Experimental and Numerical Studies of Aluminum-Alumina Composites

The preliminary goal of this study is to determine the effects of processing conditions, compositions and microstructural morphologies of the constituents on the physical and thermo-mechanical properties of alumina (Al_2O_3) reinforced aluminum (Al) composites. Composites with 0, 5, 10, 20 and 25 vol% Al_2O_3 were manufactured using powder metallurgy method. The elastic properties (Young's and shear modulus) and the coefficient of thermal expansion (CTE) of the composites were determined using Resonant Ultrasound Spectroscopy (RUS) and Thermo Mechanical Analyzer (TMA) respectively at various temperatures. Increasing compacting pressure improved relative density (or lowered porosity) of the composites. Furthermore, increasing the Al_2O_3 vol% in the composite increased the elastic moduli and reduced the CTE of the composites. Increasing the testing temperature from 25 to 450 oC, significantly reduced the elastic moduli of the composites, while the CTE of the composites changed only slightly with temperatures. Secondly, the goal of this study is to determine the effect of microstructures on the effective thermo-mechanical properties of the manufactured Al-Al_2O_3 composites using finite element (FE) method. Software OOF was used to convert the SEM micrographs of the manufactured composites to FE meshed models, which were then used to determine the effective elastic modulus and CTE. It was observed that, effective modulus dropped by 19.7% when porosity increased by 2.3%; while the effective CTE was mildly affected by the porosity. Additionally, the effect of residual stress on the effective thermo-mechanical properties was studied, and the stress free temperature of the composites was determined. Another objective of this study is to examine the stress-strain response of Al-Al_2O_3 composites due to compressive loads at various temperatures. Elastic modulus, yield stress and strain hardening parameters were determined from the stress-strain curves and their dependency on temperature, porosity and volume fraction were studied. The experimental results were compared with the numerical results. It was observed that high-localized stresses were present near the pores and at the interfaces between Al and Al_2O_3 constituents. Finally, functionally graded materials (FGMs) with varying Al_2O_3 concentration (0, 5and 10 vol%) in Al were manufactured; and their stress-strain response and CTE were determined at various temperatures

Thermoelastic Properties of Particle Reinforced Composites at the Micro and Macro Scales

Particle reinforced composites are widely used in tires, heat exchangers, thermal barrier coatings and many other applications, as they have good strength to weight ratio, excellent thermal insulation, ease of manufacturing and flexibility in design. During their service life, these composites are often subjected to harsh environments, which can degrade the thermo-mechanical properties of the constituents in the composites, affecting performance and lifetime of the composites. This study investigates performance of particle reinforced composites subjected to coupled heat conduction and thermo-elastic deformation at the macro and micro levels. A micromechanical model is used to determine the effective thermal and mechanical properties of the homogenized composite by incorporating microscopic characteristics of the composites. The constituent?s thermal conductivities of the composite are assumed to be functions of temperature and the elastic moduli to be functions of temperature and stress fields. The effective properties obtained from the micromechanical model represent average (macroscopic) properties. The effective heat conduction and thermo-elastic responses in the homogenized composites are compared with the responses of the composite with particles randomly distributed in the matrix (heterogeneous materials) which represent microscopic responses. For this purpose, two sets of finite element (FE) models are generated for composites with particle volume contents 12.5, 25, and 50%. The first FE model represents a homogenized composite panel and the effective responses from the micromechanical model are used as input for the material properties. The second FE model mimics composite microstructure with discontinuous particles randomly dispersed in a homogeneous matrix. Parametric studies on effects of conductivity ratio between particle and matrix, degree of nonlinearity, and volume fraction on the temperature distribution and steady state times have been studied. For lower volume fractions the temperature profiles of homogenized and heterogeneous composite models are in good agreement with each other. But for higher volume fractions, the detailed model showed a wavy profile whereas the effective model showed no signs of it. When the nonlinearity in thermal conductivity of the particle and matrix constituents is increased, the steady state time significantly deviates from the ones with constant constituent properties. When the volume fraction of particles in the composite increases, the steady state is reached in less time, since the thermal conductivity of particles are taken larger than that of the matrix. Effects of coefficient of thermal expansion (CTE) ratio of particle and matrix, temperature change, and volume fraction on the discontinuity of stress and strain fields at the interphase of matrix and particle have been studied. The stresses developed were more for higher CTE ratios and the magnitude of discontinuity also follows the same trend. As the volume fraction increases, the stresses developed and the magnitude of discontinuity also increase. Finally, sequentially coupled heat conduction and deformation analyses are performed on thermal barrier coating (TBC) systems to demonstrate the applicability of the micromechanical model in predicting overall thermo-elastic responses of the TBC

