Identification of systemic immune response markers through metabolomic profiling of plasma from calves given an intra-nasally delivered respiratory vaccine

Vaccination procedures within the cattle industry are important disease control tools to minimize economic and welfare burdens associated with respiratory pathogens. However, new vaccine, antigen and carrier technologies are required to combat emerging viral strains and enhance the efficacy of respiratory vaccines, particularly at the point of pathogen entry. New technologies, specifically metabolomic profiling, could be applied to identify metabolite immune-correlates representative of immune protection following vaccination aiding in the design and screening of vaccine candidates. This study for the first time demonstrates the ability of untargeted UPLC-MS metabolomic profiling to identify metabolite immune correlates characteristic of immune responses following mucosal vaccination in calves. Male Holstein Friesian calves were vaccinated with Pfizer Rispoval® PI3 + RSV intranasal vaccine and metabolomic profiling of post-vaccination plasma revealed 12 metabolites whose peak intensities differed significantly from controls. Plasma levels of glycocholic acid, N-[(3α,5β,12α)-3,12-Dihydroxy-7,24-dioxocholan-24-yl]glycine, uric acid and biliverdin were found to be significantly elevated in vaccinated animals following secondary vaccine administration, whereas hippuric acid significantly decreased. In contrast, significant upregulation of taurodeoxycholic acid and propionylcarnitine levels were confined to primary vaccine administration. Assessment of such metabolite markers may provide greater information on the immune pathways stimulated from vaccine formulations and benchmarking early metabolomic responses to highly immunogenic vaccine formulations could provide a means for rapidly assessing new vaccine formulations. Furthermore, the identification of metabolic systemic immune response markers which relate to specific cell signaling pathways of the immune system could allow for targeted vaccine design to stimulate key pathways which can be assessed at the metabolic level.

238.2T-14 Concrete Thixotropy

Fresh concrete can exhibit different rheological behavior when at rest than when flowing. This difference is due to thixotropy, which can have important consequences for formwork pressure, multi-lift casting, slip-form paving, pumping, and segregation resistance. This TechNote defines thixotropy and distinguishes it from other changes in rheological properties; discusses the origins of, test methods for measuring, and factors affecting thixotropy; and concludes with its applications.

HDAC Inhibition Overcomes Acute Resistance to MEK Inhibition in BRAF-Mutant Colorectal Cancer by Downregulation of c-FLIPL

Purpose: Activating mutations in the BRAF oncogene are found in 8% to 15% of colorectal cancer patients and have been associated with poor survival. In contrast with BRAF-mutant (MT) melanoma, inhibition of the MAPK pathway is ineffective in the majority of BRAFMT colorectal cancer patients. Therefore, identification of novel therapies for BRAFMT colorectal cancer is urgently needed.

Experimental Design: BRAFMT and wild-type (WT) colorectal cancer models were assessed in vitro and in vivo. Small-molecule inhibitors of MEK1/2, MET, and HDAC were used, overexpression and siRNA approaches were applied, and cell death was assessed by flow cytometry, Western blotting, cell viability, and caspase activity assays.

Results: Increased c-MET-STAT3 signaling was identified as a novel adaptive resistance mechanism to MEK inhibitors (MEKi) in BRAFMT colorectal cancer models in vitro and in vivo. Moreover, MEKi treatment resulted in acute increases in transcription of the endogenous caspase-8 inhibitor c-FLIPL in BRAFMT cells, but not in BRAFWT cells, and inhibition of STAT3 activity abrogated MEKi-induced c-FLIPL expression. In addition, treatment with c-FLIP–specific siRNA or HDAC inhibitors abrogated MEKi-induced upregulation of c-FLIPL expression and resulted in significant increases in MEKi-induced cell death in BRAFMT colorectal cancer cells. Notably, combined HDAC inhibitor/MEKi treatment resulted in dramatically attenuated tumor growth in BRAFMT xenografts.

Conclusions: Our findings indicate that c-MET/STAT3-dependent upregulation of c-FLIPL expression is an important escape mechanism following MEKi treatment in BRAFMT colorectal cancer. Thus, combinations of MEKi with inhibitors of c-MET or c-FLIP (e.g., HDAC inhibitors) could be potential novel treatment strategies for BRAFMT colorectal cancer.

