Contributions of MyD88-Dependent Receptors and CD11c-Positive Cells to Corneal Epithelial Barrier Function Against Pseudomonas Aeruginosa

Previously we reported that corneal epithelial barrier function against Pseudomonas aeruginosa was MyD88-dependent. Here, we explored contributions of MyD88-dependent receptors using vital mouse eyes and confocal imaging. Uninjured IL-1R (−/−) or TLR4 (−/−) corneas, but not TLR2 (−/−), TLR5 (−/−), TLR7 (−/−), or TLR9 (−/−), were more susceptible to P. aeruginosa adhesion than wild-type (3.8-fold, 3.6-fold respectively). Bacteria adherent to the corneas of IL-1R (−/−) or TLR5 (−/−) mice penetrated beyond the epithelial surface only if the cornea was superficially-injured. Bone marrow chimeras showed that bone marrow-derived cells contributed to IL-1R-dependent barrier function. In vivo, but not ex vivo, stromal CD11c+ cells responded to bacterial challenge even when corneas were uninjured. These cells extended processes toward the epithelial surface, and co-localized with adherent bacteria in superficially-injured corneas. While CD11c+ cell depletion reduced IL-6, IL-1β, CXCL1, CXCL2 and CXCL10 transcriptional responses to bacteria, and increased susceptibility to bacterial adhesion (\u3e3-fold), the epithelium remained resistant to bacterial penetration. IL-1R (−/−) corneas also showed down-regulation of IL-6 and CXCL1 genes with and without bacterial challenge. These data show complex roles for TLR4, TLR5, IL-1R and CD11c+ cells in constitutive epithelial barrier function against P. aeruginosa, with details dependent upon in vivo conditions

A Role for the Adult Fat Body in Drosophila Male Courtship Behavior

Mating behavior in Drosophila depends critically on the sexual identity of specific regions in the brain, but several studies have identified courtship genes that express products only outside the nervous system. Although these genes are each active in a variety of non-neuronal cell types, they are all prominently expressed in the adult fat body, suggesting an important role for this tissue in behavior. To test its role in male courtship, fat body was feminized using the highly specific Larval serum protein promoter. We report here that the specific feminization of this tissue strongly reduces the competence of males to perform courtship. This effect is limited to the fat body of sexually mature adults as the feminization of larval fat body that normally persists in young adults does not affect mating. We propose that feminization of fat body affects the synthesis of male-specific secreted circulating proteins that influence the central nervous system. In support of this idea, we demonstrate that Takeout, a protein known to influence mating, is present in the hemolymph of adult males but not females and acts as a secreted protein

Restoration of CFTR function in patients with cystic fibrosis carrying the F508del-CFTR mutation

<div><p>Restoration of BECN1/Beclin 1-dependent autophagy and depletion of SQSTM1/p62 by genetic manipulation or autophagy-stimulatory proteostasis regulators, such as cystamine, have positive effects on mouse models of human cystic fibrosis (CF). These measures rescue the functional expression of the most frequent pathogenic CFTR mutant, F508del, at the respiratory epithelial surface and reduce lung inflammation in <i>Cftr^F508del</i> homozygous mice. Cysteamine, the reduced form of cystamine, is an FDA-approved drug. Here, we report that oral treatment with cysteamine greatly reduces the mortality rate and improves the phenotype of newborn mice bearing the <i>F508del-CFTR</i> mutation. Cysteamine was also able to increase the plasma membrane expression of the F508del-CFTR protein in nasal epithelial cells from <i>F508del</i> homozygous CF patients, and these effects persisted for 24 h after cysteamine withdrawal. Importantly, this cysteamine effect after washout was further sustained by the sequential administration of epigallocatechin gallate (EGCG), a green tea flavonoid, both <i>in vivo</i>, in mice, and <i>in vitro</i>, in primary epithelial cells from CF patients. In a pilot clinical trial involving 10 <i>F508del-CFTR</i> homozygous CF patients, the combination of cysteamine and EGCG restored BECN1, reduced SQSTM1 levels and improved CFTR function from nasal epithelial cells <i>in vivo</i>, correlating with a decrease of chloride concentrations in sweat, as well as with a reduction of the abundance of <i>TNF/TNF-alpha (tumor necrosis factor)</i> and <i>CXCL8</i> (<i>chemokine [C-X-C motif] ligand 8</i>) transcripts in nasal brushing and TNF and CXCL8 protein levels in the sputum. Altogether, these results suggest that optimal schedules of cysteamine plus EGCG might be used for the treatment of CF caused by the <i>F508del-CFTR</i> mutation.</p></div

Normal levels of p27Xic1 are necessary for somite segmentation and determining pronephric organ size

