After silencing suppression: miRNA targets strike back.

Within the continuous tug-of-war between plants and microbes, RNA silencing stands out as a key battleground. Pathogens, in their quest to colonize host plants, have evolved a diverse arsenal of silencing suppressors as a common strategy to undermine the host's RNA silencing-based defenses. When RNA silencing malfunctions in the host, genes that are usually targeted and silenced by microRNAs (miRNAs) become active and can contribute to the reprogramming of host cells, providing an additional defense mechanism. A growing body of evidence suggests that miRNAs may act as intracellular sensors to enable a rapid response to pathogen threats. Herein we review how plant miRNA targets play a crucial role in immune responses against different pathogens.

Comparison of Synthetic Pathways for Obtaining Fluorescent Nanomaterials Based on Halloysite and Carbon Dots for Potential Biological Sensing.

Recently, fluorescent sensors have gained considerable attention due to their high sensitivity, low cost and noninvasiveness. Among the different materials that can be used for this purpose, carbon dots (CDs) represent valuable candidates for applications in sensing. These, indeed, are easily synthesized, show high quantum yield and are highly biocompatible. However, it was pointed out that the photoluminescence properties of these nanomaterials are strictly dependent on the synthetic and purification methods adopted. The presence of halloysite nanotubes (HNTs), a natural, low cost and biocompatible clay mineral, has been found to be efficient in obtaining small and highly monodispersed CDs without long and tedious purification techniques. Herein, we report the comparison of synthetic pathways for obtaining halloysite-N-doped CDs (HNTs-NCDs) that could be used in biological sensing. One was based on the synthesis of N-doped CDs by a bottom-up approach on HNTs' surface by a MW pyrolysis process; the other one was based on the post-modification of pristine N-doped CDs with halloysite derivatives. The evaluation of the best synthetic route was performed by different physico-chemical techniques. It was found that the bottom-up approach led to the formation of N-doped CDs with different functional groups onto the HNTs' surface. This evidence was also translated in the different fluorescence quantum yields and the existence of several functional groups in the obtained materials was investigated by potentiometric titrations. Furthermore, the ability of the synthesized nanomaterials as sensors for Fe3+ ions detection was assessed by spectroscopic measurements, and the cellular uptake was verified by confocal/fluorescence microscopies as well.

Electrochemical impedance spectroscopy, another arrow in the arsenal to study the biodegradability of two-dimensional materials.

Carbon nitride (C3N4) is an innovative material with a high potential in many applications including energy storage, catalysis, composites, and biomedicine. C3N4 appears remarkably interesting not only for its properties but also because its simple preparation routes involve low-cost starting materials and reagents. However, there is still a lack of information on its degradability. For this reason, in this study, we evaluate the environmental persistence of C3N4 and its oxidized form by applying the photo-Fenton reaction. The morphological and structural changes of both materials were monitored by transmission electron microscopy and Raman spectroscopy respectively. In addition, electrochemical impedance spectroscopy has been used as an original technique to validate the degradation process of C3N4.

A journey into the world of small RNAs in the arbuscular mycorrhizal symbiosis.

Arbuscular mycorrhizal (AM) symbiosis is a mutualistic interaction between fungi and most land plants that is underpinned by a bidirectional exchange of nutrients. AM development is a tightly regulated process that encompasses molecular communication for reciprocal recognition, fungal accommodation in root tissues and activation of symbiotic function. As such, a complex network of transcriptional regulation and molecular signaling underlies the cellular and metabolic reprogramming of host cells upon AM fungal colonization. In addition to transcription factors, small RNAs (sRNAs) are emerging as important regulators embedded in the gene network that orchestrates AM development. In addition to controlling cell-autonomous processes, plant sRNAs also function as mobile signals capable of moving to different organs and even to different plants or organisms that interact with plants. AM fungi also produce sRNAs; however, their function in the AM symbiosis remains largely unknown. Here, we discuss the contribution of host sRNAs in the development of AM symbiosis by considering their role in the transcriptional reprogramming of AM fungal colonized cells. We also describe the characteristics of AM fungal-derived sRNAs and emerging evidence for the bidirectional transfer of functional sRNAs between the two partners to mutually modulate gene expression and control the symbiosis.

Green synthesis of fluorescent carbon nanodots from sage leaves for selective anticancer activity on 2D liver cancer cells and 3D multicellular tumor spheroids.