Women, letter-writing and the life of the mind in England, c.1650-1750

Drawing on national and regional letter collections dating from the late seventeenth and early eighteenth centuries, this article explores women's experiences of the life of the mind through an analysis of their letter-writing. This study also highlights the shortcomings of the compartmentalised nature of scholarship on women's writing and intellectual lives and proposes the letter both as a beneficial historical source and methodological tool for research on women's mental worlds. By employing an inclusive definition of intellectual and creative life, and eschewing traditional benchmarks of achievement, this article contends that women took a full part in the cultures of knowledge of their time. 

Comet C/2012 S1 (ISON): Observations of the Dust Grains from SOFIA and of the Atomic Gas from NSO Dunn and McMath-Pierce Solar Telescopes (Invited)

Comet C/2012 S1 (ISON) is unique in that it is a dynamically new comet derived from the Oort cloud reservoir of comets with a sun-grazing orbit. Infrared (IR) and visible wavelength observing campaigns were planned on NASA's Stratospheric Observatory For Infrared Astronomy (SOFIA) and on National Solar Observatory Dunn (DST) and McMath-Pierce Solar Telescopes, respectively. We highlight our early results. SOFIA (+FORCAST [1]) mid- to far-IR images and spectroscopy (~5-35 μm) of the dust in the coma of ISON are to be obtained by the ISON-SOFIA Team during a flight window 2013 Oct 21-23 UT (r_h≈1.18 AU). Dust characteristics, identified through the 10 μm silicate emission feature and its strength [2], as well as spectral features from cometary crystalline silicates (Forsterite) at 11.05-11.2 μm, and near 16, 19, 23.5, 27.5, and 33 μm are compared with other Oort cloud comets that span the range of small and/or highly porous grains (e.g., C/1995 O1 (Hale-Bopp) [3,4,5] and C/2001 Q4 (NEAT) [6]) to large and/or compact grains (e.g., C/2007 N4 (Lulin) [7] and C/2006 P1 (McNaught) [8]). Measurement of the crystalline peaks in contrast to the broad 10 and 20 μm amorphous silicate features yields the cometary silicate crystalline mass fraction [9], which is a benchmark for radial transport in our protoplanetary disk [10]. The central wavelength positions, relative intensities, and feature asymmetries for the crystalline peaks may constrain the shapes of the crystals [11]. Only SOFIA can look for cometary organics in the 5-8 μm region. Spatially resolved measurements of atoms and simple molecules from when comet ISON is near the Sun (r_h<0.4 AU, near Nov-20--Dec-03 UT) were proposed for by the ISON-DST Team. Comet ISON is the first comet since comet Ikeya-Seki (1965f) [12,13] suitable for studying the alkalai metals Na and K and the atoms specifically attributed to dust grains including Mg, Si, Fe, as well as Ca. DST's Horizontal Grating Spectrometer (HGS) measures 4 settings: Na I, K, C2 to sample cometary organics (along with Mg I), and [O I] as a proxy for activity from water [14] (along with Si I and Fe I). State-of-the-art instruments that will also be employed include IBIS [15], which is a Fabry-Perot spectral imaging system that concurrently measures lines of Na, K, Ca II, or Fe, and ROSA (CSUN/QUB) [16], which is a rapid imager that simultaneously monitors Ca II or CN. From McMath-Pierce, the Solar-Stellar Spectrograph also will target ISON (320-900 nm, R~21,000, r_h

The 'human revolution' in lowland tropical Southeast Asia:the antiquity and behavior of anatomically modern humans at Niah Cave (Sarawak, Borneo)

