Cytosolic Phospholipase A2α and Eicosanoids Regulate Expression of Genes in Macrophages Involved in Host Defense and Inflammation

Acknowledgments: We thank Dr. Robert Barkley and Charis Uhlson for mass spectrometry analysis. Funding: This work was supported by grants from the National Institutes of Health HL34303 (to C.C.L., R.C.M. and D.L.B), DK54741 (to J.V.B.), GM5322 (to D.L.W.) and the Wellcome Trust (to N.A.R.G. and G.D.B.). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.Peer reviewedPublisher PD

NASA Technology Area 07: Human Exploration Destination Systems Roadmap

This paper gives an overview of the National Aeronautics and Space Administration (NASA) Office of Chief Technologist (OCT) led Space Technology Roadmap definition efforts. This paper will given an executive summary of the technology area 07 (TA07) Human Exploration Destination Systems (HEDS). These are draft roadmaps being reviewed and updated by the National Research Council. Deep-space human exploration missions will require many game changing technologies to enable safe missions, become more independent, and enable intelligent autonomous operations and take advantage of the local resources to become self-sufficient thereby meeting the goal of sustained human presence in space. Taking advantage of in-situ resources enhances and enables revolutionary robotic and human missions beyond the traditional mission architectures and launch vehicle capabilities. Mobility systems will include in-space flying, surface roving, and Extra-vehicular Activity/Extravehicular Robotics (EVA/EVR) mobility. These push missions will take advantage of sustainability and supportability technologies that will allow mission independence to conduct human mission operations either on or near the Earth, in deep space, in the vicinity of Mars, or on the Martian surface while opening up commercialization opportunities in low Earth orbit (LEO) for research, industrial development, academia, and entertainment space industries. The Human Exploration Destination Systems (HEDS) Technology Area (TA) 7 Team has been chartered by the Office of the Chief Technologist (OCT) to strategically roadmap technology investments that will enable sustained human exploration and support NASA s missions and goals for at least the next 25 years. HEDS technologies will enable a sustained human presence for exploring destinations such as remote sites on Earth and beyond including, but not limited to, LaGrange points, low Earth orbit (LEO), high Earth orbit (HEO), geosynchronous orbit (GEO), the Moon, near-Earth objects (NEOs), which > 95% are asteroidal bodies, Phobos, Deimos, Mars, and beyond. The HEDS technology roadmap will strategically guide NASA and other U.S. Government agency technology investments that will result in capabilities enabling human exploration missions to diverse destinations generating high returns on investments

Investigating discrepancies between experimental solid-state NMR and GIPAW calculation : NC–N 13C and OH⋯O 1H chemical shifts in pyridinium fumarates and their cocrystals

An NMR crystallography analysis is presented for four solid-state structures of pyridine fumarates and their cocrystals, using crystal structures deposited in the Cambridge Crystallographic Data Centre, CCDC. Experimental one-dimensional, one-pulse 1H and 13C cross-polarisation (CP) magic-angle spinning (MAS) nuclear magnetic resonance (NMR) and two-dimensional 14N–1H heteronuclear multiple-quantum coherence MAS NMR spectra are compared with gauge-including projector augmented wave (GIPAW) calculations of the 1H and 13C chemical shifts and the 14N shifts that additionally depend on the quadrupolar interaction. Considering the high ppm (>10 ppm) 1H resonances, while there is good agreement (within 0.4 ppm) between experiment and GIPAW calculation for the hydrogen-bonded NH moieties, the hydrogen-bonded fumaric acid OH resonances are 1.2–1.9 ppm higher in GIPAW calculation as compared to experiment. For the cocrystals of a salt and a salt formed by 2-amino-5-methylpyridinium and 2-amino-6-methylpyridinium ions, a large discrepancy of 4.2 and 5.9 ppm between experiment and GIPAW calculation is observed for the quaternary ring carbon 13C resonance that is directly bonded to two nitrogens (in the ring and in the amino group). By comparison, there is excellent agreement (within 0.2 ppm) for the quaternary ring carbon 13C resonance directly bonded to the ring nitrogen for the salt and cocrystal of a salt formed by 2,6-lutidinium and 2,5-lutidine, respectively

Isolation and characterization of the full-length cDNA encoding a member of a novel cytochrome p450 family (CYP320A1) from the tropical freshwater snail, Biomphalaria glabrata, intermediate host for Schistosoma mansoni