The Xenopus laevis cyclin dependent kinase inhibitor p27Xic1 has been shown to be involved in exit from the cell cycle and differentiation of cells into a quiescent state in the nervous system, muscle tissue, heart and retina. We show that p27Xic1 is expressed in the developing kidney in the nephrostomal regions. Using over-expression and morpholino oligonucleotide (MO) knock-down approaches we show normal levels of p27Xic1 regulate pronephros organ size by regulating cell cycle exit. Knock-down of p27Xic1 expression using a MO prevented myogenesis, as previously reported; an effect that subsequently inhibits pronephrogenesis. Furthermore, we show that normal levels of p27Xic1 are required for somite segmentation also through its cell cycle control function. Finally, we provide evidence to suggest correct paraxial mesoderm segmentation is not necessary for pronephric induction in the intermediate mesoderm. These results indicate novel developmental roles for p27Xic1, and reveal its differentiation function is not universally utilised in all developing tissues

Evidence for Smectite Clays from MSL SAM Analyses of Mudstone at Yellowknife Bay, Gale Crater, Mars

Drilled samples of mudstone from the Sheepbed unit at Yellowknife Bay were analyzed by MSL instruments including the Sample Analysis at Mars (SAM) and Chemistry and Mineralogy (CheMin) instruments in MSL's Analytical Laboratory. CheMin analyses revealed the first in situ X-ray diffraction based evidence of clay minerals on Mars, which are likely trioctahedral smectites (e.g., saponite) and comprise approx 20% of the mudstone sample (e.g., Bristow et al., this meeting). SAM analyses, which heated the mudstone samples to 1000 C and monitored volatiles evolved to perform in situ evolved gas analysis mass spectrometry (EGA-MS), resulted in a H2O trace exhibiting a wide evolution at temperatures < 500 C, and an evolution peak at higher temperatures near approx 750 C. The low temperature H2O evolution has many potential contributors, including adsorbed H2O, smectite interlayer H2O, and structural H2O/OH from bassanite and akaganeite (identified by CheMin) and H2O/OH from amorphous phases in the sample. The high temperature H2O is consistent with the evolution of H2O from the dehydroxylation of the smectite clay mineral. Comparison to EGA-MS data collected under SAM-like conditions on a variety of clay mineral reference materials indicate that a trioctahedral smectite, such as saponite, is most consistent with the high temperature H2O evolution observed. There may also be SAM EGA-MS evidence for a small high temperature H2O evolution from scoop samples from the Yellowknife Bay Rocknest sand shadow bedform. As in the mudstone samples, this evolution may indicate the detection of smectite clays, and the idea that minor clays may be present in Rocknest materials that could be expected to be at least partially derived from local sources is reasonable. But, because smectite clays were not definitively observed in CheMin analyses of Rocknest materials, they must be present at much lower abundances than the approx 20% observed in the mudstone samples. This potential detection underscores the complementary nature of the MSL CheMin and SAM instruments for investigations of martian sample mineralogy. Information on the nature of Yellowknife Bay clay minerals may also be available from the detection of H2 evolved during SAM EGA-MS at high temperature. A likely source of at least some of this H2 is H2O evolved from the smectite clays at high temperature, and it is possible these evolutions can be used in a similar fashion to high temperature H2O releases to provide constraints on the clay minerals in a sample. In addition, the D/H of this high temperature H2, as well as the H2O, can be derived from SAM MS and Tunable Laser Spectrometer (TLS) data, respectively. These D/H values may help to inform the provenance of high and low temperature water evolved from martian sample

First spectroscopic measurements of [OIII] emission from Lyman-alpha selected field galaxies at z ~ 3.1

We present the first spectroscopic measurements of the [OIII] 5007 A line in two z ~ 3.1 Lyman-alpha emitting galaxies (LAEs) using the new near-infrared instrument LUCIFER1 on the 8.4m Large Binocular Telescope (LBT). We also describe the optical imaging and spectroscopic observations used to identify these Lya emitting galaxies. Using the [OIII] line we have measured accurate systemic redshifts for these two galaxies, and discovered a velocity offset between the [OIII] and Ly-alpha lines in both, with the Lya line peaking 342 and 125 km/s redward of the systemic velocity. These velocity offsets imply that there are powerful outflows in high-redshift LAEs. They also ease the transmission of Lya photons through the interstellar medium and intergalactic medium around the galaxies. By measuring these offsets directly, we can refine both Lya-based tests for reionization, and Lya luminosity function measurements where the Lya forest affects the blue wing of the line. Our work also provides the first direct constraints on the strength of the [OIII] line in high-redshift LAEs. We find [OIII] fluxes of 7 and 36 x 10^-17 erg s^-1 cm^-2 in two z ~ 3.1 LAEs. These lines are strong enough to dominate broad-band flux measurements that include the line (in thiscase, K_s band photometry). Spectral energy distribution fits that do not account for the lines would therefore overestimate the 4000 A (and/or Balmer) break strength in such galaxies, and hence also the ages and stellar masses of such high-z galaxies.Comment: Accepted by the Astrophysical Journal. 11 pages, 3 figures, 1 table (emulateapj

Technology-Enhanced Reading Therapy for People With Aphasia: Findings From a Quasirandomized Waitlist Controlled Study.

