Search

Polysaccharide scaffolds have been successfully employed to reconstruct environments that sustain skin tissue regeneration after injuries. Three-dimensional (3D) advanced additive manufacturing technologies allow creating scaffolds with controlled and reproducible macro- and micro-structure that improve the quality of the restored tissue to favor spontaneous repair. However, when persistent inflammation occurs, the physiological tissue healing capacity is reduced, like in the presence of pathologies like diabetes, vascular diseases, chronic infection, and others. In these circumstances, the bioavailability of therapeutic adjuncts like the growth factors in addition to the standard treatments represents undoubtedly a promising strategy to accelerate the healing of skin lesions. Precisely designed polysaccharide scaffolds obtained by 3D printing represent a robust platform that can be further implemented with the controlled delivery of bioactive adjuncts. Human elastin-like polypeptides (HELPs) are stimuli-responsive biopolymers. Their structure allows the integration of domains endowed with biological functionality, making them attractive compounds to prepare composites with smart properties. In the present study, 3D-printed alginate and chitosan scaffolds were combined with the HELP components. The HELP biopolymer was fused to the epidermal growth factor (EGF) as the bioactive domain. Different constructs were prepared and the stimuli-responsive behavior as well as the biological activity were evaluated, suggesting that these smart bioactive composites are suitable to realize multifunctional dressings that sustain the local release of therapeutic adjuncts.

Inhibition of intracellular N-acylethanolamine-hydrolyzing acid amidase (NAAA) activity is a promising approach to manage the inflammatory response under disabling conditions. In fact, NAAA inhibition preserves endogenous palmitoylethanolamide (PEA) from degradation, thus increasing and prolonging its anti-inflammatory and analgesic efficacy at the inflamed site. In the present work, we report the identification of a potent, systemically available, novel class of NAAA inhibitors, featuring a pyrazole azabicyclo[3.2.1]octane structural core. After an initial screening campaign, a careful structure–activity relationship study led to the discovery of endo-ethoxymethyl-pyrazinyloxy-8-azabicyclo[3.2.1]octane-pyrazole sulfonamide 50 (ARN19689), which was found to inhibit human NAAA in the low nanomolar range (IC50 = 0.042 μM) with a non-covalent mechanism of action. In light of its favorable biochemical, in vitro and in vivo drug-like profile, sulfonamide 50 could be regarded as a promising pharmacological tool to be further investigated in the field of inflammatory conditions.

Cystic fibrosis (CF) is a life-threatening autosomal recessive disease, caused by mutations in the CF transmembrane conductance regulator (CFTR) chloride channel. CFTR modulators have been reported to address the basic defects associated with CF-causing mutations, partially restoring the CFTR function in terms of protein processing and/or channel gating. Small-molecule compounds, called potentiators, are known to ameliorate the gating defect. In this study, we describe the identification of the 2,3,4,5-tetrahydro-1H-pyrido[4,3-b]indole core as a novel chemotype of potentiators. In-depth structure-activity relationship studies led to the discovery of enantiomerically pure 39 endowed with a good efficacy in rescuing the gating defect of F508del- and G551D-CFTR and a promising in vitro druglike profile. The in vivo characterization of γ-carboline 39 showed considerable exposure levels and good oral bioavailability, with detectable distribution to the lungs after oral administration to rats. Overall, these findings may represent an encouraging starting point to further expand this chemical class, adding a new chemotype to the existing classes of CFTR potentiators.

N-(4-Nitrobenzoyl)-Phe self-assembled into a transparent supramolecular hydrogel, which displayed high fibroblast and keratinocyte cell viability. The compound showed a mild antimicrobial activity against E. coli both as a hydrogel and in solution. Single-crystal XRD data revealed packing details, including protonation of the C-terminus due to an apparent pK(a) shift, as confirmed by pH titrations. MicroRaman analysis revealed almost identical features between the gel and crystal states, although more disorder in the former. The hydrogel is thermoreversible and disassembles within a range of temperatures that can be fine-tuned by experimental conditions, such as gelator concentration. At the minimum gelling concentration of 0.63 wt %, the hydrogel disassembles in a physiological temperature range of 39-42 degrees C, thus opening the way to its potential use as a biomaterial.

Deletion of phenylalanine at position 508 (F508del) in the CFTR chloride channel is the most frequent mutation in cystic fibrosis (CF) patients. F508del impairs the stability and folding of the CFTR protein, thus resulting in mistrafficking and premature degradation. F508del-CFTR defects can be overcome with small molecules termed correctors. We investigated the efficacy and properties of VX-445, a newly developed corrector, which is one of the three active principles present in a drug (Trikafta®/Kaftrio®) recently approved for the treatment of CF patients with F508del mutation. We found that VX-445, particularly in combination with type I (VX-809, VX-661) and type II (corr-4a) correctors, elicits a large rescue of F508del-CFTR function. In particular, in primary bronchial epithelial cells of CF patients, the maximal rescue obtained with corrector combinations including VX-445 was close to 60–70% of CFTR function in non-CF cells. Despite this high efficacy, analysis of ubiquitylation, resistance to thermoaggregation, protein half-life, and subcellular localization revealed that corrector combinations did not fully normalize F508del-CFTR behavior. Our study indicates that it is still possible to further improve mutant CFTR rescue with the development of corrector combinations having maximal effects on mutant CFTR structural and functional properties.

Tissue homeostasis mainly depends on the activity of stem cells to replace damaged elements and restore tissue functions. Within this context, mesenchymal stem cells and fibroblasts are essential for maintaining tissue homeostasis in skin, in particular in the dermis. Modifications in collagen fibers are able to affect stem cell features. Skin properties can be significantly reduced after injuries or with aging, and stem cell niches, mainly comprising extracellular matrix (ECM), may be compromised. To this end, specific molecules can be administrated to prevent the aging process induced by UV exposure in the attempt to maintain a youngness phenotype. NanoPCL-M is a novel nanodevice able to control delivery of Mediterranean plant myrtle (Myrtus communis L.) extracts. In particular, we previously described that myrtle extracts, rich in bioactive molecules and nutraceuticals, were able to counteract senescence in adipose derived stem cells. In this study, we analyzed the effect of NanoPCL-M on skin stem cells (SSCs) and dermal fibroblasts in a dynamic cell culture model in order to prevent the effects of UV-induced senescence on proliferation and collagen depot. The BrdU assay results highlight the significantly positive effect of NanoPCL-M on the proliferation of both fibroblasts and SSCs. Our results demonstrate that-M is able to preserve SSCs features and collagen depot after UV-induced senescence, suggesting their capability to retain a young phenotype.