Carbon nanodots, a family of carbon-based nanomaterials, have been synthesized through different methods from various resources, affecting the properties of the resulting product and their application. Herein, carbon nanodots (CNDs) were synthesized with a green and simple hydrothermal method from sage leaves at 200 °C for 6 hours. The obtained CNDs are well dispersed in water with a negative surface charge (ζ-potential = -11 mV) and an average particle size of 3.6 nm. The synthesized CNDs showed concentration-dependent anticancer activity toward liver cancer (Hep3B) cell lines and decreased the viability of the cancer cells to 23% at the highest used concentration (250 µg ml-1 of CNDs). More interestingly, the cytotoxicity of the CNDs was tested in normal liver cell lines (LX2) revealed that the CNDs at all tested concentrations didn't affect their viability including at the highest concentration showing a viability of 86.7%. The cellular uptake mechanisms of CNDs were investigated and they are thought to be through energy-dependent endocytosis and also through passive diffusion. The main mechanisms of endocytosis were lipid and caveolae-mediated endocytosis. In addition, the CNDs have hindered the formation of 3D spheroids from the Hep3B hepatocellular carcinoma cell line. Hence, it would be concluded that the synthesized CNDs from sage are more highly selective to liver cancer cells than normal ones. The CNDs' cancer-killing ability would be referred to as the production of reactive oxygen species.

Organic Functional Group on Carbon Nanotube Modulates the Maturation of SH-SY5Y Neuronal Models.

Carbon nanotubes (CNT) have proven to be excellent substrates for neuronal cultures, showing high affinity and greatly boosting their synaptic functionality. Therefore, growing cells on CNT offers an opportunity to perform a large variety of neuropathology studies in vitro. To date, the interactions between neurons and chemical functional groups have not been studied extensively. To this end, multiwalled CNT (f-CNT) is functionalized with various functional groups, including sulfonic (-SO3 H), nitro (-NO2 ), amino (-NH2 ), and oxidized moieties. f-CNTs are spray-coated onto untreated glass substrates and are used as substrates for the incubation of neuroblastoma cells (SH-SY5Y). After 7 d, its effect is evaluated in terms of cell attachment, survival, growth, and spontaneous differentiation. Cell viability assays show quite increased proliferation on various f-CNT substrates (CNTs-NO2 > ox-CNTs ≈ CNTs-SO3 H > CNTs ≈ CNTs-NH2 ). Additionally, SH-SY5Y cells show selectively better differentiation and maturation with -SO3 H substrates, where an increased expression of ß-III tubulin is seen. In all cases, intricate cell-CNT networks are observed and the morphology of the cells adopts longer and thinner cellular processes, suggesting that the type of functionalization may have an effect of the length and thickness. Finally, a possible correlation is determined between conductivity of f-CNTs and cell-processes lengths.

Solubility-consistent force field simulations for aqueous metal carbonate systems using graphical processing units.

Crystallization of alkaline earth metal carbonates from water is important for biomineralization and environmental geochemistry. Here, large-scale computer simulations are a useful approach to complement experimental studies by providing atomistic insights and even by quantitatively determining the thermodynamics of individual steps. However, this is dependent on the existence of force field models that are sufficiently accurate while being computationally efficient enough to sample complex systems. Here, we introduce a revised force field for aqueous alkaline earth metal carbonates that reproduces both the solubilities of the crystalline anhydrous minerals, as well as the hydration free energies of the ions. The model is also designed to run efficiently on graphical processing units thereby reducing the cost of such simulations. The performance of the revised force field is compared against previous results for important properties relevant to crystallization, including ion-pairing and mineral-water interfacial structure and dynamics. This article is part of a discussion meeting issue 'Supercomputing simulations of advanced materials'.

An Electroactive and Self-Assembling Bio-Ink, based on Protein-Stabilized Nanoclusters and Graphene, for the Manufacture of Fully Inkjet-Printed Paper-Based Analytical Devices.

Hundreds of new electrochemical sensors are reported in literature every year. However, only a few of them makes it to the market. Manufacturability, or rather the lack of it, is the parameter that dictates if new sensing technologies will remain forever in the laboratory in which they are conceived. Inkjet printing is a low-cost and versatile technique that can facilitate the transfer of nanomaterial-based sensors to the market. Herein, an electroactive and self-assembling inkjet-printable ink based on protein-nanomaterial composites and exfoliated graphene is reported. The consensus tetratricopeptide proteins (CTPRs), used to formulate this ink, are engineered to template and coordinate electroactive metallic nanoclusters (NCs), and to self-assemble upon drying, forming stable films. The authors demonstrate that, by incorporating graphene in the ink formulation, it is possible to dramatically improve the electrocatalytic properties of the ink, obtaining an efficient hybrid material for hydrogen peroxide (H2 O2 ) detection. Using this bio-ink, the authors manufactured disposable and environmentally sustainable electrochemical paper-based analytical devices (ePADs) to detect H2 O2 , outperforming commercial screen-printed platforms. Furthermore, it is demonstrated that oxidoreductase enzymes can be included in the formulation, to fully inkjet-print enzymatic amperometric biosensors ready to use.

