Systematic study of constitutive cyclo-oxygenase-2 expression: role of NFκB and NFAT transcriptional pathways

Cyclooxygenase-2 (COX-2) is an inducible enzyme that drives inflammation and is the therapeutic target for widely used nonsteroidal antiinflammatory drugs (NSAIDs). However, COX-2 is also constitutively expressed, in the absence of overt inflammation, with a specific tissue distribution that includes the kidney, gastrointestinal tract, brain, and thymus. Constitutive COX-2 expression is therapeutically important because NSAIDs cause cardiovascular and renal side effects in otherwise healthy individuals. These side effects are now of major concern globally. However, the pathways driving constitutive COX-2 expression remain poorly understood. Here we show that in the kidney and other sites, constitutive COX-2 expression is a sterile response, independent of commensal microorganisms and not associated with activity of the inflammatory transcription factor NF-κB. Instead, COX-2 expression in the kidney but not other regions colocalized with nuclear factor of activated T cells (NFAT) transcription factor activity and was sensitive to inhibition of calcineurin-dependent NFAT activation. However, calcineurin/NFAT regulation did not contribute to constitutive expression elsewhere or to inflammatory COX-2 induction at any site. These data address the mechanisms driving constitutive COX-2 and suggest that by targeting transcription it may be possible to develop antiinflammatory therapies that spare the constitutive expression necessary for normal homeostatic functions, including those important to the cardiovascular-renal system

Non-palimpsested crowded Skolithos ichnofabrics in a Carboniferous tidal rhythmite: Disentangling ecological signatures from the spatio-temporal bias of outcrop

Funder: International Association of Sedimentologists; Id: http://dx.doi.org/10.13039/501100007463A heterolithic tidalite succession yielding spring–neap bundles is newly reported from a mid- Carboniferous (Serpukhovian) section of the Alston Formation of Northumberland, England. The rhythmite records deposition over an interval that can be confidently calibrated to at least 84 lunar days, and attests to a non-negligible tidal range in parts of the Northwest European Seaway in the late Mississippian. The tidalite is notable for the presence of a striking crowded Skolithos ichnofabric on both bedding planes and in vertical section. Bedding plane expressions of the ichnofabric reveal true substrates of sand piles excavated during burrow construction, in addition to an apparently remarkable equal spacing between individual burrows that is shown to be genuine through pair correlation function analysis. These characteristics show that the burrowed horizons were registered by contemporaneous ichnocoenoses, with no palimpsesting of burrows. The irregular vertical distribution of burrow horizons, despite a near-continuous semi-diurnal record of sedimentation, is suggested to be an artefact of spatial patchiness of burrowing communities in the depositional environment; imperfectly registered in a vertical profile with high-temporal, lowspatial resolution. The succession proves that burrow palimpsesting is not an inevitable ichnological conclusion of sedimentary stasis, and attests to intermittent palaeoecological fidelity of the stratigraphic record at the small spatio-temporal scales recorded a

A mechanical bottleneck explains the variation in cup growth during FcγR phagocytosis

Phagocytosis is the process by which cells internalize particulate material, and is of central importance to immunity, homeostasis and development. Here, we study the internalization of immunoglobulin G-coated particles in cells transfected with Fcγ receptors (FcγRs) through the formation of an enveloping phagocytic cup. Using confocal microscopy, we precisely track the location of fluorescently tagged FcγRs during cup growth. Surprisingly, we found that phagocytic cups growing around identical spherical particles showed great variability even within a single cell and exhibited two eventual fates: a cup either stalled before forming a half-cup or it proceeded until the particle was fully enveloped. We explain these observations in terms of a mechanical bottleneck using a simple mathematical model of the overall process of cup growth. The model predicts that reducing F-actin concentration levels, and hence the deforming force, does not necessarily lead to stalled cups, a prediction we verify experimentally. Our analysis gives a coherent explanation for the importance of geometry in phagocytic uptake and provides a unifying framework for integrating the key processes, both biochemical and mechanical, occurring during cup growth

A bioassay system of autologous human endothelial, smooth muscle cells and leucocytes for use in drug discovery, phenotyping and tissue engineering

Purpose: Blood vessels are comprised of endothelial and smooth muscle cells. Obtaining both types of cells from vessels of living donors is not possible without invasive surgery. To address this we have devised a strategy whereby human endothelial and smooth muscle cells derived from blood progenitors from the same donor could be cultured with autologous leucocytes to generate a same donor ‘vessel in a dish’ bioassay. Basic procedures: Autologous sets of blood outgrowth endothelial cells (BOECs), smooth muscle cells (BO-SMCs) and leucocytes were obtained from 4 donors. Cells were treated in mono and cumulative co-culture conditions. The endothelial specific mediator endothelin-1 along with interleukin (IL)-6, IL-8, tumour necrosis factor α, and interferon gamma-induced protein 10 were measured under control culture conditions and after stimulation with cytokines. Main findings: Co-cultures remained viable throughout. The profile of individual mediators released from cells was consistent with what we know of endothelial and smooth muscle cells cultured from blood vessels. Principle conclusions: For the first time, we report a proof of concept study where autologous blood outgrowth ‘vascular’ cells and leucocytes were studied alone and in co-culture. This novel bioassay has utility in vascular biology research, patient phenotyping, drug testing and tissue engineering

Free radical activity of PM10: iron-mediated generation of hydroxyl radicals.

