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Analgesia in burn patients is challenging given the complexity of burn pain and prolonged need beyond hospital admission. Given the risks of opioids, the impact of multimodal analgesia postdischarge needs to be further elucidated in this population. This retrospective, single-center cohort study evaluated adult burn patients who were consecutively admitted to the burn service with at least 10% total body surface area burned and subsequently followed in the burn clinic between February 2015 and September 2018. Subjects were separated into two cohorts based on discharge pain regimens: multimodal and nonmultimodal. The primary outcome was the change in opioid requirements (measured in oral morphine equivalents) between discharge and first follow-up interval. Secondary outcomes included the classes of multimodal agents utilized and a comparison of opioid requirements between the last 24 hours of admission and discharge. A total of 152 patients were included for analysis, 76 in the multimodal cohort and 76 in the nonmultimodal cohort. The multimodal cohort was noted to have increased total body surface area burned and prolonged number of days spent in the intensive care unit at baseline; however, the multimodal cohort exhibited a more significant decrease in opioid requirements from discharge to first follow-up interval when compared with the nonmultimodal cohort (106.6 vs 75.4 mg, P = .039).

In June 2017, NASA published the Ice Giants Pre-Decadal Survey Mission Study Report which took a fresh look at science priorities and mission concepts for missions to the Uranus and Neptune systems. In addition to science objectives, the team explored the state of required technologies for remote and in-situ science exploration. Notably, three of the four mission architectures considered in the study included an atmospheric probe. More recently, interest has grown within ESA for outer planet exploration. In support of this objective, ESA has performed two CFD studies (January & July 2019) which analyzed the feasibility of stand-alone elements (orbiter and probes) provided by ESA as a part of a NASA led mission to the Uranus or Neptune systems. The first study was carried out by ESA experts with active participation of NASA/JPL. ESA highlighted the necessity to deepen the knowledge characterizing the aerothermal environment of the probes. Entry environments for the NASA study were estimated using an aeroheating correlation that was calibrated to data returned from the Galileo probe entry to Jupiter. For the ESA concept study, aeroheating estimates were made using correlations employed during the design of the Galileo probe. Importantly, these correlations show large discrepancies in predicted total aeroheating (in some cases more than 100%), largely due to differences in the predicted radiative heat load. The magnitude of the disagreement is disconcerting in and of itself, but the problem is made worse by the fact that both correlations are being extrapolated from the extreme Galileo entry conditions to the (relatively) more benign Uranus and Neptune entry. It is likely that neither correlation is providing an accurate assessment of the true aeroheating loads at this time. Given that current NASA predictions are near the limits of existing TPS test capability, and that ESA predictions are more severe, improving the accuracy and associated margins of the prediction is critical to better assess mission feasibility. Recent work in NASA by Cruden (AIAA Paper No. 2015-0380) and Erb (AIAA Paper No. 2019-3360) have substantially improved our fundamental understanding of aerothermodynamics in Hydrogen-Helium atmospheres. Similar work is planned in ESA as well. However, these recent data have not been incorporated into updated design models for Outer Planet probes. In addition, this work does not address the problem of trace atmospheric constituents (such as Methane) that are known to be present in Ice Giant atmospheres and may substantially alter the resulting shock layer radiation signal by providing a ready source of free electrons to initiate excitation processes. The proposed presentation will review the current status of aerothermal modeling for Ice Giant entries and propose a path forward to reduce key uncertainties and enable optimized thermal protection system designs.

Antimicrobial therapy is often necessary for the treatment of musculoskeletal infection in patients of advanced age. Consideration of potential changes in pharmacokinetics and pharmacodynamics of antimicrobials in patients as they age is necessary to determine the most appropriate antimicrobial at the correct dose. There is significant variability among physiologic changes as patients age; therefore, patients should be assessed individually for dose adjustments and risk of adverse effects. Changes in renal function as well as distribution and metabolism of medications in elderly patients are more likely to affect the adverse effect profile of antibiotics as opposed to the efficacy of these medications. Special caution should be exercised with the use of trimethoprim/sulfamethoxazole, fluoroquinolones, and aminoglycosides in the elderly population, and vancomycin dosing and trough achievement should be monitored. Beta-lactam antibiotics are generally considered safe and effective in elderly patients.

Defects in the cystic fibrosis transmembrane conductance regulator (CFTR) chloride channel lead to viscous secretions from submucosal glands that cannot be properly hydrated and cleared by beating cilia in cystic fibrosis (CF) airways. The mechanisms by which CFTR, and the predominant epithelial sodium channel (ENaC), control the hydration and clearance of glandular secretions remain unclear. We used a proteomics approach to characterize the proteins contained in CF and non-CF submucosal gland fluid droplets and found that differentially regulated proteases (cathepsin S and H) and their antiprotease (cystatin C) influenced the equilibration of fluid on the airway surface and tracheal mucociliary clearance (MCC). Contrary to prevailing models of airway hydration and clearance, cystatin C, or raising the airway surface liquid (ASL) pH, inhibited cathepsin-dependent ENaC-mediated fluid absorption and raised the height of ASL, and yet decreased MCC velocity. Importantly, coupling of both CFTR and ENaC activities were required for effective MCC and for effective ASL height equilibration after volume challenge. Cystatin C–inhibitable cathepsins controlled initial phases of ENaC-mediated fluid absorption, whereas CFTR activity was required to prevent ASL dehydration. Interestingly, CF airway epithelia absorbed fluid more slowly owing to reduced cysteine protease activity in the ASL but became abnormally dehydrated with time. Our findings demonstrate that, after volume challenge, pH-dependent protease-mediated coupling of CFTR and ENaC activities are required for rapid fluid equilibration at the airway surface and for effective MCC. These findings provide new insights into how glandular fluid secretions may be equilibrated at the airway surface and how this process may be impaired in CF.

