Structure-activity relationships of the N-terminus of calcitonin gene-related peptide:key roles of alanine-5 and threonine-6 in receptor activation

Background and purpose - The N-terminus of calcitonin gene-related peptide (CGRP) is important for receptor activation, especially the disulphide-bonded ring (residues 1-7). However, the roles of individual amino acids within this region have not been examined and so the molecular determinants of agonism are unknown. This study has examined the role of residues 1, 3-6 and 8-9, excluding Cys-2 and Cys-7. Experimental approach - CGRP derivatives were substituted with either cysteine or alanine; further residues were introduced at position 6. Their affinity was measured by radioligand binding and their efficacy by measuring cAMP production in SK-N-MC cells and ß-arrestin 2 translocation in CHO-K1 cells at the CGRP receptor. Key results - Substitution of Ala-5 by cysteine reduced affinity 270-fold and reduced efficacy for production of cAMP in SK-N-MCs. Potency at ß-arrestin translocation was reduced by 9-fold. Substitution of Thr-6 by cysteine destroyed all measurable efficacy of both cAMP and ß-arrestin responses; substitution with either alanine or serine impaired potency. Substitutions at positions 1, 4, 8 and 9 resulted in approximately 10-fold reductions in potency at both responses. Similar observations were made at a second CGRP-activated receptor, the AMY1(a) receptor. Conclusions and implications - Ala-5 and Thr-6 are key determinants of agonist activity for CGRP. Ala-5 is also very important for receptor binding. Residues outside of the 1-7 ring also contribute to agonist activity.

Experimental characterisation of the impact of neighbouring modulation-formats on 43Gb/s P-DPSK over SSMF and LEAF

We report an experimental characterisation examining the impact of differing 50GHz neighbouring modulation formats and bit rates on the performance of 43Gb/s P-DPSK over 1300km of SSMF and LEAF types. Performance is shown to be robust for hybrid P-DPSK and OOK systems. © VDE VERLAG GMBH.

A comparison of the local authority adoption of risk management in England and Australia

This paper analyses the context and use of risk management in local authorities in England and Australia. The basic structures of risk management were found to be common across all four local authorities in both countries. However, substantial differences were found in the national context in which risk management was used. The national context in each country was compared, and a large and small local authority in each country was used for illustrative purposes. The research findings were tested against institutional, contingency, resource dependence, and political perspectives. The research finding is that each theory was necessary but not sufficient and a pluralist approach was formulated to explain the similarities and differences in risk management in local authorities across two countries.

Regulating audit beyond the crisis:a critical discussion of the EU green paper

With the European Commission making global leadership claims in the field of audit regulation, the content of its 2010 Green Paper on ‘Audit Policy: Lessons from the Crisis’ warrants careful scrutiny. Important issues raised in the Green Paper include regulatory oversight, competition in the audit market, the dangers of having very few firms with the capacity to audit global transnational corporations, professional judgement, innovative audit practices and, last but not least, social responsibility. This article analyses the principal perspectives and assumptions underpinning the construction of the Green Paper. The aims are threefold: to enhance understanding of the contemporary regulatory mindset of the European Commission, contribute to policy debate and inspire future research.

The eye of the beholder: The impact of a district implementation plan of state mandates as perceived by the implementers

The purpose of this inquiry was to investigate the impact of a large, urban school district's experience in implementing a mandated school improvement plan and to examine how that plan was perceived, interpreted, and executed by those charged with the task. The research addressed the following questions: First, by whom was the district implementation plan designed, and what factors were considered in its construction? Second, what impact did the district implementation plan have on those charged with its implementation? Third, what impact did the district plan have on the teaching and learning practices of a particular school? Fourth, what aspects of the implemention plan were perceived as most and least helpful by school personnel in achieving stated goals? Last, what were the intended and unintended consequences of an externally mandated and directed plan for improving student achievement? The implementation process was measured against Fullan's model as expounded upon in The Meaning of Educational Change (1982) and The New Meaning of Educational Change (1990). The Banya implementation model (1993), because it added a dimension not adequately addressed by Fullan, was also considered.^ A case study was used as the methodological framework of this qualitative study. Sources of data used in this inquiry included document analysis, participant observations in situ, follow-up interviews, the "long" interview, and triangulation. The study was conducted over a twelve-month period. Findings were obtained from the content analysis of interview transcripts of multiple participants. Results were described and interpreted using the Fullan and Banya models as the descriptive framework. A cross-case comparison of the multiple perspectives of the same phenomena by various participants was constructed.^ The study concluded that the school district's implementation plan to improve student achievement was closely aligned to Fullan's model, although not intentionally. The research also showed that where there was common understanding at all levels of the organization as to the expectations for teachers, level of support to be provided, and availability of resources, successful implementation occured. The areas where successful implementation did not occur were those where the complexity of the changes were underestimated and processes for dealing with unintended consequences were not considered or adequately addressed. The unique perspectives of the various participants, from the superintendent to the classroom teacher, are described. Finally, recommendations for enhancement of implementation are offered and possible topics for further research studies are postulated. ^

