Objectives/Goals: The overall goal of this project is to determine bacterial transcriptional signatures from clinical sputum and assess their potential to monitor treatment response and predict the outcome of drug therapy in patients with tuberculosis (TB). Methods/Study Population: We are developing a novel transcript capture sequencing (TC-Seq) approach to sequence the mRNA of Mycobacterium tuberculosis (Mtb) and analyze transcriptomes from clinical samples containing minimal amounts of bacterial RNA. This protocol generates single-stranded biotinylated probes from Mtb DNA. Probes are hybridized to and allow enrichment of Mtb-specific mRNA within next-generation RNA sequencing libraries. We will apply TC-Seq to sputum samples collected throughout an 18-month Phase II clinical trial investigating response to TB treatment to compare the transcriptome of Mtb between patients whose treatment results in cure or relapse. Results/Anticipated Results: We have refined a technique to generate biotinylated probes starting from DNA of lab grown Mtb. This protocol achieves robust and unbiased sampling of the Mtb transcriptome from mixed samples containing both human and Mtb RNA. Preliminary sequencing of clinical sputum collected pretreatment has generated 1–4 million Mtb-specific reads, a sequencing depth that allows examination of the entire bacterial transcriptome. We will measure differential gene expression before and during treatment as well as between cure and relapse cases. These results will allow us to characterize bacterial response to treatment and identify bacterial markers that correlate with relapse. Discussion/Significance of Impact: Understanding Mtb activity during treatment will offer new ways to assess the efficacy of different treatment regimens. Crucially, identifying clear bacterial markers that demarcate a cure or relapse outcome will have a significant impact on determining patient eligibility for shorter drug therapy.

Diets deficient in fibre are reported globally. The associated health risks of insufficient dietary fibre are sufficiently grave to necessitate large-scale interventions to increase population intake levels. The Danish Whole Grain Partnership (DWP) is a public–private enterprise model that successfully augmented whole-grain intake in the Danish population. The potential transferability of the DWP model to Slovenia, Romania and Bosnia-Herzegovina has recently been explored. Here, we outline the feasibility of adopting the approach in the UK. Drawing on the collaborative experience of DWP partners, academics from the Healthy Soil, Healthy Food, Healthy People (H3) project and food industry representatives (Food and Drink Federation), this article examines the transferability of the DWP approach to increase whole grain and/or fibre intake in the UK. Specific consideration is given to the UK’s political, regulatory and socio-economic context. We note key political, regulatory, social and cultural challenges to transferring the success of DWP to the UK, highlighting the particular challenge of increasing fibre consumption among low socio-economic status groups – which were also most resistant to interventions in Denmark. Wholesale transfer of the DWP model to the UK is considered unlikely given the absence of the key ‘success factors’ present in Denmark. However, the DWP provides a template against which a UK-centric approach can be developed. In the absence of a clear regulatory context for whole grain in the UK, fibre should be prioritised and public–private partnerships supported to increase the availability and acceptability of fibre-rich foods.

Can metaphysics yield the consolations of philosophy? One possibility, defended by Derek Parfit, is that reflection on the nature of identity and time could diminish both fear of death and grief. In this paper, I assess the prospect of such consolation, focussing especially on attempts to console a grieving third party. A shift to a reductionist view of personal identity might mean that death is less threatening. However, there is some evidence to suggest that such a shift does not necessarily translate into less death anxiety. Moreover, applied to grief at loss of another, such a perspective may be misdirected. A temporally neutral perspective offers a theoretically powerful way of reducing the sense of loss at being separated in time from a loved one. However, it is unclear whether it is psychologically possible to achieve. Even if it were possible, it may not diminish the pain of separation.

I identify a serious challenge to philosophical consolation for grief. The greater the consolation that is offered, the greater the risk of losing important attachments and the less it may be psychologically accessible.

