PHENOTYPIC AND FUNCTIONAL CHARACTERIZATION OF Ia+ AND la- SUBLINES OF THE HUMAN MONOCYTE-LIKE CELL LINE, U937

Sublines were derived from the U937 human monocytic cell line by limiting dilution techniques. Several of the sublines derived in this manner were found to constituitively express la, unlike most U937 cell lines previously examined. Thus the la+ and la- sublines were examined and characterized phenotypically and functionally. Morphologically the sublines, both la+ and la-, were found to be similar to the originally described U937 parent cell line. They were positive for production of non- specific esterase and expressed the HLA phenotype A(3,X), B(51,18), DR(2,X). The sublines were examined by immunofluorescence techniques with a large number of antibodies specific for cell surface structures and found to express MHC class I, MHC class II the major type being HLA-DR and perhaps HLA-DO in small amounts, Fc receptor, TA-1 structure typical of monocytes and T cells, endothelial antigens and CD4. The sublines and parent U937 cell line were found to express few cell surface antigens typical of mature monocytes/macrophages; however, they expressed myeloid antigens typical of early monocytic or promonocytic lineage. Expression of MHC class II by the sublines was confirmed by immunoprecipitation with monoclonal antibody to a framework determinant of human la and SDS-PAGE autoradiography which gave bands at p29:34 for Ia+ sublines but not for Ia- parent U937 cell lines. Cell surface expression of la was increased to a significant degree by treatment with gamma interferon which peaked at 24-48 hours of treatment. Functionally the parent U937 cell line and several Ia+ sublines were examined in several assays. The la- and Ia+ U937 cells were found to stimulate the generation of specific CTLs in CML assays to approximately the same degree. The Ia+ sublines were found to stimulate in MLR assay, although variable results were obtained and indicated that the U937 parent and subline cells produced factors which were found to be inherently immunosuppressive. The sublines were able to substitute for monocytes by reconstituting the C03 mediated T cell mitogenic response. The Ia+ sublines and the parent U937 cell line (also weakly Ia+) were found to present tetanus toxoid antigen to nylon-wool purified T cells following an overnight pulse with antigen, and this response was found to be significantly abrogated by addition of antibody specific for CD4 and MHC class II, but not MHC class I. Preliminary characterization of the immunosuppressive factor produced by the U937 cell line and the sublines revealed that it was strongly antiproliferative and affected lymphoid cells somewhat more than non-lymphoid cells; that its probable molecular weight was approximately 90,000, it was not inactivated by treatment with trypsin or chymotrypsin, was not immediately inactivated by freezing although dialysis and long term storage diminished its activity, and was partially inactivated by heat treatment at both 56°C and 80°C. Clear indication of soluble lL-1 production by the U937 parent cell line and the sublines was problematic due to the strong inhibition of proliferative assays by supernatants; however, partial inactivation of inhibitory activity by heat treatment as well as partial removal of the inhibitor fraction by gel filtration indicated that lL-1 or a cytokine with lL-1 activity was constituitively produced by the cells. Membrane lL-1 was detected in very low amounts in unstimulated cells and was significantly increased by treatment with phorbol esters but not other immunomodulators. Preliminary examination of Northern blots of HLA-DR alpha and HLA-DQ alpha mRNA production by the parent U937 cells and the sublines revealed that both HLA-DR alpha and HLA-DQ alpha mRNA were detectable for |a+ sublines E11, G4 and G11, that only trace amounts were detectable for relatively Ia- subline E9 and that surprisingly, HLA-DQ alpha was detectable for the 2-1 parent cell line but no HLA-DR alpha mFiNA was detectable. The results of mRNA analysis following gamma interferon treatment, as well as treatment with LPS and phorbol esters, were variable for the different sublines and indicate that in some sublines gamma interferon treatment appears to augment la specific mRNA and in others the levels are decreased indicating that the expression of MHC class II specific mRNA may be differentially regulated in the different sublines

