Chloride waters of Great Britain revisited: from subsea formation waters to onshore geothermal fluids

It has long been known that chloride-dominated saline ground waters occur at depth in the UK, not only beneath the sea but also onshore at depths of a few hundred metres. In a few places in northern England, these saline waters discharge naturally at surface in the form of springs. In recent years, however, these saline ground waters have come to be regarded as resources: as potential geothermal fluids intercepted in deep boreholes. Comparisons of the major ions and stable isotopes (δ2H, δ18O and δ34S) of these saline ground waters with North Sea oilfield formation waters, and with brines encountered in former subsea workings of coastal collieries, reveal that they are quite distinct from those found in North Sea oilfields, in that their as δ2H/δ18O signatures are distinctly “meteoric”. δ34S data preclude a significant input from evaporite dissolution – another contrast with many North Sea brines and some colliery waters. Yet, enigmatically, their total dissolved solids contents are far higher than typical meteoric waters. It is tentatively suggested that these paradoxical hydrogeochemical properties might be explained by recharge during Cenozoic uplift episodes, with high concentrations of solutes being derived by a combination of high-temperature rock–water interaction in the radiothermal granites and/or ‘freeze out’ from overlying permafrost that surely formed in this region during cold periods. Geothermometric calculations suggest these saline waters may well be representative of potentially valuable geothermal reservoirs

Quantifying physiological influences on otolith microchemistry

Trace element concentrations in fish earstones (‘otoliths’) are widely used to discriminate spatially discrete populations or individuals of marine fish, based on a commonly held assumption that physiological influences on otolith composition are minor, and thus variations in otolith elemental chemistry primarily reflect changes in ambient water chemistry. We carried out a long-term (1-year) experiment, serially sampling seawater, blood plasma and otoliths of mature and immature European plaice (Pleuronectes platessa L.) to test relationships between otolith chemistry and environmental and physiological variables. Seasonal variations in otolith elemental composition did not track seawater concentrations, but instead reflected physiological controls on metal transport and biokinetics, which are likely moderated by ambient temperature. The influence of physiological factors on otolith composition was particularly evident in Sr/Ca ratios, the most widely used elemental marker in applied otolith microchemistry studies. Reproduction also triggered specific variations in otolith and blood plasma metal chemistry, especially Zn/Ca ratios in female fish, which could potentially serve as retrospective spawning indicators. The influence of physiology on the trace metal composition of otoliths may explain the success of microchemical stock discrimination in relatively homogenous marine environments, but could complicate alternative uses for trace element compositions in biominerals of higher organism

Renewing the Exploration Approach for Mid-Enthalpy Systems: Examples from Northern England and Scotland

After a promising start in the 1970s and 80s, the UK rather fell behind other countries in the search for viable mid-enthalpy geothermal resources. This situation began to turn around in 2004, when the first of three deep geothermal exploration boreholes were drilled in northern England. What distinguished these from earlier drilling in Cornwall was the deliberate search for naturallyhigh permeability associated with major faults, especially those that have undergone strike-slip reactivation during the Cenozoic. Boreholes at Eastgate in the North Pennines targeted buried radiothermal granite, whereas the 1,821m-deep Science Central Borehole in Newcastle upon Tyne targeted a postulated deep sedimentary aquifer (the Fell Sandstones), which were inferred to be connected laterally to the granitic heat source by a major fault (the reactivation of the Iapetus geo-suture). The drilling was in both cases rewarded with impressive heat flows, and in the case of Eastgate with what is believed to be the highest permeability yet found in a deep granite batholith anywhere in the world. In parallel with these developments, a re-assessment was made of the preexisting geothermal heat flow database for the UK, applying newly-standardised correction protocols for palaeoclimatic and topographic distortions, which were found to be particularly marked in Scotland (where only shallow boreholes had been used to establish geothermal gradients in the original 1980s analysis), Similar prospects in northern England (similar to that drilled at Science Central) are now the focus of commercial exploration efforts. Appraisal of fault dispositions relative to the present-day maximum compressive stress azimuth are being used to identify the most promising areas for intersecting fault-related permeability at depth. New geophysical tools – most notably atomic dielectric resonance scanning – are also being appraised for their ability to directly detect features (such as hot brines) which are indicative of localised convection in target fault zones and aquifers