Chronic use of alcohol and tobacco cigarettes is associated to millions of deaths per year, either by direct or indirect causes. However, few studies have explored the additional risks of the combined use of these drugs. Here we assessed the effect of the combined use of alcohol and cigarette smoke on liver or kidney morphology, and on biochemical parameters in chronically treated rats. Male Wistar rats were allocated to receive 2 g/kg alcohol orally, which was followed by the inhalation of smoke from six cigarettes during 2 h (ALTB group) for 28 days. Other groups received alcohol alone (AL) or were exposed to cigarette smoke (TB) alone and were compared to control (CT) rats, which received water followed by ambient air. On day 29, rats were euthanized and blood samples were collected for aminotransferase enzymes (AST and ALT), creatinine, and urea analysis. Liver and kidney were weighted, dissected, fixed, and stained with hematoxylin and eosin for morphological analysis. Our results showed that necrosis was elevated in the AL, TB, and mainly the ALTB group in both liver and kidney of rats. Serum levels of AST and ALT were reduced by cigarette smoke exposure, independently of alcohol use. Serum creatinine levels increased after tobacco smoke exposure. On the other hand, TB and AL groups decreased serum urea levels, and their association restored that decrease. Absolute liver and kidney weights were lower in the cigarette smoke exposure rats. Lastly, body weight gain was lower in TB group and combined use restored it. Thus, we may infer that the use of alcohol, exposure to tobacco cigarette smoke or, mainly, their association promotes liver and kidney injuries, and this damage is related with biochemical changes in rats.

Chronic use of alcohol and its withdrawal impairs the delicate balance between GABAergic and glutamatergic systems. This imbalance includes changes in GABA receptors – importantly in GABAA subtypes – and glutamate receptors, especially in NMDA subtypes. A better comprehension of the different roles of GABAAR and NMDAR subunits could be helpful to define new strategies to counteract the deleterious effects observed during alcohol withdrawal. Taurine, a sulfonated amino acid, has been proposed to attenuate alcohol withdrawal symptoms due to its neuromodulatory properties. In this study, we evaluated the correlations between GABAAR and NMDAR subunits in the hippocampus of rats chronically treated with alcohol or in alcohol withdrawal, and the effects of taurine treatment on these parameters. Male Wistar rats received alcohol (2 g/kg) or water by oral gavage (control), 2 × /day, for 28 days. From day 29 to day 33, withdrawal rats received water instead of alcohol and all groups were reallocated to receive 100 mg/kg taurine or saline intraperitoneally (i.p.), once a day. On day 34, rats were euthanized and the hippocampus was dissected for GABAAR α1, α4, δ, and γ2 and NMDAR GluN2A and GluN2B subunits mRNA expression determination by RT qPCR. There were no differences between groups in the studied GABAAR and NMDA subunits. However, we observed a correlation of α1 and γ2 subunits induced by taurine, while in the alcohol group there was a correlation between α4 and GluN2A. In the group treated with alcohol and taurine, we observed an extra correlation, between α1 and GluN2A. After 5 days of withdrawal, a correlation observed in the control group, between δ and GluN2A, was reestablished. The correlation found between subunits suggests a neuroadaptation of GABAergic and glutamatergic systems in withdrawal rats. Results from this study contribute to the elucidation of the mechanisms beyond neuroadaptations observed in alcohol use and withdrawal.

The combination of a chitosan 3D-printed scaffold with a hydrogel matrix containing an elastin-like polypeptide functionalized with the epidermal growth factor (HEGF) was evaluated as a possible strategy to obtain a bioactive platform with stimuli-responsive properties. We designed a chitosan/HEGF hybrid scaffold and examined the physico-chemical properties and the in vitro behavior when in contact with simulated biological fluids. Primary human dermal fibroblasts (hDFs) were used to test the in vitro cytocompatibility. Overall, these data provide first insights into the integration of HEGF-based hydrogel with 3D-printed scaffolds, contributing towards the rational design of a new smart functional wound dressing. Graphical Abstract: [Figure not available: see fulltext.].

no abstract available

The use of graphene nanomaterials (GNMs) for biomedical applications targeted to the central nervous system is exponentially increasing, although precise information on their effects on brain cells is lacking. In this work, we addressed the molecular changes induced in cortical astrocytes by few-layer graphene (FLG) and graphene oxide (GO) flakes. Our results show that exposure to FLG/GO does not affect cell viability or proliferation. However, proteomic and lipidomic analyses unveiled alterations in several cellular processes, including intracellular Ca2+ ([Ca2+]i) homeostasis and cholesterol metabolism, which were particularly intense in cells exposed to GO. Indeed, GO exposure impaired spontaneous and evoked astrocyte [Ca2+]i signals and induced a marked increase in membrane cholesterol levels. Importantly, cholesterol depletion fully rescued [Ca2+]i dynamics in GO-treated cells, indicating a causal relationship between these GO-mediated effects. Our results indicate that exposure to GNMs alters intracellular signaling in astrocytes and may impact on astrocyte-neuron interactions., Comment: This document is the unedited Author's version of a Submitted Work that was subsequently accepted for publication in Small after peer review. To access the final edited and published work see https://onlinelibrary.wiley.com/doi/10.1002/smll.201900147 40 pages, 6 main figures and 1 supplementary figure

Fructosyl peptide oxidases (FPOXs) are enzymes currently used in enzymatic assays to measure the concentration of glycated hemoglobin and albumin in blood samples, which serve as biomarkers of diabetes. However, since FPOX are unable to work directly on glycated proteins, current enzymatic assays are based on a preliminary proteolytic digestion of the target proteins. Herein, to improve the speed and costs of the enzymatic assays for diabetes testing, we applied a rational design approach to engineer a novel enzyme with a wider access tunnel to the catalytic site, using a combination of Rosetta design and molecular dynamics simulations. Our final design, L3_35A, shows a significantly wider and shorter access tunnel, resulting from the deletion of five-amino acids lining the gate structures and from a total of 35 point mutations relative to the wild-type (WT) enzyme. Indeed, upon experimental testing, our engineered enzyme shows good structural stability and maintains significant activity relative to the WT.

