Template-Stripped Multifunctional Wedge and Pyramid Arrays for Magnetic Nanofocusing and Optical Sensing

We present large-scale reproducible fabrication of multifunctional ultrasharp metallic structures on planar substrates with capabilities including magnetic field nanofocusing and plasmonic sensing. Objects with sharp tips such as wedges and pyramids made with noble metals have been extensively used for enhancing local electric fields via the lightning-rod effect or plasmonic nanofocusing. However, analogous nanofocusing of magnetic fields using sharp tips made with magnetic materials has not been widely realized. Reproducible fabrication of sharp tips with magnetic as well as noble metal layers on planar substrates can enable straightforward application of their material and shape-derived functionalities. We use a template-stripping method to produce plasmonic-shell-coated nickel wedge and pyramid arrays at the wafer-scale with tip radius of curvature close to 10 nm. We further explore the magnetic nanofocusing capabilities of these ultrasharp substrates, deriving analytical formulas and comparing the results with computer simulations. These structures exhibit nanoscale spatial control over the trapping of magnetic microbeads and nanoparticles in solution. Additionally, enhanced optical sensing of analytes by these plasmonic-shell-coated substrates is demonstrated using surface-enhanced Raman spectroscopy. These methods can guide the design and fabrication of novel devices with applications including nanoparticle manipulation, biosensing, and magnetoplasmonics

Two Types of K⁺ Channel Subunit, Erg1 and KCNQ2/3, Contribute to the M-Like Current in a Mammalian Neuronal Cell

The potassium M current was originally identified in sympathetic ganglion cells, and analogous currents have been reported in some central neurons and also in some neural cell lines. It has recently been suggested that the M channel in sympathetic neurons comprises a heteromultimer of KCNQ2 and KCNQ3 (Wang et al., 1998) but it is unclear whether all other M-like currents are generated by these channels. Here we report that the M-like current previously described in NG108–15 mouse neuroblastoma x rat glioma cells has two components, “fast” and “slow”, that may be differentiated kinetically and pharmacologically. We provide evidence from PCR analysis and expression studies to indicate that these two components are mediated by two distinct molecular species of K+ channel: the fast component resembles that in sympathetic ganglia and is probably carried byKCNQ2/3 channels, whereas the slow component appears to be carried by merg1a channels. Thus, the channels generating M-like currents in different cells may be heterogeneous in molecular composition

Marriage and physical capability at mid to later life

Research has shown that married men and women have better physical and psychological health and greater longevity than their unmarried counterparts. However, the past 50 years have witnessed changes in the marriage and divorce rates, resulting in more people at older ages who are unmarried or with varied relationship histories. Given the strong association between marriage and health there could potentially be more people at older ages in poorer health, which may be particularly detrimental given the ageing population. Whilst there is much research looking at marriage and physical and psychological health there is little on marriage and physical capability. Physical capability is the capacity to perform the physical tasks of daily living and is predictive of mortality and future social care use. This PhD investigates the relationship between marriage and physical capability at mid to later life using two measures – grip strength and walking speed – from two nationally representative datasets of people aged 50 years and over in England and the USA. Cross-sectional associations between marriage and physical capability are investigated in a comparative analysis between England and the USA, and longitudinal associations through examining changes in walking speed over a ten year period in England. A descriptive analysis of early life circumstances and its association with entry into and exit out of marriage in England and the USA is also carried out. Findings show that married people had both higher levels of current physical capability and a slower decline in physical capability over time than their unmarried counterparts. Much of the “marriage advantage” is explained by their greater wealth, but there were some unexplained associations, particularly among widowed men. There were few gender and country differences in the association. The results of this thesis suggest that marriage is important for maintaining physical capability for people at mid to later life in England and the USA

A Simple and Non-destructive Method for Chlorophyll Quantification of Chlamydomonas Cultures Using Digital Image Analysis

Growing interest in the use of microalgae as a sustainable feedstock to support a green, circular, bio-economy has led to intensive research and development initiatives aimed at increasing algal biomass production covering a wide range of scales. At the heart of this lies a common need for rapid and accurate methods to measure algal biomass concentrations. Surrogate analytical techniques based on chlorophyll content use solvent extraction methods for chlorophyll quantification, but these methods are destructive, time consuming and require careful disposal of the resultant solvent waste. Alternative non-destructive methods based on chlorophyll fluorescence require expensive equipment and are less suitable for multiple sampling of small cultures which need to be maintained under axenic growth conditions. A simple, inexpensive and non-destructive method to estimate chlorophyll concentration of microalgal cultures in situ from digital photographs using the RGB color model is presented. Green pixel intensity and chlorophyll a, b and total chlorophyll concentration, measured by conventional means, follow a strong linear relationship (R2 = 0.985–0.988). In addition, the resulting standard curve was robust enough to accurately estimate chlorophyll concentration despite changes in sample volume, pH and low concentrations of bacterial contamination. In contrast, use of the same standard curve during nitrogen deprivation (causing the accumulation of neutral lipids) or in the presence of high quantities of bacterial contamination led to significant errors in chlorophyll estimation. The low requirement for equipment (i.e., a simple digital camera, available on smartphones) and widely available standard software for measuring pixel intensity make this method suitable for both laboratory and field-based work, particularly in situations where sample, qualified personnel and/or equipment is limited. By following the methods described here it should be possible to produce a standard curve for chlorophyll analysis in a wide range of testing conditions including different microalga cultures, culture vessel and photographic set up in any particular laboratory

Sail optimization for upwind sailing: application in a Tornado, the Olympic class catamaran.

