Dihydrotanshinone,a Natural Diterpenoid,Preserves Blood-Retinal Barrier Integrity via P2X7 Receptor

Activation of P2X7 signaling, due to high glucose levels, leads to blood retinal barrier (BRB) breakdown, which is a hallmark of diabetic retinopathy (DR). Furthermore, several studies report that high glucose (HG) conditions and the related activation of the P2X7 receptor (P2X7R) lead to the over-expression of pro-inflammatory markers. In order to identify novel P2X7R antagonists, we carried out virtual screening on a focused compound dataset, including indole derivatives and natural compounds such as caffeic acid phenethyl ester derivatives, flavonoids, and diterpenoids. Molecular Mechanics/Generalized Born Surface Area (MM/GBSA) rescoring and structural fingerprint clustering of docking poses from virtual screening highlighted that the diterpenoid dihydrotanshinone (DHTS) clustered with the well-known P2X7R antagonist JNJ47965567. A human-based in vitro BRB model made of retinal pericytes, astrocytes, and endothelial cells was used to assess the potential protective effect of DHTS against HG and 2′(3′)-O-(4-Benzoylbenzoyl)adenosine-5′-triphosphate (BzATP), a P2X7R agonist, insult. We found that HG/BzATP exposure generated BRB breakdown by enhancing barrier permeability (trans-endothelial electrical resistance (TEER)) and reducing the levels of ZO-1 and VE-cadherin junction proteins as well as of the Cx-43 mRNA expression levels. Furthermore, HG levels and P2X7R agonist treatment led to increased expression of pro-inflammatory mediators (TLR-4, IL-1β, IL-6, TNF-α, and IL-8) and other molecular markers (P2X7R, VEGF-A, and ICAM-1), along with enhanced production of reactive oxygen species. Treatment with DHTS preserved the BRB integrity from HG/BzATP damage. The protective effects of DHTS were also compared to the validated P2X7R antagonist, JNJ47965567. In conclusion, we provided new findings pointing out the therapeutic potential of DHTS, which is an inhibitor of P2X7R, in terms of preventing and/or counteracting the BRB dysfunctions elicited by HG conditions.     

Wetting,infiltration and interaction behavior of CMAS towards columnar YSZ coatings deposited by plasma spray physical vapor

Thermal barrier coatings (TBCs) produced by electron beam physical vapor deposition (EB-PVD) or plasma spray (PS) usually suffer from molten calcium-magnesium-alumino-silicate (CMAS) attack. In this study, columnar structured YSZ coatings were fabricated by plasma spray physical vapor deposition (PS-PVD). The coatings were CMAS-infiltrated at 1250?°C for short terms (1, 5, 30?min). The wetting and spreading dynamics of CMAS melt on the coating surface was in-situ investigated using a heating microscope. The results indicate that the spreading evolution of CMAS melt can be described in terms of two stages with varied time intervals and spreading velocities. Besides, the PS-PVD columnar coating (～100?μm thick) was fully penetrated by CMAS melt within 1?min. After the CMAS attack for 30?min, the original feathered-YSZ grains (tetragonal phase) in both PS-PVD and EB-PVD coatings were replaced by globular shaped monoclinic ZrO₂ grains in the interaction regions.     

Improved method for roadside barrier length of need modeling using real-world trajectories

The 2011 AASHTO Roadside Design Guide (RDG) contains perhaps the most widely used procedure for choosing an appropriate length of need (LON) for roadside barriers. However, this procedure has several limitations. The procedure uses a highly simplified model of vehicle departure, and the procedure does not allow designers to specify an explicit level of protection. A new procedure for choosing LON that addresses these limitations is presented in this paper. This new procedure is based on recent, real-world road departure trajectories and uses this departure data in a more realistic way. The new procedure also allows LON to be specified for a precisely known level of protection – a level which can be based on number of crashes, injury outcomes or even estimated crash cost – while still remaining straightforward and quick to use like the 2011 RDG procedure.     

Polymer derived ceramic materials from Si,B and MoSiB filler-loaded perhydropolysilazane precursor for oxidation protection

Silica-based coating systems were developed using polymer derived ceramics (PDCs) technology. Ceramic composites on the base of a SiO₂ and SiNO matrix and homogeneously distributed Mo₅SiB₂, SiB₆, Si and B fillers were manufactured. The coating systems have low porosity and provide a high oxidation resistance up to 100 h at 800 °C and 1100 °C in air. The influence of temperature and atmosphere of pyrolysis on the polymer precursor, the volume fraction of filler materials on the chemical composition of compacts as well as their high-temperature oxidation protection was investigated.     