Purpose This study investigated the effects of technology-enhanced reading therapy for people with reading impairments, using mainstream assistive reading technologies alongside reading strategies. Method The study used a quasirandomized waitlist controlled design. Twenty-one people with reading impairments following stroke were randomly assigned to receive 14 hr of therapy immediately or after a 6-week delay. During therapy, participants were trained to use assistive reading technology that offered a range of features to support reading comprehension. They developed skills in using the technology independently and in applying the technology to their personal reading goals. The primary outcome measure assessed reading comprehension, using Gray Oral Reading Test-Fourth Edition (GORT-4). Secondary measures were as follows: Reading Comprehension Battery for Aphasia-Second Edition, Reading Confidence and Emotions Questionnaire, Communication Activities of Daily Living-Second Edition, Visual Analog Mood Scales, and Assessment of Living With Aphasia. Matched texts were used with the GORT-4 to compare technology-assisted and unassisted reading comprehension. Mixed analyses of variance explored change between T1 and T2, when the immediate group had received therapy but the delayed group had not, thus serving as untreated controls. Pretherapy, posttherapy, and follow-up scores on the measures were also examined for all participants. Results GORT-4 results indicated that the immediately treated group improved significantly in technology-assisted reading following therapy, but not in unassisted reading. However, the data were not normally distributed, and secondary nonparametric analysis was not significant. The control group was unstable over the baseline, improving significantly in unassisted reading. The whole-group analysis showed significant gains in assisted (but not unassisted) reading after therapy that were maintained at follow-up. The Reading Confidence and Emotions Questionnaire results improved significantly following therapy, with good maintenance of change. Results on all other secondary measures were not significant. Conclusions Technology-assisted reading comprehension improved following the intervention, with treatment compensating for, rather than remediating, the reading impairment. Participants' confidence and emotions associated with reading also improved. Gains were achieved after 14 therapy sessions, using assistive technologies that are widely available and relatively affordable, meaning that this approach could be implemented in clinical practice

The Lick AGN Monitoring Project 2011: Dynamical Modeling of the Broad Line Region in Mrk 50

We present dynamical modeling of the broad line region (BLR) in the Seyfert 1 galaxy Mrk 50 using reverberation mapping data taken as part of the Lick AGN Monitoring Project (LAMP) 2011. We model the reverberation mapping data directly, constraining the geometry and kinematics of the BLR, as well as deriving a black hole mass estimate that does not depend on a normalizing factor or virial coefficient. We find that the geometry of the BLR in Mrk 50 is a nearly face-on thick disk, with a mean radius of 9.6(+1.2,-0.9) light days, a width of the BLR of 6.9(+1.2,-1.1) light days, and a disk opening angle of 25\pm10 degrees above the plane. We also constrain the inclination angle to be 9(+7,-5) degrees, close to face-on. Finally, the black hole mass of Mrk 50 is inferred to be log10(M(BH)/Msun) = 7.57(+0.44,-0.27). By comparison to the virial black hole mass estimate from traditional reverberation mapping analysis, we find the normalizing constant (virial coefficient) to be log10(f) = 0.78(+0.44,-0.27), consistent with the commonly adopted mean value of 0.74 based on aligning the M(BH)-{\sigma}* relation for AGN and quiescent galaxies. While our dynamical model includes the possibility of a net inflow or outflow in the BLR, we cannot distinguish between these two scenarios.Comment: Accepted for publication in ApJ. 8 pages, 6 figure

Low-lying fermion modes, topology and light hadrons in quenched QCD

We explore the properties of low lying eigenmodes of fermions in the quenched approximation of lattice QCD. The fermion action is a recently proposed overlap action and has exact chiral symmetry. We find that chiral zero-eigenvalue modes are localized in space and their positions correlate strongly with the locations (as defined through the density of pure gauge observables) of instantons of the appropriate charge. Nonchiral modes are also localized with peaks which are strongly correlated with the positions of both charges of instantons. These correlations slowly die away as the fermion eigenvalue rises. Correlators made of quark propagators restricted to these modes closely reproduce ordinary hadron correlators at small quark mass in many channels. Our results are in qualitative agreement with the expectations of instanton liquid models.Comment: 21 pages, Revtex, 21 postscript figures. COLO-HEP-45