Hepatic gene expression in flounder chronically exposed to multiply polluted estuarine sediment: Absence of classical exposure ‘biomarker’ signals and induction of inflammatory, innate immune and apoptotic pathways

The effects of chronic long term exposure to multiply-polluted environments on fish are not well understood, but environmental surveys suggest that such exposure may cause a variety of pathologies, including cancers. Transcriptomic profiling has recently been used to assess gene expression in European flounder (Platichthys flesus) living in several polluted and clean estuaries. However, the gene expression changes detected were not unequivocally elicited by pollution, most likely due to the confounding effects of natural estuarine ecosystem variables. In this study flounder from an uncontaminated estuary were held on clean or polluted sediments in mesocosms, allowing control of variables such as salinity, temperature, and diet. After 7 months flounder were removed from each mesocosm and hepatocytes prepared from fish exposed to clean or polluted sediments. The hepatocytes were treated with benzo(a)pyrene (BAP), estradiol (E2), copper, a mixture of these three, or with the vehicle DMSO. A flounder cDNA microarray was then used to measure hepatocyte transcript abundance after each treatment. The results show that long term chronic exposure to a multiply-polluted sediment causes increases in the expression of mRNAs coding for proteins of the endogenous apoptotic program, of innate immunity and inflammation. Contrary to expectation, the expression of mRNAs which are commonly used as biomarkers of environmental exposure to particular contaminants were not changed, or were changed contrary to expectation. However, acute treatment of hepatocytes from flounder from both clean and polluted sediments with BAP or E2 caused the expected changes in the expression of these biomarkers. Thus transcriptomic analysis of flounder exposed long-term to chronic pollution causes a different pattern of gene expression than in fish acutely treated with single chemicals, and reveals novel potential biomarkers of environmental contaminant exposure. These novel biomarkers include Diablo, a gene involved in apoptotic pathways and highly differentially regulated by both chronic and acute exposure to multiple pollutants

Utility of Survival Motor Neuron ELISA for Spinal Muscular Atrophy Clinical and Preclinical Analyses

Genetic defects leading to the reduction of the survival motor neuron protein (SMN) are a causal factor for Spinal Muscular Atrophy (SMA). While there are a number of therapies under evaluation as potential treatments for SMA, there is a critical lack of a biomarker method for assessing efficacy of therapeutic interventions, particularly those targeting upregulation of SMN protein levels. Towards this end we have engaged in developing an immunoassay capable of accurately measuring SMN protein levels in blood, specifically in peripheral blood mononuclear cells (PBMCs), as a tool for validating SMN protein as a biomarker in SMA.A sandwich enzyme-linked immunosorbent assay (ELISA) was developed and validated for measuring SMN protein in human PBMCs and other cell lysates. Protocols for detection and extraction of SMN from transgenic SMA mouse tissues were also developed.The assay sensitivity for human SMN is 50 pg/mL. Initial analysis reveals that PBMCs yield enough SMN to analyze from blood volumes of less than 1 mL, and SMA Type I patients' PBMCs show ∼90% reduction of SMN protein compared to normal adults. The ELISA can reliably quantify SMN protein in human and mouse PBMCs and muscle, as well as brain, and spinal cord from a mouse model of severe SMA.This SMN ELISA assay enables the reliable, quantitative and rapid measurement of SMN in healthy human and SMA patient PBMCs, muscle and fibroblasts. SMN was also detected in several tissues in a mouse model of SMA, as well as in wildtype mouse tissues. This SMN ELISA has general translational applicability to both preclinical and clinical research efforts

Iron Behaving Badly: Inappropriate Iron Chelation as a Major Contributor to the Aetiology of Vascular and Other Progressive Inflammatory and Degenerative Diseases

The production of peroxide and superoxide is an inevitable consequence of aerobic metabolism, and while these particular "reactive oxygen species" (ROSs) can exhibit a number of biological effects, they are not of themselves excessively reactive and thus they are not especially damaging at physiological concentrations. However, their reactions with poorly liganded iron species can lead to the catalytic production of the very reactive and dangerous hydroxyl radical, which is exceptionally damaging, and a major cause of chronic inflammation. We review the considerable and wide-ranging evidence for the involvement of this combination of (su)peroxide and poorly liganded iron in a large number of physiological and indeed pathological processes and inflammatory disorders, especially those involving the progressive degradation of cellular and organismal performance. These diseases share a great many similarities and thus might be considered to have a common cause (i.e. iron-catalysed free radical and especially hydroxyl radical generation). The studies reviewed include those focused on a series of cardiovascular, metabolic and neurological diseases, where iron can be found at the sites of plaques and lesions, as well as studies showing the significance of iron to aging and longevity. The effective chelation of iron by natural or synthetic ligands is thus of major physiological (and potentially therapeutic) importance. As systems properties, we need to recognise that physiological observables have multiple molecular causes, and studying them in isolation leads to inconsistent patterns of apparent causality when it is the simultaneous combination of multiple factors that is responsible. This explains, for instance, the decidedly mixed effects of antioxidants that have been observed, etc...Comment: 159 pages, including 9 Figs and 2184 reference