Sofosbuvir Selects for Drug-Resistant Amino Acid Variants in the Zika Virus RNA-Dependent RNA-Polymerase Complex In Vitro

The nucleotide analog sofosbuvir, licensed for the treatment of hepatitis C, recently revealed activity against the Zika virus (ZIKV) in vitro and in animal models. However, the ZIKV genetic barrier to sofosbuvir has not yet been characterized. In this study, in vitro selection experiments were performed in infected human hepatoma cell lines. Increasing drug pressure significantly delayed viral breakthrough (p = 0.029). A double mutant in the NS5 gene (V360L/V607I) emerged in 3 independent experiments at 40–80 µM sofosbuvir resulting in a 3.9 ± 0.9-fold half- maximal inhibitory concentration (IC₅₀) shift with respect to the wild type (WT) virus. A triple mutant (C269Y/V360L/V607I), detected in one experiment at 80 µM, conferred a 6.8-fold IC₅₀ shift with respect to the WT. Molecular dynamics simulations confirmed that the double mutant V360L/V607I impacts the binding mode of sofosbuvir, supporting its role in sofosbuvir resistance. Due to the distance from the catalytic site and to the lack of reliable structural data, the contribution of C269Y was not investigated in silico. By a combination of sequence analysis, phenotypic susceptibility testing, and molecular modeling, we characterized a double ZIKV NS5 mutant with decreased sofosbuvir susceptibility. These data add important information to the profile of sofosbuvir as a possible lead for anti-ZIKV drug development.     

The Critical Role of Passive Permeability in Designing Successful Drugs

Passive permeability is a key property in drug disposition and delivery. It is critical for gastrointestinal absorption, brain penetration, renal reabsorption, defining clearance mechanisms and drug-drug interactions. Passive diffusion rate is translatable across tissues and animal species, while the extent of absorption is dependent on drug properties, as well as in vivo physiology/pathophysiology. Design principles have been developed to guide medicinal chemistry to enhance absorption, which combine the balance of aqueous solubility, permeability and the sometimes unfavorable compound characteristic demanded by the target. Permeability assays have been implemented that enable rapid development of structure-permeability relationships for absorption improvement. Future advances in assay development to reduce nonspecific binding and improve mass balance will enable more accurately measurement of passive permeability. Design principles that integrate potency, selectivity, passive permeability and other ADMET properties facilitate rapid advancement of successful drug candidates to patients.     

Evolution of internal cracks and residual stress during deposition of TBC

Thermal barrier coating (TBCs) are ceramic coatings that are deposited on metallic substrates to provide high thermal resistance. Residual stress is among the critical factors that affect the performance of TBCs. It evolves during the process of coating deposition and in-service loading. High residual stresses result in significant cracking and premature delamination of the TBC layer. In the present study, a hybrid computational approach is used to predict the evolution of internal cracks and residual stress in TBC. Smooth particle hydrodynamics (SPH) is first used to model the deposition of yttria-stabilized zirconia (YSZ) layer that contains various interfaces and micropores on a steel substrate. Then, three-dimensional (3D) finite element analysis is utilized to predict the evolution of internal cracks and residual stress in the ceramic coating layer. It is found that multiple cracks emerge during the solidification of the coating layer due to the development of high tensile (quenching) stresses. The cracking density is higher at regions near the coating interface. It is also found that compressive (residual) stresses are developed when the deposited coating is cooled to room temperature. The residual stress state is equibiaxial and nonlinear across the thickness/width of the TBC layer. The residual stress profile predicted compares well with that of hole drilling experiments.     

Fabrication of Schottky barrier MOSFETs using self-assembly CoSi2 nanopatterning and spacer gate technologies

A self-assembly patterning method for generation of epitaxial CoSi₂ nanostructures was used to fabricate 50 nm channel-length MOSFETs. The transistors have either a symmetric structure with Schottky source and drain or an asymmetric structure with n⁺-source and Schottky drain. The patterning technique is based on anisotropic diffusion of Co/Si atoms in a strain field during rapid thermal oxidation. The strain field is generated along the edges of a mask consisting of 20 nm SiO₂ and 300 nm Si₃N₄. During rapid thermal oxinitridation (RTON) of the masked silicide structure, a well-defined separation of the silicide layer forms along the edge of the mask. These highly uniform gaps define the channel region of the fabricated device. The separated silicide layers act as metal source and drain. A poly-Si spacer was used as the gate contact. The asymmetric transistor was fabricated by ion implantation into the unprotected CoSi₂ layer and a subsequent out-diffusion process to form the n⁺-source. I–V characteristics of both the symmetric and asymmetric transistor structures have been investigated.     

Computational analysis of thin coating layer failure using a cohesive model and gradient plasticity

Thermal barrier coatings (TBC) are widely used to prevent transient high temperature attack and allow components high durability. Due to strong inhomogeneous material properties the TBC failure often initiates near the interface between the brittle oxide layer and the ductile substrate. A reliable prediction of the TBC failure requires detailed information about the crack tip field and the consequent fracture criteria. In the present paper both cohesive model and gradient plasticity are used to simulate the failure process and to study interdependence of the interface stress distribution with the specific fracture energies. Computations confirm that combination of the two models is able to simulate different failure mechanisms in the TBC system. The computational model has the potential to give a realistic prediction of the crack propagation process.