Genome sequence and rapid evolution of the rice pathogen Xanthomonas oryzae pv. oryzae PXO99A

Background: Xanthomonas oryzae pv. oryzae causes bacterial blight of rice (Oryza sativa L.), a major disease that constrains production of this staple crop in many parts of the world. We report here on the complete genome sequence of strain PXO99A and its comparison to two previously sequenced strains, KACC10331 and MAFF311018, which are highly similar to one another. Results: The PXO99 A genome is a single circular chromosome of 5,240,075 bp, considerably longer than the genomes of the other strains (4,941,439 bp and 4,940,217 bp, respectively), and it contains 5083 protein-coding genes, including 87 not found in KACC10331 or MAFF311018. PXO99A contains a greater number of virulence-associated transcription activator-like effector genes and has at least ten major chromosomal rearrangements relative to KACC10331 and MAFF311018. PXO99 A contains numerous copies of diverse insertion sequence elements, members of which are associated with 7 out of 10 of the major rearrangements. A rapidly-evolving CRISPR (clustered regularly interspersed short palindromic repeats) region contains evidence of dozens of phage infections unique to the PXO99A lineage. PXO99A also contains a unique, near-perfect tandem repeat of 212 kilobases close to the replication terminus. Conclusion: Our results provide striking evidence of genome plasticity and rapid evolution within Xanthomonas oryzae pv. oryzae. The comparisons point to sources of genomic variation and candidates for strain-specific adaptations of this pathogen that help to explain the extraordinary diversity of Xanthomonas oryzae pv. oryzae genotypes and races that have been isolated from around the world. © 2008 Salzberg et al; licensee BioMed Central Ltd

Alternative splicing converts STIM2 from an activator to an inhibitor of store-operated calcium channels

Store-operated calcium entry (SOCE) regulates a wide variety of essential cellular functions. SOCE is mediated by STIM1 and STIM2, which sense depletion of ER Ca2+ stores and activate Orai channels in the plasma membrane. Although the amplitude and dynamics of SOCE are considered important determinants of Ca2+-dependent responses, the underlying modulatory mechanisms are unclear. In this paper, we identify STIM2??, a highly conserved alternatively spliced isoform of STIM2, which, in contrast to all known STIM isoforms, is a potent inhibitor of SOCE. Although STIM2?? does not by itself strongly bind Orai1, it is recruited to Orai1 channels by forming heterodimers with other STIM isoforms. Analysis of STIM2?? mutants and Orai1-STIM2?? chimeras suggested that it actively inhibits SOCE through a sequence-specific allosteric interaction with Orai1. Our results reveal a previously unrecognized functional flexibility in the STIM protein family by which alternative splicing creates negative and positive regulators of SOCE to shape the amplitude and dynamics of Ca2+ signals.open

TMEM110 regulates the maintenance and remodeling of mammalian ER–plasma membrane junctions competent for STIM–ORAI signaling

The stromal interaction molecule (STIM)–ORAI calcium release-activated calcium modulator (ORAI) pathway controls store-dependent calcium entry, a major mechanism of physiological calcium signaling in mammalian cells. The core elements of the pathway are the regulatory protein STIM1, located in the endoplasmic reticulum (ER) membrane, the calcium channel ORAI1 in the plasma membrane, and sites of close contact between the ER and the plasma membrane that permit the two proteins to interact. Research on calcium signaling has centered on STIM1, ORAI1, and a few proteins that directly modulate STIM–ORAI function. However, little is known about proteins that organize ER–plasma membrane junctions for STIM–ORAI-dependent calcium signaling. Here, we report that an ER-resident membrane protein identified in a previous genome-wide RNAi screen, transmembrane protein 110 (TMEM110), regulates the long-term maintenance of ER–plasma membrane junctions and the short-term physiological remodeling of the junctions during store-dependent calcium signaling

pH-dependent interactions of coacervate-forming histidine-rich peptide with model lipid membranes