Natural cosmetic products have recently re-emerged as a novel tool able to counteract skin aging and skin related damages. In addition, recently achieved progress in nanomedicine opens a novel approach yielding from combination of modern nanotechnology with traditional treatment for innovative pharmacotherapeutics. In the present study, we investigated the antiaging effect of a pretreatment with Myrtus communis natural extract combined with a polycaprolactone nanofibrous scaffold (NanoPCL-M) on skin cell populations exposed to UV. We set up a novel model of skin on a bioreactor mimicking a crosstalk between keratinocytes, stem cells and fibroblasts, as in skin. Beta-galactosidase assay, indicating the amount of senescent cells, and viability assay, revealed that fibroblasts and stem cells pretreated with NanoPCL-M and then exposed to UV are superimposable to control cells, untreated and unexposed to UV damage. On the other hand, cells only exposed to UV stress, without NanoPCL-M pretreatment, exhibited a significantly higher yield of senescent elements. Keratinocyte-based 3D structures appeared disjointed after UV-stress, as compared to NanoPCL-M pretreated samples. Gene expression analysis performed on different senescence associated genes, revealed the activation of a molecular program of rejuvenation in stem cells pretreated with NanoPCL-M and then exposed to UV. Altogether, our results highlight a future translational application of NanoPCL-M to prevent skin aging.

The LIBRA compound library is a collection of 522 non-commercial molecules contributed by various Italian academic laboratories. These compounds have been designed and synthesized during different medicinal chemistry programs and are hosted by the Italian Institute of Technology. We report the screening of the LIBRA compound library against Trypanosoma brucei and Leishmania major pteridine reductase 1, TbPTR1 and LmPTR1. Nine compounds were active against parasitic PTR1 and were selected for cell-based parasite screening, as single agents and in combination with methotrexate (MTX). The most interesting TbPTR1 inhibitor identified was 4-(benzyloxy)pyrimidine-2,6-diamine (LIB_66). Subsequently, six new LIB_66 derivatives were synthesized to explore its Structure-Activity-Relationship (SAR) and absorption, distribution, metabolism, excretion and toxicity (ADMET) properties. The results indicate that PTR1 has a preference to bind inhibitors, which resemble its biopterin/folic acid substrates, such as the 2,4-diaminopyrimidine derivatives.

In this contribution we describe the preparation and characterization of a series of cross-linked films based on the combination of an elastin-derived biomimetic polypeptide (Human Elastin-Like Polypeptide, HELP) with alginate (ALG) to obtain a composite with enhanced properties. ALG/HELP composite films loaded with the hydrophobic natural antioxidant curcumin were prepared by solvent casting method followed by the cross-linking with calcium chloride. The compatibility between the two components as well as the final properties was evaluated. The micro-morphological study of films showed a homogeneous structure, but the film tensile strength decrease with HELP content and elongation at break was adversely affected by biopolymer addition. Spectroscopic and thermal analyses confirmed an interaction between ALG and HELP which also causes a modification in swelling kinetics and faster degradation. Moreover, the study of curcumin release showed a controlled delivery up to 10 days with a faster release rate in the presence of HELP. Human Dermal Fibroblasts (hDF) were used to test the in vitro cytocompatibility. The antioxidant activity correlated to the increase of HELP content suggested the applicability of these composites to develop smart biomaterials. Overall, these features indicated how this composite material has considerable potential as customizable platforms for various biomedical applications.

A new, bifunctional recombinant protein was expressed as the fusion product of human elastin-like polypeptide (HELP) and the bilirubin-binding protein UnaG. The engineered product displays both the HELP-specific property of forming a functional hydrogel matrix and the UnaG-specific capacity of emitting green fluorescence upon ligand binding. The new fusion protein has been proven to be effective at detecting bilirubin in complex environments with high background noise. A cell culture model of the stress response, consisting of bilirubin released in the cell culture medium, was set up to assess the bilirubin-sensing properties of the functional matrix obtained by cross-linking the HELP moiety. Our engineered protein allowed us to monitor cell induction by the release of bilirubin in the culture medium on a nanomolar scale. This study shows that elastin-like protein fusion represents a versatile platform for the development of novel and commercially viable analytical and biosensing devices.

Due to the complex and multifactorial nature of bipolar disorder (BD), single-target drugs have traditionally provided limited relief with no disease-modifying effects. In line with the polypharmacology paradigm, we attempted to overcome these limitations by devising two series of multitarget-directed ligands endowed with both a partial agonist profile at dopamine receptor D3 (D3R) and inhibitory activity against glycogen synthase kinase 3 beta (GSK-3β). These are two structurally unrelated targets that play independent, yet connected, roles in cognition and mood regulation. Two compounds (7 and 10) emerged as promising D3R/GSK-3β multitarget-directed ligands with nanomolar activity at D3R and low-micromolar inhibition of GSK-3β, thereby confirming, albeit preliminarily, the feasibility of our strategy. Furthermore, 7 showed promising drug-like properties in stability and pharmacokinetic studies.

