Investigation and development of techniques for the characterisation of the synthetic/biological interface

The purpose of this study is to increase our knowledge of the nature of the surface properties of polymeric materials and improve our understanding of how these factors influence the deposition of proteins to form a reactive biological/synthetic interface. A number of surface analytical techniques were identified as being of potential benefit to this investigation and included in a multidisciplinary research program. Cell adhesion in culture was the primary biological sensor of surface properties, and it showed that the cell response to different materials can be modified by adhesion promoting protein layers: cell adhesion is a protein-mediated event. A range of surface rugosity can be produced on polystyrene, and the results presented here show that surface rugosity does not play a major role in determining a material's cell adhesiveness. Contact angle measurements showed that surface energy (specifically the polar fraction) is important in promoting cell spreading on surfaces. The immunogold labelling technique indicated that there were small, but noticeable differences, between the distribution of proteins on a range of surfaces. This study has shown that surface analysis techniques have different sensitivities in terms of detection limits and depth probed, and these are important in determining the usefulness of the information obtained. The techniques provide information on differing aspects of the biological/synthetic interface, and the consequence of this is that a range of techniques is needed in any full study of such a complex field as the biomaterials area.

An applied linguistic analysis of the simplification of narrative texts

Simplification of texts has traditionally been carried out by replacing words and structures with appropriate semantic equivalents in the learner's interlanguage, omitting whichever items prove intractable, and thereby bringing the language of the original within the scope of the learner's transitional linguistic competence. This kind of simplification focuses mainly on the formal features of language. The simplifier can, on the other hand, concentrate on making explicit the propositional content and its presentation in the original in order to bring what is communicated in the original within the scope of the learner's transitional communicative competence. In this case, simplification focuses on the communicative function of the language. Up to now, however, approaches to the problem of simplification have been mainly concerned with the first kind, using the simplifier’s intuition as to what constitutes difficulty for the learner. There appear to be few objective principles underlying this process. The main aim of this study is to investigate the effect of simplification on the communicative aspects of narrative texts, which includes the manner in which narrative units at higher levels of organisation are structured and presented and also the temporal and logical relationships between lower level structures such as sentences/clauses, with the intention of establishing an objective approach to the problem of simplification based on a set of principled procedures which could be used as a guideline in the simplification of material for foreign students at an advanced level.

The analysis and prediction of the quantity and composition of household refuse

This thesis describes the development of a simple and accurate method for estimating the quantity and composition of household waste arisings. The method is based on the fundamental tenet that waste arisings can be predicted from information on the demographic and socio-economic characteristics of households, thus reducing the need for the direct measurement of waste arisings to that necessary for the calibration of a prediction model. The aim of the research is twofold: firstly to investigate the generation of waste arisings at the household level, and secondly to devise a method for supplying information on waste arisings to meet the needs of waste collection and disposal authorities, policy makers at both national and European level and the manufacturers of plant and equipment for waste sorting and treatment. The research was carried out in three phases: theoretical, empirical and analytical. In the theoretical phase specific testable hypotheses were formulated concerning the process of waste generation at the household level. The empirical phase of the research involved an initial questionnaire survey of 1277 households to obtain data on their socio-economic characteristics, and the subsequent sorting of waste arisings from each of the households surveyed. The analytical phase was divided between (a) the testing of the research hypotheses by matching each household's waste against its demographic/socioeconomic characteristics (b) the development of statistical models capable of predicting the waste arisings from an individual household and (c) the development of a practical method for obtaining area-based estimates of waste arisings using readily available data from the national census. The latter method was found to represent a substantial improvement over conventional methods of waste estimation in terms of both accuracy and spatial flexibility. The research therefore represents a substantial contribution both to scientific knowledge of the process of household waste generation, and to the practical management of waste arisings.

Assessment of the impact of discharges from surface water sewers on receiving water quality

A broad based approach has been used to assess the impact of discharges to rivers from surface water sewers, with the primary objective of determining whether such discharges have a measurable impact on water quality. Three parameters, each reflecting the effects of intermittent pollution, were included in a field work programme of biological and chemical sampling and analysis which covered 47 sewer outfall sites. These parameters were the numbers and types of benthic macroinvertebrates upstream and downstream of the outfalls, the concentrations of metals in sediments, and the concentrations of metals in algae upstream and downstream of the outfalls. Information on the sewered catchments was collected from Local Authorities and by observation of the time of sampling, and includes catchment areas, land uses, evidence of connection to the foul system, and receiving water quality classification. The methods used for site selection, sampling, laboratory analysis and data analysis are fully described, and the survey results presented. Statistical and graphical analysis of the biological data, with the aid of BMWP scores, showed that there was a small but persistent fall in water quality downstream of the studied outfalls. Further analysis including the catchment information indicated that initial water quality, sewered catchment size, receiving stream size, and catchment land use were important factors in determining the impact. Finally, the survey results were used to produce guidelines for the estimation of surface water sewer discharge impacts from knowledge of the catchment characteristics, so that planning authorities can consider water quality when new drainage systems are designed.

