Sinusoidal endothelial cell-derived extracellular matrix regulates basal and stimulated macrophage activation.

Fatty liver disease, be it from alcohol (ALD), obesity (NAFLD) or other sources (e.g. viral infection), share similar mechanisms of disease progression. One potential mechanism is that altered sinusoidal endothelial cell (SEC) extracellular matrix (ECM) favors a more inflammatory phenotype in resident macrophages (i.e., Kupffer cells). Here, the hypothesis was tested that SEC-derived matrices directly affect the inflammatory response of macrophages. Methods. Transformed sinusoidal endothelial cells (TSECs) were cultured for 24 or 72 hours. Culture plates were then washed with a solution that selectively removed the cells, but preserved the ECM. Cultured macrophages (RAW 264.7 cells) were then seeded on the matrix and cultured for 24 hours; the cells were then stimulated with LPS for 0, 3, 6, 12, or 24 hours (100 ng/mL) ± CycloRGDfV. Real time RT-PCR was used to measure mRNA expression of proinflammatory mediators (IL-6, IL-1β, TNF-α, and INOS) and anti-inflammatory mediators (IL-10 and TGF-β). Results. LPS stimulated the production of all mediators by macrophages; when plated on ECM from TSECS, this response was attenuated for IL-6 and IL-1β. The presence of TSEC-derived ECM increased expression of TGF-β. The addition of the small peptide antagonist CycloRGDfV produced varying effects on the expression of inflammatory mediators in the presence or absence of TSEC-derived ECM. Conclusions. These data serve as first proof-of-concept that macrophage 4 activation can be modulated by ECM produced by TSECs and identifies a new interaction between these cells that may contribute to inflammatory liver disease

Novel insight into the liver-lung axis in alcohol-enhanced acute lung injury.

Background. Individuals who chronically abuse alcohol are almost 4 times more likely to develop Acute Respiratory Distress Syndrome (ARDS), the most severe form of Acute Lung Injury (ALI), but the mechanisms by which alcohol abuse sensitizes the lung to injury are poorly understood. However, the lung appears to share many parallel mechanisms of injury with the liver- a primary target of alcohol abuse. The overarching goal of this dissertation was therefore to expand on established mechanisms of alcohol-induced liver injury to ask innovative questions about mechanisms of alcohol-enhanced acute lung injury, as well as to develop new tools that may be used to gain novel insight into the liver-lung axis of alcoholinduced injury. Methods. Male mice were exposed to ethanol containing liquid diet either chronically (6 weeks) or in a chronic + binge pattern. Some mice were administered lipopolysaccharide (LPS) to induce acute lung injury. Lung injury and inflammation were assessed. To develop an animal model by which liver-lung interactions could be investigated, tamoxifen-loaded polymer nanoparticles were administered intrasplenically to a tamoxifen-inducible, Cre-mediated, dual-fluorescent reporter construct. Results. Chapter III of this dissertation describes a mechanism by which plasminogen activator inhibitor-1 (PAI-1) is involved in alcoholenhanced acute lung injury. Specifically, it was proposed that PAI-1-mediated fibrin accumulation promotes the aggregation of platelets, thereby propagating lung injury and inflammation. Chapter IV of this dissertation characterizes a recently-developed animal model of chronic + binge alcohol exposure, finding that animals exposed to chronic + binge alcohol exposure exhibit pulmonary inflammation and airway hyperresponsiveness. Finally, Chapter V of this dissertation develops an animal model to investigate liver-lung interactions during chronic alcohol exposure. It was found that tamoxifen-loaded polymer nanoparticles, when administered intrasplenically, selectively alter the genetics of hepatic cells, while avoiding other tissues, including the lung. Discussion. The work presented in this dissertation has, in conclusion, uncovered novel mechanisms by which alcohol sensitizes the lung to a second injury, shown that ethanol alone is sufficient to cause lung inflammation, and developed a novel animal method to examine liver-lung interactions during alcohol exposure

Chronic ethanol exposure sensitizes the lung in a mouse model of endotoxemia-induced acute lung injury : potential role of plasminogen activator inhibitor-1.