On the static and dynamic properties of flax and Cordenka epoxy composites

Fibre reinforced composites have excellent specific properties and are widely sought after by engineers seeking to reduce mass. However, end of life disposal is a significant problem and so research into more sustainable natural fibre composites is extremely topical. This paper examines the applicability of natural fibre composites for high performance structural applications. Woven flax and regenerated cellulose (Cordenka) textiles were pre-impregnated with commercially available epoxy resins and consolidated into test laminates in an autoclave to determine their static (compressive, tensile, flexural) and dynamic (energy absorption) properties. The range of compressive strengths was 77.5–299.6 MPa. Tensile strengths ranged from 63 to 92.6 MPa and interlaminar shear strength (ILSS) from 10.7 to 23.3 MPa. Specific energy absorption (SEA) varied between 21.2–34.2 kJ/kg. Biotex flax combined with MTM49 resin matched the SEA of T300 carbon fibre using the same resin system and layup. This work has demonstrated that natural fibre composites have significant scope for use in structural applications but additional work is required on fibre to matrix bonding in order to maximise their properties whilst remaining an environmentally credible option

Using Extreme Value Theory for Determining the Probability of Carrington-Like Solar Flares

Space weather events can negatively affect satellites, the electricity grid, satellite navigation systems and human health. As a consequence, extreme space weather has been added to the UK and other national risk registers. By their very nature, extreme space weather events occur rarely and, therefore, statistical methods are required to determine the probability of their occurrence. Space weather events can be characterised by a number of natural phenomena such as X-ray (solar) flares, solar energetic particle (SEP) fluxes, coronal mass ejections and various geophysical indices (Dst, Kp, F10.7). In this paper extreme value theory (EVT) is used to investigate the probability of extreme solar flares. Previous work has assumed that the distribution of solar flares follows a power law. However such an approach can lead to a poor estimation of the return times of such events due to uncertainties in the tails of the probability distribution function. Using EVT and GOES X-ray flux data it is shown that the expected 150-year return level is approximately an X60 flare whilst a Carrington-like flare is a one in a 100-year event. It is also shown that the EVT results are consistent with flare data from the Kepler space telescope mission.Comment: 13 pages, 4 figures; updated content following reviewer feedbac

Extreme relativistic electron fluxes in the Earth's outer radiation belt: Analysis of INTEGRAL IREM data

Relativistic electrons (E > 500 keV) cause internal charging and are an important space weather hazard. To assess the vulnerability of the satellite fleet to these so-called “killer” electrons, it is essential to estimate reasonable worst cases, and, in particular, to estimate the flux levels that may be reached once in 10 and once in 100 years. In this study we perform an extreme value analysis of the relativistic electron fluxes in the Earth's outer radiation belt as a function of energy and L∗. We use data from the Radiation Environment Monitor (IREM) on board the International Gamma Ray Astrophysical Laboratory (INTEGRAL) spacecraft from 17 October 2002 to 31 December 2016. The 1 in 10 year flux at L∗=4.5, representative of equatorial medium Earth orbit, decreases with increasing energy ranging from 1.36 × 107 cm−2 s−1 sr−1 MeV−1 at E = 0.69 MeV to 5.34 × 105 cm−2 s−1 sr−1 MeV−1 at E = 2.05 MeV. The 1 in 100 year flux at L∗=4.5 is generally a factor of 1.1 to 1.2 larger than the corresponding 1 in 10 year flux. The 1 in 10 year flux at L∗=6.0, representative of geosynchronous orbit, decreases with increasing energy ranging from 4.35 × 106 cm−2 s−1 sr−1 MeV−1 at E = 0.69 MeV to 1.16 × 105 cm−2 s−1 sr−1 MeV−1 at E = 2.05 MeV. The 1 in 100 year flux at L∗=6.0 is generally a factor of 1.1 to 1.4 larger than the corresponding 1 in 10 year flux. The ratio of the 1 in 10 year flux at L∗=4.5 to that at L∗=6.0 increases with increasing energy ranging from 3.1 at E = 0.69 MeV to 4.6 at E = 2.05 MeV

Extreme energetic electron fluxes in low Earth orbit: Analysis of POES E > 30, E > 100 and E > 300 keV electrons