ObjectiveTo analyse the correlations between olfactory psychophysical scores and the serum levels of D-dimer, C-reactive protein, ferritin, lactate dehydrogenase, procalcitonin and neutrophil-to-lymphocyte ratio in coronavirus disease 2019 patients.MethodsPatients underwent psychophysical olfactory assessment with the Connecticut Chemosensory Clinical Research Center test, and determination of blood serum levels of the inflammatory markers D-dimer, C-reactive protein, ferritin, lactate dehydrogenase, procalcitonin and neutrophil-to-lymphocyte ratio within 10 days of the clinical onset of coronavirus disease 2019 and 60 days after.ResultsSeventy-seven patients were included in this study. D-dimer, procalcitonin, ferritin and neutrophil-to-lymphocyte ratio correlated significantly with severe coronavirus disease 2019. No significant correlations were found between baseline and 60-day Connecticut Chemosensory Clinical Research Center test scores and the inflammatory markers assessed.ConclusionOlfactory disturbances appear to have little prognostic value in predicting the severity of coronavirus disease 2019 compared to D-dimer, ferritin, procalcitonin and neutrophil-to-lymphocyte ratio. The lack of correlation between the severity and duration of olfactory disturbances and serum levels of inflammatory markers seems to further suggest that the pathogenetic mechanisms underlying the loss of smell in coronavirus disease 2019 patients are related to local rather than systemic inflammatory factors.

Exposure to cigarette smoke and ethanol are proposed to trigger neurotoxicity, apoptosis, and to impair neuronal signaling. However, it is little known how the combination of both might trigger astrogliosis and the morphological changes capable of affecting a differential susceptibility of hippocampal regions to these licit drugs. The present study investigated the chronic effects of exposure to cigarette smoke and/or ethanol on behavioral parameters, apoptosis, and alteration in immunoreactivity of glial fibrillary acid protein (GFAP) and S100β in the CA1, CA3, and dentate gyrus (DG) of the rat hippocampus. Adult male Wistar rats (n = 32) were divided into four groups: vehicle (VE, glucose 3% in water, 10 mL/kg), cigarette smoke (TOB, total 12 cigarettes per day), ethanol (ethanol, 2 g/kg), and cigarette smoke plus ethanol (TOB plus ethanol, total 12 cigarettes per day plus ethanol 2 g/kg) for 54 days. The groups were submitted to tail-flick, open-field, and inhibitory avoidance tasks. The results showed that ethanol per se worsened the short-term memory. The association between TOB and ethanol increased the immunoreactivity of cleaved caspase-3 in the CA3 and DG regions. The TOB plus ethanol group showed a lower immunoreactivity to GFAP in all regions of the hippocampus. In addition, ethanol and TOB per se also reduced the immunoreactivity for GFAP in the DG. Ethanol increased S100β immunoreactivity only in the DG. In conclusion, this study showed that only ethanol worsened short-term memory, and the DG became more susceptible to changes in the markers investigated. This evidence suggests that DG is more sensitive to neurotoxicity induced by cigarette smoke and ethanol.

Ultraviolet (UV) light exposure is the primary factor responsible for skin photoaging, affecting all the skin layers, mainly through the production of reactive oxygen species (ROS), activation of inflammatory responses, and apoptosis. In keeping with this evidence, exogenous supplementation with dietary antioxidants has been shown to provide photoprotective benefits. Moreover, oral administration of hyaluronic acid (HA) has been proved to reduce the signs of aged skin, such as wrinkles, and increase hydration and elasticity. The combination of different biologically active substances in order to slow down the onset of skin aging could represent a promising preventive strategy against photoaging. In the present study, we investigated the effects of a dietary supplement (IALUTEC® RED), consisting of high-molecular-weight HA (HMW-HA) combined with red orange extract (ROC-Red Orange Complex® ), in human fibroblasts exposed to ultra violet light B-induced oxidative stress. Our study suggests that, in fibroblasts exposed to UVB light, IALUTEC® RED is active in decreasing both the inflammatory response and the generation of ROS, two events that are involved in skin photoaging.

The polymerization of n-butyl acrylate (BA) and n-butyl methacrylate (BMA) in the presence of different amounts of a macromonomer containing vinyl aromatic terminal double bonds presents completely different behaviors. While the polymerization rate of BMA is not affected by increasing amounts of the macromonomer in the formulation (up to 40 wt%), the polymerization of BA is strongly affected; the polymerization rate decreases as the concentration of the macromonomer increases. Quantum chemical simulations of the potential reaction mechanisms for both monomer systems revealed that for BA the tertiary radical formed by the addition of a tertiary BA radical (most predominant compared to the secondary radical) to the macromonomer double bond is very stable, hence this reaction acts as a radical sink that reduces growing radicals’ concentration and polymerization rate. Furthermore, a mathematical model of the polymerization process considering the most likely mechanisms evaluated by the quantum chemical simulations was able to fit reasonably well the polymerization of the two monomers. Guidelines for the values of the unknown kinetic rate coefficients are obtained.

This work aimed to investigate the origin of different red hues of Roman copper-based red opaque glass sectilia, to shed light on the production technology behind them. This objective was achieved by the depth study of the samples of glass sectilia, which decorated the villa of co-Emperor Lucius Verus (161&ndash, 169 AD). These were selected for analysis due to their abundance, the certainty of their date and of their different red and orange hues. Using OM (optical microscopy), colourimetry and FORS (fibre optical reflectance spectroscopy) spectroscopy, four red and four orange hues were individuated. A set of representative samples for each hue was analysed by EPMA (electron probe microanalyses) to detect any correlation between colour and chemical composition. Crystalline phases were investigated through high-resolution FEG&ndash, SEM (field emission gun scanning electron microscope), &mu, Raman spectroscopy and XRD, for the identification of colouring and opacifying agents and to understand how the different hues are affected by their shape, concentration and dimension. Sub-micrometric particles of metallic copper and cuprite crystals were identified as both the colouring and opacifying agents. These were not present in the same samples and were manufactured by two distinct colouring techniques, corresponding to two different glass chemical compositions. The size and the number of the colouring particles were the main factors that distinguished one hue from another. Although produced through different colouring techniques, some red samples appeared to be very similar to each other. These data enrich a period of the Roman age through some analyses and allow the identification of the type, as well as some of the production conditions of opaque red glass produced during 2nd century AD, which could be considered to be a period of transition from one technology to another.

