Characterization of osteopontin charge forms produced by osteoblasts

Osteopontin (OPN) is a highly-phosphorylated extracellular matrix protein localized in bone, kidney, placenta, T-lymphocytes, macrophages, smooth muscle of the vascular system, milk, urine, and plasma. In ROS 17/2.8 osteoblast-like osteosarcoma cells, 1,25-dihydroxyvitamin D3 [1,25(OH)2D 3] regulates OPN at the transcriptional level resulting in increased steady state mRNA levels and increased production of OPN protein, maximal at 48 hours. Using ROS 17/2.8 cells as an osteoblast model, OPN was purified from culture medium after three hour treatments of either vehicle (ethanol) or 1,25(OH)2D3 via barium citrate precipitation followed by immunoaffinity chromatography. ^ Here, further evidence of regulation of OPN by 1,25(OH)2D 3 at the posttranslational level is presented. Prior to the up-regulation of OPN at the transcriptional level, 1,25(OH)2D3 induces a shift in OPN isoelectric point (pI) detected on two-dimensional gels from pI 4.6 to pI 5.1. Loading equal amounts of [32P]-labeled OPN recovered from ROS 17/2.8 cells exposed to 1,25(OH)2D3 or vehicle alone for three hours reveals that the shift from pI 4.6 to 5.1 is the result of reduced phosphorylation. Using structural analogs to 1,25(OH) 2D3, analog AT [25-(OH)-16-ene-23-yne-D3], which triggers Ca2+ influx through voltage sensitive Ca2+ channels but does not bind to the vitamin D receptor, mimicked the OPN pI shift while analog BT [1,25(OH)2-22-ene-24-cyclopropyl-D 3], which binds to the vitamin D receptor but does not allow Ca 2+ influx, did not. Inclusion of the Ca2+ channel blocker nifedipine also blocks the charge shift conversion of OPN. Further analysis of the signaling pathway initiated by 1,25(OH)2D3 reveals that inhibition of the cyclic 3′,5′ -adenosine monophosphate-dependent kinase, protein kinase A, or inhibition of the cyclic 3′,5′-guanine monophosphate-dependent kinase, protein kinase G, also prevents the charge shift conversion. ^ Isolation of OPN from rat femurs and tibiae provides evidence for the existence of these two OPN charge forms in vivo, evidenced by differential migration on isoelectric focusing gels and sodium dodecyl sulfate-polyacrylamide gels. Peptide sequencing of rat long bone fractions revealed the presence of a presumed dentin specific protein, dentin matrix protein-1 (DMP-1). Western blot analysis confirmed the existence of DMP-1 in these fractions. ^ Using the OPN charge forms in functional assays, it was determined that the charge forms have differential roles in both cell surface and mineralization functions. In cell attachment assays and Ca2+ influx assays using PC-3 prostate cancer cells, the pI 5.1 charge form of OPN was found to permit binding and increase intracellular Ca2+ concentrations of PC-3 cells. The increase in intracellular Ca2+ concentration was found to be integrin αvβ3-dependent. In mineralization assays, the pI 4.6 charge form of OPN promoted hydroxyapatite formation, while the pI 5.1 charge form had improved Ca2+ binding ability. ^ In conclusion, these findings suggest that 1,25(OH) 2D3 regulates OPN not only at the transcriptional level, but also plays a role in determination of the OPN phosphorylation state. The latter involves a short term (less than three hours) treatment and is associated with membrane-initiated Ca2+ influx. Functional assays utilizing the two OPN charge forms reveal the dependence of OPN post-translational state on its function. ^

Optimum Electron Temperature and Density for Short-Wavelength Plasma-Lasing from Nickel-like ions

Soft X-ray lasing across a Ni-like plasma gain-medium requires optimum electron temperature and density for attaining to the Ni-like ion stage and for population inversion in the View the MathML source3d94d1(J=0)→3d94p1(J=1) laser transition. Various scaling laws, function of operating parameters, were compared with respect to their predictions for optimum temperatures and densities. It is shown that the widely adopted local thermodynamic equilibrium (LTE) model underestimates the optimum plasma-lasing conditions. On the other hand, non-LTE models, especially when complemented with dielectronic recombination, provided accurate prediction of the optimum plasma-lasing conditions. It is further shown that, for targets with Z equal or greater than the rare-earth elements (e.g. Sm), the optimum electron density for plasma-lasing is not accessible for pump-pulses at View the MathML sourceλ=1ω=1μm. This observation explains a fundamental difficulty in saturating the wavelength of plasma-based X-ray lasers below 6.8 nm, unless using 2ω2ω pumping.