Recent research in Europe, Africa, and Southeast Asia suggests that we can no longer assume a direct and exclusive link between anatomically modern humans and behavioral modernity (the 'human revolution'), and assume that the presence of either one implies the presence of the other: discussions of the emergence of cultural complexity have to proceed with greater scrutiny of the evidence on a site-by-site basis to establish secure associations between the archaeology present there and the hominins who created it. This paper presents one such case study: Niah Cave in Sarawak on the island of Borneo, famous for the discovery in 1958 in the West Mouth of the Great Cave of a modern human skull, the 'Deep Skull,' controversially associated with radiocarbon dates of ca. 40,000 years before the present. A new chronostratigraphy has been developed through a re-investigation of the lithostratigraphy left by the earlier excavations, AMS-dating using three different comparative pre-treatments including ABOX of charcoal, and U-series using the Diffusion-Absorption model applied to fragments of bones from the Deep Skull itself. Stratigraphic reasons for earlier uncertainties about the antiquity of the skull are examined, and it is shown not to be an `intrusive' artifact. It was probably excavated from fluvial-pond-desiccation deposits that accumulated episodically in a shallow basin immediately behind the cave entrance lip, in a climate that ranged from times of comparative aridity with complete desiccation, to episodes of greater surface wetness, changes attributed to regional climatic fluctuations. Vegetation outside the cave varied significantly over time, including wet lowland forest, montane forest, savannah, and grassland. The new dates and the lithostratigraphy relate the Deep Skull to evidence of episodes of human activity that range in date from ca. 46,000 to ca. 34,000 years ago. Initial investigations of sediment scorching, pollen, palynomorphs, phytoliths, plant macrofossils, and starch grains recovered from existing exposures, and of vertebrates from the current and the earlier excavations, suggest that human foraging during these times was marked by habitat-tailored hunting technologies, the collection and processing of toxic plants for consumption, and, perhaps, the use of fire at some forest-edges. The Niah evidence demonstrates the sophisticated nature of the subsistence behavior developed by modern humans to exploit the tropical environments that they encountered in Southeast Asia, including rainforest. (c) 2006 Elsevier Ltd. All rights reserved.

Statistical Analysis of Small Ellerman Bomb Events

The properties of Ellerman bombs (EBs), small-scale brightenings in the Hα line wings, have proved difficult to establish because their size is close to the spatial resolution of even the most advanced telescopes. Here, we aim to infer the size and lifetime of EBs using high-resolution data of an emerging active region collected using the Interferometric BIdimensional Spectrometer (IBIS) and Rapid Oscillations of the Solar Atmosphere (ROSA) instruments as well as the Helioseismic and Magnetic Imager (HMI) onboard the Solar Dynamics Observatory (SDO). We develop an algorithm to track EBs through their evolution, finding that EBs can often be much smaller (around 0.3″) and shorter-lived (less than one minute) than previous estimates. A correlation between G-band magnetic bright points and EBs is also found. Combining SDO/HMI and G-band data gives a good proxy of the polarity for the vertical magnetic field. It is found that EBs often occur both over regions of opposite polarity flux and strong unipolar fields, possibly hinting at magnetic reconnection as a driver of these events.The energetics of EB events is found to follow a power-law distribution in the range of a nanoflare (1022-25 ergs).

The effect of ionization on the populations of excited levels of C IV and C V in tokamak edge plasmas

The main populating and depopulating mechanisms of the excited energy levels of ions in plasmas with densities <10²³-10²⁴ m^-3 are electron collisional excitation from the ion's ground state and radiative decay, respectively, with the majority of the electron population being in the ground state of the ionization stage. Electron collisional ionization is predominately expected to take place from one ground state to that of the next higher ionization stage. However, the question arises as to whether, in some cases, ionization can also affect the excited level populations. This would apply particularly to those cases involving transient events such as impurity influxes in a laboratory plasma. An analysis of the importance of ionization in populating the excited levels of ions in plasmas typical of those found in the edge of tokamaks is undertaken for the C IV and C V ionization stages. The emphasis is on those energy levels giving rise to transitions of most use for diagnostic purposes (n ≤ 5). Carbon is chosen since it is an important contaminant of JET plasmas; it was the dominant low Z impurity before the installation of the ITER-like wall and is still present in the plasma after its installation. Direct electron collisional ionization both from and to excited levels is considered. Distorted-wave flexible atomic code calculations are performed to generate the required ionization cross sections, due to a lack of atomic data in the literature. Employing these data, ionization from excited level populations is not found to be significant in comparison with radiative decay. However, for some energy levels, ionization terminating in the excited level has an effect in the steady-state of the order of the measurement errors (±10%). During transient events, ionization to excited levels will be of more importance and must be taken into account in the calculation of excited level populations. More accurate atomic data, including possible resonance contributions to the cross sections, would tend to increase further the importance of these effects. 

