The determinants of transverse tubular volume in resting skeletal muscle.

The transverse tubular (t)-system of skeletal muscle couples sarcolemmal electrical excitation with contraction deep within the fibre. Exercise, pathology and the composition of the extracellular fluid (ECF) can alter t-system volume (t-volume). T-volume changes are thought to contribute to fatigue, rhabdomyolysis and disruption of excitation-contraction coupling. However, mechanisms that underlie t-volume changes are poorly understood. A multicompartment, history-independent computer model of rat skeletal muscle was developed to define the minimum conditions for t-volume stability. It was found that the t-system tends to swell due to net ionic fluxes from the ECF across the access resistance. However, a stable t-volume is possible when this is offset by a net efflux from the t-system to the cell and thence to the ECF, forming a net ion cycle ECF→t-system→sarcoplasm→ECF that ultimately depends on Na(+)/K(+)-ATPase activity. Membrane properties that maximize this circuit flux decrease t-volume, including PNa(t) > PNa(s), PK(t) < PK(s) and N(t) < N(s) [P, permeability; N, Na(+)/K(+)-ATPase density; (t), t-system membrane; (s), sarcolemma]. Hydrostatic pressures, fixed charges and/or osmoles in the t-system can influence the magnitude of t-volume changes that result from alterations in this circuit flux. Using a parameter set derived from literature values where possible, this novel theory of t-volume was tested against data from previous experiments where t-volume was measured during manipulations of ECF composition. Predicted t-volume changes correlated satisfactorily. The present work provides a robust, unifying theoretical framework for understanding the determinants of t-volume.JAF was supported by a David Phillips Fellowship (BB/FO23863/1) awarded by the Biotechnology and Biological Sciences Research Council (UK). JS was supported by the Agency for Science, Technology and Research (Singapore) and a Caius Medical Association summer studentship from Gonville and Caius College, University of Cambridge.This is the final version. It was first published by Wiley at http://onlinelibrary.wiley.com/doi/10.1113/jphysiol.2014.281170/abstract

The Factor Inhibiting HIF Asparaginyl Hydroxylase Regulates Oxidative Metabolism and Accelerates Metabolic Adaptation to Hypoxia.

Animals require an immediate response to oxygen availability to allow rapid shifts between oxidative and glycolytic metabolism. These metabolic shifts are highly regulated by the HIF transcription factor. The factor inhibiting HIF (FIH) is an asparaginyl hydroxylase that controls HIF transcriptional activity in an oxygen-dependent manner. We show here that FIH loss increases oxidative metabolism, while also increasing glycolytic capacity, and that this gives rise to an increase in oxygen consumption. We further show that the loss of FIH acts to accelerate the cellular metabolic response to hypoxia. Skeletal muscle expresses 50-fold higher levels of FIH than other tissues: we analyzed skeletal muscle FIH mutants and found a decreased metabolic efficiency, correlated with an increased oxidative rate and an increased rate of hypoxic response. We find that FIH, through its regulation of oxidation, acts in concert with the PHD/vHL pathway to accelerate HIF-mediated metabolic responses to hypoxia

S-2-hydroxyglutarate regulates CD8+ T-lymphocyte fate.

R-2-hydroxyglutarate accumulates to millimolar levels in cancer cells with gain-of-function isocitrate dehydrogenase 1/2 mutations. These levels of R-2-hydroxyglutarate affect 2-oxoglutarate-dependent dioxygenases. Both metabolite enantiomers, R- and S-2-hydroxyglutarate, are detectible in healthy individuals, yet their physiological function remains elusive. Here we show that 2-hydroxyglutarate accumulates in mouse CD8+ T cells in response to T-cell receptor triggering, and accumulates to millimolar levels in physiological oxygen conditions through a hypoxia-inducible factor 1-alpha (HIF-1α)-dependent mechanism. S-2-hydroxyglutarate predominates over R-2-hydroxyglutarate in activated T cells, and we demonstrate alterations in markers of CD8+ T-cell differentiation in response to this metabolite. Modulation of histone and DNA demethylation, as well as HIF-1α stability, mediate these effects. S-2-hydroxyglutarate treatment greatly enhances the in vivo proliferation, persistence and anti-tumour capacity of adoptively transferred CD8+ T cells. Thus, S-2-hydroxyglutarate acts as an immunometabolite that links environmental context, through a metabolic-epigenetic axis, to immune fate and function

Pulmonary Thrombosis Promotes Tumorigenesis via Myeloid Hypoxia-Inducible Factors.