Effect of the Florida A+ Plan on curriculum and instruction in Title I public elementary schools

The purpose of this research was to study the effect of the Florida A+ Plan accountability program on curriculum and instruction in four Title I public elementary schools in the Miami-Dade County Public Schools system. It focused on the experiences of the school principals and the classroom teachers of the four schools as they related to curriculum and instruction. The study included an analysis of the school improvement plans in curriculum and instruction for each school during the school years 1998-2004. ^ The study was conducted in the format of interviews with the school principals and principal selected classroom teachers who taught third, fourth, or fifth grade during the first six years of the Florida A+ Plan. The analysis of the school improvement plans focused on the implementation of curriculum and instruction for each of the four schools. It focused on the goals and measurable objectives selected by each school to improve its instructional program in the academic subjects of reading, mathematics, writing, and science. ^ The findings indicated that under the pressure to improve their school grade on the Florida A+ Plan, each of the target schools, based on individual needs assessments, and restructured their instructional program each school year as documented in their school improvement plans. They altered their programs by analyzing student performance data to realign curriculum and instruction. The analysis of the interviews with the principals and the teachers showed that each school year they restructured their program to align it with the FCAT content. This realigning was a collaborative effort on the part of the administration and the instructional staff. ^

Annotated record of the detailed examination of Mn deposits from the Snellius II - Cruise 4 stations in the vicinity of the island of Misool, Eastern Indonesia

The chemical analyses of ferromanganese encrustations found on the seabed west of Misool, eastern Indonesia, indicate that these deposits formed in a way different from that of world-wide occurring manganese nodules. Ferromanganese coated pebbles and fragments that were found in the deeper parts of the study area probably originate from nearby ridges. The ferromanganese crust on the upper part of a dolomite fragment of ?30 kg is likely to be formed by hydrogenous processes, whereas that from the lower part seems to be formed by diagenetic processes mainly. These assumptions are supported by pore-water data from two box cores taken in the same area. The manganese and iron profiles versus depth in these cores indicate a high flux of these metals to the uppermost sediment layer, and possibly into the overlying bottom water. Factor analysis for the principal components of the microprobe analytical results of the mainly hydrogenous ferromanganese crust demonstrates a strong correlation of manganese with the trace metals, of iron with phosphorus and an antipathetic relationship between iron and manganese. Similar results have also been reported for abyssal manganese nodules in the world oceans. Factor analysis for the principal components of the analytical data obtained for the diagenetic ferromanganese crust results in a clear dolomite (Ca/Mg) dilution factor only.

Results of analysis and model simulation of the bed profiles

Bedforms such as dunes and ripples are ubiquitous in rivers and coastal seas, and commonly described as triangular shapes from which height and length are calculated to estimate hydrodynamic and sediment dynamic parameters. Natural bedforms, however, present a far more complicated morphology; the difference between natural bedform shape and the often assumed triangular shape is usually neglected, and how this may affect the flow is unknown. This study investigates the shapes of natural bedforms and how they influence flow and shear stress, based on four datasets extracted from earlier studies on two rivers (the Rio Paraná in Argentina, and the Lower Rhine in The Netherlands). The most commonly occurring morphological elements are a sinusoidal stoss side made of one segment and a lee side made of two segments, a gently sloping upper lee side and a relatively steep (6 to 21°) slip face. A non-hydrostatic numerical model, set up using Delft3D, served to simulate the flow over fixed bedforms with various morphologies derived from the identified morphological elements. Both shear stress and turbulence increase with increasing slip face angle and are only marginally affected by the dimensions and positions of the upper and lower lee side. The average slip face angle determined from the bed profiles is 14°, over which there is no permanent flow separation. Shear stress and turbulence above natural bedforms are higher than above a flat bed but much lower than over the often assumed 30° lee side angle.

