Magnetic resonance microscopy of the equine hoof wall: a study of resolution and potential

Reasons for performing study: Obtaining magnetic resonance images of the inner hoof wall tissue at the microscopic level would enable early accurate diagnosis of laminitis and therefore more effective therapy. Objectives: To optimise magnetic resonance imaging (MRI) parameters in order to obtain the highest possible resolution of the structures beneath the equine hoof wall. Methods: Magnetic resonance microscopy (MRM) was performed in front feet from 6 cadaver horses using T-2-weighted fast spin echo (FSE-T-2), and T-1-weighted gradient echo (GRE-T-1) sequences. Results: In T-2 weighted FSE images most of the stratum medium showed no signal, however the coronary, terminal and sole papillae were visible. The stratum lamellatum was clearly visible and primary epidermal lamellae could be differentiated from dermal lamellae. Conclusion: Most structures beneath the hoof wall were differentiated. Conventional scanners for diagnostic MRI in horses are low or high field. However this study used ultra-high field scanners currently not available for clinical use. Signal-to-noise ratio (SIN) increases as a function of field strength. An increase of spatial resolution of the image results in a decreased SIN. SIN can also be improved with better coils and the resolution of high field MRI scanners will increase as technology develops and surface array coils become more readily available. Potential relevance: Although MR images with microscopic resolution were obtained ex vivo, this study demonstrates the potential for detection of lamellar pathology as it occurs. Early recognition of the development of laminitis to instigate effective therapy at an earlier stage and may improve the outcome for laminitic horses. Clinical MR is now readily available at 3 T, while 4 T, 7 T and 9 T systems are being used for human whole body applications.

Femtosecond inscription of the first order Bragg gratings in pure fused silica

The fabrication of sub-micron periodic structures beyond diffraction limit is a major motivation for the present paper. We describe the fabrication of the periodic structure of 25 mm long with a pitch size of 260 nm which is less than a third of the wavelength used. This is the smallest reported period of the periodic structure inscribed by direct point-by-point method. A prototype of the add-drop filter, which utilizes such gratings, was demonstrated in one stage fabrication process of femtosecond inscription, in the bulk fused silica.

Femtosecond inscription of the first order Bragg gratings in pure fused silica

The fabrication of sub-micron periodic structures beyond diffraction limit is a major motivation for the present paper. We describe the fabrication of the periodic structure of 25 mm long with a pitch size of 260 nm which is less than a third of the wavelength used. This is the smallest reported period of the periodic structure inscribed by direct point-by-point method. A prototype of the add-drop filter, which utilizes such gratings, was demonstrated in one stage fabrication process of femtosecond inscription, in the bulk fused silica.

Increasing competitiveness of third party logistics with RFID

Competition in the 3PL market continues to intensify as providers compete to win and retain clients. 3PL providers are required to reduce costs while offering tailored innovative logistical solutions in order to remain competitive. 3PL providers can reduce costs through the consolidation of assets and introduction of cross-docking activities. Innovative logistical services can be tailored to each client via the introduction of real-time data updates. This paper highlights that RFID enabled RTE can assist in improvements of both these areas through increased network visibility. A framework is presented where the 3PL provider focuses on asset reduction, asset utilisation, real-time data employment and RTE cycle time reduction in order to enhance competitiveness. © 2011 IEEE.

Third sector organisations and social welfare : a study of the role of social service and charitable organisations in Taiwan

This study is about the role and operation of ‘third sector’ organisations (TSOs) within the Taiwanese social welfare context. TSOs have increased dramatically and become actively involved in social service provision. This phenomenon has not only had significant impact on the development and operation of TSOs in Taiwan but it is also of increasing interest to public policy academics. The latter are especially interested in the implications for the government-third sector relationship. This research examines the reasons why TSOs have been established, why they actively participate in social service provision, and their role and operation within the social welfare context of Taiwan. The study has both quantitative and qualitative data. It sampled ‘social service’ and ‘charitable’ organisations (SSCOs), which are the main type of TSOs in Taiwan, to examine their role, operation and interaction with government. Questionnaires were mailed to collect quantitative data first. After the quantitative data were collected and analysed, semi-structured interviews were undertaken to collect qualitative data. The study found that TSOs in Taiwan exist in a highly institutionalised environment, which is affected by traditional Confucian ideas and contemporary Western ideas such as social justice and civil rights. The rapid growth of TSOs has a strong connection with the desire to fill social service gaps left by government and family. TSOs mainly play the role of service provider rather than that of advocate. They cooperate with government in social service provision and have developed different types of symbiotic relationships with government. A ‘resonance effect’ between government and TSOs was also found as they implement social policy.

