Nanostructure Tuning of Gold Nanoparticles Films via Click Sintering.

Colloidal metal nanoparticles dispersions are commonly used to create functional printed electronic devices and they typically require time-, energy- and equipment-consuming post-treatments to improve their electrical and mechanical properties. Traditional methods, e.g. thermal, UV/IR, and microwave treatments, limit the substrate options and may require expensive equipment, not available in all the laboratories. Moreover, these processes also cause the collapse of the film (nano)pores and interstices, limiting or impeding its nanostructuration. Finding a simple approach to obtain complex nanostructured materials with minimal post-treatments remains a challenge. In this study, a new sintering method for gold nanoparticle inks that called as "click sintering" has been reported. The method uses a catalytic reaction to enhance and tune the nanostructuration of the film while sintering the metallic nanoparticles, without requiring any cumbersome post-treatment. This results in a conductive and electroactive nanoporous thin film, whose properties can be tuned by the conditions of the reaction, i.e., concentration of the reagent and time. Therefore, this study presents a novel and innovative one-step approach to simultaneously sinter gold nanoparticles films and create functional nanostructures, directly and easily, introducing a new concept of real-time treatment with possible applications in the fields of flexible electronics, biosensing, energy, and catalysis.

The effect of SARS-CoV-2 variants on the plasma oxylipins and PUFAs of COVID-19 patients.

Oxylipins are important signalling compounds that are significantly involved in the regulation of the immune system and the resolution of inflammation. Lipid metabolism is strongly activated upon SARS-CoV-2 infection, however the modulating effects of oxylipins induced by different variants remain unexplored. Here, we compare the plasma profiles of thirty-seven oxylipins and four PUFAs in subjects infected with Wild-type, Alpha (B.1.1.7), Delta (B.1.617.2), and Omicron (B.1.1.529) variants. The results suggest that oxidative stress and inflammation resulting from COVID-19 were highly dependent on the SARS-CoV-2 variant, and that the Wild-type elicited the strongest inflammatory storm. The Alpha and Delta variants induced a comparable lipid profile alteration upon infection, which differed significantly from Omicron. The latter variant increased the levels of pro-inflammatory mediators and decreased the levels of omega-3 PUFA in infected patients. We speculate that changes in therapeutics, vaccination, and prior infections may have a role in the alteration of the oxylipin profile besides viral mutations. The results shed new light on the evolution of the inflammatory response in COVID-19.

Biosensing systems for the detection and quantification of methane gas.

Climate change due to the continuous increase in the release of green-house gasses associated with anthropogenic activity has made a significant impact on the sustainability of life on our planet. Methane (CH4) is a green-house gas whose concentrations in the atmosphere are on the rise. CH4 measurement is important for both the environment and the safety at the industrial and household level. Methanotrophs are distinguished for their unique characteristic of using CH4 as the sole source of carbon and energy, due to the presence of the methane monooxygenases that oxidize CH4 under ambient temperature conditions. This has attracted interest in the use of methanotrophs in biotechnological applications as well as in the development of biosensing systems for CH4 quantification and monitoring. Biosensing systems using methanotrophs rely on the use of whole microbial cells that oxidize CH4 in presence of O2, so that the CH4 concentration is determined in an indirect manner by measuring the decrease of O2 level in the system. Although several biological properties of methanotrophic microorganisms still need to be characterized, different studies have demonstrated the feasibility of the use of methanotrophs in CH4 measurement. This review summarizes the contributions in methane biosensing systems and presents a prospective of the valid use of methanotrophs in this field. KEY POINTS: • Methanotroph environmental relevance in methane oxidation • Methanotroph biotechnological application in the field of biosensing • Methane monooxygenase as a feasible biorecognition element in biosensors.

3D Printed Platform for Impedimetric Sensing of Liquids and Microfluidic Channels.