A study of a boat’s motion is carried out in order to analyze the aerodynamic properties of the optimal sail for obtaining the maximum velocity when sailing to windward. The mechanics study shows the optimal CL and CD for a given sail and how the shape of the aerodynamic polar of the sail should be. A parametrical analysis of the aerodynamics of a sail is then carried out varying the maximum camber, position of the maximum camber in the chord direction and position of the maximum camber in the mast direction. The parametric analysis is done numerically with a vortex lattice method (VLM) and experimentally in a wind tunnel. The results show that the influence of the relevant parameters studied can be reduced to the variation of two parameters, A and B, defining the polar of the sail, CD = B + A2CL 2; and the influence of parameters A and B on the maximum VMG obtainable are calculated

Protein kinase C is essential for viability of the rice blast fungus Magnaporthe oryzae

This is the final version of the article. Available from Wiley via the DOI in this record.Protein kinase C constitutes a family of serine-threonine kinases found in all eukaryotes and implicated in a wide range of cellular functions, including regulation of cell growth, cellular differentiation and immunity. Here, we present three independent lines of evidence which indicate that protein kinase C is essential for viability of Magnaporthe oryzae. First, all attempts to generate a target deletion of PKC1, the single copy protein kinase C-encoding gene, proved unsuccessful. Secondly, conditional gene silencing of PKC1 by RNA interference led to severely reduced growth of the fungus, which was reversed by targeted deletion of the Dicer2-encoding gene, MDL2. Finally, selective kinase inhibition of protein kinase C by targeted allelic replacement with an analogue-sensitive PKC1(AS) allele led to specific loss of fungal viability in the presence of the PP1 inhibitor. Global transcriptional profiling following selective PKC inhibition identified significant changes in gene expression associated with cell wall re-modelling, autophagy, signal transduction and secondary metabolism. When considered together, these results suggest protein kinase C is essential for growth and development of M. oryzae with extensive downstream targets in addition to the cell integrity pathway. Targeting protein kinase C signalling may therefore prove an effective means of controlling rice blast disease.This work was funded by a BBSRC CASE PhD studentship to TJP with support from Syngenta and a European Research Council, Advanced Investigator Award to NJT under the European Union's Seventh Framework Programme (FP7/2007-2013)/ERC grant agreement no. 294702 GENBLAST

Fixing the Broken Phosphorus Cycle: Wastewater Remediation by Microalgal Polyphosphates

Phosphorus (P), in the form of phosphate derived from either inorganic (Pi) or organic (Po) forms is an essential macronutrient for all life. P undergoes a biogeochemical cycle within the environment, but anthropogenic redistribution through inefficient agricultural practice and inadequate nutrient recovery at wastewater treatment works have resulted in a sustained transfer of P from rock deposits to land and aquatic environments. Our present and near future supply of P is primarily mined from rock P reserves in a limited number of geographical regions. To help ensure that this resource is adequate for humanity’s food security, an energy-efficient means of recovering P from waste and recycling it for agriculture is required. This will also help to address excess discharge to water bodies and the resulting eutrophication. Microalgae possess the advantage of polymeric inorganic polyphosphate (PolyP) storage which can potentially operate simultaneously with remediation of waste nitrogen and phosphorus streams and flue gases (CO2, SOx, and NOx). Having high productivity in photoautotrophic, mixotrophic or heterotrophic growth modes, they can be harnessed in wastewater remediation strategies for biofuel production either directly (biodiesel) or in conjunction with anaerobic digestion (biogas) or dark fermentation (biohydrogen). Regulation of algal P uptake, storage, and mobilization is intertwined with the cellular status of other macronutrients (e.g., nitrogen and sulphur) in addition to the manufacture of other storage products (e.g., carbohydrate and lipids) or macromolecules (e.g., cell wall). A greater understanding of controlling factors in this complex interaction is required to facilitate and improve P control, recovery, and reuse from waste streams. The best understood algal genetic model is Chlamydomonas reinhardtii in terms of utility and shared resources. It also displays mixotrophic growth and advantageously, species of this genus are often found growing in wastewater treatment plants. In this review, we focus primarily on the molecular and genetic aspects of PolyP production or turnover and place this knowledge in the context of wastewater remediation and highlight developments and challenges in this field