Positional Differences in Pre-Season Scrimmage Performance of Division I Collegiate Football Players

This study aimed to describe the physical demands of American football players using novel performance analysis techniques. Heart rate (HR) and accelerometer-based activity levels were observed across two pre-season scrimmages in 23 Division I collegiate football players (age: 19 ± 1 y, height: 1.90 ± 0.06 m, weight: 116.2 ± 19.4 kg). Data were analyzed using a MATLAB program and inter-rater reproducibility assessed using inter-class correlations (ICC). Players were analyzed by side (offense/defense) and position (skill/non-skill). Performance variables assessed in bursts of activity included burst duration, HRmean and HRmax (bpm), and mean activity (vector magnitude units [vmu]). Exercise intensity was categorized as time spent in % HRmax in 5% increments. The burst duration (8.1±3.9 min, ICC = 0.72), HRmean (157 ± 12 bpm, ICC = 0.96) and mean activity (0.30 ± 0.05 vmu, ICC = 0.86) were reproducible. HRmean (p = 0.05) and HRmax (p = 0.001) were greater on defense. Offense spent more time at 65–70% HRmax (p = 0.01), 70–75% HRmax (p = 0.02) while defense spent more time 90–95% HRmax and ≥95% HRmax (p = 0.03). HRmean (p = 0.70) and HRpeak (p = 0.80) were not different between positions across both sides. Skilled players demonstrated greater mean activity (p = 0.02). The sport-specific analysis described HR and activity level in a reproducible manner. Automated methods of assessing HR may be useful in training and game time performance but ultimately provides support to coaching decision making

Positional differences in pre-season scrimmage performance of division i collegiate football players

This study aimed to describe the physical demands of American football players using novel performance analysis techniques. Heart rate (HR) and accelerometer-based activity levels were observed across two pre-season scrimmages in 23 Division I collegiate football players (age: 19 ± 1 y, height: 1.90 ± 0.06 m, weight: 116.2 ± 19.4 kg). Data were analyzed using a MATLAB program and inter-rater reproducibility assessed using inter-class correlations (ICC). Players were analyzed by side (offense/defense) and position (skill/non-skill). Performance variables assessed in bursts of activity included burst duration, HRmean and HRmax (bpm), and mean activity (vector magnitude units [vmu]). Exercise intensity was categorized as time spent in % HRmax in 5% increments. The burst duration (8.1±3.9 min, ICC = 0.72), HRmean (157 ± 12 bpm, ICC = 0.96) and mean activity (0.30 ± 0.05 vmu, ICC = 0.86) were reproducible. HRmean (p = 0.05) and HRmax (p = 0.001) were greater on defense. Offense spent more time at 65–70% HRmax (p = 0.01), 70–75% HRmax (p = 0.02) while defense spent more time 90–95% HRmax and ≥95% HRmax (p = 0.03). HRmean (p = 0.70) and HRpeak (p = 0.80) were not different between positions across both sides. Skilled players demonstrated greater mean activity (p = 0.02). The sport-specific analysis described HR and activity level in a reproducible manner. Automated methods of assessing HR may be useful in training and game time performance but ultimately provides support to coaching decision making

Sea TENTACLE: Track, Engage, & Neutralize Threats - Asymmetric & Conventional - in the Littoral Environment

Includes supplementary materialSea TENTACLE is a proposed afloat platform whose primary mission is to utilize the state-of-the-art technology of unmanned vehicles to monitor and neutralize all subsurface enemy threats in the littorals. This mission can be specified further as anti-submarine warfare, mine warfare and maritime surveillance. The design philosophy of Sea TENTACLE embodies the ideal of providing a multi-mission capable sea frame extending network-centric warfare into the littorals. The design goals of the TSSE team were first to develop a platform to deploy, recover, and maintain unmanned vehicle (e.g. UUVs, USVs, UAVs) and second to enableto ship to act as an afloat network operations center for distributed assets. Allowing all units to work together seamlessly to conduct focused missions in the littorals makes the Sea TENTACLE a creitical component within the network-centric environment. The versatility of its cargo hold and modular design allows Sea TENTACLE to be outfitted dynamically to complete a veriety of secondary missions including humanitarian aid, salvage and spacial operations support. Sea TENTACLE's combat management and operations system will employ the Enterprise architecture design enabling C4ISR capabilities that will meet emerging network centric warfare needs.US Navy (USN) authorsTunisian Navy authorTurkish Navy authorsApproved for public release; distribution is unlimited