Nuclear magnetic resonance studies of molecular interactions and structures

This thesis is concerned with the investigation, by nuclear magnetic resonance spectroscopy, of the molecular interactions occurring in mixtures of benzene and cyclohexane to which either chloroform or deutero-chloroform has been added. The effect of the added polar molecule on the liquid structure has been studied using spin-lattice relaxation time, 1H chemical shift, and nuclear Overhauser effect measurements. The main purpose of the work has been to validate a model for molecular interaction involving local ordering of benzene around chloroform. A chemical method for removing dissolved oxygen from samples has been developed to encompass a number of types of sample, including quantitative mixtures, and its supremacy over conventional deoxygenation technique is shown. A set of spectrometer conditions, the use of which produces the minimal variation in peak height in the steady state, is presented. To separate the general diluting effects of deutero-chloroform from its effects due to the production of local order a series of mixtures involving carbon tetrachloride, instead of deutero-chloroform, have been used as non-interacting references. The effect of molecular interaction is shown to be explainable using a solvation model, whilst an approach involving 1:1 complex formation is shown not to account for the observations. It is calculated that each solvation shell, based on deutero-chloroform, contains about twelve molecules of benzene or cyclohexane. The equations produced to account for the T1 variations have been adapted to account for the 1H chemical shift variations in the same system. The shift measurements are shown to substantiate the solvent cage model with a cage capacity of twelve molecules around each chloroform molecule. Nuclear Overhauser effect data have been analysed quantitatively in a manner consistent with the solvation model. The results show that discrete shells only exist when the mole fraction of deutero-chloroform is below about 0.08.

The making of an ideology of the third way:the evolution of christian democracy in Germany (with special reference to 1945-49)

This study investigates the search for the third way in the history of German Christian Democracy. Today, in the United Kingdom, the 'third way' is seen as a new phenomenon, a synthesis of post-war belief in the welfare state and neo-liberal conservatism. Yet it insufficiently acknowledges that the origins of third way thinking, the marriage of social justice with free market economics, of individualism with collective responsibility, are found in the early philosophies of Catholic Social Theory and Protestant Social Ethical Teaching in Germany. This study shows that in the hundred years from the 1840s to the end of the 1940s, there were Catholic and Protestant socio-ethical thinkers and political reformists in Germany who attempted to bridge the philosophical differences between liberalism and socialism, to develop a socio-economic order based on Christian moral values. It will focus on the period 1945-1949, when the CDU was founded as the first interdenominational, Christian party. The study provides the first comprehensive account of the political debates in Christian democratic groups in the Soviet, British, French and American allied occupied zones, also giving equal attention to the contribution from the Protestant wing, alongside the more widely acknowledged role of Catholics in the birth of the CDU. It examines how Christian Democrats envisaged correcting the aberrations of German history, by uniting all social classes and Christian religions in one all-embracing Volkspartei, and transforming party politics from its earlier obsession with sectarian and ideological interests towards a more pragmatic 'third way' programme. The study argues that through the making of its ideology, the CDU modified the nation's understanding of its history, re-interpreted its traditions, and redefined the meaning and perception of established political philosophies. This reveals how the ambiguity of political terminology, and the flexible practice of 'third way' politics, were an invaluable political resource in the CDU's campaign for unity, ideological legitimisation and political power.