Using the UM dynamical cores to reproduce idealised 3-D flows

This is the final version of the article. Available from the publisher via the DOI in this record.Published by Copernicus Publications on behalf of the European Geosciences UnionWe demonstrate that both the current (New Dynamics), and next generation (ENDGame) dynamical cores of the UK Met Office global circulation model, the UM, reproduce consistently, the long-term, large-scale flows found in several published idealised tests. The cases presented are the Held-Suarez test, a simplified model of Earth (including a stratosphere), and a hypothetical tidally locked Earth. Furthermore, we show that using simplifications to the dynamical equations, which are expected to be justified for the physical domains and flow regimes we have studied, and which are supported by the ENDGame dynamical core, also produces matching long-term, large-scale flows. Finally, we present evidence for differences in the detail of the planetary flows and circulations resulting from improvements in the ENDGame formulation over New Dynamics.We would like to thank Paul Ullrich and Kevin Heng for their valuable comments, when reviewing this manuscript. We would also like to thank Tom Melvin for his expert advice, and both Charline Marzin and Douglas Boyd for technical help. This work is supported by the European Research Council under the European Community’s Seventh Framework Programme (FP7/2007-2013 Grant Agreement no. 247060) and by the Consolidated STFC grant ST/J001627/1. This work is also partly supported by the Royal Society award WM090065. The calculations for this paper were performed on the DiRAC Facility jointly funded by STFC, the Large Facilities Capital Fund of BIS, and the University of Exeter

Mucin-Inspired Thermoresponsive Synthetic Hydrogels Induce Stasis in Human Pluripotent Stem Cells and Human Embryos.

Human pluripotent stem cells (hPSCs; both embryonic and induced pluripotent) rapidly proliferate in adherent culture to maintain their undifferentiated state. However, for mammals exhibiting delayed gestation (diapause), mucin-coated embryos can remain dormant for days or months in utero, with their constituent PSCs remaining pluripotent under these conditions. Here we report cellular stasis for both hPSC colonies and preimplantation embryos immersed in a wholly synthetic thermoresponsive gel comprising poly(glycerol monomethacrylate)-poly(2-hydroxypropyl methacrylate) [PGMA55-PHPMA135] diblock copolymer worms. This hydroxyl-rich mucin-mimicking nonadherent 3D gel maintained PSC viability and pluripotency in the quiescent G0 state without passaging for at least 14 days. Similarly, gel-coated human embryos remain in a state of suspended animation (diapause) for up to 8 days. The discovery of a cryptic cell arrest mechanism for both hPSCs and embryos suggests an important connection between the cellular mechanisms that evoke embryonic diapause and pluripotency. Moreover, such synthetic worm gels offer considerable utility for the short-term (weeks) storage of either pluripotent stem cells or human embryos without cryopreservation

Estrogenic mechanisms and cardiac responses following early life exposure to Bisphenol A (BPA) and Its metabolite 4-Methyl-2,4-bis(p -hydroxyphenyl)pent-1-ene (MBP) in zebrafish

This is the author accepted manuscriptEnvironmental exposure to Bisphenol A (BPA) has been associated with a range of adverse health effects, including on the cardiovascular system in humans. Lack of agreement on its mechanism(s) of action likely stem from comparisons between in vivo and in vitro test systems and potential multiple effects pathways. In rodents, in vivo, metabolic activation of BPA produces 4-methyl-2,4-bis(4-hydroxyphenyl)pent-1-ene (MBP), which is reported to be up to 1000 times more potent as an estrogen than BPA. We investigated the estrogenic effects and estrogen receptor signaling pathway(s) of BPA and MBP following early life exposure using a transgenic, estrogen responsive (ERE-TG) zebrafish and a targeted morpholino approach to knockdown the three fish estrogen receptor (ER) subtypes. The functional consequences of BPA exposure on the cardiovascular system of zebrafish larvae were also examined. The heart atrioventricular valves and the bulbus arteriosus were primary target tissues for both BPA and MBP in the ERE-TG zebrafish, and MBP was approximately 1000-fold more potent than BPA as an estrogen in these tissues. Estrogen receptor knockdown with morpholinos indicated that the estrogenic responses in the heart for both BPA and MBP were mediated via an estrogen receptor 1 (esr1) dependent pathway. At the highest BPA concentration tested (2500 μg/L), alterations in the atrial:ventricular beat ratio indicated a functional impact on the heart of 5 days post fertilization (dpf) larvae, and there was also a significantly reduced heart rate in these larvae at 14 dpf. Our findings indicate that some of the reported adverse effects on heart function associated with BPA exposure (in mammals) may act through an estrogenic mechanism, but that fish are unlikely to be susceptible to adverse effects on heart development for environmentally relevant exposures.Natural Environment Research Council (NERC)Biotechnology & Biological Sciences Research Council (BBSRC)Biotechnology & Biological Sciences Research Council (BBSRC