Fused deposition modeling 3D printing (FDM-3DP) employing electrically conductive filaments has recently been recognized as an exceptionally attractive tool for the manufacture of sensing devices. However, capabilities of 3DP electrodes to measure electric properties of materials have not yet been explored. To bridge this gap, we employ bimaterial FDM-3DP combining electrically conductive and insulating filaments to build an integrated platform for sensing conductivity and permittivity of liquids by impedance measurements. The functionality of the device is demonstrated by measuring conductivity of aqueous potassium chloride solution and bottled water samples and permittivity of water, ethanol, and their mixtures. We further implement an original idea of applying impedance measurements to investigate dimensions of 3DP channels as base structures of microfluidic devices, complemented by their optical microscopic analysis. We demonstrate that FDM-3DP allows the manufacture of microchannels of width down to 80 µm.

Biphenyl substituted lysine derivatives as recognition elements for the matrix metalloproteinases MMP-2 and MMP-9.

Matrix metalloproteinases (MMPs) are an important factor in cancer progression and metastasis, especially gelatinases MMP-2 and MMP-9. A simple methodology for their detection and monitoring is highly desirable. Molecular probes have been very widely and successfully applied to study the activity of MMPs in cellular processes in vitro. We thus synthesized a small compound library of MMP-2 and MMP-9 binding probes based on drug molecules and endowed with free amine groups for the functionalization of transducer surfaces. In this study, we combined experimental results obtained by a kinetic fluorogenic peptide substrate cleavage assay with molecular modeling studies in order to assess the ability of the probe to bind to their target enzymes. The synthesized biphenyl substituted lysine derivatives showed IC50-values in the low nanomolar concentration range against MMP-2 (ligands 3a-d: 3 nM to 8 µM, ligands 4a-d: 45 nM to 350 µM) and low micromolar range against MMP-9 (ligands 3a-d: 350 nM to 60 µM, ligands 4a-d: 5 µM to 600 µM), with a selectivity up to more than 160-fold for MMP-2. The experimental results correlated well with molecular modelling with FleXAID and X-score functions. We showed that in our compound series, the side chain remained far away from the S1' cavity and the ligand for all the docked minima. Ligands 4a-d with their free amine group on the side chain may thus be bound to transducer surfaces for the fabrication of sensors, while retaining their activity against their target enzymes.

Controlling Electroosmosis in Nanopores Without Altering the Nanopore Sensing Region.

Nanopores are powerful tools for single-molecule sensing of biomolecules and nanoparticles. The signal coming from the molecule to be analyzed strongly depends on its interaction with the narrower section of the nanopore (constriction) that may be tailored to increase sensing accuracy. Modifications of nanopore constriction have also been commonly used to induce electroosmosis, that favors the capture of molecules in the nanopore under a voltage bias and independently of their charge. However, engineering nanopores for increasing both electroosmosis and sensing accuracy is challenging. Here it is shown that large electroosmotic flows can be achieved without altering the nanopore constriction. Using continuum electrohydrodynamic simulations, it is found that an external charged ring generates strong electroosmosis in cylindrical nanopores. Similarly, for conical nanopores it is shown that moving charges away from the cone tip still results in an electroosmotic flow (EOF), whose intensity reduces increasing the diameter of the nanopore section where charges are placed. This paradigm is applied to engineered biological nanopores showing, via atomistic simulations and experiments, that mutations outside the constriction induce a relatively intense electroosmosis. This strategy provides much more flexibility in nanopore design since electroosmosis can be controlled independently from the constriction, which can be optimized to improve sensing accuracy.

Low-cost inkjet-printed nanostructured biosensor based on CRISPR/Cas12a system for pathogen detection.