Background The effects of tobacco smoke on the central nervous system are usually studied with isolated nicotine, ignoring other compounds present in cigarette smoke. The few studies that use in vivo whole-body cigarette smoke exposure are usually performed in expensive commercial apparatus. New Method We presented a feasible, safe, and low-cost apparatus for cigarette smoke exposure in rodents. Results Rats exposed to cigarette smoke in this apparatus showed cotinine levels similar to human active smokers. Additional results showed that cigarette smoke exposure increased glutamate and aspartic acid levels and decreased leucine, isoleucine, ornithine, phenylalanine, and tryptophan levels in the cerebrospinal fluid of rats. Comparison with Existing Method(s) Our apparatus is feasible, safe, and costs 67-fold less than a commercial automatized smoking machine. Beyond the low cost, it does not require specialized knowledge for building or maintenance. Conclusions We concluded that our low-cost apparatus is reliable and reproduces cigarette smoke use in humans.

We identified a small molecule that, at picomolar concentrations, rescues mutant CFTR chloride channel from protein degradation., F508del, the most frequent mutation causing cystic fibrosis (CF), results in mistrafficking and premature degradation of the CFTR chloride channel. Small molecules named correctors may rescue F508del-CFTR and therefore represent promising drugs to target the basic defect in CF. We screened a carefully designed chemical library to find F508del-CFTR correctors. The initial active compound resulting from the primary screening underwent extensive chemical optimization. The final compound, ARN23765, showed an extremely high potency in bronchial epithelial cells from F508del homozygous patients, with an EC50 of 38 picomolar, which is more than 5000-fold lower compared to presently available corrector drugs. ARN23765 also showed high efficacy, synergy with other types of correctors, and compatibility with chronic VX-770 potentiator. Besides being a promising drug, particularly suited for drug combinations, ARN23765 represents a high-affinity probe for CFTR structure-function studies.

After a skin injury, many complex metabolic events are triggered to ensure proper wound healing. Particularly for chronic, non-healing wounds or burns several risk factors such as persistent bacterial infections and fast dehydration can counteract the healing process. Intelligent wound dressings should help accelerate the healing process, while maintaining the wound bed clean and disinfected for several days at a time. Ideally, they should be self-adherent to both moist and dry skin surfaces and be transparent enough to allow prolonged wound inspection. These requirements pose challenges both in terms of materials science and pharmaceutics. Herein, we describe fabrication of a transparent bilayer construct for the sequential release and delivery of a cutaneous antiseptic and a widely used antibiotic, potentially suitable for wound dressing applications. The fabrication is a scalable waterborne and ecofriendly solution casting process. The first layer (for direct wound contact) is polyvinylpyrrolidone (PVP) containing a commercial antiseptic, Neomercurocromo® (Neo), while the second layer is a blend of hyaluronic acid (HA) and PVP containing ciprofloxacin. We show that the bilayer films have satisfactory self-adhering strength to human skin and that PVP and HA can interact via hydrogen bonds causing sustained release of the antibiotic over a period of 5 days. Biocompatibility was demonstrated on human foreskin fibroblast HFF-1 cells. Antibacterial activity was evaluated against Staphylococcus aureus, Escherichia coli and Pseudomonas aeruginosa while the wound resorption behavior was assessed through an in vivo full-thickness excisional wound healing mice model. These observations indicate that such bilayer constructs can be potentially implemented as wound care products for diverse range of skin wounds, including large area skin infections.

Dendritic cells (DCs) initiate adaptive immune responses after their migration to secondary lymphoid organs. The LXR ligands/oxysterols and the RXR ligand 9-cis Retinoic Acid (9-cis RA) were shown to dampen DC migration to lymphoid organs through the inhibition of CCR7 expression. We performed transcriptomics of DCs undergoing maturation in the presence of the LXR ligand 22R-Hydroxycholesterol (22R-HC). The analysis highlighted more than 1500 genes modulated by 22R-HC treatment, including the triggering receptor expressed on myeloid cells (TREM)-1, which was found markedly up-regulated. We tested the effect of other nuclear receptor ligands (NRL) and we reported the induction of TREM-1 following RXR, RAR and VDR activation. From a functional point of view, triggering of TREM-1 induced by retinoids increased TNFα and IL-1β release, suggesting an active role of NRL-activated TREM-1(+) DCs in inflammation-driven diseases, including cancer. Consistently with this hypothesis we detected DCs expressing TREM-1 in pleural effusions and ascites of cancer patients, an observation validated by the induction of TREM-1, LXR and RAR target genes when monocyte-DCs were activated in the presence of tumor-conditioned fluids. Finally, we observed a better control of LLC tumor growth in Trem-1(−/-) bone marrow chimera mice as compared to wild type chimera mice. Future studies will be necessary to shed light on the mechanism of TREM-1 induction by distinct NRL, and to characterize the role of TREM-1(+) DCs in tumor growth.

In recent years, many studies are focusing on the negative effects of plastic pollution, and in particular on the nanosized plastic fragments and their implications on the environment and human health. Nanoplastics in the environment interact with a great number of substances, many of which are dangerous to humans, but the interaction mechanisms, the complexes formation processes, and their biological impact are still poorly understood. Here we report a study on the interactions of polyethylene terephthalate nanoplastics, produced by laser ablation, with three different types of contaminants: glyphosate, levofloxacin and Hg2+ ions, and we demonstrate that the nanoplastics form complexes with all three contaminants through their favorable binding. Most importantly, this study highlights that to demonstrate the overall effect of the nanoplastics internalized by cells in vitro, it is important to combine alternative methodologies, such as metabolomics, with standard biological assays (i.e., cell viability and ROS production). In this way it becomes possible to better understand the body’s response to this new class of pollutants and their possible chronic toxicity. Summary: PET nanoplastics, fabricated by laser ablation, interact with aqueous pollutants forming nanoclusters. The nanoclusters affect the cells metabolism, suggesting long-term risks.