Seabirds are declining globally and are one of the most threatened groups of birds. To halt or reverse this decline they need protection both on land and at sea, requiring site-based conservation initiatives based on seabird abundance and diversity. The Important Bird and Biodiversity Area (IBA) programme is a method of identifying the most important places for birds based on globally agreed standardised criteria and thresholds. However, while great strides have been made identifying terrestrial sites, at-sea identification is lacking. The Chagos Archipelago, central Indian Ocean, supports four terrestrial IBAs (tIBAs) and two proposed marine IBAs (mIBAs). The mIBAs are seaward extensions to breeding colonies based on outdated information and, other types of mIBA have not been explored. Here, we review the proposed seaward extension mIBAs using up-to-date seabird status and distribution information and, use global positioning system (GPS) tracking from Red-footed Booby Sula sula – one of the most widely distributed breeding seabirds on the archipelago – to identify any pelagic mIBAs. We demonstrate that due to overlapping boundaries of seaward extension to breeding colony and pelagic areas of importance there is a single mIBA in the central Indian Ocean that lays entirely within the Chagos Archipelago Marine Protected Area (MPA). Covering 62,379 km2 it constitutes ~10% of the MPA and if designated, would become the 11th largest mIBA in the world and 4th largest in the Indian Ocean. Our research strengthens the evidence of the benefits of large-scale MPAs for the protection of marine predators and provides a scientific foundation stone for marine biodiversity hotspot research in the central Indian Ocean.

ABSTRACT IMPACT: We present preliminary data and an outlined approach to assess peripheral immune changes associated with PTSD in a clinical setting and in a pre-clinical rat model of PTSD. OBJECTIVES/GOALS: We report our methodology and findings indicating a relationship between CNS dopamine signaling and peripheral immune cell populations and propose to extend this methodology to a PTSD patient population to elucidate immune-brain connections in this disorder. METHODS/STUDY POPULATION: Using an IRB-approved protocol in collaboration with a board-certified psychiatrist, we will recruit PTSD patients undergoing treatment, newly diagnosed drug-naiive PTSD patients, and age-matched healthy controls. Flow cytometry will be used for immunophenotyping on blood samples from each group. To complement this data, we will also measure serum cytokine levels in each group. In order to elucidate the connection between the observed immunophenotypes in the PTSD population and CNS neurotransmitters levels, we will employ a rodent model of PTSD and high-pressure liquid chromatography to measure dopamine levels in tandem with peripheral immune changes. RESULTS/ANTICIPATED RESULTS: In both humans and rodents with low CNS dopamine, an expansion of monocyte-derived suppressor cells was observed via flow cytometry. We anticipate that human PTSD patients will exhibit a similar expansion in suppressive immune cells’‘ in agreement with existing literature suggesting a chronic inflammatory state in PTSD. Moreover, in an animal model of PTSD we anticipate an inverse correlation between the CNS dopamine levels and the size of the immune suppressor cell population. DISCUSSION/SIGNIFICANCE OF FINDINGS: Our findings will indicate whether altered dopamine neurotransmission underlies peripheral immune system changes in the context of PTSD models and human patients. Thus, these findings will provide an alternative avenue for future investigations on the role of the immune system in PTSD.

Dinosaur body fossil material is rare in Scotland, previously known almost exclusively from the Great Estuarine Group on the Isle of Skye. We report the first unequivocal dinosaur fossil from the Isle of Eigg, belonging to a Bathonian (Middle Jurassic) taxon of uncertain affinity. The limb bone NMS G.2020.10.1 is incomplete, but through a combination of anatomical comparison and osteohistology, we determine it most likely represents a stegosaur fibula. The overall proportions and cross-sectional geometry are similar to the fibulae of thyreophorans. Examination of the bone microstructure reveals a high degree of remodelling and randomly distributed longitudinal canals in the remaining primary cortical bone. This contrasts with the histological signal expected of theropod or sauropod limb bones, but is consistent with previous studies of thyreophorans, specifically stegosaurs. Previous dinosaur material from Skye and broadly contemporaneous sites in England belongs to this group, including Loricatosaurus and Sarcolestes and a number of indeterminate stegosaur specimens. Theropods such as Megalosaurus and sauropods such as Cetiosaurus are also known from these localities. Although we find strong evidence for a stegosaur affinity, diagnostic features are not observed on NMS G.2020.10.1, preventing us from referring it to any known genera. The presence of this large-bodied stegosaur on Eigg adds a significant new datapoint for dinosaur distribution in the Middle Jurassic of Scotland.

Effective management of uncertainty can lead to better, more informed decisions. However, many decision makers and their advisers do not always face up to uncertainty, in part because there is little constructive guidance or tools available to help. This paper outlines six Uncertainty Principles to manage uncertainty.

