Quantitative analysis of hypertrophic myocardium using diffusion tensor magnetic resonance imaging

Systemic hypertension is a causative factor in left ventricular hypertrophy (LVH). This study is motivated by the potential to reverse or manage the dysfunction associated with structural remodeling of the myocardium in this pathology. Using diffusion tensor magnetic resonance imaging, we present an analysis of myocardial fiber and laminar sheet orientation in ex vivo hypertrophic (6 SHR) and normal (5 WKY) rat hearts using the covariance of the diffusion tensor. First, an atlas of normal cardiac microstructure was formed using the WKY b0 images. Then, the SHR and WKY b0 hearts were registered to the atlas. The acquired deformation fields were applied to the SHR and WKY heart tensor fields followed by the preservation of principal direction (PPD) reorientation strategy. A mean tensor field was then formed from the registered WKY tensor images. Calculating the covariance of the registered tensor images about this mean for each heart, the hypertrophic myocardium exhibited significantly increased myocardial fiber derangement (p ¼ 0.017) with a mean dispersion of 38.7 deg, and an increased dispersion of the laminar sheet normal (p = 0.030) of 54.8 deg compared with 34.8 deg and 51.8 deg, respectively, in the normal hearts. Results demonstrate significantly altered myocardial fiber and laminar sheet structure in rats with hypertensive LVH

Sox9 Transcriptionally Represses Spp1 to Prevent Matrix Mineralization in Maturing Heart Valves and Chondrocytes

Sox9 is an SRY-related transcription factor required for expression of cartilaginous genes in the developing skeletal system and heart valve structures. In contrast to positively regulating cartilaginous matrix, Sox9 also negatively regulates matrix mineralization associated with bone formation. While the transcriptional activation of Sox9 target genes during chondrogenesis has been characterized, the mechanisms by which Sox9 represses osteogenic processes are not so clear. Using ChIP-on-chip and luciferase assays we show that Sox9 binds and represses transactivation of the osteogenic glycoprotein Spp1. In addition, Sox9 knockdown in post natal mouse heart valve explants and rib chondrocyte cultures promotes Spp1 expression and matrix mineralization, while attenuating expression of cartilage genes Type II Collagen and Cartilage Link Protein. Further, we show that Spp1 is required for matrix mineralization induced by Sox9 knockdown. These studies provide insights into the molecular mechanisms by which Sox9 prevents pathologic matrix mineralization in tissues that must remain cartilaginous

Fundamental insight into the effect of carbodiimide crosslinking on cellular recognition of collagen-based scaffolds

Research on the development of collagen constructs is extremely important in the field of tissue engineering. Collagen scaffolds for numerous tissue engineering applications are frequently crosslinked with 1-ethyl-3-(3-dimethylaminopropyl-carbodiimide hydrochloride (EDC) in the presence of N-hydroxy-succinimide (NHS). Despite producing scaffolds with good biocompatibility and low cellular toxicity the influence of EDC/NHS crosslinking on the cell interactive properties of collagen has been overlooked. Here we have extensively studied the interaction of model cell lines with collagen I-based materials after crosslinking with different ratios of EDC in relation to the number of carboxylic acid residues on collagen. Divalent cation-dependent cell adhesion, via integrins α$_{1}$β$_{1}$, α$_{2}$β$_{1}$, α$_{10}$β$_{1}$ and α$_{11}$β$_{1}$, were sensitive to EDC crosslinking. With increasing EDC concentration, this was replaced with cation-independent adhesion. These results were replicated using purified recombinant I domains derived from integrin α$_{1}$ and α$_{2}$ subunits. Integrin α$_{2}$β$_{1}$-mediated cell spreading, apoptosis and proliferation were all heavily influenced by EDC crosslinking of collagen. Data from this rigorous study provides an exciting new insight that EDC/NHS crosslinking is utilising the same carboxylic side chain chemistry that is vital for native-like integrin-mediated cell interactions. Due to the ubiquitous usage of EDC/NHS crosslinked collagen for biomaterials fabrication this data is essential to have a full understanding in order to ensure optimized collagen-based material performance.This work was supported by the British Heart Foundation (Grant NH/11/1/28922, RG/15/4/31268, SP/15/7/31561 and RG/09/003/27122) and the ERC Advanced Grant 320598 3D-E. D. V. Bax is funded by the Peoples Programme of the EU 7th Framework Programme (RAE no: PIIF-GA-2013-624904) and was supported by an EPSRC IKC Proof of Concept Award

SPT0346-52: Negligible AGN Activity in a Compact, Hyper-starburst Galaxy at z = 5.7