Cytochrome p450s (cyp450s) are a family of structurally related proteins, with diverse functions, including steroid synthesis and breakdown of toxins. This paper reports the full-length sequence of a novel cyp450 gene, the first to be isolated from the tropical freshwater snail Biomphalaria glabrata, an important intermediate host of Schistosoma mansoni. The nucleotide sequence is 2291 bp with a predicted amino acid sequence of 584aa. The sequence demonstrates conserved cyp450 structural motifs, but is sufficiently different from previously reported cyp450 sequences to be given a new classification, CYP320A1. Initially identified as down-regulated in partially resistant snails in response to S. mansoni infection, amplification of this gene using RT-PCR in both totally resistant or susceptible snail lines when exposed to infection, and all tissues examined, suggests ubiquitous expression. Characterization of the first cyp450 from B. glabrata is significant in understanding the evolution of these metabolically important proteins

Enabling the Participation of People with Parkinson's and Their Caregivers in Co-Inquiry around Collectivist Health Technologies

While user participation is central to HCI, co-inquiry takes this further by having participants direct and control research from conceptualisation to completion. We describe a co-inquiry, conducted over 16 months with a Parkinson's support group. We explored how the participation of members might be enabled across multiple stages of a research project, from the generation of research questions to the development of a prototype. Participants directed the research into developing alternative modes of information provision, resulting in ‘Parkinson’s Radio’ — a collectivist health information service produced and edited by members of the support group. We reflect on how we supported participation at different stages of the project and the successes and challenges faced by the team. We contribute insights into the design of collectivist health technologies for this group, and discuss opportunities and tensions for conducting co-inquiry in HCI research

Regulation of Cytosolic Phospholipase A 2 Activation and Cyclooxygenase 2 Expression in Macrophages by the β-Glucan Receptor

Phagocytosis of non-opsonized microorganisms by macrophages initiates innate immune responses for host defense against infection. Cytosolic phospholipase A(2) is activated during phagocytosis, releasing arachidonic acid for production of eicosanoids, which initiate acute inflammation. Our objective was to identify pattern recognition receptors that stimulate arachidonic acid release and cyclooxygenase 2 (COX2) expression in macrophages by pathogenic yeast and yeast cell walls. Zymosan- and Candida albicans-stimulated arachidonic acid release from resident mouse peritoneal macrophages was blocked by soluble glucan phosphate. In RAW264.7 cells arachidonic acid release, COX2 expression, and prostaglandin production were enhanced by overexpressing the beta-glucan receptor, dectin-1, but not dectin-1 lacking the cytoplasmic tail. Pure particulate (1, 3)-beta-D-glucan stimulated arachidonic acid release and COX2 expression, which were augmented in a Toll-like receptor 2 (TLR2)-dependent manner by macrophage-activating lipopeptide-2. However, arachidonic acid release and leukotriene C(4) production stimulated by zymosan and C. albicans were TLR2-independent, whereas COX2 expression and prostaglandin production were partially blunted in TLR2(-/-) macrophages. Inhibition of Syk tyrosine kinase blocked arachidonic acid release and COX2 expression in response to zymosan, C. albicans, and particulate (1, 3)-beta-D-glucan. The results suggest that cytosolic phospholipase A(2) activation triggered by the beta-glucan component of yeast is dependent on the immunoreceptor tyrosine-based activation motif-like domain of dectin-1 and activation of Syk kinase, whereas both TLR2 and Syk kinase regulate COX2 expression

The variability of the sediment plume and ocean circulation features of the Nass River Estuary, British Columbia

The Nass River discharges into Nass Bay and Iceberg Bay, which are adjoining tidal inlets located within the northern inland waters of British Columbia, Canada. After the Skeena River, the Nass River is the second longest river within northern British Columbia, which discharges directly into Canadian waters of the Pacific Ocean. It is also supports one of the most productive salmon fisheries in northern British Columbia. The Nass River discharges into the eastern end of Nass Bay. Nass Bay, in turn feeds into Portland Canal and the fresh surface waters then flows westward to the Pacific Ocean via Dixon Entrance. The tides in Northern British Columbia are very large with a tidal height range of just over 7 m. Nass Bay is a shallow inlet of less than 10 km in length with typical water depths of than 10 m or less. The existing knowledge of oceanographic processes in Nass and Iceberg Bays was rudimentary until three years ago, when the first modern oceanographic measurements were obtained. In this study, the seasonal and tidal variability of the lateral extent of the Nass River surface plume is mapped from analyses of Landsat satellite data spanning the period from 2008 to 2015. A high resolution coupled three dimensional (3D) hydrodynamic model was developed and implemented, within the widely used and accepted Delft3D modeling framework, which was forced and validated using recent 2013-2016 in-situ oceanographic measurements. The combined satellite and numerical modeling methods are used to study the physical oceanographic and sediment transport regime of Nass and Iceberg Bays and the adjoining waters of Portland Inlet and Observatory Inlet. The ocean circulation of Nass and Iceberg Bays was found to be dominated by tidal currents, and by the highly seasonal and variable Nass River freshwater discharges. Complex lateral spatial patterns in the tidal currents occur due to the opening of the southwestern side of Nass Bay onto the deeper adjoining waters of Iceberg Bay. Surface winds are limited to a secondary role in the circulation variability. The sediment dynamics of the Nass Bay system features a very prominent surface sediment plume present from the time of freshet in mid-spring through to large rainfall runoff events in the fall. The time-varying turbidity distribution and transport paths of the Nass River sediment discharges in the study area were characterized using the model results combined with an analysis of several high-resolution multi-year Landsat satellite data sets

