Modulation of endothelial cell KCa3.1 Channels during endothelium-derived hyperpolarizing factor signaling in mesenteric resistance arteries

Arterial hyperpolarization to acetylcholine (ACh) reflects coactivation of KCa3.1 (IKCa) channels and KCa2.3 (SKCa) channels in the endothelium that transfers through myoendothelial gap junctions and diffusible factor(s) to affect smooth muscle relaxation (endothelium-derived hyperpolarizing factor [EDHF] response). However, ACh can differentially activate KCa3.1 and KCa2.3 channels, and we investigated the mechanisms responsible in rat mesenteric arteries. KCa3.1 channel input to EDHF hyperpolarization was enhanced by reducing external [Ca2+]o but blocked either with forskolin to activate protein kinase A or by limiting smooth muscle [Ca2+]i increases stimulated by phenylephrine depolarization. Imaging [Ca2+]i within the endothelial cell projections forming myoendothelial gap junctions revealed increases in cytoplasmic [Ca2+]i during endothelial stimulation with ACh that were unaffected by simultaneous increases in muscle [Ca2+]i evoked by phenylephrine. If gap junctions were uncoupled, KCa3.1 channels became the predominant input to EDHF hyperpolarization, and relaxation was inhibited with ouabain, implicating a crucial link through Na+/K+-ATPase. There was no evidence for an equivalent link through KCa2.3 channels nor between these channels and the putative EDHF pathway involving natriuretic peptide receptor-C. Reconstruction of confocal z-stack images from pressurized arteries revealed KCa2.3 immunostain at endothelial cell borders, including endothelial cell projections, whereas KCa3.1 channels and Na+/K+-ATPase {alpha}2/{alpha}3 subunits were highly concentrated in endothelial cell projections and adjacent to myoendothelial gap junctions. Thus, extracellular [Ca2+]o appears to modify KCa3.1 channel activity through a protein kinase A-dependent mechanism independent of changes in endothelial [Ca2+]i. The resulting hyperpolarization links to arterial relaxation largely through Na+/K+-ATPase, possibly reflecting K+ acting as an EDHF. In contrast, KCa2.3 hyperpolarization appears mainly to affect relaxation through myoendothelial gap junctions. Overall, these data suggest that K+ and myoendothelial coupling evoke EDHF-mediated relaxation through distinct, definable pathways

Thromboxane A2 inhibition of SKCa after NO synthase block in rat middle cerebral artery

NO/prostanoid independent, EDHF-mediated hyperpolarization and dilation in rat middle cerebral arteries is mediated solely by endothelial cell IK(Ca). However, when the NO-pathway is also active, both SK(Ca) and IK(Ca) contribute to EDHF responses. As the SK(Ca) component can be inhibited by stimulation of thromboxane A(2) (TxA(2)) TP receptors and NO has the potential ability to inhibit thromboxane synthesis, we investigated whether TxA(2) might explain loss of functional input from SK(Ca) during NOS inhibition in cerebral arteries. EXPERIMENTAL APPROACH: Rat middle cerebral arteries were mounted in a wire myograph. Endothelium-dependent responses to the PAR2 agonist, SLIGRL were assessed as simultaneous changes in smooth muscle membrane potential and tension. KEY RESULTS: Responses were obtained in the presence of L-NAME as appropriate. Inhibition of TP receptors with either ICI 192,605 or SQ 29,548, did not affect EDHF mediated hyperpolarization and relaxation, but in their presence neither TRAM-34 nor apamin (to block IK(Ca) and SK(Ca) respectively) individually affected the EDHF response. However, in combination they virtually abolished it. Similar effects were obtained in the presence of the thromboxane synthase inhibitor, furegrelate, which additionally revealed an iberiotoxin-sensitive residual EDHF hyperpolarization and relaxation in the combined presence of TRAM-34 and apamin. CONCLUSIONS AND IMPLICATIONS: In the rat middle cerebral artery, inhibition of NOS leads to a loss of the SK(Ca) component of EDHF responses. Either antagonism of TP receptors or block of thromboxane synthase restores an input through SK(Ca). These data indicate that NO normally enables SK(Ca) activity in rat middle cerebral arteries

Nitric oxide suppresses cerebral vasomotion by sGC-independent effects on ryanodine receptors and voltage-gated calcium channels