Structurally distinct mitoviruses: are they an ancestral lineage of the Mitoviridae exclusive to arbuscular mycorrhizal fungi (Glomeromycotina)?

Mitoviruses in the family Mitoviridae are the mitochondria-replicating "naked RNA viruses" with genomes encoding only the replicase RNA-dependent RNA polymerase (RdRp) and prevalent across fungi, plants, and invertebrates. Arbuscular mycorrhizal fungi in the subphylum Glomeromycotina are obligate plant symbionts that deliver water and nutrients to the host. We discovered distinct mitoviruses in glomeromycotinian fungi, namely "large duamitovirus," encoding unusually large RdRp with a unique N-terminal motif that is endogenized in some host genomes. More than 400 viral sequences similar to the large duamitoviruses are present in metatranscriptome databases. They are globally distributed in soil ecosystems, consistent with the cosmopolitan distribution of glomeromycotinian fungi, and formed the most basal clade of the Mitoviridae in phylogenetic analysis. Given that glomeromycotinian fungi are the only confirmed hosts of these viruses, we propose the hypothesis that large duamitoviruses are the most ancestral lineage of the Mitoviridae that have been maintained exclusively in glomeromycotinian fungi.

Ultrasensitive detection of SARS-CoV-2 spike protein by graphene field-effect transistors.

COVID-19, caused by the severe acute respiratory syndrome-coronavirus 2 (SARS-CoV-2), originated a global health crisis, causing over 2 million casualties and altering human daily life all over the world. This pandemic emergency revealed the limitations of current diagnostic tests, highlighting the urgency to develop faster, more precise and sensitive sensors. Graphene field effect transistors (GFET) are analytical platforms that enclose all these requirements. However, the design of a sensitive and robust GFET is not a straightforward objective. In this work, we report a GFET array biosensor for the detection of SARS-CoV-2 spike protein using the human membrane protein involved in the virus internalisation: angiotensin-converting enzyme 2 (ACE2). By finely controlling the graphene functionalisation, by tuning the Debye length, and by deeply characterising the ACE2-spike protein interactions, we have been able to detect the target protein with an extremely low limit of detection (2.94 aM). This work set the basis for a new class of analytical platforms, based on human membrane proteins, with the potential to detect a broad variety of pathogens, even before their isolation, being a powerful tool in the fight against future pandemics.

The Covalent Functionalization of Surface-Supported Graphene: An Update.

In the last decade, the use of graphene supported on solid surfaces has broadened its scope and applications, and graphene has acquire a promising role as a major component of high-performance electronic devices. In this context, the chemical modification of graphene has become essential. In particular, covalent modification offers key benefits, including controllability, stability, and the facility to be integrated into manufacturing operations. In this Review, we critically comment on the latest advances in the covalent modification of supported graphene on substrates. We analyze the different chemical modifications with special attention to radical reactions. In this context, we review the latest achievements in reactivity control, tailoring electronic properties, and introducing active functionalities. Finally, we extended our analysis to other emerging 2D materials supported on surfaces, such as transition metal dichalcogenides, transition metal oxides, and elemental analogs of graphene.

Factors affecting duration of post-surgical orthodontics in the Surgery First/Early Approach: A retrospective study.

