A model of intervertebral disc degeneration using combined cyclic overloading and enzyme digestion

This is the author accepted manuscript

A novel in-vitro model of intervertebral disc degeneration using hyperphysiological loading.

This is the final version. Available from Elsevier via the DOI in this record. Intervertebral disc (IVD) degeneration includes changes in tissue biomechanics, physical attributes, biochemical composition, disc microstructure, and cellularity, which can all affect the normal function of the IVD, and ultimately may lead to pain. The purpose of this research was to develop an in-vitro model of degeneration that includes the evaluation of physical, biomechanical, and structural parameters, and that does so over several load/recovery periods. Hyperphysiological loading was used as the degenerative initiator with three experimental groups employed using bovine coccygeal IVD specimens: Control; Single-Overload; and Double-Overload. An equilibrium stage comprising a static load followed by two load/recovery periods was followed by six further load/recovery periods. In the Control group all load/recovery periods were the same, comprising physiological cyclic loading. The overload groups differed in that hyperphysiological loading was applied during the 4th loading period (Single-Overload), or the 4th and 5th loading period (Double-Overload). Overloading led to a significant reduction in disc height compared to the Control group, which was not recovered in subsequent physiological load/recovery periods. However, there were no significant changes in stiffness. Overloading also led to significantly more microstructural damage compared to the Control group. Taking all outcome measures into account, the overload groups were evaluated as replicating clinically relevant aspects of moderate IVD degeneration. Further research into a potential dose-effect, and how more severe degeneration can be replicated would provide a model with the potential to evaluate new treatments and interventions for different stages of IVD degeneration

Experimental assessment of the mechanical performance of graphene nanoplatelets coated polymers

This is the author accepted manuscript. The final version is available on open access from Wiley via the DOI in this recordThis study presents the effect of spray-coated graphene nanoplatelets on the mechanical response of various polymers to cyclic loadings. The substrates material (three polymers) and the coating (various numbers of coating layers) are assessed. The experimental results suggest that the compressive stiffness, compressive modulus, damping and energy dissipation of the samples coated with graphene nanoplatelets improve with respect to the uncoated samples. The outcomes of this experimental research highlight the feasibility of utilising films of graphene nanoplatelets to improve the mechanical properties of polymers for vibration isolation, foreseeing application in various environments for instance in buildings and infrastructures (bridges, railways) for seismic and acoustic isolation.Engineering and Physical Sciences Research Council (EPSRC

Polyamines are required for normal growth in sinorhizobium meliloti

Polyamines (PAs) are ubiquitous polycations derived from basic L-amino acids whose physiological roles are still being defined. Their biosynthesis and functions in nitrogen-fixing rhizobia such as Sinorhizobium meliloti have not been extensively investigated. Thin layer chromatographic and mass spectrometric analyses showed that S. meliloti Rm8530 produces the PAs, putrescine (Put), spermidine (Spd) and homospermidine (HSpd), in their free forms and norspermidine (NSpd) in a form bound to macromolecules. The S. meliloti genome encodes two putative ornithine decarboxylases (ODC) for Put synthesis. Activity assays with the purified enzymes showed that ODC2 (SMc02983) decarboxylates both ornithine and lysine. ODC1 (SMa0680) decarboxylates only ornithine. An odc1 mutant was similar to the wild-type in ODC activity, PA production and growth. In comparison to the wild-type, an odc2 mutant had 45% as much ODC activity and its growth rates were reduced by 42, 14 and 44% under non-stress, salt stress or acid stress conditions, respectively. The odc2 mutant produced only trace levels of Put, Spd and HSpd. Wild-type phenotypes were restored when the mutant was grown in cultures supplemented with 1mM Put or Spd or when the odc2 gene was introduced in trans. odc2 gene expression was increased under acid stress and reduced under salt stress and with exogenous Put or Spd. An odc1 odc2 double mutant had phenotypes similar to the odc2 mutant. These results indicate that ODC2 is the major enzyme for Put synthesis in S. meliloti and that PAs are required for normal growth in vitro

Validation of the SCID-hu Thy/Liv mouse model with four classes of licensed antiretrovirals.