The activation of the CGRP receptor

The CGRP (calcitonin gene-related peptide) receptor is a family B GPCR (G-protein-coupled receptor). It consists of a GPCR, CLR (calcitonin receptor-like receptor) and an accessory protein, RAMP1 (receptor activity modifying protein 1). RAMP1 is needed for CGRP binding and also cell-surface expression of CLR. CLR is an example of a family B GPCR. Unlike family A GPCRs, little is known about how these receptors are activated by their endogenous ligands. This review considers what is known about the activation of family B GPCRs and then considers how this might be applied to CLR, particularly in light of new knowledge of the crystal structures of family A GPCRs.

A formal model of the Semantic Web Service Ontology (WSMO)

Semantic Web Service, one of the most significant research areas within the Semantic Web vision, has attracted increasing attention from both the research community and industry. The Web Service Modelling Ontology (WSMO) has been proposed as an enabling framework for the total/partial automation of the tasks (e.g., discovery, selection, composition, mediation, execution, monitoring, etc.) involved in both intra- and inter-enterprise integration of Web services. To support the standardisation and tool support of WSMO, a formal model of the language is highly desirable. As several variants of WSMO have been proposed by the WSMO community, which are still under development, the syntax and semantics of WSMO should be formally defined to facilitate easy reuse and future development. In this paper, we present a formal Object-Z formal model of WSMO, where different aspects of the language have been precisely defined within one unified framework. This model not only provides a formal unambiguous model which can be used to develop tools and facilitate future development, but as demonstrated in this paper, can be used to identify and eliminate errors present in existing documentation.

Investigation of the activation mechanism and structural characterization of the CGRP receptor

The calcitonin gene-related peptide (CGRP) receptor is an unusual G protein-coupled receptor (GPCR) in that it comprises the calcitonin receptor-like receptor (CLR), receptor activity modifying protein 1 (RAMP1) and the receptor component protein (RCP). The RAMP1 has two other homologues – RAMP2 and RAMP3. The endogenous ligand for this receptor is CGRP, a 37 amino acid neuropeptide that act as a vasodilator. This peptide has been implicated in the aetiology of health conditions such as inflammation, Reynaud’s disease and migraine. A clear understanding of the mode of activation of this receptor could be key in developing therapeutic agents for associated health conditions. Although the crystal structure of the N-terminal extracellular domain (ECD) of this receptor (in complex with an antagonist) has been published, the details of receptor-agonist interactions at this domain, and so ultimately the mechanism of receptor activation, are still unclear. Also, the C-terminus of the CLR (in the CGRP receptor), especially around the presumed helix 8 (H8) region, has not been well studied for its role in receptor signalling. This research project investigated these questions. In this study, certain residues making up the putative N-terminal ligand-binding core of the CLR (in the CGRP receptor) were mapped out and found to be crucial for receptor signalling. They included W69 and D70 of the WDG motif in family B GPCRs, as well as Y91, F92, D94 and F95 in loop 2 of CLR N-terminus. Also, F163 at the cytoplasmic end of TM1 and certain residues spanning H8 and associated C-terminal region of CLR were found to be required for CGRP receptor signalling. These residues were investigated by site-directed mutagenesis where they were mutated to alanine (or other residues in specific cases) and the effect of the mutations on receptor pharmacology assessed by evaluating cAMP production, cell surface expression, total cell expression and aCGRP-mediated receptor internalization. Moreover, the N-terminal ECDs of the CLR and RAMPs (RAMP1, RAMP2 and RAMP3) were produced in a yeast host strain (Pichia pastoris) for the purpose of structural interaction study by surface plasmon resonance (SPR). Following expression and purification, these receptor proteins were found to individually retain their secondary structures when analysed by circular dichroism (CD). Results were analysed and interpreted with the knowledge of the secretin family receptor paradigm. The research described in this thesis has produced novel data that contributes to a clearer understanding of CGRP receptor pharmacology. The study on CLR and RAMPs ECDs could be a useful tool in determining novel interacting GPCR partners of RAMPs.