BACKGROUND: Surgery First Approach (SFA) and Surgery Early (SE) are considered promising alternatives, compared to the conventional three-stages orthodontic-surgical approach, for treatment of dento-maxillofacial deformities. However, many features need further study, like the role of the orthodontist. Aim of the study was to analyse the clinical characteristics of patients who underwent SFA and SE, and if differences in duration of orthodontics could be influenced by clinical features. METHODS: A retrospective research was performed on patients who met the inclusion criteria for SFA (31) and SE (12), of the total of patients affected by dento-facial deformities in our Unit (191) in the period 2012-2017. After collection of clinical data, duration of orthodontics, age, pre-treatment PAR Index, ANB angle, amount of the curve of Spee were compared. A regression analysis evaluated if these clinical parameters, together with type of bracket and type of intervention, could influence the duration of post-surgical orthodontics. RESULTS: All patients who performed the SFA/SE were affected by class III, II and active Unilateral Condylar Hyperplasia (22% of total population). Pre-treatment mean differences of age (p = 0.0518), PAR Index (p = 0.0916), curve of Spee (p = 0.1006) between groups were not statistically significant. A statically significant difference was found for the overall duration of therapy, for the significant shorter duration of pre-surgical orthodontics, while the difference of post-surgical orthodontics duration was not significant (p = 0.4753). Type of bracket (rho=-0.19039, p = 0.266) and intervention performed (rho=-0.11522, p = 0.5034) were not correlated with duration of post-surgical orthodontics, as well as pre-treatment PAR Index, ANB angle and depth of the curve of Spee. CONCLUSIONS: Surgery First/Early Approach is a therapeutic choice that could be performed only in patients affected by specific malocclusions and who exactly meet indications. Protocol and post-surgical occlusal stability are factors that should influence the duration of therapy more than clinical characteristics.

Broad-Spectrum Antibacterial Activity of Synthesized Carbon Nanodots from d-Glucose.

Carbon nanodots, a class of carbon nano-allotropes, have been synthesized through different routes and methods from a wide range of precursors. The selected precursor, synthetic method, and conditions can strongly alter the physicochemical properties of the resulting material and their intended applications. Herein, carbon nanodots (CNDs) have been synthesized from d-glucose by combining pyrolysis and chemical oxidation methods. The effect of the pyrolysis temperature, equivalents of oxidizing agent, and refluxing time were studied on the product and quantum yield. In the optimum conditions (pyrolysis temperature of 300 °C, 4.41 equiv of H2O2, 90 min of reflux) CNDs were obtained with 40% and 3.6% of product and quantum yields, respectively. The obtained CNDs are negatively charged (ζ-potential = -32 mV), excellently dispersed in water, with average diameter of 2.2 nm. Furthermore, ammonium hydroxide (NH4OH) was introduced as dehydrating and/or passivation agent during CNDs synthesis resulting in significant improvement of both product and quantum yields of about 1.5 and 3.76-fold, respectively. The synthesized CNDs showed a broad spectrum of antibacterial activities toward different Gram-positive and Gram-negative bacteria strains. Both synthesized CNDs caused highly colony forming unit reduction (CFU), ranging from 98% to 99.99% for most of the tested bacterial strains. However, CNDs synthesized in the absence of NH4OH, due to a negatively charged surface enriched in oxygenated groups, performed better in zone inhibition and minimum inhibitory concentration. The elevated antibacterial activity of high-oxygen-containing carbon nanodots is directly correlated to their ROS formation ability.

Obtaining Consistent Free Energies for Ion Binding at Surfaces from Solution: Pathways versus Alchemy for Determining Kink Site Stability.

Ion incorporation or removal from a solid at the interface with solution is a fundamental part of crystal growth. Despite this, there have been few quantitative determinations of the thermodynamics for such processes from atomistic molecular dynamics due to the associated technical challenges. In this study, we compute the free energies for ion removal from kink sites at the interface between NaCl and water as an illustrative example. To examine the influence of the free energy technique used, we compare methods that follow an explicit pathway for dissolution with those that focus on the thermodynamics of the initial and final states using metadynamics and free energy perturbation, respectively. While the initial results of the two approaches are found to be completely different, it is demonstrated that the thermodynamics can be reconciled with appropriate corrections for the standard states, thus illustrating the need for caution in interpreting raw free energy curves for ion binding as widely found in the literature. In addition, a new efficient approach is introduced to correct for the system size dependence of kink site energies both due to the periodic interaction of charges in an inhomogeneous dielectric system and due to the dipolar interactions between pairs of kinks along a row. Ultimately, it is shown that with suitable care, both classes of free energy techniques are capable of producing kink site stabilities that are consistent with the solubility of the underlying bulk solid. However, the precise values for individual kink sites exhibit a small systematic offset, which can be ascribed to the contribution of the interfacial potential to the pathway-based results. For the case of NaCl, the free energies of the kink sites relative to a 1 M aqueous solution for Na+ and Cl- are found to be surprisingly different and of opposite sign, despite the ions having very similar hydration free energies.

The era of nano-bionic: 2D materials for wearable and implantable body sensors.