Pacific Northwest National Laboratory has recently opened a shallow underground laboratory intended for measurement of low-concentration levels of radioactive isotopes in samples collected from the environment. The development of a low-background liquid scintillation counter is currently underway to further augment the measurement capabilities within this underground laboratory. Liquid scintillation counting is especially useful for measuring charged particle (e.g., $\beta$, $\alpha$) emitting isotopes with no (or very weak) gamma-ray yields. The combination of high-efficiency detection of charged particle emission in a liquid scintillation cocktail coupled with the low-background environment of an appropriately-designed shield located in a clean underground laboratory provides the opportunity for increased-sensitivity measurements of a range of isotopes. To take advantage of the 35 meters-water-equivalent overburden of the underground laboratory, a series of simulations have evaluated the scintillation counter's shield design requirements to assess the possible background rate achievable. This report presents the design and background evaluation for a shallow underground, low background liquid scintillation counter design for sample measurements.

Liquid scintillation counting (LSC) supports a range of environmental science measurements. At Pacific Northwest National Laboratory, we are constructing an LSC system with an expected background reduction of 10–100 relative to values reported in the literature. In this paper, a number of current measurement applications of LSC have been considered with an emphasis on determining which aspects of such measurements would gain the greatest benefit: improved minimum detectable activity (MDA), reduction in sample size, and reduction in total analysis time.

The aberrant expression of androgen receptor (AR)-dependent transcriptional programs is a defining pathology of the development and progression of prostate cancers. Transcriptional cofactors that bind AR are critical determinants of prostate tumorigenesis. To gain a deeper understanding of the proteins linked to AR-dependent gene transcription, we performed a DNA-affinity chromatography-based proteomic screen designed to identify proteins involved in AR-mediated gene transcription in prostate tumor cells. Functional experiments validated the coregulator roles of known AR-binding proteins in AR-mediated transcription in prostate tumor cells. More importantly, novel coregulatory functions were detected in components of well-established cell surface receptor-dependent signal transduction pathways. Further experimentation demonstrated that components of the TNF, TGF-β, IL receptor, and epidermal growth factor signaling pathways modulated AR-dependent gene transcription and androgen-dependent proliferation in prostate tumor cells. Collectively, our proteomic dataset demonstrates that the cell surface receptor- and AR-dependent pathways are highly integrated, and provides a molecular framework for understanding how disparate signal-transduction pathways can influence AR-dependent transcriptional programs linked to the development and progression of human prostate cancers.

The covalent attachment of ubiquitin onto proteins can elicit a variety of downstream consequences. Attachment is mediated by a large array of E3 ubiquitin ligases, each thought be subject to regulatory control and to have a specific repertoire of substrates. Assessing the biological roles of ligases, and in particular, identifying their biologically relevant substrates has been a persistent yet challenging question. In this study, we describe tools that may help achieve both of these goals. We describe a strategy whereby the activity of a ubiquitin ligase has been enzymatically reversed, accomplished by fusing it to a catalytic domain of an exogenous deubiquitinating enzyme. We present a library of 72 "anti-ligases" that appear to work in a dominant-negative fashion to stabilize their cognate substrates against ubiquitin-dependent proteasomal and lysosomal degradation. We then used the ligase-deubiquitinating enzyme (DUb) library to screen for E3 ligases involved in post-Golgi/endosomal trafficking. We identify ligases previously implicated in these pathways (Rsp5 and Tul1), in addition to ligases previously localized to endosomes (Pib1 and Vps8). We also document an optimized workflow for isolating and analyzing the "ubiquitome" of yeast, which can be used with mass spectrometry to identify substrates perturbed by expression of particular ligase-DUb fusions.

Protein:protein interactions are fundamental in living organism homeostasis. Here we introduce VHH6, a junctional epitope antibody capable of specifically recognizing a neo-epitope when two proteins interact, albeit transiently, to form a complex. Orthogonal biophysical techniques have been used to prove the “junctional epitope” nature of VHH6, a camelid single domain antibody recognizing the IL-6–gp80 complex but not the individual components alone. X-ray crystallography, HDX-MS and SPR analysis confirmed that the CDR regions of VHH6 interact simultaneously with IL-6 and gp80, locking the two proteins together. At the cellular level, VHH6 was able to alter the response of endothelial cells to exogenous IL-6, promoting a sustained STAT3 phosphorylation signal, an accumulation of IL-6 in vesicles and an overall pro-inflammatory phenotype supported further by transcriptomic analysis. Junctional epitope antibodies, like VHH6, not only offer new opportunities in screening and structure-aided drug discovery, but could also be exploited as therapeutics to modulate complex protein:protein interactions.

Decreases of up to 50% in the moisture uptake of polycyanurate networks based on 2,2-bis(4-cyanatophenyl)propane (BADCy) and 1,1-bis(4-cyanatophenyl)ethane (LECy) were observed when analogous netwo...

In gamma spectroscopy, a compromise must be made between energy resolution and event-rate capability. Some foreseen nuclear material safeguards applications require a spectrometer with energy resolution typical of high purity germanium (HPGe) detectors, operated at event rates up to and exceeding ${10^6}$ per second. We report the performance of an HPGe spectrometer system adapted to run under such conditions. Our system consists of a commercial semi-coaxial HPGe detector, a modified high-voltage-rail, resistive-feedback, charge-sensitive preamplifier and a continuous waveform digitizer. Digitized waveforms are analyzed offline with a novel time-variant trapezoidal filter algorithm. Several time-invariant trapezoidal filters are run in parallel and the slowest one not rejected by instantaneous pileup conditions is used to measure each pulse height. We have attained full-width-at-half-maximum energy resolution approximately 8 keV measured at 662 keV with $1.03 \times {10^6}$ per second incoming event rate and 39% throughput. An additional constraint on the width of the fast trigger filter removes a significant amount of rising edge pileup that passes the first pileup cut, reducing throughput to 25%. While better resolution has been reported by other authors, our throughput is an order of magnitude higher than any other reported HPGe system operated at such an event rate.