We present Chandra ACIS-S and ATCA radio continuum observations of the strongly lensed dusty, star-forming galaxy SPT-S J034640-5204.9 (hereafter SPT0346-52) at $z$ = 5.656. This galaxy has also been observed with ALMA, HST, Spitzer, Herschel, APEX, and the VLT. Previous observations indicate that if the infrared (IR) emission is driven by star formation, then the inferred lensing-corrected star formation rate ($\sim$ 4500 M_{\sun} yr$^{-1}$) and star formation rate surface density $\Sigma_{\rm SFR}$ ($\sim$ 2000 M_{\sun} {yr^{-1}} {kpc^{-2}}) are both exceptionally high. It remained unclear from the previous data, however, whether a central active galactic nucleus (AGN) contributes appreciably to the IR luminosity. The {\it Chandra} upper limit shows that SPT0346-52 is consistent with being star-formation dominated in the X-ray, and any AGN contribution to the IR emission is negligible. The ATCA radio continuum upper limits are also consistent with the FIR-to-radio correlation for star-forming galaxies with no indication of an additional AGN contribution. The observed prodigious intrinsic IR luminosity of (3.6 $\pm$ 0.3) $\times$ 10$^{13}$ L_{\sun} originates almost solely from vigorous star formation activity. With an intrinsic source size of 0.61 $\pm$ 0.03 kpc, SPT0346-52 is confirmed to have one of the highest $\Sigma_{SFR}$ of any known galaxy. This high $\Sigma_{SFR}$, which approaches the Eddington limit for a radiation pressure supported starburst, may be explained by a combination of very high star formation efficiency and gas fraction.Comment: 8 pages, 6 figures, accepted for publication in Ap

Highly sensitive and specific protein detection via combined capillary isoelectric focusing and proximity ligation

Detection and quantification of proteins and their post-translational modifications are crucial to decipher functions of complex protein networks in cell biology and medicine. Capillary isoelectric focusing together with antibody-based detection can resolve and identify proteins and their isoforms with modest sample input. However, insufficient sensitivity prevents detection of proteins present at low concentrations and antibody cross-reactivity results in unspecific detection that cannot be distinguished from bona fide protein isoforms. By using DNA-conjugated antibodies enhanced signals can be obtained via rolling circle amplification (RCA). Both sensitivity and specificity can be greatly improved in assays dependent on target recognition by pairs of antibodies using in situ proximity ligation assays (PLA). Here we applied these DNA-assisted RCA techniques in capillary isoelectric focusing to resolve endogenous signaling transducers and isoforms along vascular endothelial growth factor (VEGF) signaling pathways at concentrations too low to be detected in standard assays. We also demonstrate background rejection and enhanced specificity when protein detection depended on binding by pairs of antibodies using in situ PLA, compared to assays where each antibody preparation was used on its own.</p

Highly sensitive and specific protein detection via combined capillary isoelectric focusing and proximity ligation

Detection and quantification of proteins and their post-translational modifications are crucial to decipher functions of complex protein networks in cell biology and medicine. Capillary isoelectric focusing together with antibody-based detection can resolve and identify proteins and their isoforms with modest sample input. However, insufficient sensitivity prevents detection of proteins present at low concentrations and antibody cross-reactivity results in unspecific detection that cannot be distinguished from bona fide protein isoforms. By using DNA-conjugated antibodies enhanced signals can be obtained via rolling circle amplification (RCA). Both sensitivity and specificity can be greatly improved in assays dependent on target recognition by pairs of antibodies using in situ proximity ligation assays (PLA). Here we applied these DNA-assisted RCA techniques in capillary isoelectric focusing to resolve endogenous signaling transducers and isoforms along vascular endothelial growth factor (VEGF) signaling pathways at concentrations too low to be detected in standard assays. We also demonstrate background rejection and enhanced specificity when protein detection depended on binding by pairs of antibodies using in situ PLA, compared to assays where each antibody preparation was used on its own.</p

The Rest-Frame Submillimeter Spectrum of High-Redshift, Dusty, Star-Forming Galaxies

We present the average rest-frame spectrum of high-redshift dusty, star-forming galaxies from 250-770GHz. This spectrum was constructed by stacking ALMA 3mm spectra of 22 such sources discovered by the South Pole Telescope and spanning z=2.0-5.7. In addition to multiple bright spectral features of 12CO, [CI], and H2O, we also detect several faint transitions of 13CO, HCN, HNC, HCO+, and CN, and use the observed line strengths to characterize the typical properties of the interstellar medium of these high-redshift starburst galaxies. We find that the 13CO brightness in these objects is comparable to that of the only other z>2 star-forming galaxy in which 13CO has been observed. We show that the emission from the high-critical density molecules HCN, HNC, HCO+, and CN is consistent with a warm, dense medium with T_kin ~ 55K and n_H2 >~ 10^5.5 cm^-3. High molecular hydrogen densities are required to reproduce the observed line ratios, and we demonstrate that alternatives to purely collisional excitation are unlikely to be significant for the bulk of these systems. We quantify the average emission from several species with no individually detected transitions, and find emission from the hydride CH and the linear molecule CCH for the first time at high redshift, indicating that these molecules may be powerful probes of interstellar chemistry in high-redshift systems. These observations represent the first constraints on many molecular species with rest-frame transitions from 0.4-1.2mm in star-forming systems at high redshift, and will be invaluable in making effective use of ALMA in full science operations.Comment: 19 pages, 10 figures (2 in appendices); accepted for publication in Ap