Nano-bionics have the potential of revolutionizing modern medicine. Among nano-bionic devices, body sensors allow to monitor in real-time the health of patients, to achieve personalized medicine, and even to restore or enhance human functions. The advent of two-dimensional (2D) materials is facilitating the manufacturing of miniaturized and ultrathin bioelectronics, that can be easily integrated in the human body. Their unique electronic properties allow to efficiently transduce physical and chemical stimuli into electric current. Their flexibility and nanometric thickness facilitate the adaption and adhesion to human body. The low opacity permits to obtain transparent devices. The good cellular adhesion and reduced cytotoxicity are advantageous for the integration of the devices in vivo. Herein we review the latest and more significant examples of 2D material-based sensors for health monitoring, describing their architectures, sensing mechanisms, advantages and, as well, the challenges and drawbacks that hampers their translation into commercial clinical devices.

Assessment of maxillary canting on cone beam computed tomography and digital models: A retrospective study and proposal of a method.

OBJECTIVES: The mounting of the plaster casts on articulator procedure is routinely performed in orthognathic surgery to assess canting of the maxillary occlusal plane, but the currently used protocols and reference plane could be source of errors which affect reliability. Nowadays the assessment of canting of the maxillary occlusal plane could be also performed with an entirely digital protocol. Aim of the study was to propose a method to evaluate canting in patients affected by Unilateral Condylar Hyperplasia, comparing the measurements performed on digital models matched on CBCT with those made on traditional articulator. MATERIALS AND METHODS: A retrospective cross-sectional study was designed on 20 patients affected by vertical Unilateral Condylar Hyperplasia treated in the Units of Orthodontics and Maxillo-Facial Surgery. The canting of the maxillary occlusal plane was measured on plaster casts mounted on the conventional articulator and the measures were compared with those made on digital models matched on CBCT, according the protocol developed in our Unit. Molar, canine and basal difference were measured. To compare the two protocols and to test the agreement, we performed descriptive statistics, comparison between means and Bland Altman analysis. P value was set at 0.05. RESULTS: Statistic comparison demonstrated agreement between measurements performed with the digital protocol and conventional physical method. CONCLUSION: Measurements of canting with digital protocol are comparable to the physical standard method. A total digital protocol allows faster availability and storage of patient's data and better communication between orthodontist and maxillo-facial surgeon, especially in patients affected by three-dimensional malocclusions.

A Universal Force Field for Materials, Periodic GFN-FF: Implementation and Examination.

In this study, the adaption of the recently published molecular GFN-FF for periodic boundary conditions (pGFN-FF) is described through the use of neighbor lists combined with appropriate charge sums to handle any dimensionality from 1D polymers to 2D surfaces and 3D solids. Numerical integration over the Brillouin zone for the calculation of π bond orders of periodic fragments is also included. Aside from adapting the GFN-FF method to handle periodicity, improvements to the method are proposed in regard to the calculation of topological charges through the inclusion of a screened Coulomb term that leads to more physical charges and avoids a number of pathological cases. Short-range damping of three-body dispersion is also included to avoid collapse of some structures. Analytic second derivatives are also formulated with respect to both Cartesian and strain variables, including prescreening of terms to accelerate the dispersion/coordination number contribution to the Hessian. The modified pGFN-FF scheme is then applied to a wide range of different materials in order to examine how well this universal model performs.

Global Performance Index for Integrated Management System: GPI-IMS.

BACKGROUND: The present work starts from a literature review of the evolution of Integrated Management Systems (IMSs), considering different points of view and standards: quality, environmental, occupational health and safety, sustainability and social issues. Even if the benefits are possible, there is not a common approach and a clear link between the integration of management systems and business performance, in particular considering safety performance. METHODS: The present study analyzes the application of Risk Assessment in order to realize the integration of management systems. The main objective is to provide a tool for an integrated evaluation of all company performances, starting from the definition of some Key Performance Indicators-KPIs-proposed for a particular case study, even if their choice is not the core of the paper. The assessment team members on the basis of their knowledge, experience and useful literature, could choose the right KPIs for the specific application, able to take a picture of the current state and to suggest a possible recommended action of improving. The proposed Risk Assessment approach is an integration of modern management techniques: Integrated Management System and Improving Cycle DMAIC. RESULTS: The new method, called the Global Performance Index for Integrated Management System-GPI-IMS, has been applied to a real case study in the logistic field in order to evaluate its goodness and possible generalization. CONCLUSIONS: The proposed method allows to define the requirements that any company must have to perform the best. The role of the assessment team is very important to evaluate the global performance of the company and to suggest the corrective actions to be adopted.