Single B cell technologies, which avoid traditional hybridoma fusion and combinatorial display, provide a means to interrogate the naturally-selected antibody repertoire of immunized animals. Many methods enable the sampling of memory B cell subsets, but few allow for the direct interrogation of the plasma cell repertoire, i.e., the subset of B cells responsible for producing immunoglobulin in serum. Here, we describe the use of a robust and simple fluorescence-based technique, called the fluorescent foci method, for the identification and isolation of antigen-specific IgG-secreting cells, such as plasma cells, from heterogeneous bone marrow preparations. Following micromanipulation of single cells, cognate pairs of heavy and light chain variable region genes were recovered by reverse transcription (RT)-polymerase chain reaction (PCR). During the PCR, variable regions were combined with a promoter fragment and a relevant constant region fragment to produce two separate transcriptionally-active PCR (TAP) fragments that were directly co-transfected into a HEK-293F cell line for recombinant antibody expression. The technique was successfully applied to the generation of a diverse panel of high-affinity, functional recombinant antibodies to human tumor necrosis factor (TNF) receptor 2 and TNF derived from the bone marrow of immunized rabbits and rats, respectively. Progression from a bone marrow sample to a panel of functional recombinant antibodies was possible within a 2-week timeframe.

Both LARGE1 (formerly LARGE) and its paralog LARGE2 are bifunctional glycosyltransferases with xylosy- and glucuronyltransferase activities, and are capable of synthesizing polymers composed of a repeating disaccharide [-3Xylα1,3GlcAβ1-]. Post-translational modification of the O-mannosyl glycan of α-dystroglycan (α-DG) with the polysaccharide is essential for it to act as a receptor for ligands in the extracellular matrix (ECM), and both LARGE paralogs contribute to the modification in vivo. LARGE1 and LARGE2 have different tissue distribution profiles and enzymatic properties; however, the functional difference of the homologs remains to be determined, and α-DG is the only known substrate for the modification by LARGE1 or LARGE2. Here we show that LARGE2 can modify proteoglycans (PGs) with the laminin-binding glycan. We found that overexpression of LARGE2, but not LARGE1, mediates the functional modification on the surface of DG-/-, Pomt1-/- and Fktn-/- embryonic stem cells. We identified a heparan sulfate-PG glypican-4 as a substrate for the LARGE2-dependent modification by affinity purification and subsequent mass spectrometric analysis. Furthermore, we showed that LARGE2 could modify several additional PGs with the laminin-binding glycan, most likely within the glycosaminoglycan (GAG)-protein linkage region. Our results indicate that LARGE2 can modify PGs with the GAG-like polysaccharide composed of xylose and glucuronic acid to confer laminin binding. Thus, LARGE2 may play a differential role in stabilizing the basement membrane and modifying its functions by augmenting the interactions between laminin globular domain-containing ECM proteins and PGs.

The large-conductance, voltage-gated, calcium (Ca(2+))-activated potassium channel (BKCa) plays an important role in regulating Ca(2+)signaling and is implicated in the maintenance of uterine quiescence during pregnancy. We used immunopurification and mass spectrometry to identify proteins that interact with BKCain myometrium samples from term pregnant (≥37 wk gestation) women. From this screen, we identified alpha-2-macroglobulin (α2M). We then used immunoprecipitation followed by immunoblot and the proximity ligation assay to confirm the interaction between BKCaand both α2M and its receptor, low-density lipoprotein receptor-related protein 1 (LRP1), in cultured primary human myometrial smooth muscle cells (hMSMCs). Single-channel electrophysiological recordings in the cell-attached configuration demonstrated that activated α2M (α2M*) increased the open probability of BKCain an oscillatory pattern in hMSMCs. Furthermore, α2M* caused intracellular levels of Ca(2+)to oscillate in oxytocin-primed hMSMCs. The initiation of oscillations required an interaction between α2M* and LRP1. By using Ca(2+)-free medium and inhibitors of various Ca(2+)signaling pathways, we demonstrated that the oscillations required entry of extracellular Ca(2+)through store-operated Ca(2+)channels. Finally, we found that the specific BKCablocker paxilline inhibited the oscillations, whereas the channel opener NS11021 increased the rate of these oscillations. These data demonstrate that α2M* and LRP1 modulate the BKCachannel in human myometrium and that BKCaand its immunomodulatory interacting partners regulate Ca(2+)dynamics in hMSMCs during pregnancy.

Mutations in genes encoding components of the sarcolemmal dystrophin-glycoprotein complex (DGC) are responsible for a large number of muscular dystrophies. As such, molecular dissection of the DGC is expected to both reveal pathological mechanisms, and provides a biological framework for validating new DGC components. Establishment of the molecular composition of plasma-membrane protein complexes has been hampered by a lack of suitable biochemical approaches. Here we present an analytical workflow based upon the principles of protein correlation profiling that has enabled us to model the molecular composition of the DGC in mouse skeletal muscle. We also report our analysis of protein complexes in mice harboring mutations in DGC components. Bioinformatic analyses suggested that cell-adhesion pathways were under the transcriptional control of NFκB in DGC mutant mice, which is a finding that is supported by previous studies that showed NFκB-regulated pathways underlie the pathophysiology of DGC-related muscular dystrophies. Moreover, the bioinformatic analyses suggested that inflammatory and compensatory mechanisms were activated in skeletal muscle of DGC mutant mice. Additionally, this proteomic study provides a molecular framework to refine our understanding of the DGC, identification of protein biomarkers of neuromuscular disease, and pharmacological interrogation of the DGC in adult skeletal muscle https://www.mda.org/disease/congenital-muscular-dystrophy/research.