The escalating global incidence of infectious diseases caused by pathogenic bacteria, especially in developing countries, emphasises the urgent need for rapid and portable pathogen detection devices. This study introduces a sensitive and specific electrochemical biosensing platform utilising cost-effective electrodes fabricated by inkjet-printing gold and silver nanoparticles on a plastic substrate. The biosensor exploits the CRISPR/Cas12a system for detecting a specific DNA sequence selected from the genome of the target pathogen. Upon detection, the trans-activity of Cas12a/gRNA is triggered, leading to the cleavage of rationally designed single-strand DNA reporters (linear and hairpin) labelled with methylene blue (ssDNA-MB) and bound to the electrode surface. In principle, this sensing mechanism can be adapted to any bacterium by choosing a proper guide RNA to target a specific sequence of its DNA. The biosensor's performance was assessed for two representative pathogens (a Gram-negative, Escherichia coli, and a Gram-positive, Staphylococcus aureus), and results obtained with inkjet-printed gold electrodes were compared with those obtained by commercial screen-printed gold electrodes. Our results show that the use of inkjet-printed nanostructured gold electrodes, which provide a large surface area, in combination with the use of hairpin reporters containing a poly-T loop can increase the sensitivity of the assay corresponding to a signal variation of 86%. DNA targets amplified from various clinically isolated bacteria, have been tested and demonstrate the potential of the proposed platform for point-of-need applications.

Salivary lipid mediators: Key indexes of inflammation regulation in heart failure disease.

Cardiovascular diseases (CVDs) are the leading cause of premature death and disability in humans and their incidence continues to increase. Oxidative stress and inflammation have been recognized as key pathophysiological factors in cardiovascular events. The targeted modulation of the endogenous mechanisms of inflammation, rather than its simple suppression, will become key in treating chronic inflammatory diseases. A comprehensive characterization of the signalling molecules involved in inflammation, such as endogenous lipid mediators, is thus needed. Here, we propose a powerful MS-based platform for the simultaneous quantitation of sixty salivary lipid mediators in CVD samples. Saliva, which represents a non-invasive and painless alternative to blood, was collected from patients suffering from acute and chronic heart failure (AHF and CHF, respectively), obesity and hypertension. Of all the patients, those with AHF and hypertension showed higher levels of isoprostanoids, which are key indexes of oxidant insult. Compared to the obese population, AHF patients showed lower levels (p < 0.02) of antioxidant omega-3 fatty acids, in line with the "malnutrition-inflammation complex syndrome" typical of HF patients. At hospital admission, AHF patients showed significantly higher levels (p < 0.001) of omega-3 DPA and lower levels (p < 0.04) of lipoxin B4 than CHF patients, suggesting a lipid rearrangement typical of the failing heart during acute decompensation. If confirmed, our results highlight the potential use of lipid mediators as predictive markers of re-acutisation episodes, thus providing opportunities for preventive intervention and a reduction in hospitalizations.

Emerging Biosensing Technologies towards Early Sepsis Diagnosis and Management.

Sepsis is defined as a systemic inflammatory dysfunction strictly associated with infectious diseases, which represents an important health issue whose incidence is continuously increasing worldwide. Nowadays, sepsis is considered as one of the main causes of death that mainly affects critically ill patients in clinical settings, with a higher prevalence in low-income countries. Currently, sepsis management still represents an important challenge, since the use of traditional techniques for the diagnosis does not provide a rapid response, which is crucial for an effective infection management. Biosensing systems represent a valid alternative due to their characteristics such as low cost, portability, low response time, ease of use and suitability for point of care/need applications. This review provides an overview of the infectious agents associated with the development of sepsis and the host biomarkers suitable for diagnosis and prognosis. Special focus is given to the new emerging biosensing technologies using electrochemical and optical transduction techniques for sepsis diagnosis and management.

Effects of long-term vegan diet on breath composition.

