Layer by layer assembly of polyelectrolytes with proteins is a convenient tool for the development of functional biomaterials. Most of the studies presented in the literature are based on the electrostatic interaction between components of opposite charges, limiting the assembly possibilities. However, this process can be tuned by modifying the environment where the main constituents are dissolved. In this work, the electron transfer behavior between an electroactive polyelectrolyte (polyallylamine derivatized with an osmium complex) and a redox enzyme (glucose oxidase) is studied by assembling them in the presence of phosphate ions at different ionic strengths. Our results show that the environment from which the assembly is constructed has a significant effect on the electrochemical response. Notably, the polyelectrolyte dissolved in the presence of phosphate at high ionic strength presents a globular structure which is preserved after adsorption with substantial effects on the buildup of the multilayer system, improving the electron transfer process through the film.

Biologically friendly room temperature ionic liquids and nanomaterials for the development of innovative enzymatic biosensors: Part II.

A newly modified electrode based on glassy carbon (GC) has been prepared and characterized electrochemically for application in electroanalytical chemistry. In particular, a GC screen-printed electrode (SPE) has been modified with nanostructures, namely multi-walled carbon nanotubes (MWCNTs), and TiO2 nanoparticles, and combined with a new generation of eco-friendly room-temperature ionic liquids (RTILs). The green RTILs here used are suitable for the immobilization of enzymes on the electrode surface and, additionally, facilitate the kinetics of electron transfer due to their intrinsic electrical conductivity. Upon evaluation of these newly modified electrodes we found an improvement in terms of electrochemically active area (Aea) with respect to the electrodes we previously reported. The modified SPEs were then used as substrates for the construction of two enzymatic biosensors for analytical applications: the first is an enzymatic biosensor based on alcohol dehydrogenase (ADH) for the analysis of ethyl alcohol; the second biosensor is based on lipase enzyme and has been tested for the analysis and the classification of Extra Virgin Olive Oil (EVOO). The performances of the here projected sensors appear comparable with biosensors having similar finalities. It is here envisaged that such a kind of electrodes could represent the starting tool for the construction and the definition of new portable devices for screening and field analyses.