The goal of this project is to characterize a new mouse model of alcohol-enhanced acute lung injury (ALI) and to determine the role of plasminogen activator inhibitor-1 (PAI-1) in this model. Male mice (WT and PAI-1-/-) were exposed to ethanol-containing Lieber-DeCarli diet or pair-fed control diet for 6 weeks; some animals were administered intraperitoneal lipopolysaccharide (LPS) prior to sacrifice. Chronic alcohol feeding enhanced induction of the chemokines MIP-2 and KC (murine IL-8 homologues) after LPS injection in wild type animals. This enhanced chemokine expression did not correlate with enhanced pulmonary neutrophil infiltration, however animals exposed to chronic ethanol showed sustained alveolar septal thickening and enhanced 4-HNE staining, indicative of inflammatory damage. Septal thickening was completely attenuated in PAI-1-/- animals. This work has developed a new mouse model which can be used to elucidate the mechanisms of alcohol-enhanced ALI. A potential role of PAI-1 in alcohol-enhanced ALI has also been identified

Arm Positioning and Walking Styles of Children During First 5 Months of Independent Walking

Title: ARM POSITIONING AND WALKING STYLE OF CHILDREN DURING FIRST 5 MONTHS OF INDEPENDENT WALKING Authors: Poole, Lauren; Brett, Katrina; Gunselman, Alison; Schrempf, Bernadette; Looper, Julia Affiliation: Physical Therapy, University of Puget Sound, Tacoma, WA, United States. Purpose: Arm position during initial independent walking (IW) affects upper trunk biomechanics with gait. Arm positioning plays a role in forward propulsion and in maintaining upright position. Previous research has not established time lines or percentages of time spent in each arm position as IW progresses in typically developing infants. This study describes arm positioning across the initial 5 months of IW in typically developing infants. We hypothesized that high to middle guard positioning would be dominant during early visits with a progression to low guard by later visits. We did not expect to see consistent flexed arm movement or reciprocal arm swing within this time period. Subjects : Eight typically-developing children (5 M, 3 F). Materials/Methods : In this secondary analysis, researchers evaluated and coded video from trials at 1–5 months of IW. Researchers coded three trials per visit using arm positioning descriptions adapted from Ledebt (2000), including reciprocal arm swing (RAS), flexed movement (FM), high guard(HG), middle guard(MG), and low guard (LG). Time in each arm position for each arm in seconds is the dependent variable. Researchers performed descriptive statistics on the dependent variable for each visit. Other statistics performed include a 2(hand)x5(visit) ANOVA with repeated measures and post hoc analysis with Bonferonni correction. Results : The mean percentage of time in each arm position during the first visit was 2.6% RAS, 6.3% FM, 20.7% LG, 52.2% MG, and 18.5% HG. During the fifth visit, the mean percentages were 10.7% RAS, 76.4% FM, 8.1% LG, 3.5% MG, and 1.7% HG. For the fifth visit, the means were 1.7% high guard, 3.5% middle guard, 8.1% low guard, 76.4% flex movement, and 10.7% reciprocal arm swing. A 5 (visits) x 2 (sides) ANOVA with repeated measures revealed a left-to-right difference for HG (p=.035) as well as a significant side-by-visit interaction for HG (p=.023) and MG (p=.018). There was also a significant visit effect between visits 1 and 3 and between visits 1 and 5 for MG (p=.024, .003, respectively) and FM (p=.015, .005, respectively) with a trend toward significance for low guard (p=.055). Conclusions : Infants demonstrate a transition from stable arm positions to more dynamic arm movement with more walking experience. Dynamic arm positioning development begins early in IW, and provides a foundation for the emergence of RAS and adult-like upper extremity mechanics during gait. Clinical Relevance : This study describes changes in arm position over time – from stable upper extremity strategies to more mobile, exploration-facilitating strategies. Therapists should therefore monitor the transition from guarding to dynamic strategies during the first few months of IW for typically developing infants. If the transition does not occur, closer observation may be needed in order to facilitate early intervention. KEYWORDS: High Guard, Middle Guard, Low Guard, Flexed Movement, Reciprocal Arm Swing, Gait, Arm Position

Transitional Remodeling of the Hepatic Extracellular Matrix in Alcohol-Induced Liver Injury

Alcohol consumption is a common custom worldwide, and the toxic effects of alcohol on several target organs are well understood. The liver is the primary site of alcohol metabolism and is therefore the major target of alcohol toxicity. Alcoholic liver disease is a spectrum of disease states, ranging from simple steatosis (fat accumulation), to inflammation, and eventually to fibrosis and cirrhosis if untreated. The fibrotic stage of ALD is primarily characterized by robust accumulation of extracellular matrix (ECM) proteins (collagens) which ultimately impairs the function of the organ. The role of the ECM in early stages of ALD is poorly understood, but recent research has demonstrated that a number of changes in the hepatic ECM in prefibrotic ALD not only are present, but may also contribute to disease progression. The purpose of this review is to summarize the established and proposed changes to the hepatic extracellular matrix (ECM) that may contribute to earlier stages of ALD development and to discuss potential mechanisms by which these changes may mediate the progression of the disease