The composition of exhaled breath derives from an intricate combination of normal and abnormal physiological processes that are modified by the consumption of food and beverages, circadian rhythms, bacterial infections, and genetics as well as exposure to xenobiotics. This complexity, which results wide intra- and inter-individual variability and is further influenced by sampling conditions, hinders the identification of specific biomarkers and makes it difficult to differentiate between pathological and nominally healthy subjects. The identification of a 'normal' breath composition and the relative influence of the aforementioned parameters would make breath analyses much faster for diagnostic applications. We thus compared, for the first time, the breath composition of age-matched volunteers following a vegan and a Mediterranean omnivorous diet in order to evaluate the impact of diet on breath composition. Mixed breath was collected from 38 nominally healthy volunteers who were asked to breathe into a 2 l handmade Nalophan bag. Exhalation flow rate and carbon dioxide values were monitored during breath sampling. An aliquot (100 ml) of breath was loaded into a sorbent tube (250 mg of Tenax GR, 60/80 mesh) before being analyzed by thermal desorption-gas chromatography-mass spectrometry (TD-GC-MS). Breath profiling using TD-GC-MS analysis identified five compounds (methanol, 1-propanol, pentane, hexane, and hexanal), thus enabling differentiation between samples collected from the different group members. Principal component analysis showed a clear separation between groups, suggesting that breath analysis could be used to study the influence of dietary habits in the fields of nutrition and metabolism. Surprisingly, one Italian woman and her brother showed extremely low breath isoprene levels (about 5 pbv), despite their normal lipidic profile and respiratory data, such as flow rate and pCO2. Further investigations to reveal the reasons behind low isoprene levels in breath would help reveal the origin of isoprene in breath.

MS-based targeted profiling of oxylipins in COVID-19: A new insight into inflammation regulation.

The key role of inflammation in COVID-19 induced many authors to study the cytokine storm, whereas the role of other inflammatory mediators such as oxylipins is still poorly understood. IMPRECOVID was a monocentric retrospective observational pilot study with COVID-19 related pneumonia patients (n = 52) admitted to Pisa University Hospital between March and April 2020. Our MS-based analytical platform permitted the simultaneous determination of sixty plasma oxylipins in a single run at ppt levels for a comprehensive characterisation of the inflammatory cascade in COVID-19 patients. The datasets containing oxylipin and cytokine plasma levels were analysed by principal component analysis (PCA), computation of Fisher's canonical variable, and a multivariate receiver operating characteristic (ROC) curve. Differently from cytokines, the panel of oxylipins clearly differentiated samples collected in COVID-19 wards (n = 43) and Intensive Care Units (ICUs) (n = 27), as shown by the PCA and the multivariate ROC curve with a resulting AUC equal to 0.92. ICU patients showed lower (down to two orders of magnitude) plasma concentrations of anti-inflammatory and pro-resolving lipid mediators, suggesting an impaired inflammation response as part of a prolonged and unsolvable pro-inflammatory status. In conclusion, our targeted oxylipidomics platform helped shedding new light in this field. Targeting the lipid mediator class switching is extremely important for a timely picture of a patient's ability to respond to the viral attack. A prediction model exploiting selected lipid mediators as biomarkers seems to have good chances to classify patients at risk of severe COVID-19.

A label-free impedance biosensing assay based on CRISPR/Cas12a collateral activity for bacterial DNA detection.

The rapid and selective identification in the clinical setting of pathogenic bacteria causing healthcare associated infections (HAIs) and in particular blood stream infections (BSIs) is a major challenge, as the number of people affected worldwide and the associated mortality are on the rise. In fact, traditional laboratory techniques such culture and polymerase chain reaction (PCR)-based methodologies are often associated to long turnaround times, which justify the pressing need for the development of rapid, specific and portable point of care devices. The recently discovered clustered regularly interspaced short palindromic repeat loci (CRISPR) and the new class of programmable endonuclease enzymes called CRISPR associated proteins (Cas) have revolutionised molecular diagnostics. The use of Cas proteins in optical and electrochemical biosensing devices has significantly improved the detection of nucleic acids in clinical samples. In this study, a CRISPR/Cas12a system was coupled with electrochemical impedance spectroscopy (EIS) measurements to develop a label-free biosensing assay for the detection of Escherichia coli and Staphylococcus aureus, two bacterial species commonly associated to BSI infections. The programmable Cas12a endonuclease activity, induced by a specific guide RNA (gRNA), and the triggered collateral activity were assessed in in vitro restriction analyses, and evaluated thanks to impedance measurements using a modified gold electrode. The Cas12a/gRNA system was able to specifically recognize amplicons from different clinical isolates of E. coli and S. aureus with a limit of detection of 3 nM and a short turnaround time approximately of 1.5 h. To the best of our knowledge, this is the first biosensing device based on CRISPR/Cas12a label free impedance assay.