Induction of protein degradation in skeletal muscle by a phorbol ester involves upregulation of the ubiquitin-proteasome proteolytic pathway

Although muscle atrophy is common to a number of disease states there is incomplete knowledge of the cellular mechanisms involved. In this study murine myotubes were treated with the phorbol ester 12-0-tetradecanoylphorbol-13-acetate (TPA) to evaluate the role of protein kinase C (PKC) as an upstream intermediate in protein degradation. TPA showed a parabolic dose-response curve for the induction of total protein degradation, with an optimal effect at a concentration of 25 nM, and an optimal incubation time of 3 h. Protein degradation was attenuated by co-incubation with the proteasome inhibitor lactacystin (5 μM), suggesting that it was mediated through the ubiquitin-proteasome proteolytic pathway. TPA induced an increased expression and activity of the ubiquitin-proteasome pathway, as evidenced by an increased functional activity, and increased expression of the 20S proteasome α-subunits, the 19S subunits MSS1 and p42, as well as the ubiquitin conjugating enzyme E214k, also with a maximal effect at a concentration of 25 nM and with a 3 h incubation time. There was also a reciprocal decrease in the cellular content of the myofibrillar protein myosin. TPA induced activation of PKC maximally at a concentration of 25 nM and this effect was attenuated by the PKC inhibitor calphostin C (300 nM), as was also total protein degradation. These results suggest that stimulation of PKC in muscle cells initiates protein degradation through the ubiquitin-proteasome pathway. TPA also induced degradation of the inhibitory protein, I-κBα, and increased nuclear accumulation of nuclear factor-κB (NF-κB) at the same time and concentrations as those inducing proteasome expression. In addition inhibition of NF-κB activation by resveratrol (30 μM) attenuated protein degradation induced by TPA. These results suggest that the induction of proteasome expression by TPA may involve the transcription factor NF-κB. © 2005 Elsevier Inc. All rights reserved.

The structure of the rare-earth phosphate glass(Sm2O3)0.205(P2O5)0.795 studied by anomalous dispersion neutron diffraction

The role of the Sm3+ ions in the structure of vitreous Sm2O3•4P2O5 has been investigated using the neutron diffraction anomalous dispersion technique, which employs the wavelength dependence of the real and imaginary parts of the neutron scattering length close to an absorption resonance. The data described here represent the first successful complete neutron anomalous dispersion study on an amorphous material. This experimental methodology permits one to determine exclusively the closest Sm• •• Sm separation. Knowledge of the R•••R (R = rare-earth) pairwise correlation is key to understanding the optical and magnetic properties of rare-earth phosphate glasses. The anomalous difference correlation function, ΔT''(r), shows a dominant feature pertaining to a Sm•••Sm separation, centred at 4.8 Å. The substantial width and marked asymmetry of this peak indicates that the minimum approach of Sm3+ ions could be as close as 4 Å. Information on other pairwise correlations is also revealed via analysis of T (r) and ΔT (r) correlation functions: Sm3+ ions display an average co-ordination number, n Sm(O), of 7, with a mean Sm–O bond length of 2.375(5) Å whilst the PO4 tetrahedra have a mean P–O bond length of 1.538(2) Å. Second- and third-neighbour correlations are also identified. These results corroborate previous findings. Such consistency lends support to the application of the anomalous dispersion technique to determine separations.

Laypersons' awareness of the role and function of the nurse practitioner

The purpose of this descriptive study was to document the level of awareness that laypersons have regarding the role and function of the nurse practitioner (NP). An instrument developed for a similar study, conducted in 1994, comparing patients' and physicians' level of knowledge of the role and function of the NP, had a low reliability. Revision of the instrument was required before further use could be justified.^ A pilot study of 25 laypersons was conducted to ensure that the revised tool was reliable prior to conducting a study. Reliability for the pilot sample was 0.84.^ The study results indicated that the majority of the sample (83%), (n = 100) knew that a NP was a registered nurse who was qualified to diagnose and treat minor illnesses. The level of layperson awareness was limited regarding prescriptive privileges, well-woman exams, and the NPs' ability to perform physical exams, and interpret lab results and x-rays. ^

Student perceptions of the school physical fitness testing program: The Fitnessgram