In recent years, a number of drugs have been approved for the treatment of cystic fibrosis (CF). Among them, newly released Trikafta, a combination of 3 drugs (VX-661/VX-445/VX-770), holds great promise to radically improve the quality of life for a large portion of patients with CF carrying 1 copy of F508del, the most frequent CF transmembrane conductance regulator (CFTR) mutation. Currently available disease-modifying CF drugs work by rescuing the function of the mutated CFTR anion channel. Recent research has shown that membrane lipids, and the cell lipidome in general, play a significant role in the mechanism of CFTR-defective trafficking and, on the other hand, its rescue. In this paper, by using untargeted lipidomics on CFBE41o- cells, we identified distinctive changes in the bronchial epithelial cell lipidome associated with treatment with Trikafta and other CF drugs. Particularly interesting was the reduction of levels of ceramide, a known molecular player in the induction of apoptosis, which appeared to be associated with a decrease in the susceptibility of cells to undergo apoptosis. This evidence could account for additional beneficial roles of the triple combination of drugs on CF phenotypes., Untargeted lipidomics reveals unique signatures associated with cystic fibrosis drugs.

Taurine is an amino acid usually added to energy drinks. In rodents, acute taurine administration decreases voluntary alcohol intake and sub chronic administration restores different behavioral features impaired by alcohol withdrawal. In the present study, we evaluated the effects of chronic taurine treatment on voluntary alcohol consumption and changes in behavioral parameters in rats. Adult Male Wistar rats were divided into two groups and left free to choose from two bottles containing alcohol 20% or saccharin 0.08% (vehicle solution), or two bottles containing vehicle, 24 h per day for five weeks. After three weeks, rats received 100 mg/kg taurine (TAU) or saline (SAL) intraperitoneally once a day for two weeks, and daily alcohol consumption was monitored. On days 22 and 33, rats were tested in the open-field, and on day 34, they were exposed to the light/dark task (LDT). Our results show for the first time that chronic taurine treatment enhanced voluntary alcohol intake and preference in rats, and these changes were accompanied by an anxiolytic-like phenotype in alcohol-treated rats, possibly due to its synergistic effect with alcohol on the dopaminergic and GABAergic systems.

Mixed-metal olivine cathode materials are one of the most promising systems in the field of lithium batteries. Their main drawback is their low electronic conductivity, which limits the current rate at which they can be cycled. Herein, the effect on electrochemical properties of vicariant high-valence transition metals (HVTMs) inserted in the olivine crystal structure has been studied. The strategy adopted herein consists of doping a LiFe1/3Ni1/15Co3/5PO4 cathode by substituting part of the cobalt(II) ions with vanadium(IV), niobium(V), or tantalum(V) HVTMs. The WAXD studies reveal that small HVTMs in molar quantities (HVTM/P = 0.04) are vicariant into the olivine crystal lattice. Single-cell tests show that the Nb-doped cathode has a high-voltage (ca. 4.5 V) and a specific capacity of 149 mA h g−1 (656 mW h g−1). Thus, the insertion of HVTMs in the cathode structure exceptionally improved the electric conductivity of the material and yielded an improved cyclability and rate capability owing to its electro-compensation effect.

Strong reduction of multiple scattering for channeled particles has been observed in an experiment on the deflection of a 180 GeV/c π+-meson beam by bent silicon crystals. The RMS deflections due to multiple scattering for the channeled particles were about six times smaller than for non-channeled ones. It was shown that the approach suggested recently for the description of multiple scattering for channeled particles using the experimental data for random crystal orientations gives fair agreement with the experiment. Keywords: Crystal, Channeling, Multiple scattering, Dechanneling, Beam deflection

Visceral leishmaniasis is a serious zoonosis, and has a high incidence in Brazil. The ubiquitous presence of sand flies is a major player in the high incidence of leishmaniasis in urban areas of Brazil. Prevention and control or canine leishmaniasis is important to decrease the incidence of the zoonosis. Control is done with topical use of products with antiparasitic, repellent and insecticide action. This study evaluated the repellent efficacy of line-on application of a topical permethrin-dinotefuran-pyriproxyfen formulation in dogs. Six dogs were treated with the product and six did not receive treatment. All dogs were sedated and exposed to sand flies of the species Lutzomyia longipalpis weekly for 28 days. The result showed that the product had an efficacy of 90.4% in repelling sand flies. There was statistical significance (p ≤ 0,05) difference in the number (mean) of live engorged sand fly females between the two groups in all evaluations. The product showed repellent efficacy against Lutzomyia longipalpis for 28 days.

Objective: The use of a hyaluronic acid-based product has the potential to improve the hydration status of the mucous membranes, especially in the oral cavity. This study has evaluated the in vivo adhesiveness of a product based on hyaluronic acid combined with xanthan gum and carbomer to the oral cavity mucosa in human healthy subjects. The product was administered to 10 healthy volunteers during two tests: in the first participants were asked to abstain from drinking and eating for the duration of the test; in the second participants were free to follow their daily routine. In both tests, samples were taken by brushing the oral cavity immediately after administration of the product, and then after each hour, up to 4 hours. A qualitative and quantitative analysis of the drug was carried out by observation of the samples stained with Alcian Blue at pH 2.5. Results: In both tests the analysis demonstrated the presence of the product in all collected samples, up to 4 hours after administration of the product, confirming that the combination of hyaluronic acid with xanthan gum and carbomer can provide a long-lasting, durable mucoadhesive hydrogel.Keywords: Adhesiveness, carbomer, hyaluronic acid, oral cavity mucosa, xanthan gum.

Taurine, an amino acid with antioxidant and osmoregulatory properties, has been studied for its possible antidiabetic properties in type 1 and type 2 diabetic animals. In type 2 diabetic mice, taurine decreases blood glucose through increased insulin secretion and insulin receptor sensitization. However, insulin is absent in type 1 diabetic individuals. The aim of this study was to evaluate the effects of taurine on parameters related to the energy balance that could explain the metabolic action of this amino acid in type 1 diabetic rats. Control and streptozotocin-induced diabetic rats received saline or taurine (100 mg/kg/day), intraperitoneally, for 30 days. Parameters such as palatable food intake, gastrointestinal transit rate, serum glucose, insulin, leptin, and glucagon levels were measured 60 min after the last taurine administration. Liver, kidneys, heart, and retroperitoneal fat were dissected and weighted. Glycogen levels were measured in the liver and soleus muscle. Our results showed that acute taurine administration decreased glycemia. It also decreased food intake in diabetic rats, without affecting other metabolic parameters. Altogether, our results suggest that in type 1 diabetic rats, taurine decreases blood glucose by a non-insulin-dependent mechanism. Due to the safety profile of taurine, and its effect on glycemia, this amino acid may help to design new drugs to add benefit to insulin therapy in type 1 diabetic individuals.

