Quantitative nucleotide level analysis of regulation of translation in response to depolarization of cultured neural cells

Studies on regulation of gene expression have contributed substantially to understanding mechanisms for the long-term activity-dependent alterations in neural connectivity that are thought to mediate learning and memory. Most of these studies, however, have focused on the regulation of mRNA transcription. Here, we utilized high-throughput sequencing coupled with ribosome footprinting to globally characterize the regulation of translation in primary mixed neuronal-glial cultures in response to sustained depolarization. We identified substantial and complex regulation of translation, with many transcripts demonstrating changes in ribosomal occupancy independent of transcriptional changes. We also examined sequence-based mechanisms that might regulate changes in translation in response to depolarization. We found that these are partially mediated by features in the mRNA sequence—notably upstream open reading frames and secondary structure in the 5′ untranslated region—both of which predict downregulation in response to depolarization. Translationally regulated transcripts are also more likely to be targets of FMRP and include genes implicated in autism in humans. Our findings support the idea that control of mRNA translation plays an important role in response to neural activity across the genome

Impaired Hyperemic Response to Exercise Post Stroke

Individuals with chronic stroke have reduced perfusion of the paretic lower limb at rest; however, the hyperemic response to graded muscle contractions in this patient population has not been examined. This study quantified blood flow to the paretic and non-paretic lower limbs of subjects with chronic stroke after submaximal contractions of the knee extensor muscles and correlated those measures with limb function and activity. Ten subjects with chronic stroke and ten controls had blood flow through the superficial femoral artery quantified with ultrasonography before and immediately after 10 second contractions of the knee extensor muscles at 20, 40, 60, and 80% of the maximal voluntary contraction (MVC) of the test limb. Blood flow to the paretic and non-paretic limb of stroke subjects was significantly reduced at all load levels compared to control subjects even after normalization to lean muscle mass. Of variables measured, increased blood flow after an 80% MVC was the single best predictor of paretic limb strength, the symmetry of strength between the paretic and non-paretic limbs, coordination of the paretic limb, and physical activity. The impaired hemodynamic response to high intensity contractions was a better predictor of lower limb function than resting perfusion measures. Stroke-dependent weakness and atrophy of the paretic limb do not explain the reduced hyperemic response to muscle contraction alone as the response is similarly reduced in the non-paretic limb when compared to controls. These data may suggest a role for perfusion therapies to optimize rehabilitation post stroke

Two Weeks of Ischemic Conditioning Improves Walking Speed and Reduces Neuromuscular Fatigability in Chronic Stroke Survivors

This pilot study examined whether ischemic conditioning (IC), a noninvasive, cost-effective, and easy-to-administer intervention, could improve gait speed and paretic leg muscle function in stroke survivors. We hypothesized that 2 wk of IC training would increase self-selected walking speed, increase paretic muscle strength, and reduce neuromuscular fatigability in chronic stroke survivors. Twenty-two chronic stroke survivors received either IC or IC Sham on their paretic leg every other day for 2 wk (7 total sessions). IC involved 5-min bouts of ischemia, repeated five times, using a cuff inflated to 225 mmHg on the paretic thigh. For IC Sham, the cuff inflation pressure was 10 mmHg. Self-selected walking speed was assessed using the 10-m walk test, and paretic leg knee extensor strength and fatigability were assessed using a Biodex dynamometer. Self-selected walking speed increased in the IC group (0.86 ± 0.21 m/s pretest vs. 1.04 ± 0.22 m/s posttest, means ± SD; P\u3c 0.001) but not in the IC Sham group (0.92 ± 0.47 m/s pretest vs. 0.96 ± 0.46 m/s posttest; P= 0.25). Paretic leg maximum voluntary contractions were unchanged in both groups (103 ± 57 N·m pre-IC vs. 109 ± 65 N·m post-IC; 103 ± 59 N·m pre-IC Sham vs. 108 ± 67 N·m post-IC Sham; P = 0.81); however, participants in the IC group maintained a submaximal isometric contraction longer than participants in the IC Sham group (278 ± 163 s pre-IC vs. 496 ± 313 s post-IC, P = 0.004; 397 ± 203 s pre-IC Sham vs. 355 ± 195 s post-IC Sham; P = 0.46). The results from this pilot study thus indicate that IC training has the potential to improve walking speed and paretic muscle fatigue resistance poststroke

Macrophage TNF-α mediates parathion-induced airway hyperreactivity in guinea pigs.