The purpose of this study was to determine fifth grade students' perceptions of the Fitnessgram physical fitness testing program. This study examined if the Fitnessgram physical fitness testing experience promotes an understanding of the health-related fitness components and examined the relationship between individual fitness test scores and time spent participating in out-of-school physical activity. Lastly, students' thoughts and feelings concerning the Fitnessgram experience were examined. ^ The primary participant population for the study was 110 fifth grade students at Redland Elementary School, a Miami-Dade County Public School (M-DCPS). Data were collected over the course of 5 months. Multiple sources of data allowed for triangulation. Data sources included Fitnessgram test scores, questionnaires, document analysis, and in-depth interviews. ^ Interview data were analyzed qualitatively for common broad themes, which were identified and defined. Document analysis included analyzing student fitness test scores and student questionnaire data. This information was analyzed to determine if the Fitnessgram test scores have an impact on student views about the school fitness-testing program. Data were statistically analyzed using analysis of frequency, crosstabulations (Bryman & Duncan, 1997), and Somers'd Correlation (Bryman & Duncan, 1997). The results of the analysis of data on student knowledge of the physical fitness components tested by each Fitnessgram test revealed students do not understand the health-related fitness components. ^ The results of determining a relationship between individuals' fitness test scores and time spent in out-of-school physical activity revealed a significant positive relationship for 2 of the 6 Fitnessgram tests. ^ The results of examining students' thoughts and feelings about each Fitnessgram test focused around 2 broad themes: (a) these children do not mind the physical fitness testing and (b) how they felt about the experience was directly related to how they thought they had performed. ^ If the goal of physical fitness was only to get children fit, this test may be appropriate. However, the ultimate goal of physical fitness is to encourage students to live active and healthy lifestyles. Findings suggest the Fitnessgram as implemented by M-DCPS may not be the most suitable measurement instrument when assessing attitudinal changes that affect a healthy lifelong lifestyle. ^

High-resolution cyclostratigraphy of the Cenomanian-Turonian interval: Paleoceanographic implications, a comparison between deep sea drilling project site 386 and 144

The purpose of this study was to determine the extent to which oceanic anoxic events (OAE's) are recorded in deep-water deposits of the former western Tethyan Sea, by investigating the Cenomanian-Turonian time interval characterized by the worldwide OAE 2 event. The study improved our knowledge of the possible controlling mechanisms that triggered this event at these sites, and furthered our understanding of this global phenomenon. This was examined by high-resolution, multi-proxy analyses of sediments at DSDP Sites 386 and 144, including sedimentology, scanning electron microscopy, stable isotopes, bulk and clay mineralogy, major and trace element geochemistry, biomarkers, and paleontological data. ^ The results provide a better stratigraphic resolution for the Cenomanian-Turonian, which allowed for more precise determination of chronologic boundaries, sedimentation rates at DSDP Site 386, and a more accurate calculation of the frequency of the cycles recorded in the sequence, which fall predominantly within the precession (∼23 kyr) and short eccentricity (∼100 kyr) ranges. The combined proxies allow assessment of the correlation of δ13Corg, and major and trace elements with the predominance of cyanobacteria. These organisms were the main producers of the organic matter during the dysoxic and euxinic conditions of OAE 2 at DSDP Site 386. A huge amount of microcrystalline quartz of eolian origin is also associated with OAE 2. The geochemical proxies further provide evidence that OAE 2 was linked to increased volcanism outside the deep water of the proto-Atlantic Ocean. The clays in the Turonian sediments are terrigenous and derived predominantly from eolian transport. Comparing DSDP Site 386 and 144 with stratotype sections, the δ13C org and TOC data indicate that OAE 2 seems diachronous throughout the proto-Atlantic Ocean. ^ This study concludes that the development of anoxic conditions in the deep water of the Atlantic during the latest Cenomanian-Turonian resulted from a combination of factors related to local oceanic setting and mitigated by global tectonism and climate. The data provide a more comprehensive view of the interacting factors that led to sustained high productivity of the cyanobacteria and photosynthetic protists that produced organic-carbon-rich deposits in the world's oceans. ^

Re -"interpreting" the conquest: European and Amerindian translators and go -betweens in the colonization of the Americas, 1492--1675