BackgroundThe SCID-hu Thy/Liv mouse model of HIV-1 infection is a useful platform for the preclinical evaluation of antiviral efficacy in vivo. We performed this study to validate the model with representatives of all four classes of licensed antiretrovirals.Methodology/principal findingsEndpoint analyses for quantification of Thy/Liv implant viral load included ELISA for cell-associated p24, branched DNA assay for HIV-1 RNA, and detection of infected thymocytes by intracellular staining for Gag-p24. Antiviral protection from HIV-1-mediated thymocyte depletion was assessed by multicolor flow cytometric analysis of thymocyte subpopulations based on surface expression of CD3, CD4, and CD8. These mice can be productively infected with molecular clones of HIV-1 (e.g., the X4 clone NL4-3) as well as with primary R5 and R5X4 isolates. To determine whether results in this model are concordant with those found in humans, we performed direct comparisons of two drugs in the same class, each of which has known potency and dosing levels in humans. Here we show that second-generation antiretrovirals were, as expected, more potent than their first-generation predecessors: emtricitabine was more potent than lamivudine, efavirenz was more potent than nevirapine, and atazanavir was more potent than indinavir. After interspecies pharmacodynamic scaling, the dose ranges found to inhibit viral replication in the SCID-hu Thy/Liv mouse were similar to those used in humans. Moreover, HIV-1 replication in these mice was genetically stable; treatment of the mice with lamivudine did not result in the M184V substitution in reverse transcriptase, and the multidrug-resistant NY index case HIV-1 retained its drug-resistance substitutions.ConclusionGiven the fidelity of such comparisons, we conclude that this highly reproducible mouse model is likely to predict clinical antiviral efficacy in humans

The LCES HIRES/Keck Precision Radial Velocity Exoplanet Survey

This document is the Accepted Manuscript version of the following article: R. Paul Butler, et al, The LCES HIRES/Keck Precision Radial Velocity Exoplanet Survey, The Astronomical Journal, Vol 153 (5), 19 pp., published 13 April 2017. The Version of Record is available online at doi: https://doi.org/10.3847/1538-3881/aa66ca. Paper data available at: http://home.dtm.ciw.edu/ebps/data/. © 2017. The American Astronomical Society. All rights reserved.We describe a 20-year survey carried out by the Lick-Carnegie Exoplanet Survey Team (LCES), using precision radial velocities from HIRES on the Keck-I telescope to find and characterize extrasolar planetary systems orbiting nearby F, G, K, and M dwarf stars. We provide here 60,949 precision radial velocities for 1,624 stars contained in that survey. We tabulate a list of 357 significant periodic signals that are of constant period and phase, and not coincident in period and/or phase with stellar activity indices. These signals are thus strongly suggestive of barycentric reflex motion of the star induced by one or more candidate exoplanets in Keplerian motion about the host star. Of these signals, 225 have already been published as planet claims, 60 are classified as significant unpublished planet candidates that await photometric follow-up to rule out activity-related causes, and 54 are also unpublished, but are classified as "significant" signals that require confirmation by additional data before rising to classification as planet candidates. Of particular interest is our detection of a candidate planet with a minimum mass of 3.9 Earth masses and an orbital period of 9.9 days orbiting Lalande 21185, the fourth-closest main sequence star to the Sun. For each of our exoplanetary candidate signals, we provide the period and semi-amplitude of the Keplerian orbital fit, and a likelihood ratio estimate of its statistical significance. We also tabulate 18 Keplerian-like signals that we classify as likely arising from stellar activity.Peer reviewedFinal Accepted Versio

Involvement of Noradrenergic Neurotransmission in the Stress- but not Cocaine-Induced Reinstatement of Extinguished Cocaine-Induced Conditioned Place Preference in Mice: Role for β-2 Adrenergic Receptors