The redshift distribution of dusty star forming galaxies from the SPT survey

We use the Atacama Large Millimeter/submillimeter Array (ALMA) in Cycle 1 to determine spectroscopic redshifts of high-redshift dusty star-forming galaxies (DSFGs) selected by their 1.4mm continuum emission in the South Pole Telescope (SPT) survey. We present ALMA 3mm spectral scans between 84-114GHz for 15 galaxies and targeted ALMA 1mm observations for an additional eight sources. Our observations yield 30 new line detections from CO, [CI] , [NII] , H_2O and NH_3. We further present APEX [CII] and CO mid-J observations for seven sources for which only a single line was detected in spectral-scan data from ALMA Cycle 0 or Cycle 1. We combine the new observations with previously published and new mm/submm line and photometric data of the SPT-selected DSFGs to study their redshift distribution. The combined data yield 39 spectroscopic redshifts from molecular lines, a success rate of >85%. Our sample represents the largest data set of its kind today and has the highest spectroscopic completeness among all redshift surveys of high-z DSFGs. The median of the redshift distribution is z=3.9+/-0.4, and the highest-redshift source in our sample is at z=5.8. We discuss how the selection of our sources affects the redshift distribution, focusing on source brightness, selection wavelength, and strong gravitational lensing. We correct for the effect of gravitational lensing and find the redshift distribution for 1.4mm-selected sources with a median redshift of z=3.1+/-0.3. Comparing to redshift distributions selected at shorter wavelengths from the literature, we show that selection wavelength affects the shape of the redshift distribution

The Dragonfly Galaxy: II. ALMA unveils a triple merger and gas exchange in a hyper-luminous radio galaxy at z = 2

The Dragonfly Galaxy (MRC 0152-209), at redshift z ~  2, is one of the most vigorously star-forming radio galaxies in the Universe. What triggered its activity? We present ALMA Cycle 2 observations of cold molecular CO(6−5) gas and dust, which reveal that this is likely a gas-rich triple merger. It consists of a close double nucleus (separation ~4 kpc) and a weak CO-emitter at ~10  kpc distance, all of which have counterparts in HST/NICMOS imagery. The hyper-luminous starburst and powerful radio-AGN were triggered at this precoalescent stage of the merger. The CO(6−5) traces dense molecular gas in the central region, and complements existing CO(1−0) data, which reveal more widespread tidal debris of cold gas. We also find ~1010 M☉ of molecular gas with enhanced excitation at the highest velocities. At least 20−50% of this high-excitation, high-velocity gas shows kinematics that suggests it is being displaced and redistributed within the merger, although with line-of-sight velocities of |v| < 500 km s-1, this gas will probably not escape the system. The processes that drive the redistribution of cold gas are likely related to either the gravitational interaction between two kpc-scale discs, or starburst/AGN-driven outflows. We estimate that the rate at which the molecular gas is redistributed is at least [Ṁentity!#x2009!]~ 1200 ± 500 M☉ yr-1, and could perhaps even approach the star formation rate of ~3000 ± 800 M☉ yr-1. The fact that the gas depletion and gas redistribution timescales are similar implies that dynamical processes can be important in the evolution of massive high-z galaxies

The Role of Repeated Exposure to Multimodal Input in Incidental Acquisition of Foreign Language Vocabulary

Prior research has reported incidental vocabulary acquisition with complete beginners in a foreign language (FL), within 8 exposures to auditory and written FL word forms presented with a picture depicting their meaning. However, important questions remain about whether acquisition occurs with fewer exposures to FL words in a multimodal situation and whether there is a repeated exposure effect. Here we report a study where the number of exposures to FL words in an incidental learning phase varied between 2, 4, 6, and 8 exposures. Following the incidental learning phase, participants completed an explicit learning task where they learned to recognize written translation equivalents of auditory FL word forms, half of which had occurred in the incidental learning phase. The results showed that participants performed better on the words they had previously been exposed to, and that this incidental learning effect occurred from as little as 2 exposures to the multimodal stimuli. In addition, repeated exposure to the stimuli was found to have a larger impact on learning during the first few exposures and decrease thereafter, suggesting that the effects of repeated exposure on vocabulary acquisition are not necessarily constant