Arginine 125 Is an Essential Residue for the Function of MRAP2

MRAP2 is a small simple transmembrane protein arranged in a double antiparallel topology on the plasma membrane. It is expressed in the paraventricular nucleus of the hypothalamus, where it interacts with various G protein-coupled receptors, such as the prokineticin receptors, and regulates energy expenditure and appetite. The aim of this work was to analyze the functional role of the specific arginine residue at position 125 of MRAP2, which affects protein conformation, dimer formation, and PKR2 binding. Results obtained with the MRAP2 mutants R125H and R125C, which are found in human patients with extreme obesity, and mouse MRAP2, in which arginine 125 is normally replaced by histidine, were compared with those obtained with human MRAP2. Understanding the mechanism by which MRAP2 regulates G protein-coupled receptors helps in elucidating the metabolic pathways involved in metabolic dysfunction and in developing new drugs as specific targets of the MRAP2-PKR2 complex.     

Advanced ultra‐high molecular weight polyethylene/antioxidant‐functionalized carbon nanotubes nanocomposites with improved thermo‐oxidative resistance

Multiwalled carbon nanotubes (CNTs) functionalized with hindered phenol moieties are dispersed in ultra‐high molecular weight polyethylene (UHMWPE), and the stabilizing action of the antioxidant (AO) functionalized CNTs (AO‐f‐CNTs) is studied through a combination of rheological and spectroscopic (FT‐IR) analyses. The effectiveness of two alternative compounding methods, namely hot compaction (HC) and melt mixing (MM), is compared. The combination of high temperature and mechanical stress experienced during MM brings about noticeable degradation phenomena of the matrix already in the course of the compounding step. Differently, the milder conditions of the HC process preserve the stability of the polymer, making this method preferable when dealing with highly viscous matrices. In addition, HC guarantees a better CNT dispersion, allowing for the maximization of the stabilizing action of the AO grafted on the nanotubes. As a result, the HC samples exhibit improved thermo‐oxidative resistance despite the very low amount of AO grafted onto the CNTs. Besides demonstrating the effectiveness of our AO‐f‐CNTs as stabilizers for polymer matrices, our results prove that CNTs can serve as a support on which grafting specific functional molecules to be dispersed in a host polymer matrix. © 2015 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2015 , 132, 42420.     

Variability in Immunohistochemical Detection of Programmed Death Ligand 1 (PD-L1) in Cancer Tissue Types

In normal cell physiology, programmed death 1 (PD-1) and its ligand, PD-L1, play an immunoregulatory role in T-cell activation, tolerance, and immune-mediated tissue damage. The PD-1/PD-L1 pathway also plays a critical role in immune escape of tumor cells and has been demonstrated to correlate with a poor prognosis of patients with several types of cancer. However, recent reports have revealed that the immunohistochemical (IHC) expression of the PD-L1 in tumor cells is not uniform for the use of different antibodies clones, with variable specificity, often doubtful topographical localization, and with a score not uniquely defined. The purpose of this study was to analyze the IHC expression of PD-L1 on a large series of several human tumors to correctly define its staining in different tumor tissues.     

How sensory sensitivity to graininess could be measured?

Considering the lack of the literature and the need for developing a valid method to measure the texture sensitivity, in this study, we investigated the individual sensitivity to discriminate among different levels of graininess. With this purpose, five samples of cocoa-based creams were prepared, by changing the refining time. Samples were first characterized in terms of particles size distribution, by means of laser diffraction. Then, 59 subjects evaluated the cream sample graininess intensity, by using gLM scales, and their liking, by using LAM scales. The graininess scores of each subject were fitted with a power model, already observed with the instrumental results, estimating both the power law exponent and the R² coefficient, and using them as clustering parameters. Subjects were then clustered into three groups: high sensitivity; moderate sensitivity; low sensitivity. First, as it was hypothesized, results showed a significant difference between the three groups in terms of perceived graininess. Second, even though results showed a significant difference between the three groups in terms of perceived graininess, only little differences were found in terms of liking scores. Indeed, all the samples were equally liked for both the moderate and low sensitivity groups, whereas a significant trend was observed for the highly sensitive subjects who liked more the most refined samples. No significant relationships were found with age, and only a little trend was observed with gender: females seemed to be more sensitive than males. Texture attributes discrimination ability, as for example sensitivity to graininess, could affect food rejection or preferences and choice, but it is poorly investigated. This exploratory study, proposes a method to cluster consumers, based on their sensitivity. A statistical methodology has been developed to discriminate among consumer sensitivity levels. The results provide useful information about graininess sensitivity suggesting that the used methodologies could be applied to other texture properties resulting in a valid tool for the industry in the development and optimization of tailored new products.     