Background Most authors have evaluated the location of lower leg arterial perforators, but little is still known about the relationship between the arterial network and great saphenous vein (GSV) and saphenous nerve (SN). The aim of this study is to evaluate the relationship between the arterial network of the posterior tibial artery perforators, the cutaneous nerves, and the superficial venous system in the lower one third of the leg. Methods Eighteen lower limbs from cadavers were used for this study. The arterial and venous compartment were selectively injected with a mixture of barium sulfate and epoxy. The specimen were CT scanned and the superficial veins, nerves, and the arterial perforators were dissected. Results A large perforator of the posterior tibial artery was found at a mean distance of 6.23 cm ± 0.88, with a 95% CI: 5.79-6.67, from the medial malleolus. The average diameter was 0.9 mm ± 0.17, with a 95% CI: 0.81-0.99. In 67% the connection of the venae comitantes to the superficial venous system was established with the GSV, in the other cases, with Leonardo's vein. Both dissection and imaging studies showed perineural interperforator connections along the branches of SN in all the specimens examined. Conclusions The distribution pattern of posterior tibial artery perforators followed the superficial nerves in this region. There is an interperforator anastomotic network along the SN. The various patterns of the venous drainage system, in relationship to the distribution of the branches of posterior tibial artery perforators, have been clarified.

We report the observation of the steering of 855 MeV electrons by bent silicon and germanium crystals at the MAinzer MIkrotron. 15 $\mu $m long crystals, bent along (111) planes, were exploited to investigate orientational coherent effects. By using a piezo-actuated mechanical holder, which allowed to remotely change the crystal curvature, it was possible to study the steering capability of planar channeling and volume reflection vs. the curvature radius and the atomic number, Z. For silicon, the channeling efficiency exceeds 35 %, a record for negatively charged particles. This was possible due to the realization of a crystal with a thickness of the order of the dechanneling length. On the other hand, for germanium the efficiency is slightly below 10 % due to the stronger contribution of multiple scattering for a higher-Z material. Nevertheless this is the first evidence of negative beam steering by planar channeling in a Ge crystal. Having determined for the first time the dechanneling length, one may design a Ge crystal based on such knowledge providing nearly the same channeling efficiency of silicon. The presented results are relevant for crystal-based beam manipulation as well as for the generation of e.m. radiation in bent and periodically bent crystals., Comment: 12 pages, 10 figures, submitted to Eur. Phys. J. C

We propose a new QSRR model based on a Kernel-based partial least-squares method for predicting UPLC retention times in reversed phase mode. The model was built using a combination of classical (physicochemical and topological) and nonclassical (fingerprints) molecular descriptors of 1383 compounds, encompassing different chemical classes and structures and their accurately measured retention time values. Following a random splitting of the data set into a training and a test set, we tested the ability of the model to predict the retention time of all the compounds. The best predicted/experimental R(2) value was higher than 0.86, while the best Q(2) value we observed was close to 0.84. A comparison of our model with traditional and simpler MLR and PLS regression models shows that KPLS better performs in term of correlation (R(2)), prediction (Q(2)), and support to MetID peak assignment. The KPLS model succeeded in two real-life MetID tasks by correctly predicting elution order of Phase I metabolites, including isomeric monohydroxylated compounds. We also show in this paper that the model's predictive power can be extended to different gradient profiles, by simple mathematical extrapolation using a known equation, thus offering very broad flexibility. Moreover, the current study includes a deep investigation of different types of chemical descriptors used to build the structure-retention relationship.

Cell encapsulation technology is still a challenging issue. Innovative methodologies such as additive manufacturing, and alternative bioprocesses, such as cell therapeutic delivery, where cell encapsulation is a key tool are rapidly gaining importance for their potential in regenerative medicine. Responsive materials such as elastin-based recombinant expression products have features that are particularly attractive for cell encapsulation. They can be designed and tailored to meet desired requirements. Thus, they represent promising candidates for the development of new concept-based materials that can be employed in this field. Areas covered: An overview of the design and employment of elastin-like polypeptides for cell encapsulation is given to outline the state of the art. Special attention is paid to the design of the macromolecule employed as well as to the method of matrix formation and the biological system involved. Expert opinion: As a result of recent progress in regenerative medicine there is a compelling need for materials that provide specific properties and demonstrate defined functional features. Rationally designed materials that may adapt according to applied external stimuli and that are responsive to biological systems, such as elastin-like polypeptides, belong to this class of smart material. A run through the components described to date represents a good starting point for further advancement in this area. Employment of these components in cell encapsulation application will promote its advance toward 'smart cell encapsulation technology'.

Direct antivirals are available for treating recurrent hepatitis C (RHC). This study reported outcomes of 424 patients with METAVIR F3–F4 RHC who were treated for 24 weeks with sofosbuvir/ribavirin and followed for 12 weeks within the Italian sofosbuvir compassionate use program. In 55 patients, daclatasvir or simeprevir were added. Child–Pugh class and model of end stage liver disease (MELD) scores were evaluated at baseline and 36 weeks after the start of therapy. The sustained viral response (SVR) was 86.7% (316/365) in patients who received sofosbuvir/ribavirin and 98.3% (58/59) in patients who received a second antiviral (P < 0.01). In patients treated with sofosbuvir/ribavirin, a significant difference in SVR was observed between patients diagnosed with METAVIR F4 (211/250; 84.4%), METAVIR F3 (95/105; 90.5%) and fibrosing cholestatic hepatitis (10/10; 100%) (P = 0.049). A significant association was found between patients who worsened from Child–Pugh class A and who experienced viral relapse (4/26 vs. 8/189, P = 0.02). In patients with a baseline MELD score