Nutrients of pore water in sediments of the central equatorial Pacific

Shipboard whole-core squeezing was used to measure pore water concentration vs depth profiles of [NO3]-, O2 and SiO2 at 12 stations in the equatorial Pacific along a transect from 15°S to 11°N at 135°W. The [NO3]- and SiO2 profiles were combined with fine-scale resistivity and porosity measurements to calculate benthic fluxes. After using O2 profiles, coupled with the [NO3]- profiles, to constrain the C:N of the degrading organic matter, the [NO3]- fluxes were converted to benthic organic carbon degradation rates. The range in benthic organic carbon degradation rates is 7-30 ?mol cm**-2 y**-1, with maximum values at the equator and minimum values at the southern end of the transect. The zonal trend of benthic degradation rates, with its equatorial maximum and with elevated values skewed to the north of the equator, is similar to the pattern of primary production observed in the region. Benthic organic carbon degradation is 1-2% of primary production. The range of benthic biogenic silica dissolution rates is 6.9-20 µmol cm**-2 y**-1, representing 2.5-5% of silicon fixation in the surface ocean of the region. Its zonal pattern is distinctly different from that of organic carbon degradation: the range in the ratio of silica dissolution to carbon degradation along the transect is 0.44-1.7 mol Si mol C**-1, with maximum values occurring between 12°S and 2°S, and with fairly constant values of 0.5-0.7 north of the equator. A box model calculation of the average lifetime of the organic carbon in the upper 1 cm of the sediments, where 80 +/- 11% of benthic organic carbon degradation occurs, indicates that it is short: from 3.1 years at high flux stations to 11 years at low flux stations. The reactive component of the organic matter must have a shorter lifetime than this average value. In contrast, the average lifetime of biogenic silica in the upper centimeter of these sediments is 55 +/- 28 years, and shows no systematic variations with benthic flux.

Targeted sequencing for high-resolution evolutionary analyses following genome duplication in salmonid fish : Proof of concept for key components of the insulin-like growth factor axis

Acknowledgements This study was funded by a Natural Environment Research Council grant (NERC, project code: NBAF704). FML is funded by a NERC Doctoral Training Grant (Project Reference: NE/L50175X/1). RLS was an undergraduate student at the University of Aberdeen and benefitted from financial support from the School of Biological Sciences. DJM is indebted to Dr. Steven Weiss (University of Graz, Austria), Dr. Takashi Yada (National Research Institute of Fisheries Science, Japan), Dr. Robert Devlin (Fisheries and Oceans Canada, Canada), Prof. Samuel Martin (University of Aberdeen, UK), Mr. Neil Lincoln (Environment Agency, UK) and Prof. Colin Adams/Mr. Stuart Wilson (University of Glasgow, UK) for providing salmonid material or assisting with its sampling. We are grateful to staff at the Centre for Genomics Research (University of Liverpool, UK) (i.e. NERC Biomolecular Analysis Facility – Liverpool; NBAF-Liverpool) for performing sequence capture/Illumina sequencing and providing us with details on associated methods that were incorporated into the manuscript. Finally, we are grateful to the organizers of the Society of Experimental Biology Satellite meeting 'Genome-powered perspectives in integrative physiology and evolutionary biology' (held in Prague, July 2015) for inviting us to contribute to this special edition of Marine Genomics and hosting a really stimulating meeting.

Non-prescription medicine misuse, abuse and dependence : a cross-sectional survey of the UK general population

This study was supported by the Society of the Study of Addiction in the form of a PhD studentship awarded to NF.

Efficacy, safety, and immunogenicity of the human papillomavirus 16/18 AS04-adjuvanted vaccine in women older than 25 years : 7-year follow-up of the phase 3, double-blind, randomised controlled VIVIANE study

Acknowledgments The VIVIANE study was funded and coordinated by GlaxoSmithKline Biologicals SA, which also covered all costs associated with development and publication of this report. We thank all study participants and their families. We gratefully acknowledge the work of the central and local study coordinators, and staff members of the sites who participated in this study. Writing support services were provided by Mary Greenacre (An Sgriobhadair, Isle of Barra, UK), on behalf of GSK Vaccines; editing and publication coordination services were provided by Jérôme Leemans (Keyrus Biopharma, Lasne, Belgium), Stéphanie Delval (XPE Pharma and Science, Wavre, Belgium), and Matthieu Depuydt (Business Decision Life Sciences, Brussels, Belgium), on behalf of GSK Vaccines

Variability of the IFN-γ ELISpot assay in the context of proficiency testing and bridging studies.