The responsiveness of central noradrenergic systems to stressors and cocaine poses norepinephrine as a potential common mechanism through which drug re-exposure and stressful stimuli promote relapse. This study investigated the role of noradrenergic systems in the reinstatement of extinguished cocaine-induced conditioned place preference by cocaine and stress in male C57BL/6 mice. Cocaine- (15 mg/kg, i.p.) induced conditioned place preference was extinguished by repeated exposure to the apparatus in the absence of drug and reestablished by a cocaine challenge (15 mg/kg), exposure to a stressor (6-min forced swim (FS); 20–25°C water), or administration of the α-2 adrenergic receptor (AR) antagonists yohimbine (2 mg/kg, i.p.) or BRL44408 (5, 10 mg/kg, i.p.). To investigate the role of ARs, mice were administered the nonselective β-AR antagonist, propranolol (5, 10 mg/kg, i.p.), the α-1 AR antagonist, prazosin (1, 2 mg/kg, i.p.), or the α-2 AR agonist, clonidine (0.03, 0.3 mg/kg, i.p.) before reinstatement testing. Clonidine, prazosin, and propranolol failed to block cocaine-induced reinstatement. The low (0.03 mg/kg) but not high (0.3 mg/kg) clonidine dose fully blocked FS-induced reinstatement but not reinstatement by yohimbine. Propranolol, but not prazosin, blocked reinstatement by both yohimbine and FS, suggesting the involvement of β-ARs. The β-2 AR antagonist ICI-118551 (1 mg/kg, i.p.), but not the β-1 AR antagonist betaxolol (10 mg/kg, i.p.), also blocked FS-induced reinstatement. These findings suggest that stress-induced reinstatement requires noradrenergic signaling through β-2 ARs and that cocaine-induced reinstatement does not require AR activation, even though stimulation of central noradrenergic neurotransmission is sufficient to reinstate

Herschelobservations of the W3 GMC (II): clues to the formation of clusters of high-mass stars

The W3 giant molecular cloud is a prime target for investigating the formation of high-mass stars and clusters. This second study of W3 within the HOBYS Key Program provides a comparative analysis of subfields within W3 to further constrain the processes leading to the observed structures and stellar population. Probability density functions (PDFs) and cumulative mass distributions (CMDs) were created from dust column density maps, quantified as extinction AV. The shape of the PDF, typically represented with a lognormal function at low Av “breaking” to a power-law tail at high Av, is influenced by various processes including turbulence and selfgravity. The breaks can also be identified, often more readily, in the CMDs. The PDF break from lognormal (Av(SF)» 6–10 mag) appears to shift to higher Av by stellar feedback, so that high-mass star-forming regions tend to have higher PDF breaks. A second break at Av> 50 mag traces structures formed or influenced by a dynamic process. Because such a process has been suggested to drive high-mass star formation in W3, this second break might then identify regions with potential for hosting high-mass stars/clusters. Stellar feedback appears to be a major mechanism driving the local evolution and state of regions within W3. A high initial star formation efficiency in a dense medium could result in a self-enhancing process, leading to more compression and favorable star formation conditions (e.g., colliding flows), a richer stellar content, and massive stars. This scenario would be compatible with the “convergent constructive feedback” model introduced in our previous Herschel study

Biochemical characterization of the skeletal matrix of the massive coral, Porites australiensis – The saccharide moieties and their localization

To construct calcium carbonate skeletons of sophisticated architecture, scleractinian corals secrete an extracellular skeletal organic matrix (SOM) from aboral ectodermal cells. The SOM, which is composed of proteins, saccharides, and lipids, performs functions critical for skeleton formation. Even though polysaccharides constitute the major component of the SOM, its contribution to coral skeleton formation is poorly understood. To this end, we analyzed the SOM of the massive colonial coral, Porites australiensis, the skeleton of which has drawn great research interest because it records environmental conditions throughout the life of the colony. The coral skeleton was extensively cleaned, decalcified with acetic acid, and organic fractions were separated based on solubility. These fractions were analyzed using various techniques, including SDS-PAGE, FT-IR, in vitro crystallization, CHNS analysis, chromatography analysis of monosaccharide and enzyme-linked lectin assay (ELLA). We confirmed the acidic nature of SOM and the presence of sulphate, which is thought to initiate CaCO3 crystallization. In order to analyze glycan structures, we performed ELLA on the soluble SOM for the first time and found that it exhibits strong specificity to Datura stramonium lectin (DSL). Furthermore, using biotinylated DSL with anti-biotin antibody conjugated to nanogold, in situ localization of DSL-binding polysaccharides in the P. australiensis skeleton was performed. Signals were distributed on the surfaces of fiber-like crystals of the skeleton, suggesting that polysaccharides may modulate crystal shape. Our study emphasizes the importance of sugar moieties in biomineralization of scleractinian corals