In September 2009, the University of Iowa Libraries embarked on an experiment with patron-driven acquisition (PDA) of e-books with ebrary and YBP. An e-book–only PDA plan was initiated, entirely unmediated and with instantaneous access to the content. MARC records were loaded for each title, determined by our YBP approval profile and other limitations, for a total of 12,000 PDA records. Usage, cost, subject, and publisher data were analyzed for 850 purchased PDA e-books and thousands of other ebrary subscription titles. Results indicate that PDA can be a useful and effective tool for meeting user needs and building the local collection, but the role of PDA in the library’s collection management program presents challenges as well as opportunities.

Kapton polyimde is extensively used in solar arrays, spacecraft thermal blankets, and space inflatable structures. Upon exposure to atomic oxygen in low Earth orbit (LEO), Kapton is severely eroded. An effective approach to prevent this erosion is to incorporate polyhedral oligomeric silsesquioxane (POSS) into the polyimide matrix by copolymerizing POSS monomers with the polyimide precursor. The copolymerization of POSS provides Si and O in the polymer matrix on the nano level. During exposure of POSS polyimide to atomic oxygen, organic material is degraded, and a silica passivation layer is formed. This silica layer protects the underlying polymer from further degradation. Laboratory and space-flight experiments have shown that POSS polyimides are highly resistant to atomic-oxygen attack, with erosion yields that may be as little as 1% those of Kapton. The results of all the studies indicate that POSS polyimide would be a space-survivable replacement for Kapton on spacecraft that operate in the LEO environment.

A new scriptedMonte Carlo analysis software tool has been developed for use with the fully implicit ablation and thermal response program thermal-protection-system-sizing code. This paper describes the new software tool and gives examples of its increased utility and useful insights onmany aeroshell heatshield design considerations, such as maximum bond line temperature, recession, isotherm depth, surface temperature, and char depth. Finally, results are presented that illustrate the effect of uncertainties in key parameters that were used in the Mars Science Laboratory aeroshell thermal protection system design. The parameters having the greatest influence on bond line temperature dispersion for theMars Science Laboratory backshell are virginmaterial thermal density (34%), initial material temperature (20%), virgin material conductivity (17%), and char density (13%).

[Abstract] Aerothermal analyses of the Phoenix entry vehicle in the Martian Atmosphere are presented. Two Navier-Stokes flow solvers are used to predict the environments experienced by the vehicle along several design trajectories. The majority of the solutions are computed as axisymmetric in order to yield higher, and therefore more conservative, heating rates. Several three-dimensional cases are also computed to assess the heating enhancement caused by surface singularities and the complex aftbody vortices. When the thermal protection system was assessed in a separate analysis in response to the computed aeroheating environments, the prebuilt design is found perform successfully.

A new and highly efficient method for determining relative carboxyl group distribution in carboxyl-terminated polybutadiene has been developed using practical synthetic and analytical techniques. Using oxalyl chloride, samples of carboxyl-terminated polybutadiene were rapidly transformed to acid chlorides that were then chemically derivatized with benzyl alcohol, 4-nitrobenzyl alcohol, and 3,5-dinitrobenzyl alcohol. This provided quick and quantitative conversion to the corresponding benzyl ester derivatives. Each new derivative was fully characterized by nuclear magnetic resonance and Fourier transform infrared spectroscopy. The benzyl ester modified polymers were investigated in detail to determine their relative carboxyl group concentrations. To do this, gel permeation chromatography combined with ultra violet/refractive index dual detection was employed. The 4-nitrobenzyl ester, having the highest extinction factor at 270 nm provided the best UV data for analysis. The ultra violet/refractive index data of four separate polymer samples were plotted as a function of molecular weight. The data were compared with a theoretical plot (carboxyl group = two for all molecular weights) to illustrate the relative carboxyl concentration over the entire molecular weight range. Supplemental characterization of the 4-nitrobenzyl modified polymer was carried out using matrix-assisted laser desorption ionization coupled with time of flight mass spectrometry. © 2011 Wiley Periodicals, Inc. J Appl Polym Sci, 2011

Localized chromatin modifications of histone tails play an important role in regulating gene transcription, and aberration of these processes leads to carcinogenesis. Methylated histone lysine residues, a key player in chromatin remodeling, are demethylated by the JmjC class of enzymes. Here we show that JMJD5 (now renamed KDM8), a JmjC family member, demethylates H3K36me2 and is required for cell cycle progression. Chromatin immunoprecipitation assays applied to human genome tiling arrays in conjunction with RNA microarray revealed that KDM8 occupies the coding region of cyclin A1 and directly regulates transcription. Mechanistic analyses showed that KDM8 functioned as a transcriptional activator by inhibiting HDAC recruitment via demethylation of H3K36me2, an epigenetic repressive mark. Tumor array experiments revealed KDM8 is overexpressed in several types of cancer. In addition, loss-of-function studies in MCF7 cells leads to cell cycle arrest. These studies identified KDM8 as an important cell cycle regulator.