Assays that assess cellular mediated immune responses performed under Good Clinical Laboratory Practice (GCLP) guidelines are required to provide specific and reproducible results. Defined validation procedures are required to establish the Standard Operating Procedure (SOP), include pass and fail criteria, as well as implement positivity criteria. However, little to no guidance is provided on how to perform longitudinal assessment of the key reagents utilized in the assay. Through the External Quality Assurance Program Oversight Laboratory (EQAPOL), an Interferon-gamma (IFN-γ) Enzyme-linked immunosorbent spot (ELISpot) assay proficiency testing program is administered. A limit of acceptable within site variability was estimated after six rounds of proficiency testing (PT). Previously, a PT send-out specific within site variability limit was calculated based on the dispersion (variance/mean) of the nine replicate wells of data. Now an overall 'dispersion limit' for the ELISpot PT program within site variability has been calculated as a dispersion of 3.3. The utility of this metric was assessed using a control sample to calculate the within (precision) and between (accuracy) experiment variability to determine if the dispersion limit could be applied to bridging studies (studies that assess lot-to-lot variations of key reagents) for comparing the accuracy of results with new lots to results with old lots. Finally, simulations were conducted to explore how this dispersion limit could provide guidance in the number of replicate wells needed for within and between experiment variability and the appropriate donor reactivity (number of antigen-specific cells) to be used for the evaluation of new reagents. Our bridging study simulations indicate using a minimum of six replicate wells of a control donor sample with reactivity of at least 150 spot forming cells per well is optimal. To determine significant lot-to-lot variations use the 3.3 dispersion limit for between and within experiment variability.

Lithium concentration in manganese nodules of the Pacific Ocean (Hakurei Maru Cruise GH77-1, January-March, 1977)

Various manganese nodules donated to the Scripps Institution of Oceanography from the Hakurei Maru Cruise GH77-1, January-March, 1977, in the Central Pacific Basin have been analysed for their lithium content by J. Korkish from the Institute of Analytical Chemistry, Analysis of Nuclear Raw Materials Division, University of Vienna, Austria. The author has used a Perkin-Elmer atomic-absorption spectrometer 303 after speration by dissolution in hydrochloric acid.

Coccolithophores in surface sediments of the Arabian Sea (TAble 1)

One hundred surface sediment samples of the Arabian Sea (Indian Ocean) were investigated and relative abundances of coccoliths were compared to mean annual gradients of temperature, salinity, chlorophyll, PO4 and mixed layer depth. Total coccolith concentrations ranged from 42*10**6/g sediment in coastal areas to more than 19000*10**6/g sediment in oceanic regions. The general distribution does not seem to be dependent on coccolithophore productivity in surface waters alone, but also on the diluting input of terrigenous material. A total of 27 taxa were identified. The main species dominating the assemblages were Gephyrocapsa oceanica, Emiliania huxleyi and Florisphaera profunda with a combined average abundance of more than 70%. Several species and species groups reflect with their distribution the environmental parameters of the overlying water masses and may be successfully used to improve palaeoclimatic reconstructions, e.g. (a) F. profunda exhibits a high similarity or even positive correlation to the mean annual mixed layer depth, (b) calciosolenids can be described as coastal or shelf species. While temperature and salinity gradients do not seem to be crucial for coccolithophores in this region, the mean mixed layer depth as well as the PO4 concentration (representative for total nutrient availability) may control in part the coccolithophore assemblages. According to the results of a cluster analysis and the distribution pattern of all species, it was possible to differentiate three main coccolithophore assemblages. A G. oceanica dominated assemblage mainly occurs in the northern part of the study area and can be described as 'high nutrient assemblage'. The second assemblage, dominated by F. profunda, may be typical for oligotrophic and stable conditions in open ocean waters. A third assemblage, with high amounts of 'coastal species', characterises coastal conditions on the shelves.