Covalent immobilisation of VEGF on plasma-coated electrospun scaffolds for tissue engineering applications.

Recent findings in the field of biomaterials and tissue engineering provide evidence that surface immobilised growth factors display enhanced stability and induce prolonged function. Cell response can be regulated by material properties and at the site of interest. To this end, we developed scaffolds with covalently bound vascular endothelial growth factor (VEGF) and evaluated their mitogenic effect on endothelial cells in vitro. Nano- (254±133 nm) or micro-fibrous (4.0±0.4 μm) poly(ɛ-caprolactone) (PCL) non-wovens were produced by electrospinning and coated in a radio frequency (RF) plasma process to induce an oxygen functional hydrocarbon layer. Implemented carboxylic acid groups were converted into amine-reactive esters and covalently coupled to VEGF by forming stable amide bonds (standard EDC/NHS chemistry). Substrates were analysed by X-ray photoelectron spectroscopy (XPS), enzyme-linked immuno-assays (ELISA) and immunohistochemistry (anti-VEGF antibody and VEGF-R2 binding). Depending on the reaction conditions, immobilised VEGF was present at 127±47 ng to 941±199 ng per substrate (6mm diameter; concentrations of 4.5 ng mm(-2) or 33.3 ng mm(-2), respectively). Immunohistochemistry provided evidence for biological integrity of immobilised VEGF. Endothelial cell number of primary endothelial cells or immortalised endothelial cells were significantly enhanced on VEGF-functionalised scaffolds compared to native PCL scaffolds. This indicates a sustained activity of immobilised VEGF over a culture period of nine days. We present a versatile method for the fabrication of growth factor-loaded scaffolds at specific concentrations.

Backscattered energetic neutral atoms from the Moon in the Earth's plasma sheet observed by Chandarayaan-1/Sub-keV Atom Reflecting Analyzer instrument

We present the observations of energetic neutral atoms (ENAs) produced at the lunar surface in the Earth's magnetotail. When the Moon was located in the terrestrial plasma sheet, Chandrayaan-1 Energetic Neutrals Analyzer (CENA) detected hydrogen ENAs from the Moon. Analysis of the data from CENA together with the Solar Wind Monitor (SWIM) onboard Chandrayaan-1 reveals the characteristic energy of the observed ENA energy spectrum (the e-folding energy of the distribution function) ∼100 eV and the ENA backscattering ratio (defined as the ratio of upward ENA flux to downward proton flux) <∼0.1. These characteristics are similar to those of the backscattered ENAs in the solar wind, suggesting that CENA detected plasma sheet particles backscattered as ENAs from the lunar surface. The observed ENA backscattering ratio in the plasma sheet exhibits no significant difference in the Southern Hemisphere, where a large and strong magnetized region exists, compared with that in the Northern Hemisphere. This is contrary to the CENA observations in the solar wind, when the backscattering ratio drops by ∼50% in the Southern Hemisphere. Our analysis and test particle simulations suggest that magnetic shielding of the lunar surface in the plasma sheet is less effective than in the solar wind due to the broad velocity distributions of the plasma sheet protons.

Impact of platelet rich plasma and adipose stem cells on lymphangiogenesis in a murine tail lymphedema model.

BACKGROUND Lymphedema is an underdiagnosed pathology which in industrialized countries mainly affects cancer patients that underwent lymph node dissection and/or radiation. Currently no effective therapy is available so that patients' life quality is compromised by swellings of the concerned body region. This unfortunate condition is associated with body imbalance and subsequent osteochondral deformations and impaired function as well as with an increased risk of potentially life threatening soft tissue infections. METHODS The effects of PRP and ASC on angiogenesis (anti-CD31 staining), microcirculation (Laser Doppler Imaging), lymphangiogenesis (anti-LYVE1 staining), microvascular architecture (corrosion casting) and wound healing (digital planimetry) are studied in a murine tail lymphedema model. RESULTS Wounds treated by PRP and ASC healed faster and showed a significantly increased epithelialization mainly from the proximal wound margin. The application of PRP induced a significantly increased lymphangiogenesis while the application of ASC did not induce any significant change in this regard. CONCLUSIONS PRP and ASC affect lymphangiogenesis and lymphedema development and might represent a promising approach to improve regeneration of lymphatic vessels, restore disrupted lymphatic circulation and treat or prevent lymphedema alone or in combination with currently available lymphedema therapies.