Peptide-based liquid droplets (coacervates) produced by spontaneous liquid-liquid phase separation (LLPS), have emerged as a promising class of drug delivery systems due to their high entrapping efficiency and the simplicity of their formulation. However, the detailed mechanisms governing their interaction with cell membranes and cellular uptake remain poorly understood. In this study, we investigated the interactions of peptide coacervates composed of HBpep—peptide derived from the histidine-rich beak proteins (HBPs) of the Humboldt squid—with model cellular membranes in the form of supported lipid bilayers (SLBs). We employed quartz crystal microbalance with dissipation monitoring (QCM-D), neutron reflectometry (NR) and atomistic molecular dynamics (MD) simulations to reveal the nature of these interactions in the absence of fluorescent labels or tags. HBpep forms small oligomers at pH 6 whereas it forms µm-sized coacervates at physiological pH. Our findings reveal that both HBpep oligomers and HBpep-coacervates adsorb onto SLBs at pH 6 and 7.4, respectively. At pH 6, when the peptide carries a net positive charge, HBpep oligomers insert into the SLB, facilitated by the peptide’s interactions with the charged lipids and cholesterol. Importantly, however, HBpep coacervate adsorption at physiological pH, when it is largely uncharged, is fully reversible, suggesting no significant lipid bilayer rearrangement. HBpep coacervates, previously identified as efficient drug delivery vehicles, do not interact with the lipid membrane in the same manner as traditional cationic drug delivery systems or cell-penetrating peptides. Based on our findings, HBpep coacervates at physiological pH cannot cross the cell membrane by a simple passive mechanism and are thus likely to adopt a non-canonical cell entry pathway

Development of Brain Targeting Peptide Based MMP-9 Inhibiting Nanoparticles for the Treatment of Brain Diseases with Elevated MMP-9 Activity.

Latent and active levels of cerebral matrix metalloproteinase 9 (MMP-9) are elevated in neurological diseases and brain injuries, contributing to neurological damage and poor clinical outcomes. This study aimed developing peptide-based nanoparticles with ability to cross the blood-brain-barrier (BBB) and inhibit MMP-9. Three amphiphilic peptides were synthesised containing brain-targeting ligands (HAIYPRH or CKAPETALC) conjugated with MMP-9 inhibiting peptide (CTTHWGFTLC) linked by glycine (spacer) at the N-terminus, and the peptide sequences were conjugated at the N- terminus to cholesterol. 19F NMR assay was developed to measure MMP-9 inhibition. Cell toxicity was evaluated by the LDH assay, and dialysis studies were conducted with/without fetal bovine serum. An in vitro model was employed to evaluate the ability of nanoparticles crossing the BBB. The amphiphilic peptide (Cholesterol-GGGCTTHWGFTLCHAIYPRH) formed nanoparticles (average size of 202.8 nm) with ability to cross the BBB model. MMP-9 inhibiting nanoparticles were non-toxic to cells, and reduced MMP-9 activity from kobs of 4.5 × 10-6s-1 to complete inhibition. Dialysis studies showed that nanoparticles did not disassemble by extreme dilution (40 folds), but gradually hydrolysed by serum enzymes. In conclusion, the MMP-9 inhibiting nanoparticles reduced the activity of MMP-9, with acceptable serum stability, minimal cell toxicity and ability to cross the in vitro BBB model

Peptide nanovesicles formed by the self-assembly of branched amphiphilic peptides

