Prevalence of Substance Abuse Among Trauma Patients in Rural West Virginia.

BACKGROUND:  Substance abuse poses considerable clinical, economic, and social challenges. West Virginia is hailed as the epicenter of the substance abuse in the United States, the prevalence and pattern of different trauma mechanisms in a rural context or in patients with different forms of substance abuse remain unclear. OBJECTIVE:  We performed the following analysis to understand the prevalence of substance abuse in patients with different trauma mechanisms in the rural setting with high substance abuse in the West Virginia. METHODS:  We performed a cross-sectional retrospective analysis of adult trauma patients (motor vehicle, fall, assault, firearm suicide, brawl/rape and machinery) hospitalized in two tertiary care hospitals in West Virginia between 2006 and 2016. We identified all patients who had a urine drug screen (UDS) test and extracted the data related to the substance and trauma. RESULTS:  Among 8734 patients screened using UDS, 5940 (68.1%) patients were tested positive for the substance. Opiates, alcohol, benzodiazepines, and cannabis were the four most common substances identified in trauma victims. In all instances, the prescribed drug was less than 20%. Fatal outcome was observed in 366 patients in the sample, with 44% (n=162) testing positive for UDS, 12% (n=45) testing positive for only alcohol, and 15% (n=56) testing positive for both alcohol and UDS. Regarding the trauma mechanism, the motor vehicle accident (MVA) was the most prominent with a clear association of substance abuse with fatal outcome. CONCLUSION:  The most prevalent trauma mechanism was a MVA, with a strong link between drug usage and mortality. Due to the high incidence of positive substance abuse screens, UDS tests may need to be more widely implemented in trauma in the West Virginia region. The findings of this study might help in establishing regional or national policies to reduce acute substance abuse.

Chemistry of aqueous silica nanoparticle surfaces and the mechanism of selective peptide adsorption.

Control over selective recognition of biomolecules on inorganic nanoparticles is a major challenge for the synthesis of new catalysts, functional carriers for therapeutics, and assembly of renewable biobased materials. We found low sequence similarity among sequences of peptides strongly attracted to amorphous silica nanoparticles of various size (15-450 nm) using combinatorial phage display methods. Characterization of the surface by acid base titrations and zeta potential measurements revealed that the acidity of the silica particles increased with larger particle size, corresponding to between 5% and 20% ionization of silanol groups at pH 7. The wide range of surface ionization results in the attraction of increasingly basic peptides to increasingly acidic nanoparticles, along with major changes in the aqueous interfacial layer as seen in molecular dynamics simulation. We identified the mechanism of peptide adsorption using binding assays, zeta potential measurements, IR spectra, and molecular simulations of the purified peptides (without phage) in contact with uniformly sized silica particles. Positively charged peptides are strongly attracted to anionic silica surfaces by ion pairing of protonated N-termini, Lys side chains, and Arg side chains with negatively charged siloxide groups. Further, attraction of the peptides to the surface involves hydrogen bonds between polar groups in the peptide with silanol and siloxide groups on the silica surface, as well as ion-dipole, dipole-dipole, and van-der-Waals interactions. Electrostatic attraction between peptides and particle surfaces is supported by neutralization of zeta potentials, an inverse correlation between the required peptide concentration for measurable adsorption and the peptide pI, and proximity of cationic groups to the surface in the computation. The importance of hydrogen bonds and polar interactions is supported by adsorption of noncationic peptides containing Ser, His, and Asp residues, including the formation of multilayers. We also demonstrate tuning of interfacial interactions using mutant peptides with an excellent correlation between adsorption measurements, zeta potentials, computed adsorption energies, and the proposed binding mechanism. Follow-on questions about the relation between peptide adsorption on silica nanoparticles and mineralization of silica from peptide-stabilized precursors are raised.

Rapid, multiplexed microfluidic phage display.