Critical Structural and Functional Roles for the N-Terminal Insertion Sequence in Surfactant Protein B Analogs

Surfactant protein B (SP-B; 79 residues) belongs to the saposin protein superfamily, and plays functional roles in lung surfactant. The disulfide cross-linked, N- and C-terminal domains of SP-B have been theoretically predicted to fold as charged, amphipathic helices, suggesting their participation in surfactant activities. Earlier structural studies with Mini-B, a disulfide-linked construct based on the N- and C-terminal regions of SP-B (i.e., approximately residues 8-25 and 63-78), confirmed that these neighboring domains are helical; moreover, Mini-B retains critical in vitro and in vivo surfactant functions of the native protein. Here, we perform similar analyses on a Super Mini-B construct that has native SP-B residues (1-7) attached to the N-terminus of Mini-B, to test whether the N-terminal sequence is also involved in surfactant activity.FTIR spectra of Mini-B and Super Mini-B in either lipids or lipid-mimics indicated that these peptides share similar conformations, with primary alpha-helix and secondary beta-sheet and loop-turns. Gel electrophoresis demonstrated that Super Mini-B was dimeric in SDS detergent-polyacrylamide, while Mini-B was monomeric. Surface plasmon resonance (SPR), predictive aggregation algorithms, and molecular dynamics (MD) and docking simulations further suggested a preliminary model for dimeric Super Mini-B, in which monomers self-associate to form a dimer peptide with a "saposin-like" fold. Similar to native SP-B, both Mini-B and Super Mini-B exhibit in vitro activity with spread films showing near-zero minimum surface tension during cycling using captive bubble surfactometry. In vivo, Super Mini-B demonstrates oxygenation and dynamic compliance that are greater than Mini-B and compare favorably to full-length SP-B.Super Mini-B shows enhanced surfactant activity, probably due to the self-assembly of monomer peptide into dimer Super Mini-B that mimics the functions and putative structure of native SP-B

Association of peripheral blood DNA methylation level with Alzheimer’s disease progression

Background: Identifying biomarkers associated with Alzheimer's disease (AD) progression may enable patient enrichment and improve clinical trial designs. Epigenome-wide association studies have revealed correlations between DNA methylation at cytosine-phosphate-guanine (CpG) sites and AD pathology and diagnosis. Here, we report relationships between peripheral blood DNA methylation profiles measured using Infinium® MethylationEPIC BeadChip and AD progression in participants from the Alzheimer's Disease Neuroimaging Initiative (ADNI) cohort. Results: The rate of cognitive decline from initial DNA sampling visit to subsequent visits was estimated by the slopes of the modified Preclinical Alzheimer Cognitive Composite (mPACC; mPACCdigit and mPACCtrailsB) and Clinical Dementia Rating Scale Sum of Boxes (CDR-SB) plots using robust linear regression in cognitively normal (CN) participants and patients with mild cognitive impairment (MCI), respectively. In addition, diagnosis conversion status was assessed using a dichotomized endpoint. Two CpG sites were significantly associated with the slope of mPACC in CN participants (P < 5.79 × 10-8 [Bonferroni correction threshold]); cg00386386 was associated with the slope of mPACCdigit, and cg09422696 annotated to RP11-661A12.5 was associated with the slope of CDR-SB. No significant CpG sites associated with diagnosis conversion status were identified. Genes involved in cognition and learning were enriched. A total of 19, 13, and 5 differentially methylated regions (DMRs) associated with the slopes of mPACCtrailsB, mPACCdigit, and CDR-SB, respectively, were identified by both comb-p and DMRcate algorithms; these included DMRs annotated to HOXA4. Furthermore, 5 and 19 DMRs were associated with conversion status in CN and MCI participants, respectively. The most significant DMR was annotated to the AD-associated gene PM20D1 (chr1: 205,818,956 to 205,820,014 [13 probes], Sidak-corrected P = 7.74 × 10-24), which was associated with both the slope of CDR-SB and the MCI conversion status. Conclusion: Candidate CpG sites and regions in peripheral blood were identified as associated with the rate of cognitive decline in participants in the ADNI cohort. While we did not identify a single CpG site with sufficient clinical utility to be used by itself due to the observed effect size, a biosignature composed of DNA methylation changes may have utility as a prognostic biomarker for AD progression