This dissertation aims to recover the lives and careers of those Amerindians and Europeans who voluntarily or involuntarily took on the role of intercultural interpreters in the contact, conquest, and early colonial period in the Americas between 1492 and 1675. It intends to prove that these so-called “marginal” figures assumed roles that went far beyond those of linguistic and cultural translators, and often had a decisive impact on early Indian-colonial relations. ^ In the course of my research, I consulted hundreds of published sixteenth- and seventeenth-century chronicles, narratives, and memoirs in my search for references to interpreters. I augmented these accounts with information derived from unpublished archival documents, drawn primarily from the Archivo General de Indias, in Seville, Spain. ^ I organized my findings in theme-driven chapters that begin with a consideration of the historiography of that subject. Each chapter is further subdivided into chronologically-arranged historical vignettes that focus on the interpreters who mediated between the Spanish, Portuguese, French, English and Dutch and the various Native American polities and cultures. ^ I found that colonial authorities and Amerindian communities alike recognized the absolute necessity of recruiting competent and loyal interpreters and go-betweens, and that both sides tried to secure their loyal service by means both fair and foul. Although pressured, pushed, and pulled in contrary directions, most interpreters recognized the pivotal position they held in cross-cultural negotiations and rarely remained passive pawns in the contests between the forces of domination and defense. ^ All across the Americas, interpreters used their linguistic and diplomatic skills, and their intimate knowledge of the “other” not simply to facilitate conquest or spearhead the opposition, but to transform themselves from “culture brokers” into “power brokers.” Many of the decisive events that shaped colonial-Indian relations turned on the actions of these culturally-ambiguous individuals, a fact bemoaned and begrudgingly acknowledged by most of the contemporary conquistadors, chroniclers, and colonial founders, and recognized by this author. ^

A precise measurement of the two -photon exchange effect

The two-photon exchange phenomenon is believed to be responsible for the discrepancy observed between the ratio of proton electric and magnetic form factors, measured by the Rosenbluth and polarization transfer methods. This disagreement is about a factor of three at Q 2 of 5.6 GeV2. The precise knowledge of the proton form factors is of critical importance in understanding the structure of this nucleon. The theoretical models that estimate the size of the two-photon exchange (TPE) radiative correction are poorly constrained. This factor was found to be directly measurable by taking the ratio of the electron-proton and positron-proton elastic scattering cross sections, as the TPE effect changes sign with respect to the charge of the incident particle. A test run of a modified beamline has been conducted with the CEBAF Large Acceptance Spectrometer (CLAS) at Thomas Jefferson National Accelerator Facility. This test run demonstrated the feasibility of producing a mixed electron/positron beam of good quality. Extensive simulations performed prior to the run were used to reduce the background rate that limits the production luminosity. A 3.3 GeV primary electron beam was used that resulted in an average secondary lepton beam of 1 GeV. As a result, the elastic scattering data of both lepton types were obtained at scattering angles up to 40 degrees for Q2 up to 1.5 GeV2. The cross section ratio displayed an &epsis; dependence that was Q2 dependent at smaller Q2 limits. The magnitude of the average ratio as a function of &epsis; was consistent with the previous measurements, and the elastic (Blunden) model to within the experimental uncertainties. Ultimately, higher luminosity is needed to extend the data range to lower &epsis; where the TPE effect is predicted to be largest.

On-screen pre-deblurring of digital images using the wavefront aberration function of the human eye to improve computer access for the visually impaired

Traditional Optics has provided ways to compensate some common visual limitations (up to second order visual impairments) through spectacles or contact lenses. Recent developments in wavefront science make it possible to obtain an accurate model of the Point Spread Function (PSF) of the human eye. Through what is known as the "Wavefront Aberration Function" of the human eye, exact knowledge of the optical aberration of the human eye is possible, allowing a mathematical model of the PSF to be obtained. This model could be used to pre-compensate (inverse-filter) the images displayed on computer screens in order to counter the distortion in the user's eye. This project takes advantage of the fact that the wavefront aberration function, commonly expressed as a Zernike polynomial, can be generated from the ophthalmic prescription used to fit spectacles to a person. This allows the pre-compensation, or onscreen deblurring, to be done for various visual impairments, up to second order (commonly known as myopia, hyperopia, or astigmatism). The technique proposed towards that goal and results obtained using a lens, for which the PSF is known, that is introduced into the visual path of subjects without visual impairment will be presented. In addition to substituting the effect of spectacles or contact lenses in correcting the loworder visual limitations of the viewer, the significance of this approach is that it has the potential to address higher-order abnormalities in the eye, currently not correctable by simple means.