Energetic electrons are an important space weather hazard. Electrons with energies less than about 100 keV cause surface charging while higher energy electrons can penetrate materials and cause internal charging. In this study we conduct an extreme value analysis of the maximum 3-hourly flux of E> 30 keV, E> 100 keV and E> 300 keV electrons in low Earth orbit as a function of L∗, for geomagnetic field lines that map to the outer radiation belt, using data from the National Oceanic and Atmospheric Administration (NOAA) Polar Operational Environmental Satellites (POES) from July 1998 to June 2014. The 1 in 10 year flux of E> 30 keV electrons shows a general increasing trend with distance ranging from 1.8×107 cm−2s−1sr−1 at L∗ = 3.0 to 6.6×107 cm−2s−1sr−1 at L∗ = 8.0. The 1 in 10 year flux of E> 100 keV electrons peaks at L∗= 4.5 - 5.0 at 1.9×107 cm−2s−1sr−1 decreasing to minima of 7.1×106 and 8.7×106 cm−2s−1sr−1 at L∗ = 3.0 and 8.0 respectively. In contrast to the E> 30 keV electrons, the 1 in 10 year flux of E> 300 keV electrons shows a general decreasing trend with distance, ranging from 2.4×106 cm−2s−1sr−1 at L∗ = 3.0 to 1.2×105 cm−2s−1sr−1 at L∗= 8.0. Our analysis suggests that there is a limit to the E> 30 keV electrons with an upper bound in the range 5.1×107- 8.8×107 cm−2s−1sr−1. However, the results suggest that there is no upper bound for the E> 100 keV and E> 300 keV electrons

The use of the political categories of Brexiter and Remainer in online comments about the EU referendum

In June 2016, the UK held a referendum on EU membership; 52% of those who voted, voted to leave, and 48% voted to remain. During the referendum campaign two identities emerged: ‘Brexiter’ and ‘Remainer’, which remained salient post-referendum. This study explores how the categories of Brexiter and Remainer were deployed by posters online. Data comprises comment threads collected from four online newspapers both during the campaign and after the vote, which focus on the Brexit campaign promise: ‘We send £350m a week to the EU. Let’s fund our NHS instead’. We draw on membership categorisation analysis and discursive psychology to analyse when categories were made salient and what responses to the invocation of categories were. Analysis revealed that posters explicitly categorise the outgroup and in doing so implicitly define their group. Posters resisted other political identities when attributed to them in relation to the referendum. The analysis shows how Brexiter and Remainer are new, albeit contested, political categories and identities in their own right, with other political identities resisted when used. The paper highlights implications for the political system in the UK and for social divisions within UK society

Hypoxia-mediated upregulation of MCT1 expression supports the glycolytic phenotype of glioblastomas

Background: Glioblastomas (GBM) present a high cellular heterogeneity with conspicuous necrotic regions associated with hypoxia, which is related to tumor aggressiveness. GBM tumors exhibit high glycolytic metabolism with increased lactate production that is extruded to the tumor microenvironment through monocarboxylate transporters (MCTs). While hypoxia-mediated regulation of MCT4 has been characterized, the role of MCT1 is still controversial. Thus, we aimed to understand the role of hypoxia in the regulation of MCT expression and function in GBM, MCT1 in particular. Methods: Expression of hypoxia-and glycolytic-related markers, as well as MCT1 and MCT4 isoforms was assessed in in vitro and in vivo orthotopic glioma models, and also in human GBM tissues by immunofluorescence/immunohistochemistry and Western blot. Following MCT1 inhibition, either pharmacologically with CHC (a-cyano-4-hydroxynnamic acid) or genetically with siRNAs, we assessed GBM cell viability, proliferation, metabolism, migration and invasion, under normoxia and hypoxia conditions. Results: Hypoxia induced an increase in MCT1 plasma membrane expression in glioma cells, both in in vitro and in vivo models. Additionally, treatment with CHC and downregulation of MCT1 in glioma cells decreased lactate production, cell proliferation and invasion under hypoxia. Moreover, in the in vivo orthotopic model and in human GBM tissues, there was extensive co-expression of MCT1, but not MCT4, with the GBM hypoxia marker CAIX. Conclusion: Hypoxia-induced MCT1 supports GBM glycolytic phenotype, being responsible for lactate efflux and an important mediator of cell survival and aggressiveness. Therefore, MCT1 constitutes a promising therapeutic target in GBM.This study was supported by Projecto Estratégico- LA 26 – 2013-2014 (PEst-C/SAL/LA0026/2013) and ON.2 SR&TD Integrated Program (NORTE-07-0124FEDER-000017)” co-funded by Programa Operacional Regional do Norte (ON.2- O Novo Norte), Quadro de Referência Estratégico Nacional (QREN), through Fundo Europeu de Desenvolvimento Regional (FEDER), as well as MCTI/CNPq Nº 73/2013 (Brazil). VMG received a fellowship from Fundação para a Ciência e Tecnologia (FCT) ref. SFRH/BD/51997/2012.info:eu-repo/semantics/publishedVersio