Background/Aims: In cerebral arteries, nitric oxide (NO) release plays a key role in suppressing vasomotion. Our aim was to establish the pathways affected by NO in rat middle cerebral arteries. Methods: In isolated segments of artery, isometric tension and simultaneous measurements of either smooth muscle membrane potential or intracellular [Ca 2+ ] ([Ca 2+ ] SMC ) changes were recorded. Results: In the absence of L -NAME, asynchronous propagating Ca 2+ waves were recorded that were sensitive to block with ryanodine, but not nifedipine. L -NAME stimulated pronounced vasomotion and synchronous Ca 2+ oscillations with close temporal coupling between membrane potential, tone and [Ca 2+ ] SMC . If nifedipine was applied together with L -NAME, [Ca 2+ ] SMC decreased and synchronous Ca 2+ oscillations were lost, but asynchronous propagating Ca 2+ waves persisted. Vasomotion was similarly evoked by either iberiotoxin, or by ryanodine, and to a lesser extent by ODQ. Exogenous application of NONOate stimulated endothelium-independent hyperpolarization and relaxation of either L -NAME-induced or spontaneous arterial tone. NO-evoked hyperpolarization involved activation of BK Ca channels via ryanodine receptors (RYRs), with little involvement of sGC. Further, in whole cell mode, NO inhibited current through L-type voltage-gated Ca 2+ channels (VGCC), which was independent of both voltage and sGC. Conclusion: NO exerts sGC-independent actions at RYRs and at VGCC, both of which normally suppress cerebral artery myogenic tone

Microplastic in Riverine Fish is Connected to Species Traits

Microplastic is a contaminant of concern worldwide. Rivers are implicated as major pathways of microplastic transport to marine and lake ecosystems, and microplastic ingestion by freshwater biota is a risk associated with microplastic contamination, but there is little research on microplastic ecology within freshwater ecosystems. Microplastic uptake by fish is likely affected by environmental microplastic abundance and aspects of fish ecology, but these relationships have rarely been addressed. We measured the abundance and composition of microplastic in fish and surface waters from 3 major tributaries of Lake Michigan, USA. Microplastic was detected in fish and surface waters from all 3 sites, but there was no correlation between microplastic concentrations in fish and surface waters. Rather, there was a significant effect of functional feeding group on microplastic concentration in fish. Neogobius melanostomus (round goby, a zoobenthivore) had the highest concentration of gut microplastic (19 particles fish−1) compared to 10 other fish taxa measured, and had a positive linear relationship between body size and number of microplastic particles. Surface water microplastic concentrations were lowest in the most northern, forested watershed, and highest in the most southern, agriculturally dominated watershed. Results suggest microplastic pollution is common in river food webs and is connected to species feeding characteristics. Future research should focus on understanding the movement of microplastic from point-source and diffuse sources and into aquatic ecosystems, which will support pollution management efforts on inland waters

Preliminary evaluation of predicted peak release rates from the engineered barrier system for a potential repository at Yucca Mountain, Nevada

Any potential repository for the ultimate disposal of the nation`s high-level radioactive wastes is subject to meeting post-closure regulatory requirements as specified by the NRC. Three NRC sub-system performance measures are relevant to the evaluation of the Yucca Mountain site and possible engineered barriers. These performance requirements are specified in 10 CFR 60. These include the substantially complete containment requirement, the engineered barrier system (EBS) release requirement, and the pre-waste emplacement groundwater travel time requirement. The present paper documents an initial evaluation of the peak EBS release rates. A number of key factors significantly impact the maximum release rate from the engineered barrier system. The authors have conducted four simulations to approximate the effects of delaying and spreading out the failure distribution that are based on different thermal loads and criteria for the initiation of aqueous corrosion. Using an assumed outer barrier of 10 cm and an inner barrier of 0.95 cm and the Stahl model for aqueous pitting corrosion, they have analyzed the EBS release rates for thermal loads of 28.5, 57 and 83 kW/Ac using temperature as the corrosion limiting factor and at 57 kW/Ac for saturation limiting the initiation of corrosion. The later had the earliest failures and the most rapid failure rates observed in the TSPA-1993 analyses so provides the upper bound on the release rates

Oncologist-led BRCA ‘mainstreaming’ in the ovarian cancer clinic: A study of 255 patients and its impact on their management