Regulation of Cytosolic Phospholipase a\u3csub\u3e2\u3c/sub\u3e Activation and Cyclooxygenase 2 Expression in Macrophages by the β-Glucan Receptor

Phagocytosis of non-opsonized microorganisms bymacrophages initiates innate immune responses for host defense against infection. Cytosolic phospholipase A2 is activated during phagocytosis, releasing arachidonic acid for production of eicosanoids, which initiate acute inflammation. Our objective was to identify pattern recognition receptors that stimulate arachidonic acid release and cyclooxygenase 2 (COX2) expression in macrophages by pathogenic yeast and yeast cell walls. Zymosan- and Candida albicans-stimulated arachidonic acid release from resident mouse peritoneal macrophages was blocked by soluble glucan phosphate. In RAW264.7 cells arachidonic acid release, COX2 expression, and prostaglandin production were enhanced by overexpressing the β-glucan receptor, dectin-1, but not dectin-1 lacking the cytoplasmic tail. Pure particulate (1, 3)-β-D-glucan stimulated arachidonic acid release and COX2 expression, which were augmented in a Toll-like receptor 2 (TLR2)-dependent manner by macrophage-activating lipopeptide-2. However, arachidonic acid release and leukotriene C4 production stimulated by zymosan and C. albicans were TLR2-independent, whereas COX2 expression and prostaglandin production were partially blunted in TLR2-/- macrophages. Inhibition of Syk tyrosine kinase blocked arachidonic acid release and COX2 expression in response to zymosan, C. albicans, and particulate (1, 3)-β-D-glucan. The results suggest that cytosolic phospholipase A2 activation triggered by the β-glucan component of yeast is dependent on the immunoreceptor tyrosine-based activation motif-like domain of dectin-1 and activation of Syk kinase, whereas both TLR2 and Syk kinase regulate COX2 expression

Pathways Regulating Cytosolic Phospholipase a\u3csub\u3e2\u3c/sub\u3e Activation and Eicosanoid Production in Macrophages by Candida Albicans

Resident tissue macrophages are activated by the fungal pathogen Candida albicans to release eicosanoids, which are important modulators of inflammation and immune responses. Our objective was to identify the macrophage receptors engaged by C. albicans that mediate activation of group IVA cytosolic phospholipase A2 (cPLA2α), a regulatory enzyme that releases arachidonic acid (AA) for production of prostaglandins and leukotrienes. A comparison of peritoneal macrophages from wild type and knock-out mice demonstrates that the β-glucan receptor Dectin-1 and MyD88 regulate early release of AA and eicosanoids in response to C. albicans. However, cyclooxygenase 2 (COX2) expression and later phase eicosanoid production are defective in MyD88-/- but not Dectin-1-/- macrophages. Furthermore, C. albicans-stimulated activation of MAPK and phosphorylation of cPLA2α on Ser-505 are regulated by MyD88 and not Dectin-1. In contrast, Dectin-1 mediates MAPK activation, cPLA 2α phosphorylation, and COX2 expression in response to particulate β-glucan suggesting that other receptors engaged by C. albicans preferentially mediate these responses. Results also implicate the mannan-binding receptor Dectin-2 in regulating cPLA2α. C. albicans-stimulated MAPK activation and AA release are blocked by D-mannose and Dectin-2-specific antibody, and overexpression of Dectin-2 in RAW264.7 macrophages enhances C. albicans-stimulated MAPK activation, AA release, and COX2 expression. In addition, calcium mobilization is enhanced in RAW264.7 macrophages overexpressing Dectin-1 or -2. The results demonstrate that C. albicans engages both β-glucan and mannan-binding receptors on macrophages that act with MyD88 to regulate the activation of cPLA2α and eicosanoid production