An effective method for producing high density fuel candidates from pinenes is provided. MMT-K10 is an efficient catalyst for the reaction, although significant amounts of p-cymene and camphene produced as byproducts limit the overall yield to about 80%. Nafion is also an excellent catalyst for pinene dimerization and was capable of producing dimers in up to 90% yield. Pinene dimers synthesized with these heterogenous catalysts have a density and net heat of combustion comparable to JP-10. It is emphasized that this abstract is provided to comply with the rules requiring an abstract that will allow a searcher or other reader to quickly ascertain the subject matter of the technical disclosure. It is submitted with the understanding that it will not be used to interpret or limit the scope of the claims.

Quantitative conversion of 1-butene to a Schultz−Flory distribution of oligomers has been accomplished by use of Group 4 transition-metal catalysts in the presence of methylaluminoxane (MAO). The o...

Novel hybrid polymers with thiophenylanilino and furanylanilino backbones and substituted phenyl side groups are reported. The new monomers bis-(4-heterocyclic-2-yl-phenyl)-aryl-amine (heterocyclic = thiophen with aryl = 4-benzoyl (2a), 4-nitro-phenyl (2b) and furan with aryl = 4-benzoyl-phenyl (3a), 4-nitro-phenyl (3b)) were prepared by monosubstituting triphenylamine under electrophilic aromatic conditions affording 4-nitrotriphenylamine and 4-benzoyltriphenylamine. Di(bromination) of the latter compounds followed by Stille cross-coupling reactions with 2-tributylstannylthiophene or 2-tributylstannylfuran produces the new monomers 2a–b and 3a–b in high yield. The monomers are electrochemically polymerized at relatively low potentials (

The preparation of polyimides containing side-chain chromophores and the long-term aging performance of poled films are described. These materials were compared to guest–host polycarbonate films. Mach–Zehnder optical interferometers were fabricated from these polymers that contained CLD- and FTC-type chromophores. Changes in optical properties were monitored for months at four temperatures ranging from ambient to 110 °C. The isothermal relaxation data were modeled using both a stretched exponential equation and a power law in time equation. The temperature dependency of the time constants of these equations was modeled using a new activation-energy equation: ln( τ / τ p ) = E R (1 + tanh[( T c − T )/ D ])/2 RT + E p / RT where T c is the central temperature of the transition zone, D is the breadth of the zone, and E s are the activation energies of rigid and pliable materials. Multi-year high-temperature stability of the poled guest–host and side-chain materials was predicted.

andM. Joseph RobertsNAVAIR Research Department, ChemistryDiVision-NAWCWD, China Lake, California 93555ReceiVed February 9, 2007ReVised Manuscript ReceiVed March 23, 2007Polyhedral oligomeric silsesquioxanes (POSS) are a familyof nanoscale chemical structures that contain a silicon/oxygencore based on (SiO

We have developed a novel androgen receptor (AR) expression system in the 293 human embryonic kidney cell line that recapitulates AR biochemical activity as a steroid hormone receptor in prostate cancer cells. We used this system to identify putative AR-binding proteins in the cytosolic and nuclear compartments of mammalian cells using a large scale co-immunoprecipitation strategy coupled to quantitative mass spectrometry. For example, the heat shock 70 and 90 chaperones, which are known regulators of steroid hormone receptor, were identified as AR-binding proteins. AR purification enriched for proteins involved in RNA processing, protein transport, and cytoskeletal organization, suggesting a functional link between AR and these protein modules in mammalian cells. For example, AR purification in the nuclear compartment led to the specific enrichment of α-actinin-4, clathrin heavy chain, and serine-threonine protein kinase C δ. Short interfering RNA knockdown studies and co-transcriptional reporter assays revealed that clathrin heavy chain possessed co-activator activity during AR-mediated transcription, whereas α-actinin-4 and protein kinase C δ displayed both co-activator and co-repressor activity during AR-mediated transcription that was dependent upon their relative expression levels. Lastly immunohistochemical staining of prostate tissue showed that α-actinin-4 levels decreased in the nucleus of high grade cancerous prostate samples, suggesting its possible deregulation in advanced prostate cancers as previously observed in late stage metastatic breast cancers. Taken together, these findings suggest AR binds to specific protein modules in mammalian cells and that these protein modules may provide a molecular framework for interrogating AR function in normal and cancerous prostate epithelial cells.

The National Science Foundation (NSF) funded Center for Biophotonics Science and Technology (CBST) has created various high school biophotonics research academies for both students and teachers from diverse socioeconomic backgrounds. These academies engage diverse students for 10 hours to over 350 hours per year for multiple years with an emphasis on learning the basics of biophotonics and then conducting original, team-based research. We have developed three versions of the academy, one focused on biology and biophotonics, one on cancer and biophotonics, and a third on plants and biophotonics. A fourth emphasis on biomedical engineering and biophotonics is planned. We have conducted one of these academies for three years and have had very good student retention and science fair winners. As part of our program we also have a summer academy for training teachers. Challenges have arisen amongst the various levels of Academies, chief among them sustainability. In the future, more extensive evaluation, curriculum consolidation, and widespread dissemination are critical.

Liquid scintillation counters measure charged particle-emitting radioactive isotopes and are used for environmental studies, nuclear chemistry, and life science. Alpha and beta emissions arising from the material under study interact with the scintillation cocktail to produce light. The prototypical liquid scintillation counter employs low-level photon-counting detectors to measure the arrival of the scintillation. For reliable operation, the counting instrument must convey the scintillation light to the detectors efficiently and predictably. Current best practices employ the use of two or more detectors for coincidence processing to discriminate true scintillation events from background events due to instrumental effects such as photomultiplier tube dark rates, tube flashing, or other light emission not generated in the scintillation cocktail vial. In low-background liquid scintillation counters, additional attention is paid to shielding the scintillation cocktail from naturally occurring radioactive material present in the laboratory and within the instrument’s construction materials. Low-background design is generally at odds with optimal light collection. This study presents the evolution of a light collection design for liquid scintillation counting (LSC) in a low-background shield. The basic approach to achieve both good light collection and a low-background measurement is described. The baseline signals arising from the scintillation vial are modeled and methods to efficiently collect scintillation light are presented as part of the development of a customized low-background, high-sensitivity LSC system.