Although guidelines recommend BRCA testing for all women with non-mucinous epithelial ovarian cancer, there is significant variability in access to testing across the UK. A germline BRCA mutation (BRCAm) in ovarian cancer patients provides prognostic and predictive information and influences clinical management, such as the use of PARP inhibitors, which have demonstrated a progression-free survival benefit in the BRCAm cohort. Additionally, the finding of a BRCAm has significant implications for patients and their families in terms of cancer risk and prevention. We studied the impact of a newly-formed, oncologist-led ‘mainstreaming’ germline BRCA testing pathway in 255 ovarian cancer patients at Imperial College NHS Trust. Prior to the establishment of ‘mainstreaming’, uptake of germline BRCA testing was 14% with a mean turnaround time of 148.2 calendar days. The ‘mainstreaming’ approach led to a 95% uptake of germline BRCA testing and a mean turnaround time of 20.6 days. Thirty-four (13.33%) BRCAm patients were identified. At the time of data collection nine BRCAm patients had received a PARP inhibitor off-trial, three had entered a PARP inhibitor trial and 5 were receiving platinum-based chemotherapy with a plan to receive PARP inhibitor maintenance. This study provides further evidence of the impact of oncologist-led ‘mainstreaming’ programs

Total System Performance Assessment, 1993: An evaluation of the potential Yucca Mountain repository, B00000000-01717-2200-00099, Rev. 01

Total System Performance Assessments are an important component in the evaluation of the suitability of Yucca Mountain, Nevada as a potential site for a mined geologic repository for the permanent disposal of high-level radioactive wastes in the United States. The Total System Performance Assessments are conducted iteratively during the site characterization to identify issues which should be addressed by the characterization and design activities as well as providing input to regulatory/licensing and programmatic decisions. During fiscal years 1991 and 1992, the first iteration of Total System Performance Assessment (hereafter referred to as TSPA 1991) was completed by Sandia National Laboratories and Pacific Northwest Laboratory. Beginning in fiscal year 1993, the Civilian Radioactive Waste Management System Management and Operating Contractor was assigned the responsibility to plan, coordinate, and contribute to the second iteration of Total System Performance Assessment (hereafter referred to as TSPA 1993). This document presents the objectives, approach, assumptions, input, results, conclusions, and recommendations associated with the Management and Operating Contractor contribution to TSPA 1993. A parallel effort was conducted by Sandia National Laboratories and is reported in Wilson et al. (1994, in press)

Sensitivity of the engineered barrier system (EBS) release rate to alternative conceptual models of advective release from waste packages under dripping fractures

The first model assumed that dripping water directly contacts the waste form inside the ``failed`` waste package and radionuclides are released from the EBS by advection. The second model assumed that dripping water is diverted around the package (because of corrosion products plugging the perforations), thereby being prevented from directly contacting the waste form. In the second model, radionuclides were assumed to diffuse through the perforations, and, once outside the waste package, to be released from the EBS by advection. For the case with the second EBS release model, most radionuclides had lower peak EBS release rates than with the first model. Impacts of the alternative EBS release models were greater for the radionuclides with low solubility. The analysis indicated that the EBS release model representing advection through a ``failed`` waste package (the first model) may be too conservative; thus a ``failed`` waste package container with multiple perforations may still be an important barrier to radionuclide release

Thinking beyond the hybrid:“actually-existing” cities “after neoliberalism” in Boyle <i>et al.</i>

In their article, ‘The spatialities of actually existing neoliberalism in Glasgow, 1977 to present’, Mark Boyle, Christopher McWilliams and Gareth Rice (2008) usefully problematise our current understanding of neoliberal urbanism. Our response is aimed at developing a sympathetic but critical approach to Boyle et al's understanding of neoliberal urbanism as illustrated by the Glasgow example. In particular, the counterposing by Boyle et al of a 'hybrid, mutant' model to a 'pure' model of neoliberalism for us misrepresents existing models of neoliberalism as a perfectly finished object rather than a roughly mottled process. That they do not identify any ‘pure’ model leads them to create a straw construct against which they can claim a more sophisticated, refined approach to the messiness of neoliberal urbanism. In contrast, we view neoliberalism as a contested and unstable response to accumulation crises at various scales of analysis