Stability of volatile organic compounds in sorbent tubes following SARS-CoV-2 inactivation procedures.

COVID-19 is a highly transmissible respiratory illness that has rapidly spread all over the world causing more than 115 million cases and 2.5 million deaths. Most epidemiological projections estimate that the Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) virus causing the infection will circulate in the next few years and raise enormous economic and social issues. COVID-19 has a dramatic impact on health care systems and patient management, and is delaying or stopping breath research activities due to the risk of infection to the operators following contact with patients, potentially infected samples or contaminated equipment. In this scenario, we investigated whether virus inactivation procedures, based on a thermal treatment (60 °C for 1 h) or storage of tubes at room temperature for 72 h, could be used to allow the routine breath analysis workflow to carry on with an optimal level of safety during the pandemic. Tests were carried out using dry and humid gaseous samples containing about 100 representative chemicals found in exhaled breath and ambient air. Samples were collected in commercially available sorbent tubes, i.e. Tenax GR and a combination of Tenax TA, Carbograph 1TD and Carboxen 1003. Our results showed that all compounds were stable at room temperature up to 72 h and that sample humidity was the key factor affecting the stability of the compounds upon thermal treatment. Tenax GR-based sorbent tubes were less impacted by the thermal treatment, showing variations in the range 20%-30% for most target analytes. A significant loss of aldehydes and sulphur compounds was observed using carbon molecular sieve-based tubes. In this case, a dry purge step before inactivation at 60 °C significantly reduced the loss of the target analytes, whose variations were comparable to the method variability. Finally, a breath analysis workflow including a SARS-CoV-2 inactivation treatment is proposed.

Three-Dimensional (3D) Laser-Induced Graphene: Structure, Properties, and Application to Chemical Sensing.

Notwithstanding its relatively recent discovery, graphene has gone through many evolution steps and inspired a multitude of applications in many fields, from electronics to life science. The recent advancements in graphene production and patterning, and the inclusion of two-dimensional (2D) graphenic materials in three-dimensional (3D) superstructures, further extended the number of potential applications. In this Review, we focus on laser-induced graphene (LIG), an intriguing 3D porous graphenic material produced by direct laser scribing of carbonaceous precursors, and on its applications in chemical sensors and biosensors. LIG can be shaped in different 3D forms with a high surface-to-volume ratio, which is a valuable characteristic for sensors that typically rely on phenomena occurring at surfaces and interfaces. Herein, an overview of LIG, including synthesis from various precursors, structure, and characteristic properties, is first provided. The discussion focuses especially on transport and surface properties, and on how these can be controlled by tuning the laser processing. Progresses and trends in LIG-based chemical sensors are then reviewed, discussing the various transduction mechanisms and different LIG functionalization procedures for chemical sensing. A comparative evaluation of sensors performance is then provided. Finally, sensors for glucose detection are reviewed in more detail, since they represent the vast majority of LIG-based chemical sensors.

Advances in biosensing: The CRISPR/Cas system as a new powerful tool for the detection of nucleic acids.

A main challenge in the development of biosensing devices for the identification and quantification of nucleic acids is to avoid the amplification of the genetic material from the sample by polymerase chain reaction (PCR), which is at present necessary to enhance sensitivity and selectivity of assays. PCR has undoubtedly revolutionized genetic analyses, but it requires careful purification procedures that are not easily implemented in point of care (POC) devices. In recent years, a new strategy for nucleic acid detection based on clustered regularly interspaced short palindromic repeats (CRISPR) and associated protein systems (Cas) seems to offer unprecedented possibilities. The coupling of the CRISPR/Cas system with recent isothermal amplification methods is fostering the development of innovative optical and electrochemical POC devices. In this review, the mechanisms of action of several new CRISRP/Cas systems are reported together with their use in biosensing of nucleic acids.