The development of a method for high-throughput, automated proteomic screening could impact areas ranging from fundamental molecular interactions to the discovery of novel disease markers and therapeutic targets. Surface display techniques allow for efficient handling of large molecular libraries in small volumes. In particular, phage display has emerged as a powerful technology for selecting peptides and proteins with enhanced, target-specific binding affinities. Yet, the process becomes cumbersome and time-consuming when multiple targets are involved. Here we demonstrate for the first time a microfluidic chip capable of identifying high affinity phage-displayed peptides for multiple targets in just a single round and without the need for bacterial infection. The chip is shown to be able to yield well-established control consensus sequences while simultaneously identifying new sequences for clinically important targets. Indeed, the confined parameters of the device allow not only for highly controlled assay conditions but also introduce a significant time-reduction to the phage display process. We anticipate that this easily-fabricated, disposable device has the potential to impact areas ranging from fundamental studies of protein, peptide, and molecular interactions, to applications such as fully automated proteomic screening.

Preferential binding of peptides to graphene edges and planes.

Peptides identified from combinatorial peptide libraries have been shown to bind to a variety of abiotic surfaces. Biotic-abiotic interactions can be exploited to create hybrid materials with interesting electronic, optical, or catalytic properties. Here we show that peptides identified from a combinatorial phage display peptide library assemble preferentially to the edge or planar surface of graphene and can affect the electronic properties of graphene. Molecular dynamics simulations and experiments provide insight into the mechanism of peptide binding to the graphene edge.

Chemical functionalization of graphene enabled by phage displayed peptides.

The development of a general approach for the nondestructive chemical and biological functionalization of graphene could expand opportunities for graphene in both fundamental studies and a variety of device platforms. Graphene is a delicate single-layer, two-dimensional network of carbon atoms whose properties can be affected by covalent modification. One method for functionalizing materials without fundamentally changing their inherent structure is using biorecognition moieties. In particular, oligopeptides are molecules containing a broad chemical diversity that can be achieved within a relatively compact size. Phage display is a dominant method for identifying peptides that possess enhanced selectivity toward a particular target. Here, we demonstrate a powerful yet benign approach for chemical functionalization of graphene via comprehensively screened phage displayed peptides. Our results show that graphene can be selectively recognized even in nanometer-defined strips. Further, modification of graphene with bifunctional peptides reveals both the ability to impart selective recognition of gold nanoparticles and the development of an ultrasensitive graphene-based TNT sensor. We anticipate that these results could open exciting opportunities in the use of graphene in fundamental biochemical recognition studies, as well as applications ranging from sensors to energy storage devices.

Bioassembled layered silicate-metal nanoparticle hybrids.

Here we report on the bioenabled assembly of layered nanohybrids using peptides identified with regard to their affinity to the nanoparticle surface. A dodecamer peptide termed M1, determined from a phage peptide display library, was found to bind to the surface of a layered aluminosilicate (montmorillonite, MMT). Fusion of a metal binding domain to the M1 peptide or the M1 peptide by itself was able to direct the growth of metal nanoparticles, such as gold and cobalt-platinum, respectively, on the MMT. This method of producing hybrid nanoclay materials will have utility in catalytic, optical, biomedical, and composite materials applications.

Biomimetic synthesis of Pd nanocatalysts for the Stille coupling reaction.

Here we report on the biomimetic synthesis of Pd nanoparticles for use as models of green catalytic systems. The nanomaterials are synthesized using peptides isolated via phage-display techniques that are specific to Pd surfaces. Using this synthetic strategy, peptide-functionalized Pd nanoparticles of 1.9 +/- 0.3 nm in diameter are produced, which are soluble and stable in aqueous solutions. Once characterized, these biobased materials were then used as catalysts to drive the formation of C-C bonds using the Stille coupling reaction. Under the conditions of an aqueous solvent at room temperature, quantitative product yields were achieved within 24.0 h employing catalyst loadings of > or = 0.005 mol % of Pd. Additionally, high TOF values of 3207 +/- 269 mol product x (mol Pd x h)(-1) have been determined for these materials. The catalytic reactivity was then examined over a set of substrates with substitutions for both functional group and halide substituents, demonstrating that the peptide-based Pd nanoparticles are reactive toward a variety of functionalities. Taken together, these bioinspired materials represent unique model systems for catalytic studies to elucidate ecologically friendly reactive species and conditions.

Rapid bioenabled formation of ferroelectric BaTiO3 at room temperature from an aqueous salt solution at near neutral pH.