Background Identifying cellular signaling pathways that become corrupted in the presence of androgens that increase the metastatic potential of organ-confined tumor cells is critical to devising strategies capable of attenuating the metastatic progression of hormone-naïve, organ-confined tumors. In localized prostate cancers, gene fusions that place ETS-family transcription factors under the control of androgens drive gene expression programs that increase the invasiveness of organ-confined tumor cells. C-X-C chemokine receptor type 4 (CXCR4) is a downstream target of ERG, whose upregulation in prostate-tumor cells contributes to their migration from the prostate gland. Recent evidence suggests that CXCR4-mediated proliferation and metastasis of tumor cells is regulated by CXCR7 through its scavenging of chemokine CXCL12. However, the role of androgens in regulating CXCR4-mediated motility with respect to CXCR7 function in prostate-cancer cells remains unclear. Methods Immunocytochemistry, western blot, and affinity-purification analyses were used to study how androgens influenced the expression, subcellular localization, and function of CXCR7, CXCR4, and androgen receptor (AR) in LNCaP prostate-tumor cells. Moreover, luciferase assays and quantitative polymerase chain reaction (qPCR) were used to study how chemokines CXCL11 and CXCL12 regulate androgen-regulated genes (ARGs) in LNCaP prostate-tumor cells. Lastly, cell motility assays were carried out to determine how androgens influenced CXCR4-dependent motility through CXCL12. Results Here we show that, in the LNCaP prostate-tumor cell line, androgens coordinate the expression of CXCR4 and CXCR7, thereby promoting CXCL12/CXCR4-mediated cell motility. RNA interference experiments revealed functional interactions between AR and CXCR7 in these cells. Co-localization and affinity-purification experiments support a physical interaction between AR and CXCR7 in LNCaP cells. Unexpectedly, CXCR7 resided in the nuclear compartment and modulated AR-mediated transcription. Moreover, androgen-mediated cell motility correlated positively with the co-localization of CXCR4 and CXCR7 receptors, suggesting that cell migration may be linked to functional CXCR4/CXCR7 heterodimers. Lastly, CXCL12-mediated cell motility was CXCR7-dependent, with CXCR7 expression required for optimal expression of CXCR4 protein. Conclusions Overall, our results suggest that inhibition of CXCR7 function might decrease the metastatic potential of organ-confined prostate cancers. Electronic supplementary material The online version of this article (doi:10.1186/s12885-015-1201-5) contains supplementary material, which is available to authorized users.

The role at admission of nasal polymerase chain reaction (PCR) for patients with methicillin-resistant Staphylococcus aureus (MRSA) in guiding antibiotic therapy for lower respiratory tract infection is unknown.To determine whether nasal MRSA PCR at admission can predict the absence of MRSA in lower respiratory tract secretions.We performed a retrospective study of adult patients admitted to a large urban hospital. Patients had a nasal MRSA PCR test and a lower respiratory tract culture obtained within 48 hours of admission and the culture yielded S aureus.Sensitivity, specificity, and positive and negative predictive values.Our results showed high sensitivity (93.3%) and negative predictive value (95.2%) of nasal PCR for MRSA in the lower respiratory tract.With its high sensitivity and negative predictive value, a nasal MRSA PCR test performed within 48 hours of hospital admission could help guide the discontinuation of MRSA-directed empiric antibiotic therapy in patients who are unlikely to be infected with this organism. A prospective study is needed to confirm these findings.

A new cyanate ester monomer was prepared from bis(4-cyanatophenyl)dimethylsilane (SiMCy) and fully characterized by analytical and spectroscopic techniques. The monomer was found to have a melting point about 20 °C lower than that of the commercial bis(4-cyanatophenyl)dimethylmethane (BADCy) with similar melt viscosity, curing kinetics, and postcure glass transition temperature. Analysis of the single-crystal molecular structure by X-ray diffraction showed that intermolecular packing was dominated by weak hydrogen-bonding attractions between the aromatic rings and the −OCN nitrogen atoms. In contrast, the packing interactions found in BADCy are dominated by dipole−dipole interactions of the OCN groups. These differences may explain the 50% reduction in moisture uptake observed in SiMCy as compared to BADCy during exposure to boiling water. In addition, thermogravimetric analysis revealed that SiMCy exhibited a significantly higher char yield in air than BADCy, presumably due to the formation of silicates ...

This paper gives an overview of recent work on new organic π-electron-conjugated materials prepared and tested in our laboratories. Second-order nonlinear optical polymers for electro-optic modulation were investigated, both guest–host and side-chain chromophoric materials. Chromophores and polymers were synthesized, thin films were prepared, and the following properties were measured: electro-optic coefficient, optical absorption loss and thermal stability. It was concluded that optical modulators fabricated from these materials could be viable components in optical communication systems. Work on novel polyphenylenevinylenes is summarized. Besides their low electrical resistivity, photoluminescent, and electro-active properties, these materials have interesting photoconductive properties that may prove useful in optical control of photonic logic devices.