Geochemistry and oxygen and iron isotope ratios of mafic rocks

Recycling of oceanic crust into the deep mantle via subduction is a widely accepted mechanism for creating compositional heterogeneity in the upper mantle and for explaining the distinct geochemistry of mantle plumes. The oxygen isotope ratios (d18O) of some ocean island basalts (OIB) span values both above and below that of unmetasomatised upper mantle (5.5 ± 0.4 per mil) and provide support for this hypothesis, as it is widely assumed that most variations in d18O are produced by near-surface low-temperature processes. Here we show a significant linear relationship between d18O and stable iron isotope ratios (d57Fe) in a suite of pristine eclogite xenoliths. The d18O values of both bulk samples and garnets range from values within error of normal mantle to significantly lighter values. The observed range and correlation between d18O and d57Fe is unlikely to be inherited from oceanic crust, as d57Fe values determined for samples of hydrothermally altered oceanic crust do not differ significantly from the mantle value and show no correlation with d18O. It is proposed that the correlated d57Fe and d18O variations in this particular eclogite suite are predominantly related to isotopic fractionation by disequilibrium partial melting although modification by melt percolation processes cannot be ruled out. Fractionation of Fe and O isotopes by removal of partial melt enriched in isotopically heavy Fe and O is supported by negative correlations between bulk sample d57Fe and Cr content and bulk sample and garnet d18O and Sc contents, as Cr and Sc are elements that become enriched in garnet- and pyroxene-bearing melt residues. Melt extraction could take place either during subduction, where the eclogites represent the residues of melted oceanic lithosphere, or could take place during long-term residence within the lithospheric mantle, in which case the protoliths of the eclogites could be of either crustal or mantle origin. This modification of both d57Fe and d18O by melting processes and specifically the production of low-d18O signatures in mafic rocks implies that some of the isotopically light d18O values observed in OIB and eclogite xenoliths may not necessarily reflect near-surface processes or components.

Geochemistry of Indian Ocean tephra and Afro-Arabian lavas

Widespread silicic pyroclastic eruptions of the Oligocene Afro-Arabian flood volcanic province (ignimbrites and airfall tuffs) produced up to 20% of the total flood volcanic stratigraphy (>6*10**4 km**3). Volumes of individual ignimbrites and tuffs exposed on land range from ~150 to >2000 km**3 and eight major units (15-100 m thick) were erupted in <2 Myr, placing these amongst the largest-magnitude silicic pyroclastic eruptions on Earth. They are compositionally distinctive time-stratigraphic markers which were deposited as co-ignimbrite ashfall deposits on a near-global scale around the time of the Oi2 cooling anomaly at ~30 Ma. Two ignimbrites from the lower part of the flood volcanic succession in Yemen have been correlated to: (a) the conjugate rifted margin of Ethiopia (>500 km distant); and (b) to two deep sea ash layers sampled by ODP Leg 115 in the Indian Ocean ~2700 km to the southeast. This correlation is based on whole rock analyses of silicic units for isotope ratios (Pb, Nd) and rare earth element compositions, in conjunction with novel in situ Pb isotope laser ablation multicollector inductively coupled plasma mass spectroscopy analysis of groundmass and glass shards. Compositional diversity preserved on the scale of individual ash shards in these deep sea tephra layers record chemical heterogeneity present in the silicic magma chambers that is not evident in the welded on-land deposits. Ages of the ash layers can be established by correlation to precisely dated on-land ignimbrites, and current evidence suggests that while these eruptions may have exacerbated already changing climatic conditions, they both marginally post-date the Oi2 global cooling anomaly.