Biocompatible polymeric electrospun matrices: Micro–nanotopography effect on cell behavior

This work reports a preliminary biological study performed on nanofibrous biocompatible polylactidecopolycaprolactone (PLA-PCL) scaffolds intended for tissue regeneration. The aim was to evaluate how matrix surface topography affects cell adhesion and proliferation. Scaffolds prepared by electrospinning either equipped with plane or rotating mandrel collectors, were characterized for their surface topography and nanofiber size. Cell culture studies were carried out using mouse embryonic fibroblast cells lines (NIH-3T3), as model for skin, murine neuroblastoma neuro-2α cell line, as model for neuronal tissue, and mouse mesenchymal stem cells (MSCs), because of their differentiation ability. Imaging analysis by scanning electron microscope and laser scanning confocal microscopy together with cell viability (MTT, L 3-(4,5-dymethiltiazol-2-y)-2,5 diphenyltetrazolium bromide) test, were performed on cell cultures at fixed time laps. The results showed that electrospun nanofibers supported growth and proliferation of the tested cell lines, but electrospun matrices obtained with rotating mandrel showed significantly higher cell viability that follows the orientation of electrospun nanofibers.     

Reorganization of Thalamic Inputs to Lesioned Cortex Following Experimental Traumatic Brain Injury

Traumatic brain injury (TBI) disrupts thalamic and cortical integrity. The effect of post-injury reorganization and plasticity in thalamocortical pathways on the functional outcome remains unclear. We evaluated whether TBI causes structural changes in the thalamocortical axonal projection terminals in the primary somatosensory cortex (S1) that lead to hyperexcitability. TBI was induced in adult male Sprague Dawley rats with lateral fluid-percussion injury. A virus carrying the fluorescent-tagged opsin channel rhodopsin 2 transgene was injected into the ventroposterior thalamus. We then traced the thalamocortical pathways and analyzed the reorganization of their axonal terminals in S1. Next, we optogenetically stimulated the thalamocortical relays from the ventral posterior lateral and medial nuclei to assess the post-TBI functionality of the pathway. Immunohistochemical analysis revealed that TBI did not alter the spatial distribution or lamina-specific targeting of projection terminals in S1. TBI reduced the axon terminal density in the motor cortex by 44% and in S1 by 30%. A nematic tensor-based analysis revealed that in control rats, the axon terminals in layer V were orientated perpendicular to the pial surface (60.3°). In TBI rats their orientation was more parallel to the pial surface (5.43°, difference between the groups p < 0.05). Moreover, the level of anisotropy of the axon terminals was high in controls (0.063) compared with TBI rats (0.045, p < 0.05). Optical stimulation of the sensory thalamus increased alpha activity in electroencephalography by 312% in controls (p > 0.05) and 237% (p > 0.05) in TBI rats compared with the baseline. However, only TBI rats showed increased beta activity (33%) with harmonics at 5 Hz. Our findings indicate that TBI induces reorganization of thalamocortical axonal terminals in the perilesional cortex, which alters responses to thalamic stimulation.     

Photosensitive acrylates containing bio-based epoxy-acrylate soybean oil for 3D printing application

Stereolithography is a 3D-printing process that is rapidly shifting from being an expensive and limited technology to an affordable, precise, and fast method of mass production. However, most of the current resins are petroleum-based, which makes them toxic, non-degradable and with poor biocompatibility. In this study, a standard petroleum-based resin containing urethane acrylate and acrylic monomers was combined with epoxy-acrylate soybean oil (EASO), aiming to reduce its impact on the environment. Ratios varying from 10 to 50 wt% of EASO were incorporated into the commercial resin while maintaining the viscosity low, between 0.27 and 1.06 Pa s. The printed samples showed good quality and complete integration between the layers. The addition of 50 wt% of EASO increased the samples elongation at break by 108% (from 2.3% to 4.8%) and decreased the contact angle by 26.4% (from 72 to 53°). Moreover, the mixture showed good thermal and swelling stability and tensile strength in the range of other commercial cured systems. The addition of EASO may significantly contribute to the exploitation of greener materials, which well matches today's circular economy concept.     

Exploring free-energy profiles through ion channels: Comparison on a test case

The calculation of free-energy profiles in proteins, and, more specifically, in ion channels, is a challenge for modern numerical simulations due both to the convergence problems associated with the electrostatics of the environment and to the difficulties in modeling the fields acting on the permeating ions. The present study is aims at comparing three different simulation techniques available in the literature on a nanometric channel protein chosen as a test case, with the purpose of establishing their real predictivity and limits.     

Total reducing capacity of fresh sweet peppers and five different Italian pepper recipes

Processing and preparation methods are generally believed to result in a depletion of naturally occurring antioxidants in food. To evaluate the antioxidant properties of fresh sweet peppers (Capsicum annuum) and five different Italian recipes based on sweet peppers (pickled; “peperonata”; grilled; in sour–sweet condiment; salted), water- and lipid-soluble extracts from fresh and processed peppers were analysed using high-performance liquid chromatography coupled with an electrochemical detector. Total reducing capacity (TRC) and contributions of hydrophilic reducing capacity (HRC) and lipophilic reducing capacity (LRC) to the TRC were determined in all the samples. Three important antioxidant compounds were measured: ascorbic acid, β-carotene and lycopene. The contribution of these individual compounds to TRC was estimated.