Successful treatment of multiple cancer types requires early detection and identification of reliable biomarkers present in specific cancer tissues. To test the feasibility of identifying proteins from archival cancer tissues, we have developed a methodology, termed direct tissue proteomics (DTP), which can be used to identify proteins directly from formalin-fixed paraffin-embedded prostate cancer tissue samples. Using minute prostate biopsy sections, we demonstrate the identification of 428 prostate-expressed proteins using the shotgun method. Because the DTP method is not quantitative, we employed the absolute quantification method and demonstrate picogram level quantification of prostate-specific antigen. In depth bioinformatics analysis of these expressed proteins affords the categorization of metabolic pathways that may be important for distinct stages of prostate carcinogenesis. Furthermore, we validate Wnt-3 as an upregulated protein in cancerous prostate cells by immunohistochemistry. We propose that this general strategy provides a roadmap for successful identification of critical molecular targets of multiple cancer types.

A series of four new end-capped and hydroxymethyl-functionalized polyimides were prepared. Through a two-step chemical modification process (3-aminopropyl)(hepta-i-butyl) polyhedral oligomeric silsesquioxane (POSS) was covalently attached to the polymer backbone. POSS loading levels as high as 36 wt % could be obtained while maintaining excellent processability and optical clarity of thin films. Concurrent attachment of either a cyanate ester or hydroxyethyl methacrylate (HEMA) group afforded processable POSS-polyimides that underwent thermal curing to yield solvent-resistant films, both having final Tg's of 251 °C. Kinetic analysis of the cure reactions yielded energy of activations of 93 kJ/mol (cyanate ester) and 103 kJ/mol (HEMA). Exposure of a POSS-polyimide containing ∼31 wt % POSS to atomic oxygen displayed no measurable level of erosion relative to a Kapton H standard.

Highly crystalline phenyl 2,5-dichlorobenzenesulfonate (PDBS, Tmelt = 86–87 °C) and pentafluorophenyl 2,5-dichlorobenzenesulfonate (FPDBS, Tmelt = 120–122 °C) were synthesized. Single-crystal X-ray molecular structure determinations show that both compounds have similar three-dimensional molecular structures; however, PDBS crystals are thin platelets and FPDBS crystals form hexagonal tube-like structures that are predominately hollow at one end. PDBS crystals exhibit offset π-stacking of the phenoxy-rings that form complete two-dimensional layers each two molecules thick. Hydrogen-bonding interactions are calculated at ∼3.2 A between the C6-hydrogen and the sulfonyl-oxygen of a neighboring molecule. On the other hand, for FPDBS, π-stacking of the dichloro-substituted ring as well as dipole–dipole interactions of the fluorinated-phenoxy rings appears to be the predominate intermolecular interactions. Neither structure exhibits any kind of side-on interaction of the phenyl rings. PDBS and FPDBS exhibit melting point depressions of 26 and 40 °C, respectively, in the presence of supercritical CO2. Although both sulfonates exhibit high solubility in CO2, much lower pressures are needed to dissolve FPDBS compared to PDBS. For example, at 100 °C FPDBS dissolves at 4750 psia and PDBS dissolves at 11,000 psia. The solubility data reinforce the observation that fluorinating a compound can significantly lower the conditions needed to dissolve that compound in CO2.

Data are reported on the phase behavior of hydrocarbon and semifluoroinated octyl ester benzoate dimers in CO 2 to temperatures of 100 °C and pressures of 1 600 bar. The experimental data at 75 °C demonstrate that the non-fluorinated head-to-head (H–H) dimer dissolves in CO 2 at ∼400 bar lower pressures than the non-fluorinated tail-to-tail (T–T) dimer. Even though partially fluorinating the octyl tails of the H–H and T–T dimers renders them soluble in CO 2 at pressures near 200 bar, it still takes ∼40 bar more pressure to dissolve the fluorinated T–T dimer as compared to the H–H dimer. The difference in pressures needed to dissolve these dimers is attributed to steric constraints on the coplanarity of the benzene rings imposed by the H–H regiochemistry that do not exist with T–T dimers. Semi-empirical quantum mechanics calculations suggest that the H–H dimer has a twisted, non-coplanar conformation due to the steric effect of the octyl ester groups while the T–T dimer has a less twisted conformation. Steric hindrance in the H–H dimer reduces considerably resonance or conjugation between the π electrons of the aromatic groups which also reduces the dipole moments of the H–H dimers compared to those of the T–T dimers.

Two new monomers were prepared of the formula 3,X-bis(4-aminophenoxy)benzenemethanol (where X = 5, 3a; X = 4, 3b) and each copolymerized with 2,2-bis(4-aminophenyl)hexafluoropropane, 6-FDA, and 6 mol % of phthalic anhydride as an end-cap to afford polyimides 4 and 5, respectively. The CH2OH group in the polymers could be modified with excess 1,6-hexanediisocyanate to afford 100% modification of polymer 6 concomitant with formation of a pendent and reactive isocyanate group. This latter polymer was shown to react with disperse red 1 (DR-1) to produce a polyimide-supported dye (7, Tg 199 °C) via formation of a urethane linkage. Treatment of the polymers 4 and 5 with excess adipoyl chloride afforded polymers 8 and 9, respectively, with a pendent acid chloride functional group. Further reaction with disperse red 1 (DR-1) in the presence of DMAP led to rapid esterification, yielding high-Tg dye-attached polyimides 11 (Tg 168 °C) and 12 (Tg 175 °C). Treatment of polymer 4 with sebacoyl chloride (a longer spacer...