Properties of seawater and particulate matter from a WETLabs ALFA hyperspectral laser spectrofluorometer mounted on the continuous surface water sampling system during the Tara Oceans expedition 2009-2013

The Tara Oceans Expedition (2009-2013) sampled the world oceans on board a 36 m long schooner, collecting environmental data and organisms from viruses to planktonic metazoans for later analyses using modern sequencing and state-of-the-art imaging technologies. Tara Oceans Data are particularly suited to study the genetic, morphological and functional diversity of plankton. The present data set provides continuous measurements made with an Aquatic Laser Fluorescence Analyzer (ALFA) (Chekalyuk et al., 2014), connected in-line to the TARA flow through system during 2013. The ALFA instrument provides dual-wavelength excitation (405 and 514 nm) of laser-stimulated emission (LSE) for spectral and temporal analysis. It offers in vivo fluorescence assessments of phytoplankton pigments, biomass, photosynthetic yield (Fv/Fm), phycobiliprotein (PBP)-containing phytoplankton groups, and chromophoric dissolved organic matter (CDOM) (Chekalyuk and Hafez, 2008; 2013A). Spectral deconvolution (SDC) is used to assess the overlapped spectral bands of aquatic fluorescence constituents and water Raman scattering (R). The Fv/Fm measurements are spectrally corrected for non-chlorophyll fluorescence background produced by CDOM and other constituents (Chekalyuk and Hafez, 2008). The sensor was cleaned weakly following the manufacturer recommended protocol.

Laser ablation data of benthic foraminifera from surface sediments collected from the muddy intertidal flats near Dorum, Northwestern Germany in Autumn 2008

Biological activity introduces variability in element incorporation during calcification and thereby decreases the precision and accuracy when using foraminifera as geochemical proxies in paleoceanography. This so-called 'vital effect' consists of organismal and environmental components. Whereas organismal effects include uptake of ions from seawater and subsequent processing upon calcification, environmental effects include migration- and seasonality-induced differences. Triggering asexual reproduction and culturing juveniles of the benthic foraminifer Ammonia tepida under constant, controlled conditions allow environmental and genetic variability to be removed and the effect of cell-physiological controls on element incorporation to be quantified. Three groups of clones were cultured under constant conditions while determining their growth rates, size-normalized weights and single-chamber Mg/Ca and Sr/Ca using laser ablation-inductively coupled plasma-mass spectrometry (LA-ICP-MS). Results show no detectable ontogenetic control on the incorporation of these elements in the species studied here. Despite constant culturing conditions, Mg/Ca varies by a factor of similar to 4 within an individual foraminifer while intra-individual Sr/Ca varies by only a factor of 1.6. Differences between clone groups were similar to the intra-clone group variability in element composition, suggesting that any genetic differences between the clone-groups studied here do not affect trace element partitioning. Instead, variability in Mg/Ca appears to be inherent to the process of bio-calcification itself. The variability in Mg/Ca between chambers shows that measurements of at least 6 different chambers are required to determine the mean Mg/Ca value for a cultured foraminiferal test with a precision of <= 10%

Bloche-Maxwell treatment of amplification of high harmonic seed in soft x-ray laser amplifiers in both direct and chirped amplifications

Seeding plasma-based softx-raylaser (SXRL) demonstrated diffraction-limited, fully coherent in space and in time beam but with energy not exceeding 1 μJ per pulse. Quasi-steady-state (QSS) plasmas demonstrated to be able to store high amount of energy and then amplify incoherent SXRL up to several mJ. Using 1D time-dependant Bloch–Maxwell model including amplification of noise, we demonstrated that femtosecond HHG cannot be efficiently amplified in QSS plasmas. However, using Chirped Pulse Amplification concept on HHG seed allows to extract most of the stored energy, reaching up to 5 mJ in fully coherent pulses that can be compressed down to 130 fs.

Comparison of natural and forced amplification regimes in plasma-based soft-x-ray lasers seeded by high-order harmonic

The amplification of high-order harmonics (HOH) in a plasma-based amplifier is a multiscale, temporal phenomenon that couples plasma hydrodynamics, atomic processes, and HOH electromagnetic fields. We use a one-dimensional, time-dependent Maxwell-Bloch code to compare the natural amplification regime and another regime where plasma polarization is constantly forced by the HOH. In this regime, a 10-MW (i.e., 100 times higher than current seeded soft x-ray laser power), 1.5-μJ, 140-fs pulse free from the parasitic temporal structures appearing on the natural amplification regime can be obtained.