• Face up to uncertainty

• Deconstruct the problem

• Don’t be fooled (un/intentional biases)

• Models can be helpful, but also dangerous

• Think about adaptability and resilience

• Bring people with you

The European Commission is an increasingly important source of funding for international research projects and is due to announce its Framework 5 program early in 1999. The Outcomes of Depression International Network (ODIN), funded from the current EC Biomed 2 program, is a case study in European academic co-operation. Its organization has three key elements. First, engaging the principal investigators: this has involved identifying potential partners, ensuring reciprocity of interests, effective co-ordination, `dividing the spoils' in advance, and setting up good personal and electronic communication systems. Second, an esprit de corps has been created amongst the researchers, maintaining contact and consistency, and promoting higher degrees. Third, ongoing problems including difficulties in negotiations with the EC, divergence of detailed study methods, and isolation and demoralization amongst researchers, have been addressed. ODIN may provide a useful model for researchers wishing to set up international collaborative groups.

Examination of a number of satellite and aerial images of the Milesian peninsula has allowed the mapping of a large number of apparently ancient linear features across the landscape. These are here interpreted, for the most part, as relicts of agro-economic field systems of unknown date, but most plausibly established during the Archaic, Hellenistic or late antique periods and perhaps used for centuries after, before the economic decline of the region in the second millennium AD. While earlier survey work has noted the existence of terracing and rural divisions at certain points in the landscape, the new remote-sensing data have provided an unprecedented large-scale insight into the extent and variety of forms of division, as well as documenting the stripping of macquis overgrowth by modern farming practices, which has, on the one hand, exposed these ancient landscapes but also, on the other, poses a threat to their preservation. The extent of the linear features suggests a high degree of land use on the peninsula at certain points in the past. Further investigation of these important features has the potential to provide critical insights into the economic history of rural and urban Miletos over the last 2,000 to 5,000 years.

The particle size of the forage has been proposed as a key factor to ensure a healthy rumen function and maintain dairy cow performance, but little work has been conducted on ryegrass silage (GS). To determine the effect of chop length of GS and GS:maize silage (MS) ratio on the performance, reticular pH, metabolism and eating behaviour of dairy cows, 16 multiparous Holstein-Friesian cows were used in a 4×4 Latin square design with four periods each of 28-days duration. Ryegrass was harvested and ensiled at two mean chop lengths (short and long) and included at two ratios of GS:MS (100:0 or 40:60 dry matter (DM) basis). The forages were fed in mixed rations to produce four isonitrogenous and isoenergetic diets: long chop GS, short chop GS, long chop GS and MS and short chop GS and MS. The DM intake (DMI) was 3.2 kg/day higher (P<0.001) when cows were fed the MS than the GS-based diets. The short chop length GS also resulted in a 0.9 kg/day DM higher (P<0.05) DMI compared with the long chop length. When fed the GS:MS-based diets, cows produced 2.4 kg/day more (P<0.001) milk than when fed diets containing GS only. There was an interaction (P<0.05) between chop length and forage ratio for milk yield, with a short chop length GS increasing yield in cows fed GS but not MS-based diets. An interaction for DM and organic matter digestibility was also observed (P<0.05), where a short chop length GS increased digestibility in cows when fed the GS-based diets but had little effect when fed the MS-based diet. When fed the MS-based diets, cows spent longer at reticular pH levels below pH 6.2 and pH 6.5 (P<0.01), but chop length had little effect. Cows when fed the MS-based diets had a higher (P<0.05) milk fat concentration of C18 : 2n-6 and total polyunsaturated fatty acids compared with when fed the GS only diets. In conclusion, GS chop length had little effect on reticular pH, but a longer chop length reduced DMI and milk yield but had little effect on milk fat yield. Including MS reduced reticular pH, but increased DMI and milk performance irrespective of the GS chop length.

Sandstones of the Palaeocene Montrose Group were deposited in a deepwater fan environment, and form a major oil reservoir in the North Sea. Calcite concretions occur commonly within thick-bedded and structureless sandstones. These concretions have been identified by sonic logs and well reports, and were cross-checked with available core data. Regionally, 101 wells have been examined and carbonate concretions form 0.6–7.2% of the core. Concretions are most abundant along the flank of the Fladen Ground Spur, the north Witch Ground Graben (WGG), the east south Viking Graben and East Central Graben (ECG). Concretions of the ECG formed at deep burial, with C from decarboxylation. Geochemical inheritance of Mn and Sr from Cretaceous chalk clasts may occur. Concretion growth may also have been influenced by vertical expulsion of fluids (leak-off) localized above salt tectonics. Isotopic and petrographic evidence indicates that much carbonate C in the WGG was derived from biodegradation of migrating oil in meteoric water at shallow depth. The locations of abundant carbonate with characteristic negative C isotope signatures can be used as shallow exploration guides to leak-off points located above deep overpressured structures.