Formalin-Fixed Paraffin-Embedded (FFPE) samples are not a beneficial replacement for frozen tissues in fetal membrane microbiota research

Formalin-Fixed Paraffin-Embedded (FFPE) tissues are routinely collected, archived, and used for clinical diagnosis, including maternal and neonatal health. Applying FFPE samples to microbiota research would be beneficial to reduce preparation, storage and costs associated with limited available frozen samples. This research aims to understand if FFPE fetal membrane samples are comparable to frozen tissues, which are the current gold standard for DNA microbiota analysis. Extracted DNA from nine matched paired patients were sequenced by Illumina sequencing of the V4 16S rRNA gene region. This included duplicate frozen amnion and chorion fetal membrane rolls or FFPE combined amniochorionic samples. Negative controls of surrounding wax blocks and DNA extraction reagents were processed alongside samples using identical methods. DNA quality and quantity was assessed by NanoDrop, agarose gel electrophoresis and Bioanalyzer. Decontam and SourceTracker were integrated into microbiota analysis to identify the presence of contaminating sources. The bacterial profile and nine genera differed between FFPE and frozen fetal membranes. There were no differences in bacterial profiles between FFPE samples and corresponding wax negative controls, with 49% of bacteria in FFPE fetal membrane samples matched to the source origin of paraffin wax, and 40% originating from DNA extraction reagent sources. FFPE samples displayed high fragmentation and low quantity of extracted DNA compared to frozen samples. The microbiota of FFPE fetal membrane samples is influenced by processing methods, with the inability to differentiate between the microbiota of the tissue sample and the surrounding wax block. Illumina sequencing results of FFPE and frozen fetal membrane samples should not be compared using the methods employed here. Variation could be influenced by limitations including storage time, DNA extraction and purification methods. To utilise FFPE fetal membrane samples in microbiota research then contamination prevention and detection methods must be included into optimised and standardised protocols, with recommendations presented here

Soil biogeochemistry across Central and South American tropical dry forests

The availability of nitrogen (N) and phosphorus (P) controls the flow of carbon (C) among plants, soils, and the atmosphere, thereby shaping terrestrial ecosystem responses to global change. Soil C, N, and P cycles are linked by drivers operating at multiple spatial and temporal scales: landscape-level variation in macroclimate and soil geochemistry, stand-scale heterogeneity in forest composition, and microbial community dynamics at the soil pore scale. Yet in many biomes, we do not know at which scales most of the biogeochemical variation emerges, nor which processes drive cross-scale feedbacks. Here, we examined the drivers and spatial/temporal scales of variation in soil biogeochemistry across four tropical dry forests spanning steep environmental gradients. To do so, we quantified soil C, N, and P pools, extracellular enzyme activities, and microbial community structure across wet and dry seasons in 16 plots located in Colombia, Costa Rica, Mexico, and Puerto Rico. Soil biogeochemistry exhibited marked heterogeneity across the 16 plots, with total organic C, N, and P pools varying fourfold, and inorganic nutrient pools by an order of magnitude. Most soil characteristics changed more across space (i.e., among sites and plots) than over time (between dry and wet season samplings). We observed stoichiometric decoupling among C, N, and P cycles, which may reflect their divergent biogeochemical drivers. Organic C and N pool sizes were positively correlated with the relative abundance of ectomycorrhizal trees and legumes. By contrast, the distribution of soil P pools was driven by soil geochemistry, with larger inorganic P pools in soils with P-rich parent material. Most earth system models assume that soils within a texture class operate similarly, and ignore subgrid cell variation in soil properties. Here we reveal that soil nutrient pools and fluxes exhibit as much variation among four Neotropical dry forests as is observed across terrestrial ecosystems at the global scale. Soil biogeochemical patterns are driven not only by regional differences in soil parent material and climate, but also by local-scale variation in plant and microbial communities. Thus, the biogeochemical patterns we observed across the Neotropical dry forest biome challenge representation of soil processes in ecosystem models