An overview on armor research for the laser fusion project HiPER

During the current preparatory phase of the European laser fusion project HiPER, an intensive effort has being placed to identify an armour material able to protect the internal walls of the chamber against the high thermal loads and high fluxes of x-rays and ions produced during the fusion explosions. This poster addresses the different threats and limitations of a poly-crystalline Tungsten armour. The analysis is carried out under the conditions of an experimental chamber hypothetically constructed to demonstrate laser fusion in a repetitive mode, subjected to a few thousand 48MJ shock ignition shots during its entire lifetime. If compared to the literature, an extrapolation of the thermomechanical and atomistic effects obtained from the simulations of the experimental chamber to the conditions of a Demo reactor (working 24/7 at hundreds of MW) or a future power plant (producing GW) suggests that “standard” tungsten will not be a suitable armour. Thus, new materials based on nano-structured W and C are being investigated as possible candidates. The research programme launched by the HiPER material team is introduced.

Stochastic homogenization of the laser intensity to improve the irradiation uniformity of capsules directly driven by thousands laser beams

Illumination uniformity of a spherical capsule directly driven by laser beams has been assessed numerically. Laser facilities characterized by ND = 12, 20, 24, 32, 48 and 60 directions of irradiation with associated a single laser beam or a bundle of NB laser beams have been considered. The laser beam intensity profile is assumed super-Gaussian and the calculations take into account beam imperfections as power imbalance and pointing errors. The optimum laser intensity profile, which minimizes the root-mean-square deviation of the capsule illumination, depends on the values of the beam imperfections. Assuming that the NB beams are statistically independents is found that they provide a stochastic homogenization of the laser intensity associated to the whole bundle, reducing the errors associated to the whole bundle by the factor  , which in turn improves the illumination uniformity of the capsule. Moreover, it is found that the uniformity of the irradiation is almost the same for all facilities and only depends on the total number of laser beams Ntot = ND × NB.

Optimization of laser-firing processes for silicon-heterojunction solar-cell back contacts

One of the key steps to achieve high efficiencies in amorphous/crystalline silicon photovoltaic structures is to design low-ohmic-resistance backcontacts with good passivation in the rear part of the cell. A well known approach to achieve this goal is to use laser-fired contact (LFC) processes in which a metal layer is fired through the dielectric to define good contacts with the semiconductor. However, and despite the fact that this approach has demonstrated to be extremely successful, there is still enough room for process improvement with an appropriate optimization. In this paper, a study focused on the optimal adjustment of the irradiation parameters to produce laser-fired contacts in a-Si:H/c-Si heterojunctionsolarcells is presented. We used samples consisting of crystalline-silicon (c-Si) wafers together with a passivation layer of intrinsic hydrogenated amorphous silicon (a-Si:H(i)) deposited by plasma-enhanced chemical deposition (PECVD). Then, an aluminum layer was evaporated on both sides, the thickness of this layer varied from 0.2 to 1 μm in order to identify the optimal amount of Al required to create an appropriate contact. A q-switched Nd:YVO4laser source, λ = 532 nm, was used to locally fire the aluminum through the thin a-Si:H(i)-layers to form the LFC. The effects of laser fluences were analyzed using a comprehensive morphological and electrical characterization.

Plasma–wall interaction in laser inertial fusion reactors: novel proposals for radiation tests of first wall materials

Dry-wall laser inertial fusion (LIF) chambers will have to withstand strong bursts of fast charged particles which will deposit tens of kJ m−2 and implant more than 1018 particles m−2 in a few microseconds at a repetition rate of some Hz. Large chamber dimensions and resistant plasma-facing materials must be combined to guarantee the chamber performance as long as possible under the expected threats: heating, fatigue, cracking, formation of defects, retention of light species, swelling and erosion. Current and novel radiation resistant materials for the first wall need to be validated under realistic conditions. However, at present there is a lack of facilities which can reproduce such ion environments. This contribution proposes the use of ultra-intense lasers and high-intense pulsed ion beams (HIPIB) to recreate the plasma conditions in LIF reactors. By target normal sheath acceleration, ultra-intense lasers can generate very short and energetic ion pulses with a spectral distribution similar to that of the inertial fusion ion bursts, suitable to validate fusion materials and to investigate the barely known propagation of those bursts through background plasmas/gases present in the reactor chamber. HIPIB technologies, initially developed for inertial fusion driver systems, provide huge intensity pulses which meet the irradiation conditions expected in the first wall of LIF chambers and thus can be used for the validation of materials too.