Freshwater reservoir effect (FRE) on re-dating of the prehistoric Eurasian Steppe cultures

There is a major problem with dating human skeletal material in Eurasian
steppe, possibly due to the freshwater reservoir effects (FRE). The goal of the
project is to examine the extent of the FRE in key areas of Siberia and
Kazakhstan through AMS 14C dating and stable isotope (δ13C, δ15N, δ34S)
analysis of c. 150 archaeological and modern samples. Here, we present the
very first results of the tests. We hope that within next years the study will
provide advanced knowledge of FRE across the region; improved 14C sampling
protocols for 14C and isotopic studies; new data on the chronology, diet and
isotopic ecology of particular archaeological sites, and a database on the
extent and variability of the FRE in the region

Using radiocarbon and stable isotopes to identify sediment carbon sources in an alkaline, humic lake.

We present a pilot study that uses the radiocarbon (∆14C) method to determine the source of carbon buried in the surface sediment of Lough Erne, a humic, alkaline lake in northwest Ireland. ∆14C, δ13C and δ15N values were measured from phytoplankton, dissolved inorganic, dissolved organic and particulate organic carbon. A novel radiocarbon method, Stepped Combustion1 was used to estimate the degree of the burial of terrestrial carbon in surface sediment. The ∆14C values of the low temperature fractions were comparable to algal ∆14C, while the high temperature fractions were 14C-depleted (older than bulk sediment). The ∆14C end-member model indicated that ~64% of carbon in surface sediment was derived from detrital terrestrial carbon. The use of ∆14C in conjunction with stepped combustion allows the quantification of the pathways of terrestrial carbon in the system, which has implications for regional and global carbon burial.
1McGeehin, J., Burr, G.S., Jull, A.J.T., Reines, D., Gosse, J., Davis, P.T., Muhs, D., and Southon, J.R., 2001, Stepped-combustion C-14 dating of sediment: A comparison with established techniques: Radiocarbon, v. 43, p. 255-261.

Freshwater Reservoir Effect on Re-Dating of Eurasian Steppe Cultures: First Results for Eneolithic and Early Bronze Age North-East Kazakhstan

Freshwater reservoir effects (FRE) can cause a major problem with radiocarbon dating human skeletal material in the Eurasian steppe. We present the first results of research into the extent of the FRE in the sites of Borly 4 (Eneolithic), and Shauke 1 and 8b (Early Bronze Age), North-Eastern Kazakhstan. AMS 14C dating and stable isotope (δ13C, δ15N) analysis of associated groups of samples (32 samples, 11 groups in total) demonstrate that: a) the diet of the humans and fauna analysed was based on the C3 foodchain with no evidence of a C4 plant (such as millet) contribution; aquatic resources apparently were a continuous dietary feature for the humans; b) the first 14C dates obtained for the Upper and Middle Irtysh River region attribute the Eneolithic period of the area to the 34th-30th c. BC, and the Early Bronze Age – to the 25th-20th c. BC; there is a ca. 450 years hiatus between the two periods; c) the maximum fish-herbivore freshwater reservoir offset observed equals 301±47 14C yrs. As such, 14C dates from aquatic and human samples from the area need to be interpreted with caution as they are likely to be affected by the offset (i.e. appear older).
The paper also discusses the effect of a sodium hydroxide (NaOH) wash on δ13C, δ15N, C:Natomic levels and collagen yields of the bone samples. Our results indicate a minor but significant effect of NaOH treatment only on C:Natomic ratios of the samples.

A sedimentologic and 14C dating study of five eastern Australian upper continental slope submarine landslides

Sedimentologic and AMS 14C age data are reported for calcareous hemipelagic mud samples taken from gravity cores collected at sites within, or adjacent to five submarine landslides identified with multibeam bathymetry data on the Nerrang Plateau segment and surrounding canyons of eastern Australia's continental slope (Bribie Bowl, Coolangatta-2, Coolangatta-1, Cudgen and Byron). Sediments are comprised of mixtures of calcareous and terrigenous clay (10-20%), silt (50-65%) and sand (15-40%) and are generally uniform in appearance. Their carbonate contents vary between and 17% and 22% by weight while organic carbon contents are less than 10% by weight. Dating of conformably deposited material identified in ten of the twelve cores indicates a range of sediment accumulation rates between 0.017mka-1 and 0.2 mka-1 which are consistent with previous estimates reported for this area. One slide-adjacent core, and four within-landslide cores present depositional hiatus surfaces located at depths of 0.8 to 2.2 meters below the present-day seafloor and identified by a sharp, colour-change boundary; discernable but small increases in sediment stiffness; and a slight increase in sediment bulk density of 0.1 gcm-3. Distinct gaps in AMS 14C age of at least 20ka are recorded across these boundary surfaces. Examination of sub-bottom profiler records of transects through three of the within-slide core-sites and their nearby landslide scarps available for the Coolangatta-1 and Cudgen slides indicate that: 1) the youngest identifiable sediment layer reflectors upslope of these slides, terminate on and are truncated by slide rupture surfaces; and 2) there is no obvious evidence in the sub-bottom profiles for a post-slide sediment layer draped over or otherwise burying slide ruptures or exposed slide detachment surfaces. This suggests that both these submarine landslides are geologically recent and suggests that the hiatus surfaces identified in Coolangatta-1's and Cudgen's within-slide cores are either: a) erosional features that developed after the occurrence of the landslide in which case the hiatus surface age provides a minimum age for landslide occurrence or b) detachment surfaces from which slabs of near-surface sediment were removed during landsliding in which case the post-hiatus sediment dates indicates approximately when landsliding occurred. In either case, it is reasonable to suggest that these two spatially adjacent slides occurred penecontemporaneously approximately 20,000 years ago.