The invisible plan: how English teachers develop their expertise and the special place of adapting the skills of lesson planning

This paper analyses how English teachers learn to become expert designers of learning and why sharing that expertise is increasingly vital. Its conceptual framework is the widely recognised, empirically tested, five-stage developmental Dreyfus model of skill acquisition, exemplifying the development of teacher expertise, constituted by the “milestone” [m] and “transitory” [t] phases connecting with the five stages of: Novice [m], Advanced Beginner [t], Competent [m], Proficient [t] and Expert [m]. Teacher planning is analysed as one key tacit or non-tangible component of developing expertise. Focusing specifically on English teachers as key participants in this pioneer teacher cognition study, the defining characteristics of milestone stages of expertise development are explored with specific attention to the remarkably under-researched area of planning. We introduce three new categories, defining modes of planning: (i) visible practical planning, (ii) external reflective planning and (iii) internal reflective planning, demonstrating their role in teacher development through the Dreyfus five stages. English is a subject which suffers from frequent disruptive changes to curriculum and assessment: new learning designs are constantly demanded, making planning an ongoing challenge. The implications for practice include the importance of an explicit understanding of how teachers’ planning moves through the three phases from the very “visible” novice phase to the internal relatively “automatic” competent teacher and finally the seemingly “invisible” expert phase. Further research is needed to explore how English teachers can share planning expertise between the three phases to improve teachers’ skills and student learning

Terminal osteoblast differentiation, mediated by runx2 and p27KIP1, is disrupted in osteosarcoma

The molecular basis for the inverse relationship between differentiation and tumorigenesis is unknown. The function of runx2, a master regulator of osteoblast differentiation belonging to the runt family of tumor suppressor genes, is consistently disrupted in osteosarcoma cell lines. Ectopic expression of runx2 induces p27KIP1, thereby inhibiting the activity of S-phase cyclin complexes and leading to the dephosphorylation of the retinoblastoma tumor suppressor protein (pRb) and a G1 cell cycle arrest. Runx2 physically interacts with the hypophosphorylated form of pRb, a known coactivator of runx2, thereby completing a feed-forward loop in which progressive cell cycle exit promotes increased expression of the osteoblast phenotype. Loss of p27KIP1 perturbs transient and terminal cell cycle exit in osteoblasts. Consistent with the incompatibility of malignant transformation and permanent cell cycle exit, loss of p27KIP1 expression correlates with dedifferentiation in high-grade human osteosarcomas. Physiologic coupling of osteoblast differentiation to cell cycle withdrawal is mediated through runx2 and p27KIP1, and these processes are disrupted in osteosarcoma

Isolation and characterization of resident endogenous c-Kit⁺ cardiac stem cells from the adult mouse and rat heart

This protocol describes the isolation of endogenous c-Kit (also known as CD117)-positive (c-Kit⁺), CD45-negative (CD45⁻) cardiac stem cells (eCSCs) from whole adult mouse and rat hearts. The heart is enzymatically digested via retrograde perfusion of the coronary circulation, resulting in rapid and extensive breakdown of the whole heart. Next, the tissue is mechanically dissociated further and cell fractions are separated by centrifugation. The c-Kit⁺ CD45⁻ eCSC population is isolated by magnetic-activated cell sorting technology and purity and cell numbers are assessed by flow cytometry. This process takes ∼4 h for mouse eCSCs or 4.5 h for rat eCSCs. We also describe how to characterize c-Kit⁺ CD45⁻ eCSCs. The c-Kit⁺ CD45⁻eCSCs exhibit the defining characteristics of stem cells: they are self-renewing, clonogenic and multipotent. This protocol also describes how to differentiate eCSCs into three main cardiac lineages: functional, beating cardiomyocytes, smooth muscle, and endothelial cells. These processes take 17-20 d