The biochemical properties of muscle extracellular matrix are essential for stem cell adhesion, motility, proliferation and myogenic development. Recombinant elastin-like polypeptides are synthetic polypeptides that, besides maintaining some properties of the native protein, can be tailored by fusing bioactive sequences to their C-terminal. Our laboratory synthesized several Human Elastin-Like Polypeptides (HELP) derived from the sequence of human tropoelastin. Here, we developed a novel HELP family member by fusing the elastin-like backbone to the sequence of human Epidermal Growth Factor. We employed this synthetic protein, named HEGF, either alone or in combination with other proteins of the HELP family carrying RGD-integrin binding sites, as adhesion substrate for C2C12 myoblasts and satellite cells primary cultures. Adhesion of myoblasts to HEGF-based substrates induced scattering, decreased adhesion and cytoskeleton assembly; the concomitant presence of the RGD motifs potentiated all these effects. Recombinant substrates induced myoblasts proliferation, differentiation and the development of multinucleated myotubes, thus favoring myoblasts expansion and preserving their myogenic potential. The effects induced by adhesion substrates were inhibited by AG82 (Tyrphostin 25) and herbimycin A, indicating their dependence on the activation of both the EGF receptor and the tyrosine kinase c-src. Finally, HEGF increased the number of muscle stem cells (satellite cells) derived from isolated muscle fibers in culture, thus highlighting its potential as a novel substrate for skeletal muscle regeneration strategies.

Stimuli-responsive hydrogel matrices are inspiring manifold applications in controlled delivery of bioactive compounds. Elastin-derived polypeptides form hydrogel matrices that may release bioactive moieties as a function of local increase of active elastases, as it would occur in several processes like inflammation. In view of the development of a patch for healing wounds, recombinant elastin-based polypeptides were combined with a proteolysis-resistant scaffold, made of electrospun poly-L-lactic acid (PLLA) fibers. The results of this study demonstrated the compatibility of these two components. An efficient procedure to obtain a composite material retaining the main features of each component was established. The release of the elastin moiety was monitored by means of a simple protocol. Our data showed that electrospun PLLA can form a composite with fusion proteins bound to elastin-derived polypeptides. Therefore, our approach allows designing a therapeutic agent delivery platform to realize devices capable of responding and interacting with biological systems at the molecular level.

Graphene-based materials are the focus of intense research efforts to devise novel theranostic strategies for targeting the central nervous system. In this work, we have investigated the consequences of long-term exposure of primary rat astrocytes to pristine graphene (GR) and graphene oxide (GO) flakes. We demonstrate that GR/GO interfere with a variety of intracellular processes as a result of their internalization through the endo-lysosomal pathway. Graphene-exposed astrocytes acquire a more differentiated morphological phenotype associated with extensive cytoskeletal rearrangements. Profound functional alterations are induced by GO internalization, including the upregulation of inward-rectifying K+ channels and of Na+-dependent glutamate uptake, which are linked to the astrocyte capacity to control the extracellular homeostasis. Interestingly, GO-pretreated astrocytes promote the functional maturation of co-cultured primary neurons by inducing an increase in intrinsic excitability and in the density of GABAergic synapses. The results indicate that graphene nanomaterials profoundly affect astrocyte physiology in vitro, with consequences for neuronal network activity. This work supports the view that GO-based materials could be of great interest to address pathologies of the central nervous system associated to astrocyte dysfunctions., Comment: This document is the unedited Author's version of a Submitted Work that was subsequently accepted for publication in Nano Letters. To access the final edited and published work see https://pubs.acs.org/articlesonrequest/AOR-wvjrggcBp7kC8NwEImXr

In the present study, we investigated the oxidative biotransformation of 2,2',4,4'-tetrabromodiphenyl ether (BDE-47) and 2,2',4,4',5-pentabromodiphenyl ether (BDE-99) by liver microsomes from wild lesser snow geese (Chen caerulescens caerulescens) and domesticated Japanese quail (Coturnix japonica). Formation of hydroxy-metabolites was analyzed using an ultra-high performance liquid chromatography-tandem mass spectrometry-based method. Incubation of BDE-47 with avian liver microsomes produced sixteen hydroxy-metabolites, eight of which were identified using authentic standards. The major metabolites formed by liver microsomes from individual lesser snow geese were 4-hydroxy-2,2',3,4'-tetrabromodiphenyl ether (4-OH-BDE-42), 3-hydroxy-2,2',4,4'-tetrabromodiphenyl ether (3-OH-BDE-47), and 4'-hydroxy-2,2',4,5'-tetrabromodiphenyl ether (4'-OH-BDE-49). By comparison, 4-OH-BDE-42 and 4'-OH-BDE-49, but not 3-OH-BDE-47, were major metabolites of Japanese quail liver microsomes. Unidentified metabolites included monohydroxy- and dihydroxy-tetrabromodiphenyl ethers. Incubation of BDE-99 with avian liver microsomes produced seventeen hydroxy-metabolites, twelve of which were identified using authentic standards. The major metabolites formed by lesser snow goose liver microsomes were 2,4,5-tribromophenol, 3-OH-BDE-47, 4'-OH-BDE-49, 4-hydroxy-2,2',3,4',5-pentabromodiphenyl ether (4-OH-BDE-90), and 5'-hydroxy-2,2',4,4',5-pentabromodiphenyl ether (5'-OH-BDE-99). By comparison, the major metabolites produced by liver microsomes from Japanese quail included 6-hydroxy-2,2',4,4'-tetrabromodiphenyl ether (6-OH-BDE-47) and 2-hydroxy-2',3,4,4',5-pentabromodiphenyl ether (2-OH-BDE-123), but not 3-OH-BDE-47. Unidentified metabolites consisted of monohydroxy-pentabromodiphenyl ethers, monohydroxy-tetrabromodiphenyl ethers and dihydroxy-tetrabromodiphenyl ethers. Another difference between the two species was that formation rates of BDE-47 and BDE-99 metabolites were greater with liver microsomes from male than female Japanese quail, but a sex difference was not observed with lesser snow geese.