Prehabilitation for adults diagnosed with cancer: A systematic review of long‐term physical function, nutrition and patient‐reported outcomes

Objective: Prehabilitation is increasingly being used to mitigate treatment-related complications and enhance recovery. An individual's state of health at diagnosis, including obesity, physical fitness and comorbidities, are influencing factors for the occurrence of adverse effects. This review explores whether prehabilitation works in improving health outcomes at or beyond the initial 30 days post-treatment and considers the utility of prehabilitation before cancer treatment. Methods: A database search was conducted for articles published with prehabilitation as a pre-cancer treatment intervention between 2009 and 2017. Studies with no 30 days post-treatment data were excluded. Outcomes post-prehabilitation were extracted for physical function, nutrition and patient-reported outcomes. Results: Sixteen randomised controlled trials with a combined 2017 participants and six observational studies with 289 participants were included. Prehabilitation interventions provided multi-modality components including exercise, nutrition and psychoeducational aspects. Prehabilitation improved gait, cardiopulmonary function, urinary continence, lung function and mood 30 days post-treatment but was not consistent across studies. Conclusion: When combined with rehabilitation, greater benefits were seen in 30-day gait and physical functioning compared to prehabilitation alone. Large-scale randomised studies are required to translate what is already known from feasibility studies to improve overall health and increase long-term cancer patient outcomes

Detection of toxins and harmful algal bloom cells in shellfish hatcheries and efforts toward removal

As the start of the supply chain for the aquaculture industry, hatcheries are a crucial component in the success of oyster and northern quahog (hard clam) aquaculture on the East Coast of the US. Intermittent failures in hatchery production slow industry growth and reduce profits. To begin investigations into the possible role of algal toxins in hatchery production failure, post-treatment hatchery water from one research and four commercial hatcheries in lower Chesapeake Bay, USA, was sampled for (1) toxin presence and (2) harmful algal bloom (HAB) cell enumeration. Overall, seven toxin classes, likely produced by six different HAB species, were detected in post- treatment hatchery water, despite a lack of visually identifiable HAB cells within the facility. Toxins detected include pectenotoxin-2, goniodomin A, karlotoxin-1 and karlotoxin-3, okadaic acid and dinophysistoxin-1, azaspiracid-1 and azaspiracid-2, brevetoxin-2, and microcystin-LR. In a second, more targeted study, two batches of source water were followed and sampled at each step of a water-treatment process in the VIMS Aquaculture Genetics and Breeding Technology Center research hatchery in Gloucester Point, Virginia, USA. Two treatment steps showed particular promise for decreasing the concentrations of the three toxins detected in the source water, 24-h circulation through sand filters and activated charcoal filtration. Toxin concentrations of pectenotoxin-2, 3.53 ± 0.56 pg m

Methane Emission in a Specific Riparian-Zone Sediment Decreased with Bioelectrochemical Manipulation and Corresponded to the Microbial Community Dynamics

Dissimilatory metal-reducing bacteria are widespread in terrestrial ecosystems, especially in anaerobic soils and sediments. Thermodynamically, dissimilatory metal reduction is more favorable than sulfate reduction and methanogenesis but less favorable than denitrification and aerobic respiration. It is critical to understand the complex relationships, including the absence or presence of terminal electron acceptors, that govern microbial competition and coexistence in anaerobic soils and sediments, because subsurface microbial processes can effect greenhouse gas emissions from soils, possibly resulting in impacts at the global scale. Here, we elucidated the effect of an inexhaustible, ferrous-iron and humic-substance mimicking terminal electron acceptor by deploying potentiostatically poised electrodes in the sediment of a very specific stream riparian zone in Upstate New York state. At two sites within the same stream riparian zone during the course of six weeks in the spring of 2013, we measured CH4 and N2/N2O emissions from soil chambers containing either poised or unpoised electrodes, and we harvested biofilms from the electrodes to quantify microbial community dynamics. At the upstream site, which had a lower vegetation cover and highest soil temperatures, the poised electrodes inhibited CH4 emissions by ~45% (when normalized to remove temporal effects). CH4 emissions were not significantly impacted at the downstream site. N2/N2O emissions were generally low at both sites and were not impacted by poised electrodes. We did not find a direct link between bioelectrochemical treatment and microbial community membership; however, we did find a correspondence between environment/function and microbial community dynamics