Cancer patients have a greater risk of thrombosis than individuals without cancer. Conversely, thrombosis is a diagnostic predictor of cancer, but the mechanisms by which thrombosis promotes tumor propagation are incompletely understood. Our previous studies showed that hypoxia-inducible factors (HIF) 1α and HIF2α are stabilized in myeloid cells of murine thrombi. We also previously showed that pulmonary thrombosis increases the levels of HIF1α and HIF2α in murine lungs, enhances the levels of tumorigenic factors in the circulation, and promotes pulmonary tumorigenesis. In this study, we aimed to investigate the regulation of thrombosis-induced tumorigenesis by myeloid cell-specific HIFs (i.e., HIF1 and HIF2 in neutrophils and macrophages). Our in vitro studies showed that multiple tumorigenic factors are upregulated in the secretome of hypoxic versus normoxic neutrophils and macrophages, which promotes lung cancer cell proliferation and migration in a myeloid-HIF-dependent manner. Next, we used a mouse model of pulmonary microvascular occlusion to study the impact of pulmonary thrombosis and myeloid HIFs on lung tumorigenesis. Experiments on mice lacking either HIF1α or HIF2α in myeloid cells demonstrated that loss of either factor eliminates the advantage given to pulmonary tumor formation by thrombotic insult. The myeloid HIF-dependent and tumorigenic impact of pulmonary thrombosis on tumor burden may be partly driven by paracrine thymidine phosphorylase (TP), given that TP levels were increased by hypoxia in neutrophil and macrophage supernates, and that plasma TP levels were positively correlated with multiple measures of tumor progression in wild type mice but not myeloid cell-specific HIF1α or HIF2α knockout mice. These data together demonstrate the importance of thrombotic insult in a model of pulmonary tumorigenesis and the essential role of myeloid HIFs in mediating tumorigenic success

Direct membrane filtration of municipal wastewater: linking periodical physical cleaning with fouling mechanisms

Direct membrane filtration (DMF) is a promising alternative secondary wastewater treatment process in Iceland, where biological treatment is not effective due to low strength wastewater nature and low temperature. This study aims to investigate membrane fouling mechanisms and mitigation approaches during DMF of municipal wastewater using a crossflow membrane filtration system integrated with an optical coherence tomography (OCT) imaging system. During DMF of wastewater, it was observed that intermediate pore blocking was dominant during the early stage of fouling, followed by cake filtration. Multi-filtration cycles were performed under different conditions, and the results revealed that (1) elevating flushing water temperature from 25 to 50 °C greatly reduced the intermediate pore blocking constant accompanied with a decreased physically-irreversible fouling; (2) increasing both filtration and flushing crossflow velocities did not influence the pore blocking constant, but caused a lower cake filtration constant with reducing both physically-reversible and irreversible fouling; (3) extending filtration-flushing duration interval appeared to slightly lower the pore blocking constant; (4) with extending filtration cycles, a shift of reversible fouling to irreversible fouling was noticed and associated with the compression of the tightly attached cake layer that was not readily removed by periodical flushing. A combination of periodical physical flushing with short term chemical-enhanced cleaning was employed and sustainable long-term operation of DMF was achieved. Furthermore, the foulants autopsy indicated that biofouling combined with organic/inorganic fouling influenced the cake fouling development.This work was supported by the University of Iceland Research Fund

Thermally Triggered Mechanically Destructive Electronics Based On Electrospun Poly(epsilon-caprolactone) Nanofibrous Polymer Films

Electronics, which functions for a designed time period and then degrades or destructs, holds promise in medical implants, reconfigurable electronic devices and/or temporary functional systems. Here we report a thermally triggered mechanically destructive device, which is constructed with an ultrathin electronic components supported by an electrospun poly(epsilon-caprolactone) nanofibrous polymer substrate. Upon heated over the melting temperature of the polymer, the pores of the nanofibers collapse due to the nanofibers&apos; microscopic polymer chain relaxing and packing. As a result, the polymer substrate exhibits approximately 97.5% area reduction. Ultra-thin electronic components can therefore be destructed concurrently. Furthermore, by integrating a thin resistive heater as the thermal trigger of Joule heating, the device is able to on-demand destruct. The experiment and analytical results illustrate the essential aspects and theoretical understanding for the thermally triggered mechanical destructive devices. The strategy suggests a viable route for designing destructive electronics