Citation: Gudlur S, Sukthankar P, Gao J, Avila LA, Hiromasa Y, Chen J, et al. (2012) Peptide Nanovesicles Formed by the Self-Assembly of Branched Amphiphilic Peptides. PLoS ONE 7(9): e45374. https://doi.org/10.1371/journal.pone.0045374Peptide-based packaging systems show great potential as safer drug delivery systems. They overcome problems associated with lipid-based or viral delivery systems, vis-a-vis stability, specificity, inflammation, antigenicity, and tune-ability. Here, we describe a set of 15 & 23-residue branched, amphiphilic peptides that mimic phosphoglycerides in molecular architecture. These peptides undergo supramolecular self-assembly and form solvent-filled, bilayer delimited spheres with 50–200 nm diameters as confirmed by TEM, STEM and DLS. Whereas weak hydrophobic forces drive and sustain lipid bilayer assemblies, these all-peptide structures are stabilized potentially by both hydrophobic interactions and hydrogen bonds and remain intact at low micromolar concentrations and higher temperatures. A linear peptide lacking the branch point showed no self-assembly properties. We have observed that these peptide vesicles can trap fluorescent dye molecules within their interior and are taken up by N/N 1003A rabbit lens epithelial cells grown in culture. These assemblies are thus potential drug delivery systems that can overcome some of the key limitations of the current packaging systems

Branched oligopeptides form nano-capsules with lipid vesicle characteristics

In a recent article (Gudlur et al. PLOS ONE, 2012, 7 (9) e45374), we described the special properties of a mixed branched peptide assembly in which equimolar bis(FLIVI)-K-KKKK and bis(FLIVIGSII)-K-KKKK self-associate to form bilayer delimited capsules capable of trapping solutes. These polycationic vesicle-like capsules are readily taken up by epithelial cells in culture, escape or evade the endocytic pathway, and accumulate in the perinuclear region where they persist without any apparent degradation. In this report, we examine the lipidlike properties of this system including initial assembly; solute encapsulation and washing; fusion and resizing by membrane extrusion through polycarbonate filters with defined pore sizes. The resized peptide capsules have uniform diameters in nm size ranges. Once resized, the capsules can be maintained at the new size by storing them at 4 °C. Having the ability to prepare stable uniform nanoscale capsules of desired sizes makes them potentially attractive as biocompatible delivery vehicles for various solutes/drugs

Peptide nanovesicles: supramolecular assembly of branched amphiphilic peptides

Doctor of PhilosophyDepartment of BiochemistryJohn M. TomichPeptide-based delivery systems show great potential as safer drug delivery vehicles. They overcome problems associated with lipid-based or viral delivery systems, vis-a-vis stability, specificity, inflammation, antigenicity, and tune-ability. We have designed and synthesized a set of 15 and 23-residue branched, amphiphilic peptides that mimic phosphoglycerides in molecular architecture. They undergo supramolecular self-assembly and form solvent-filled, bilayer delineated spheres with 50-150 nm diameters (confirmed by TEM and DLS). Whereas weak hydrophobic forces drive and sustain lipid bilayer assemblies, these structures are further stabilized by β-sheet hydrogen bonding and are stable at very low concentrations and even in the presence of SDS, urea and trypsin as confirmed by circular dichroism spectroscopy. Given sufficient time, they fuse together to form larger assemblies and trap compounds of different sizes within the enclosed space. They are prepared using a protocol that is similar to preparing lipid vesicles. We have shown that different concentrations of the fluorescent dye, 5(6)-Carboxyfluorescein can be encapsulated in these assemblies and delivered into human lens epithelial cells and MCF-7 cells grown on coverslips. Besides fluorescent dyes, we have delivered the plasmid (EGFP-N3, 4.7kb) into N/N 1003A lens epithelial cells and observed expression of EGFP (in the presence and absence of a selection media). In the case of large molecules like DNA, these assemblies act as nanoparticles and offer some protection to DNA against certain nucleases. Linear peptides that lacked a branching point and other branched peptides with their sequences randomized did not show any of the lipid-like properties exhibited by the branched peptides. The peptides can be chemically decorated with target specific sequences for use as DDS for targeted delivery