Interacting Supernovae and Supernova Impostors: SN 2009ip, is this the End?

We report the results of a three-year-long dedicated monitoring campaign of a restless luminous blue variable (LBV) in NGC 7259. The object, named SN 2009ip, was observed photometrically and spectroscopically in the optical and near-infrared domains. We monitored a number of erupting episodes in the past few years, and increased the density of our observations during eruptive episodes. In this paper, we present the full historical data set from 2009 to 2012 with multi-wavelength dense coverage of the two high-luminosity events between 2012 August and September. We construct bolometric light curves and measure the total luminosities of these eruptive or explosive events. We label them the 2012a event (lasting ~50 days) with a peak of 3 × 1041 erg s-1, and the 2012b event (14 day rise time, still ongoing) with a peak of 8 × 1042 erg s-1. The latter event reached an absolute R-band magnitude of about -18, comparable to that of a core-collapse supernova (SN). Our historical monitoring has detected high-velocity spectral features (~13,000 km s-1) in 2011 September, one year before the current SN-like event. This implies that the detection of such high-velocity outflows cannot, conclusively, point to a core-collapse SN origin. We suggest that the initial peak in the 2012a event was unlikely to be due to a faint core-collapse SN. We propose that the high intrinsic luminosity of the latest peak, the variability history of SN 2009ip, and the detection of broad spectral lines indicative of high-velocity ejecta are consistent with a pulsational pair-instability event, and that the star may have survived the last outburst. The question of the survival of the LBV progenitor star and its future fate remain open issues, only to be answered with future monitoring of this historically unique explosion.

The pyrrolo-1,5-benzoxazepine, PBOX-6, inhibits the growth of breast cancer cells in vitro independent of estrogen receptor status and inhibits breast tumour growth in vivo

Members of a novel series of pyrrolo-1,5-benzoxazepine (PBOX) compounds have been shown to induce apoptosis in a number of human leukemia cell lines of different haematological lineage, suggesting their potential as anti-cancer agents. In this study, we sought to determine if PBOX-6, a well characterised member of the PBOX series of compounds, is also an effective inhibitor of breast cancer growth. Two estrogen receptor (ER)-positive (MCF-7 and T-47-D) and two ER-negative (MDA-MB-231 and SK-BR-3) cell lines were examined. The 3,4,5-dimethylthiazol-2-yl-2,5-diphenyl-tetrazolium bromide (MTT) assay was used to determine reduction in cell viability. PBOX-6 reduced the cell viability of all four cell lines tested, regardless of ER status, with IC(50) values ranging from 1.0 to 2.3 microM. PBOX-6 was most effective in the SK-BR-3 cells, which express high endogenous levels of the HER-2 oncogene. Overexpression of the HER-2 oncogene has been associated with aggressive disease and resistance to chemotherapy. The mechanism of PBOX-6-induced cell death was due to apoptosis, as indicated by the increased proportion of cells in the pre-G1 peak and poly(ADP-ribose) polymerase (PARP) cleavage. Moreover, intratumoural administration of PBOX-6 (7.5 mg/kg) significantly inhibited tumour growth in vivo in a mouse mammary carcinoma model (p=0.04, n=5, Student's t-test). Thus, PBOX-6 could be a promising anti-cancer agent for both hormone-dependent and -independent breast cancers.