Stabilization of dense Antarctic water supply to the Atlantic Ocean overturning circulation

The lower limb of the Atlantic overturning circulation is resupplied by the sinking of dense Antarctic Bottom Water (AABW) that forms via intense air–sea–ice interactions next to Antarctica, especially in the Weddell Sea. In the last three decades, AABW has warmed, freshened and declined in volume across the Atlantic Ocean and elsewhere, suggesting an ongoing major reorganization of oceanic overturning. However, the future contributions of AABW to the Atlantic overturning circulation are unclear. Here, using observations of AABW in the Scotia Sea, the most direct pathway from the Weddell Sea to the Atlantic Ocean, we show a recent cessation in the decline of the AABW supply to the Atlantic overturning circulation. The strongest decline was observed in the volume of the densest layers in the AABW throughflow from the early 1990s to 2014; since then, it has stabilized and partially recovered. We link these changes to variability in the densest classes of abyssal waters upstream. Our findings indicate that the previously observed decline in the supply of dense water to the Atlantic Ocean abyss may be stabilizing or reversing and thus call for a reassessment of Antarctic influences on overturning circulation, sea level, planetary-scale heat distribution and global climate

State of the Antarctic and Southern Ocean Climate System

This paper reviews developments in our understanding of the state of the Antarctic and Southern Ocean climate and its relation to the global climate system over the last few millennia. Climate over this and earlier periods has not been stable, as evidenced by the occurrence of abrupt changes in atmospheric circulation and temperature recorded in Antarctic ice core proxies for past climate. Two of the most prominent abrupt climate change events are characterized by intensification of the circumpolar westerlies (also known as the Southern Annular Mode) between ∼6000 and 5000 years ago and since 1200–1000 years ago. Following the last of these is a period of major trans-Antarctic reorganization of atmospheric circulation and temperature between A.D. 1700 and 1850. The two earlier Antarctic abrupt climate change events appear linked to but predate by several centuries even more abrupt climate change in the North Atlantic, and the end of the more recent event is coincident with reorganization of atmospheric circulation in the North Pacific. Improved understanding of such events and of the associations between abrupt climate change events recorded in both hemispheres is critical to predicting the impact and timing of future abrupt climate change events potentially forced by anthropogenic changes in greenhouse gases and aerosols. Special attention is given to the climate of the past 200 years, which was recorded by a network of recently available shallow firn cores, and to that of the past 50 years, which was monitored by the continuous instrumental record. Significant regional climate changes have taken place in the Antarctic during the past 50 years. Atmospheric temperatures have increased markedly over the Antarctic Peninsula, linked to nearby ocean warming and intensification of the circumpolar westerlies. Glaciers are retreating on the peninsula, in Patagonia, on the sub-Antarctic islands, and in West Antarctica adjacent to the peninsula. The penetration of marine air masses has become more pronounced over parts of West Antarctica. Above the surface, the Antarctic troposphere has warmed during winter while the stratosphere has cooled year-round. The upper kilometer of the circumpolar Southern Ocean has warmed, Antarctic Bottom Water across a wide sector off East Antarctica has freshened, and the densest bottom water in the Weddell Sea has warmed. In contrast to these regional climate changes, over most of Antarctica, near-surface temperature and snowfall have not increased significantly during at least the past 50 years, and proxy data suggest that the atmospheric circulation over the interior has remained in a similar state for at least the past 200 years. Furthermore, the total sea ice cover around Antarctica has exhibited no significant overall change since reliable satellite monitoring began in the late 1970s, despite large but compensating regional changes. The inhomogeneity of Antarctic climate in space and time implies that recent Antarctic climate changes are due on the one hand to a combination of strong multidecadal variability and anthropogenic effects and, as demonstrated by the paleoclimate record, on the other hand to multidecadal to millennial scale and longer natural variability forced through changes in orbital insolation, greenhouse gases, solar variability, ice dynamics, and aerosols. Model projections suggest that over the 21st century the Antarctic interior will warm by 3.4° ± 1°C, and sea ice extent will decrease by ∼30%. Ice sheet models are not yet adequate enough to answer pressing questions about the effect of projected warming on mass balance and sea level. Considering the potentially major impacts of a warming climate on Antarctica, vigorous efforts are needed to better understand all aspects of the highly coupled Antarctic climate system as well as its influence on the Earth\u27s climate and oceans