A 12-mer peptide, identified through phage display biopanning, has been used for the first time to induce the rapid formation of ferroelectric (tetragonal) nanocrystalline BaTiO3 at room temperature from an aqueous salt precursor solution at near neutral pH. BaTiO3 is widely used in capacitors, thermistors, displays, and sensors owing to its attractive dielectric, ferroelectric, pyroelectric, optical, and electrochemical properties. Two 12-mer peptides (BT1 and BT2) were selected from a phage-displayed peptide library via binding to tetragonal BaTiO3 powder. While these peptides possessed various types of amino acids, 8 of the 12 amino acids were common to both peptides. Each of these peptides induced the formation of faceted nanoparticles (50-100 nm diameter) from an aqueous precursor solution. X-ray diffraction and selected area electron diffraction patterns obtained from these faceted nanoparticles were consistent with the BaTiO3 compound. Rietveld analyses of the X-ray diffraction patterns yielded good fits to tetragonal crystal structures, with the BaTiO3 formed in the presence of the BT2 peptide exhibiting the most tetragonal character. A coating of the latter BaTiO3 nanoparticles exhibited polarization hysteresis (a well-known characteristic of ferroelectric materials) at room temperature and a relative permittivity of 2200. Such rapid, peptide-induced precipitation at room temperature provides new opportunities for direct BaTiO3 formation on low-melting or reactive materials (e.g., plastics, cloths, bio-organics) and the low temperature integration of BaTiO3 into electronic devices (e.g., on silicon or flexible polymer substrates).

Characterization of microbial contamination in United States Air Force aviation fuel tanks.

Bacteria and fungi, isolated from United States Air Force (USAF) aviation fuel samples, were identified by gas chromatograph fatty acid methyl ester (GC-FAME) profiling and 16S or 18S rRNA gene sequencing. Thirty-six samples from 11 geographically separated USAF bases were collected. At each base, an above-ground storage tank, a refueling truck, and an aircraft wing tank were sampled at the lowest sample point, or sump, to investigate microbial diversity and dispersion within the fuel distribution chain. Twelve genera, including four Bacillus species and two Staphylococcus species, were isolated and identified. Bacillus licheniformis, the most prevalent organism isolated, was found at seven of the 11 bases. Of the organisms identified, Bacillus sp., Micrococcus luteus, Sphinogmonas sp., Staphylococcus sp., and the fungus Aureobasidium pullulans have previously been isolated from aviation fuel samples. The bacteria Pantoea ananatis, Arthrobacter sp., Alcaligenes sp., Kocuria rhizophilia, Leucobacter komagatae, Dietza sp., and the fungus Discophaerina fagi have not been previously reported in USAF aviation fuel. Only at two bases were the same organisms isolated from all three sample points in the fuel supply distribution chain. Isolation of previously undocumented organisms suggests either, changes in aviation fuel microbial community in response to changes in aviation fuel composition, additives and biocide use, or simply, improvements in isolation and identification techniques.

Biomolecular stress-sensitive gauges: surface-mediated immobilization of mechanosensitive membrane protein.

We report the observation of structural reorganizations associated with unique, stress-assisted gating of mechanosensitive (MscL) membrane protein on a silicon surface modified with alkyl-terminated monolayers. We observed that the shape of MscL membrane proteins changed dramatically depending upon the packing density of alkyl tails and the surface tension of the supporting organic layer. High-resolution atomic force microscopy confirmed a transition from an elongated, prolate shape of MscL molecules within a monolayer with low surface tension to a flattened, oblate shape with a wide central opening within a monolayer with high surface tension. These observations are consistent with the conformation reorganizations associated with the two-stage, "iris"-like expansion proposed for the gating of the MscL molecules.

Biomimetic synthesis and patterning of silver nanoparticles.

The creation of nanoscale materials for advanced structures has led to a growing interest in the area of biomineralization. Numerous microorganisms are capable of synthesizing inorganic-based structures. For example, diatoms use amorphous silica as a structural material, bacteria synthesize magnetite (Fe3O4) particles and form silver nanoparticles, and yeast cells synthesize cadmium sulphide nanoparticles. The process of biomineralization and assembly of nanostructured inorganic components into hierarchical structures has led to the development of a variety of approaches that mimic the recognition and nucleation capabilities found in biomolecules for inorganic material synthesis. In this report, we describe the in vitro biosynthesis of silver nanoparticles using silver-binding peptides identified from a combinatorial phage display peptide library.