The purpose of this study was to test the hypothesis that particulate matter < or = 10 microns in aerodynamic diameter (PM10) particles have the ability to generate free radical activity at their surface. We collected PM10 filters from the Edinburgh, United Kingdom, Enhanced Urban Network sampling site, removed particles from the filter, and tested their ability to cause free radical damage to supercoiled plasmid DNA. We found that the PM10 particles did cause damage to the DNA that was mediated by hydroxyl radicals, as shown by inhibition of the injury with mannitol. The PM10-associated hydroxyl radical activity was confirmed using a high-performance liquid chromatography-based assay to measure the hydroxyl radical adduct of salicylic acid. Desferrioxamine abolished the hydroxyl radical-mediated injury, which suggests that iron was involved. Analysis of PM10 filters confirmed the presence of large amounts of iron and leaching studies confirmed that the PM10 samples could release substantial amounts of Fe(III) and lesser amounts of Fe(II). To investigate the size of the particles involved in the hydroxyl radical injury, we centrifuged the suspension of PM10 to clarity, tested the clear supernatant, and found that it had all of the suspension activity. We conclude, therefore, that the free radical activity is derived either from a fraction that is not centrifugeable on a bench centrifuge, or that the radical generating system is released into solution

Robust increase in population exposure to heat stress with increasing global warming

This is the final version. Available on open access from IOP Publishing via the DOI in this recordData availability statement: No new data were created or analysed in this study.Extreme heat, particularly if combined with humidity, poses a severe risk to human health. To estimate future global risk of extreme heat with humidity on health, we calculate indicators of heat stress that have been commonly used: The Heat Index, the Wet-Bulb Globe Temperature and the Wet-Bulb Temperature, from the latest Climate Model Intercomparison Project (CMIP6) projections. We analyse how and where different levels of heat stress hazards will change, from severe to deadly, and how results are sensitive to the choice of the index used. We evaluate this risk at country-level and use population and GDP | PPP growth scenario to estimate the vulnerability of each nation. Consistent with previous studies, we find that South and East Asia, and the Middle-East, are highly exposed to heat stress hazards, and that this exposure increases by 20%-60% with global mean temperature change from 1.5 to 3 âC. However, we also find substantial increases in heat health risk for some vulnerable countries with less adaptive capacity, such as West Africa, and Central and South America. For these regions, about 20 to more than 50% of the population could be exposed to severe heat stress each year on average, independent of the index used. For global warming of 3â, European countries and the USA will also be exposed several times per year to conditions with daily mean heat stress level equal to the maximum heat stress of the 2003 heat wave.Natural Environment Research Council (NERC

Oral nitrate supplementation improves cardiovascular risk markers in COPD: ON-BC a randomised controlled trial

BACKGROUND: Short term studies suggest that dietary nitrate supplementation may improve cardiovascular risk profile, lowering blood pressure (BP) and enhancing endothelial function. It is not clear if these beneficial effects are sustained and whether they apply in people with COPD, who have a worse cardiovascular profile than those without COPD. Nitrate-rich beetroot juice BRJ (NR-BRJ) is a convenient dietary source of nitrate. METHODS: The ON-BC trial was a randomized, double-blind, placebo-controlled parallel group study in stable COPD patients with home systolic BP (SBP) measurement ≥130 mmHg. Participants were randomly allocated (1:1) using computer-generated, block randomization to either 70 mL of NR-BRJ (400 mg NO3 -) (n=40) or an otherwise identical nitrate-depleted placebo juice Pl-BRJ (0 mg NO3 -) (n=41), once daily for 12 weeks. The primary endpoint was between group change in home SBP measurement. Secondary outcomes included change in 6-minute walking distance (6MWD) and measures of endothelial function (reactive hyperaemia index (RHI) and augmentation index (AIx75)) using an EndoPAT device. Plasma nitrate and platelet function were also measured. RESULTS: Compared to placebo, active treatment lowered SBP (Hodges-Lemman treatment effect MD[95% CI]; -4.5[-3.0 to -5.9] improved 6MWD (+30.0 m [15.7 to 44.2], p<0.001), RHI +0.34 (0.03 to 0.63) p=0.03, and AIx75 -7.61% [-14.3 to -0.95], p=0.026. CONCLUSIONS: In people with COPD, prolonged dietary nitrate supplementation in the form of beetroot juice produces a sustained reduction in BP, associated with an improvement in endothelial function and exercise capacity

Exercise hyperthermia induces greater changes in gastrointestinal permeability than equivalent passive hyperthermia

Hyperthermia and exertional heat illness increase gastrointestinal (GI) permeability, although whether the latter is only via hyperthermia is unclear. The aim of this pilot study was to determine whether different changes in GI permeability, characterized by an increased plasma lactulose:rhamnose concentration ratio ([L:R]), occurred in exercise hyperthermia in comparison to equivalent passive hyperthermia. Six healthy adult male participants (age 25 ± 5 years, mass 77.0 ± 6.7 kg, height 181 ± 6 cm, peak oxygen uptake [urn:x-wiley:2051817X:media:phy214945:phy214945-math-0001] 48 ± 8 ml.kg−1.min−1) underwent exercise under hot conditions (Ex-Heat) and passive heating during hot water immersion (HWI). Heart rate (HR), rectal temperature (TCORE), rating of perceived exertion (RPE), and whole-body sweat loss (WBSL) were recorded throughout the trials. The L:R ratio, peak HR, change in HR, and change in RPE were higher in Ex-Heat than HWI, despite no differences in trial duration, peak core temperature or WBSL. L:R was strongly correlated (p < 0.05) with HR peak (r = 0.626) and change in HR (r = 0.615) but no other variable. The greater L:R in Ex-Heat, despite equal TCORE responses to HWI, indicates that increased cardiovascular strain occurred during exercise, and exacerbates hyperthermia-induced GI permeability at the same absolute temperature