Isopleths and solution densities, from ∼25 to 100 °C, are reported for mixtures of CO 2 with semi-fluorinated and nonfluorinated propyl, butyl, octyl, and decyl 2,5-dichlorobenzoates. The maxima in the pressure–composition ( P – x ) isotherms for the alkyl 2,5-dichlorobenzoates range from 70 to 600 bar and the maxima increase with increasing alkyl chain length at each temperature. When the alkyl side chains are semi-fluorinated, the P – x maxima range from 60 to 175 bar over the same temperature range as with the nonfluorinated analogs. The P – x maxima of the semi-fluorinated 2,5-dichlorobenzoates in CO 2 , first decrease as the benzoate alkyl chain is increased from propyl to butyl, but then increase as the alkyl chain length is further increased from butyl to octyl to decyl, although the differences in the maxima at a given temperature between these four semi-fluorinated benzoates are not as great as that observed with the nonfluorinated analogs. Also, the alkyl 2,5-dichlorobenzoate–CO 2 mixtures exhibit three-phase, liquid–liquid–vapor (LLV), equilibria near the vapor pressure curve and critical point of CO 2 whereas three phases are not observed for the semi-fluorinated analogs.

Androgen-ablation therapy is an effective method for treating prostate cancer. However, prostate tumors that survive long-term androgen-ablation therapy are classified as androgen-independent as they proliferate in the absence of androgens, and they tend to be enriched for neuroendocrine (NE) cells. Androgen withdrawal causes androgen-dependent prostate cancer cells to adopt a pronounced NE phenotype, suggesting that androgen receptor (AR) represses an intrinsic NE transdifferentiation process in prostate cancer cells. In this report we show that short interfering RNA-induced AR silencing induced a NE phenotype that manifested itself in the growth of dendritic-like processes in both the androgen-dependent LNCaP and androgen-independent LNCaP-AI human prostate cancer cells. Western blot analysis revealed that neuronal-specific enolase, a marker of the neuronal lineage, was increased by AR knockdown in LNCaP cells. The expression levels of the neuronal-specific cytoskeletal proteins beta-tubulin III, nestin, and glial acidic fibrillary protein were also characterized in AR knockdown cells. Most interestingly, AR silencing induced beta-tubulin III expression in LNCaP cells, while AR knockdown increased glial acidic fibrillary protein levels in both LNCaP and LNCaP-AI cells. Lastly, AR silencing reduced the proliferative capacity of LNCaP and LNCaP-AI cells. Our data demonstrate that AR actively represses an intrinsic NE transdifferentiation process in androgen-responsive prostate cancer cells and suggest a potential link between AR inactivation and the increased frequency of NE cells in androgen-independent tumors.

Computational fluid dynamics (CFD) is used to simulate the wake flow and afterbody heating of the Project Fire II ballistic reentry to Earth at 11.4 km/sec. Laminar results are obtained over a portion of the trajectory between the initial heat pulse and peak afterbody heating. Although non-catalytic forebody convective heating results are in excellent agreement with previous computations, initial predictions of afterbody heating were about a factor of two below the experimental values. Further analysis suggests that significant catalysis may be occurring on the afterbody heat shield. Computations including finite-rate catalysis on the afterbody surface are in good agreement with the data over the early portion of the trajectory, but are conservative near the peak afterbody heating point, especially on the rear portion of the conical frustum. Further analysis of the flight data from Fire II shows that peak afterbody heating occurs before peak forebody heating, a result that contradicts computations and flight data from other entry vehicles. This result suggests that another mechanism, possibly pyrolysis, may be occurring during the later portion of the trajectory, resulting in less total heat transfer than the current predictions.

Monomers of the structure 2,5-dichloro-1,4(RO 2 C) 2 benzene, where R = CH 2 CF 2 CF 3 (la), -(CH 2 ) 2 CH 3 (1b), -CH 2 CH 2 (CF 2 ) 5 CF 3 (1c), and -(CH 2 ) 7 CH 3 (1d), were prepared and characterized in the study. Polymerization by the Ni-catalyzed/Zn-mediated homocoupling methodology afforded poly(p-phenylenes) 2a-2d. The polymers obtained after precipitation into methanol showed molecular weights (M n , relative to polystyrene standards) in the range 7000-13 000. The regiochemistry in the polymer was deduced to be approximately 70% head-to-tail with the remainder head-to-head. The polymers displayed excellent solubility in common polar organic solvents such at THF and chloroform. The poly-(p-phenylenes) 2a and 2b showed no solubility in supercritical ethane, propane, butane, and carbon dioxide even at 190 °C and 35 000 psia. Polymer 2d was remarkably soluble in the supercritical hydrocarbon solvents at low pressures (1000 psia). However, 2d does not dissolve in supercritical carbon dioxide. The fluorinated poly(p-phenylene) 2c dissolved in the supercritical hydrocarbons at pressures below 6000 psia, and it dissolved in carbon dioxide at pressures as low as 1000 psia at 25 °C.

The first series of conjugated organic polymers with the formula {1,4-[2-(RO)-5-(R‘O)C6H2]NNC6H4C⋮C−}n, where R = methyl, R‘ = hexyl (7a) and R = hexyl, R‘ = SiMe2-t-Bu (8), were produced in palladium-catalyzed cross-coupling and homocoupling reactions of selected bis(bromo) and bis(alkynyl) azo monomers. The molecular weights (Mn) for the conjugated polymers ranged from 5500 to 22 000. Degenerate four wave mixing (DFWM) with 780 nm femtosecond pulses was used to investigate electronic third-order nonlinearities for azo monomer 3a and conjugated azo polymer 7a. Both compounds had ultrafast nonlinearities with τ1 = 100 fs response time. The real part of 7a second hyperpolarizability, γ7a, is negative, Re γ7a = −(4.3 ± 0.3) × 10-46 m5 V-2. Polymer 7a's second hyperpolarizability also has an imaginary component, Im γ7a = (1.7 ± 0.3) × 10-46 m5 V-2. Calculated per repeat unit, polymer |γ7a| = (4.6 ± 0.5) × 10-46 m5 V-2 and is about 780 times larger than that for monomer 3a. γ7a was analyzed with the three-lev...