Fouling and mitigation mechanisms during direct microfiltration and ultrafiltration of primary wastewater

Direct membrane filtration (DMF) has recently gained attention as an alternative secondary biological wastewater treatment process. This study evaluated direct microfiltration (MF) and ultrafiltration (UF) performance and cleaning protocols during crossflow DMF of primary municipal wastewater effluent. Several types of MF and UF membranes were examined by threshold flux determination, and two types of membranes (MF, 0.08 μm; UF, 100 kDa) were chosen for exploring membrane fouling mechanisms at different feed pressures via both fouling resistance analysis and optical coherence tomography (OCT) observation. The results revealed that both MF and UF displayed three-stage fouling behaviors, i.e., initial intermediate pore blocking followed by two-stage cake filtration. Increasing feed pressure from 8 kPa to 50 kPa could accelerate physically reversible fouling rate (consistent with simulated cake filtration constant). During physical flushing, the cake layer was more readily removed from the UF membrane; while residual porous cake layer was present on the MF membrane, regardless of the feed pressure. With extending filtration-cleaning cycle, shortening filtration duration and elevating cleaning solution temperature to from 25 °C to 50 °C benefited for irreversible fouling alleviation. At 50 °C, the geothermal water performed similar cleaning behaviors as clean water, facilitating lower reversible and irreversible fouling than the geothermal brine. This study shed light on the feasibility of using high temperature geothermal water for periodic physical cleaning during DMF of wastewater under Icelandic scenario.Economic Development Board (EDB)This work was supported by the University of Iceland Research Fund. The Student Innovation Fund from Rannís in Iceland was acknowledged for providing summer research grant to Dagmar Olafsd ´ ottir. The Economic Development Board (EDB) of Singapore is acknowledged for funding the Singapore MembraneTechnology Centre (SMTC) , Nanyang Technological University

Evaluation of Ceramics Adsorption Filter as a Pretreatment for Seawater Reverse-Osmosis Desalination

Seawater reverse osmosis (SWRO) is the most energy-efficient process for desalination to produce drinking water from seawater. However, its sustainability is still challenged by membrane fouling. Appropriate feed water quality is one of the crucial prerequisites for SWRO operation. In the current study, a ceramic adsorption filter (CAF), which was predominantly coated with an aluminum-based adsorbent (i.e., Alumina, Al2O3), was employed to enhance the pretreatment performance of SWRO. The fouling performance of SWRO pre-treated with a CAF was evaluated by feeding with real ultrafiltration (UF)-filtrated seawater collected from a seawater desalination R&amp;D facility in Singapore. The flux decline profile showed that the presence of CAF after UF could mitigate around 10&ndash;30% of SWRO fouling. Based on the autopsy of the fouled SWRO membranes, it was observed that SWRO with CAF pre-treatment and daily regeneration could alleviate around 77.5% of Ca-induced inorganic fouling as well as 76% of lower biofouling. The present work highlights the potential of applying adsorption technology to enhance pre-treatment performance to extend the lifespan of SWRO membranes. Coupling the adsorbents on a ceramic filter should be a useful way to ease their implementation, i.e., inline adsorption and re-generation

The FIH (Factor Inhibiting HIF) asparaginyl hydroxyls regulates oxidative metabolism and accelerates adaptation to hypoxia

Animals require an immediate response to oxygen availability to allow rapid shifts between oxidative and glycolytic metabolism. These metabolic shifts are highly regulated by the HIF transcription factor. The Factor Inhibiting HIF (FIH) is an asparaginyl hydroxylase that controls HIF transcriptional activity in an oxygen-dependent manner. We show here that FIH loss increases oxidative metabolism, while also increasing glycolytic capacity, and that this gives rise to an increase in oxygen consumption. We further show that the loss of FIH acts to accelerate the cellular metabolic response to hypoxia. Skeletal muscle expresses 50-fold higher levels of FIH than other tissues: we analyzed skeletal muscle FIH mutants, and found a decreased metabolic efficiency, correlated with an increased oxidative rate and an increased rate of hypoxic response. We find that FIH, through its regulation of oxidation, acts in concert with the PHD/VHL pathway to accelerate HIF-mediated metabolic responses to hypoxia