Organophosphorus pesticides (OPs) are implicated in human asthma. We previously demonstrated that, at concentrations that do not inhibit acetylcholinesterase activity, the OP parathion causes airway hyperreactivity in guinea pigs as a result of functional loss of inhibitory M2 muscarinic receptors on parasympathetic nerves. Because macrophages are associated with asthma, we investigated whether macrophages mediate parathion-induced M2 receptor dysfunction and airway hyperreactivity. Airway physiology was measured in guinea pigs 24 h after a subcutaneous injection of parathion. Pretreatment with liposome-encapsulated clodronate induced alveolar macrophage apoptosis and prevented parathion-induced airway hyperreactivity in response to electrical stimulation of the vagus nerves. As determined by qPCR, TNF-α and IL-1β mRNA levels were increased in alveolar macrophages isolated from parathion-treated guinea pigs. Parathion treatment of alveolar macrophages ex vivo did not significantly increase IL-1β and TNF-α mRNA but did significantly increase TNF-α protein release. Consistent with these data, pretreatment with the TNF-α inhibitor etanercept but not the IL-1β receptor inhibitor anakinra prevented parathion-induced airway hyperreactivity and protected M2 receptor function. These data suggest a novel mechanism of OP-induced airway hyperreactivity in which low-level parathion activates macrophages to release TNF-α-causing M2 receptor dysfunction and airway hyperreactivity. These observations have important implications regarding therapeutic approaches for treating respiratory disease associated with OP exposures

Ischemic Conditioning Increases Strength and Volitional Activation of Paretic Muscle in Chronic Stroke: A Pilot Study

7siIschemic conditioning (IC) on the arm or leg has emerged as an intervention to improve strength and performance in healthy populations, but the effects on neurologic populations are unknown. The purpose of this study was to quantify the effects of a single session of IC on knee extensor strength and muscle activation in chronic stroke survivors. Maximal knee extensor torque measurements and surface EMG were quantified in 10 chronic stroke survivors (>1 year post-stroke) with hemiparesis before and after a single session of IC or Sham on the paretic leg. IC consisted of five minutes of compression with a proximal thigh cuff (inflation pressure = 225 mmHg for IC or 25 mmHg for Sham) followed by five minutes of rest. This was repeated five times. Maximal knee extensor strength, EMG magnitude, and motor unit firing behavior were measured before and immediately after IC or Sham. IC increased paretic leg strength by 10.6plus minus8.5 Nm while no difference was observed in the Sham group (change in Sham = 1.3plus minus2.9 Nm; p = 0.001 IC vs. Sham). IC-induced increases in strength were accompanied by a 31plus minus15% increase in the magnitude of muscle EMG during maximal contractions and a 5% decrease in motor unit recruitment thresholds during sub-maximal contractions. Individuals who had the most asymmetry in strength between their paretic and non-paretic legs had the largest increases in strength (r2= 0.54). This study provides evidence that a single session of IC can increase strength through improved muscle activation in chronic stroke survivors.openembargoed_20190204Hyngstrom, Allison S; Murphy, Spencer A; Nguyen, Jennifer; Schmit, Brian D; Negro, Francesco; Gutterman, David D; Durand, Matthew JHyngstrom, Allison S; Murphy, Spencer A; Nguyen, Jennifer; Schmit, Brian D; Negro, Francesco; Gutterman, David D; Durand, Matthew

A controlled study of cold dust content in galaxies from z=0−2z=0-2

At $z=1-3$, the formation of new stars is dominated by dusty galaxies whose far-IR emission indicates they contain colder dust than local galaxies of a similar luminosity. We explore the reasons for the evolving IR emission of similar galaxies over cosmic time using: 1) Local galaxies from GOALS $(L_{\rm IR}=10^{11}-10^{12}\,L_\odot)$; 2) Galaxies at $z\sim0.1-0.5$ from the 5MUSES ($L_{\rm IR}=10^{10}-10^{12}\,L_\odot$); 3) IR luminous galaxies spanning $z=0.5-3$ from GOODS and Spitzer xFLS ($L_{\rm IR}>10^{11}\,L_\odot$). All samples have Spitzer mid-IR spectra, and Herschel and ground-based submillimeter imaging covering the full IR spectral energy distribution, allowing us to robustly measure $L_{\rm IR}^{\rm\scriptscriptstyle SF}$, $T_{\rm dust}$, and $M_{\rm dust}$ for every galaxy. Despite similar infrared luminosities, $z>0.5$ dusty star forming galaxies have a factor of 5 higher dust masses and 5K colder temperatures. The increase in dust mass is linked with an increase in the gas fractions with redshift, and we do not observe a similar increase in stellar mass or star formation efficiency. $L_{160}^{\rm\scriptscriptstyle SF}/L_{70}^{\rm\scriptscriptstyle SF}$, a proxy for $T_{\rm dust}$, is strongly correlated with $L_{\rm IR}^{\rm\scriptscriptstyle SF}/M_{\rm dust}$ independently of redshift. We measure merger classification and galaxy size for a subsample, and there is no obvious correlation between these parameters and $L_{\rm IR}^{\rm \scriptscriptstyle SF}/M_{\rm dust}$ or $L_{160}^{\rm\scriptscriptstyle SF}/L_{70}^{\rm\scriptscriptstyle SF}$. In dusty star forming galaxies, the change in $L_{\rm IR}^{\rm\scriptscriptstyle SF}/M_{\rm dust}$ can fully account for the observed colder dust temperatures, suggesting that any change in the spatial extent of the interstellar medium is a second order effect.Comment: Accepted for publication in ApJ. 21 pages, 11 figure