Biosensors for Detecting Lymphocytes and Immunoglobulins.

Lymphocytes (B, T and natural killer cells) and immunoglobulins are essential for the adaptive immune response against external pathogens. Flow cytometry and enzyme-linked immunosorbent (ELISA) kits are the gold standards to detect immunoglobulins, B cells and T cells, whereas the impedance measurement is the most used technique for natural killer cells. For point-of-care, fast and low-cost devices, biosensors could be suitable for the reliable, stable and reproducible detection of immunoglobulins and lymphocytes. In the literature, such biosensors are commonly fabricated using antibodies, aptamers, proteins and nanomaterials, whereas electrochemical, optical and piezoelectric techniques are used for detection. This review describes how these measurement techniques and transducers can be used to fabricate biosensors for detecting lymphocytes and the total content of immunoglobulins. The various methods and configurations are reported, along with the advantages and current limitations.

Fluorinated vs. Zwitterionic-Polymer Grafted Surfaces for Adhesion Prevention of the Fungal Pathogen Candida albicans.

Fluorinated (F6) and zwitterionic, as well as phosphorylcholine (MPC) and sulfobetaine (MSA), copolymers containing a low amount (1 and 5 mol%) of 3-(trimethoxysilyl)propyl methacrylate (PTMSi) were prepared and covalently grafted to glass slides by using the trimethoxysilyl groups as anchorage points. Glass-surface functionalization and polymer-film stability upon immersion in water were proven by contact angle and angle-resolved X-ray photoelectron spectroscopy (AR-XPS) measurements. Antifouling performance of the grafted films was assayed against the yeast Candida albicans, the most common Candida species, which causes over 80% of candidiasis. Results revealed that the F6 fluorinated, hydrophobic copolymers performed much better in reducing the adhesion of C. albicans, with respect to both corresponding zwitterionic, hydrophilic MPC and MSA counterparts, and were similar to the glass negative control, which is well-known to inhibit the adhesion of C. albicans. A composition-dependent activity was also found, with the films of copolymer with 99 mol% F6 fluorinated co-units performing best.

Entrepreneurship and Nursing: integrative review.

OBJECTIVE: To identify the knowledge produced on business entrepreneurship in Nursing. METHOD: Integrative literature review in the following databases: Cumulative Index to Nursing and Allied Health Literature (CINAHL), EMBASE, SCOPUS, Web of Science, PubMed, Medline, Latin American and Caribbean Literature in Health Sciences (LILACS), Nursing Database (BDENF), Index Psychology and National Information Center of Medical Sciences of Cuba (CUMED). We included available studies in their totality in the period from 2007 to 2017. RESULTS: 22 articles were included. The categories that emerged from the study are the following: Concepts of entrepreneurship in nursing, Profile of the entrepreneur nurse, Business Diversity, Business Management, Barriers to business entrepreneurship in nursing, Support to entrepreneurial nurses and Entrepreneurship in Nursing Undergraduate. CONCLUSION: There is a need to prepare nurses with adequate skills to increase the capacity to integrate into the labor market and to improve their own well-being and that of society.

Sampling Methodology of Alphitobius diaperinus (Coleptera: Tenebrionidae) Population in Poultry Houses

Abstract This paper describes a methodology to evaluate Alphitobius diaperinus (Darkling beetle) population in order to estimate the effectiveness of control methods, consisting of direct sample collections from the poultry litter in 18 points, counting of adults and larvae, and subsequent comparison of the values obtained before and after of control strategies. It allows estimating the total population, population in a known area, as well to analyze the variations in different points all over the aviary. The method has shown to be very versatile, being indicated for population studies and efficacy evaluations, once it has already been validated in different experimental conditions.