Samples of granulated slag from Scunthorpe, Humberside, collected over several years, vary somewhat in chemical composition reflecting differences in iron-making practice. All contain some melilite and oldhamite crystals and minor amounts of iron along with the glass. Granulated slag from Usinor, Dunkirk, France, has a little more CaO than the Scunthorpe material with crystals of merwinite and oldhamite. Pelletised slag from Redcar, Teeside, is much more vescicular than the granulated material and contains melilite and oldhamite.

The melilite crystals contain many inclusions of oldhamite and iron. They span a wide range within the akermanite-gehlenite series and are non-stoichiometric in composition. The merwinite may have formed largely by quench crystallization. Oldhamite is probably the first phase to form in all the slags. The silicate mineralogy can be explained in terms of the phase relationships within the CaO-SiO2-Al2O3-MgO quaternary system. The small difference in composition of the French slags is sufficient for them to fall into the primary phase field of merwinite rather than melilite. Assuming that only a minor proportion of crystals is acceptable in slags for use in Portland Blastfurnace Cement, the present low Al (< 11 % Al2O3) British and French slags are approaching an optimum composition.

Recent experimental studies have suggested that colloidal silica can form in high-T (300 to >700°C) hydrothermal fluids (Wilkinson et al., 1996). Natural evidence in support of this was found by Williamson et al. (1997) who proposed a colloidal (gel) silica origin for <50 μm irregularly-shaped inclusions of quartz contained in greisen topaz from southwest England. Confocal and microprobe studies, presented here, strengthen this argument although rather than forming a gel in the hydrothermal fluid, it is suggested that the colloidal silica aggregated as a viscous coagulated colloid, with much of its volume (<10 to 30 vol.%) consisting of metal (mainly Fe) -rich particles. This is evident from the largely solid nature of metal-rich shrinkage bubbles contained at the margins of the inclusions of quartz which shows that the material forming the inclusions contained much less liquid than would be expected in a silica gel. These findings may have important implications for models of ore formation since the precipitation of a coagulated colloid could inhibit hydrothermal fluid transport and cause co-deposition of silica and entrained ore-forming elements. The mode of formation of the colloidal silica and further implications of the study are discussed.

The Kabwe Zn-Pb deposit (central Zambia) consists of a cluster of mixed sulfide and non-sulfide orebodies. The sulfide ores comprise sphalerite, galena, pyrite, chalcopyrite and accessory Ge-sulfides (±Ga and In). The non-sulfide ores comprise: (1) willemite-dominated zones encasing massive sulfide orebodies and (2) oxide-dominated alteration bands, overlying both the sulfide and Zn-silicate orebodies. This study focuses on the Ge, In and Ga distribution in the non-sulfide mineralization, and was carried out on a suite of Kabwe specimens, housed in the Natural History Museum Ore Collection (London). Petrography confirmed that the original sulfides were overprinted by at least two contrasting oxidation stages dominated by the formation of willemite (W1 and W2), and a further event characterized by weathering-related processes. Oxygen isotopic analyses have shown that W1 and W2 are unrelated genetically and furthermore not related to supergene Zn-Pb-carbonates in the oxide-dominated assemblage. The δ18O composition of 13.9–15.7‰ V-SMOW strongly supports a hydrothermal origin for W1. The δ18O composition of W2 (−3.5‰ to 0‰ V-SMOW) indicates that it precipitated from groundwaters of meteoric origin in either a supergene or a low-T hydrothermal environment. Gallium and Ge show a diversity of distribution among the range of Zn-bearing minerals. Gallium has been detected at the ppm level in W1, sphalerite, goethite and hematite. Germanium occurs at ppm levels in W1 and W2, and in scarcely detectable amounts in hemimorphite, goethite and hematite. Indium has low concentrations in goethite and hematite. These different deportments among the various phases are probably due to the different initial Ga, In and Ge abundances in the mineralization, to the different solubilities of the three elements at different temperatures and pH values, and finally to their variable affinities with the various minerals formed.