Nanostructured Polymer Lithography for Photovoltaic Applications

The self-assembly of diblock copolymers into ordered domains holds great potential to furthering the efficiency of photovoltaic devices. Solutions containing polystyrene-block-poly(ethylene oxide) (PS-b-PEO) and poly(methyl methacrylate) (PMMA) were applied to silicon wafers from toluene solutions. Hexagonally ordered domains, with pore sizes ranging from 10-30 nm, were obtained by annealing films in solvent vapor, with the best results produced from a humidified benzene environment. Exposing the films to UV light cross-linked the polystyrene matrix and degraded the PMMA. Removal of the PMMA and PEO produced an ordered polystyrene template, which can be used for nanolithography for the deposition of quantum dots onto the wafers. Details of the film preparation, annealing times and conditions, and characterization will be presented

CD28 and the Tyrosine Kinase Lck Stimulate Mitogen-Activated Protein Kinase Activity in T Cells via Inhibition of the Small G Protein Rap1

Proliferation of T cells via activation of the T-cell receptor (TCR) requires concurrent engagement of accessory costimulatory molecules to achieve full activation. The best-studied costimulatory molecule, CD28, achieves these effects, in part, by augmenting signals from the TCR to the mitogen-activated protein (MAP) kinase cascade. We show here that TCR-mediated stimulation of MAP kinase extracellular-signal-regulated kinases (ERKs) is limited by activation of the Ras antagonist Rap1. CD28 increases ERK signaling by blocking Rap1 action. CD28 inhibits Rap1 activation because it selectively stimulates an extrinsic Rap1 GTPase activity. The ability of CD28 to stimulate Rap1 GTPase activity was dependent on the tyrosine kinase Lck. Our results suggest that CD28-mediated Rap1 GTPase-activating protein activation can help explain the augmentation of ERKs during CD28 costimulation

γδ T Cells Are Required for Pulmonary IL-17A Expression after Ozone Exposure in Mice: Role of TNFα

Ozone is an air pollutant that causes pulmonary symptoms. In mice, ozone exposure causes pulmonary injury and increases bronchoalveolar lavage macrophages and neutrophils. We have shown that IL-17A is important in the recruitment of neutrophils after subacute ozone exposure (0.3 ppm for 24–72 h). We hypothesized that γδ T cells are the main producers of IL-17A after subacute ozone. To explore this hypothesis we exposed wildtype mice and mice deficient in γδ T cells (TCRδ−/−) to ozone or room air. Ozone-induced increases in BAL macrophages and neutrophils were attenuated in TCRδ−/− mice. Ozone increased the number of γδ T cells in the lungs and increased pulmonary Il17a mRNA expression and the number of IL-17A+ CD45+ cells in the lungs and these effects were abolished in TCRδ−/− mice. Ozone-induced increases in factors downstream of IL-17A signaling, including G-CSF, IL-6, IP-10 and KC were also decreased in TCRδ−/− versus wildtype mice. Neutralization of IL-17A during ozone exposure in wildtype mice mimicked the effects of γδ T cell deficiency. TNFR2 deficiency and etanercept, a TNFα antagonist, also reduced ozone-induced increases in Il17a mRNA, IL-17A+ CD45+ cells and BAL G-CSF as well as BAL neutrophils. TNFR2 deficient mice also had decreased ozone-induced increases in Ccl20, a chemoattractant for IL-17A+ γδ T cells. Il17a mRNA and IL-17A+ γδ T cells were also lower in obese Cpefat versus lean WT mice exposed to subacute ozone, consistent with the reduced neutrophil recruitment observed in the obese mice. Taken together, our data indicate that pulmonary inflammation induced by subacute ozone requires γδ T cells and TNFα-dependent recruitment of IL-17A+ γδ T cells to the lung