Editorial overview:new protein production tools for structural biology

Full text: The idea of producing proteins from recombinant DNA hatched almost half a century ago. In his PhD thesis, Peter Lobban foresaw the prospect of inserting foreign DNA (from any source, including mammalian cells) into the genome of a λ phage in order to detect and recover protein products from Escherichia coli [ 1 and 2]. Only a few years later, in 1977, Herbert Boyer and his colleagues succeeded in the first ever expression of a peptide-coding gene in E. coli — they produced recombinant somatostatin [ 3] followed shortly after by human insulin. The field has advanced enormously since those early days and today recombinant proteins have become indispensable in advancing research and development in all fields of the life sciences. Structural biology, in particular, has benefitted tremendously from recombinant protein biotechnology, and an overwhelming proportion of the entries in the Protein Data Bank (PDB) are based on heterologously expressed proteins. Nonetheless, synthesizing, purifying and stabilizing recombinant proteins can still be thoroughly challenging. For example, the soluble proteome is organized to a large part into multicomponent complexes (in humans often comprising ten or more subunits), posing critical challenges for recombinant production. A third of all proteins in cells are located in the membrane, and pose special challenges that require a more bespoke approach. Recent advances may now mean that even these most recalcitrant of proteins could become tenable structural biology targets on a more routine basis. In this special issue, we examine progress in key areas that suggests this is indeed the case. Our first contribution examines the importance of understanding quality control in the host cell during recombinant protein production, and pays particular attention to the synthesis of recombinant membrane proteins. A major challenge faced by any host cell factory is the balance it must strike between its own requirements for growth and the fact that its cellular machinery has essentially been hijacked by an expression construct. In this context, Bill and von der Haar examine emerging insights into the role of the dependent pathways of translation and protein folding in defining high-yielding recombinant membrane protein production experiments for the common prokaryotic and eukaryotic expression hosts. Rather than acting as isolated entities, many membrane proteins form complexes to carry out their functions. To understand their biological mechanisms, it is essential to study the molecular structure of the intact membrane protein assemblies. Recombinant production of membrane protein complexes is still a formidable, at times insurmountable, challenge. In these cases, extraction from natural sources is the only option to prepare samples for structural and functional studies. Zorman and co-workers, in our second contribution, provide an overview of recent advances in the production of multi-subunit membrane protein complexes and highlight recent achievements in membrane protein structural research brought about by state-of-the-art near-atomic resolution cryo-electron microscopy techniques. E. coli has been the dominant host cell for recombinant protein production. Nonetheless, eukaryotic expression systems, including yeasts, insect cells and mammalian cells, are increasingly gaining prominence in the field. The yeast species Pichia pastoris, is a well-established recombinant expression system for a number of applications, including the production of a range of different membrane proteins. Byrne reviews high-resolution structures that have been determined using this methylotroph as an expression host. Although it is not yet clear why P. pastoris is suited to producing such a wide range of membrane proteins, its ease of use and the availability of diverse tools that can be readily implemented in standard bioscience laboratories mean that it is likely to become an increasingly popular option in structural biology pipelines. The contribution by Columbus concludes the membrane protein section of this volume. In her overview of post-expression strategies, Columbus surveys the four most common biochemical approaches for the structural investigation of membrane proteins. Limited proteolysis has successfully aided structure determination of membrane proteins in many cases. Deglycosylation of membrane proteins following production and purification analysis has also facilitated membrane protein structure analysis. Moreover, chemical modifications, such as lysine methylation and cysteine alkylation, have proven their worth to facilitate crystallization of membrane proteins, as well as NMR investigations of membrane protein conformational sampling. Together these approaches have greatly facilitated the structure determination of more than 40 membrane proteins to date. It may be an advantage to produce a target protein in mammalian cells, especially if authentic post-translational modifications such as glycosylation are required for proper activity. Chinese Hamster Ovary (CHO) cells and Human Embryonic Kidney (HEK) 293 cell lines have emerged as excellent hosts for heterologous production. The generation of stable cell-lines is often an aspiration for synthesizing proteins expressed in mammalian cells, in particular if high volumetric yields are to be achieved. In his report, Buessow surveys recent structures of proteins produced using stable mammalian cells and summarizes both well-established and novel approaches to facilitate stable cell-line generation for structural biology applications. The ambition of many biologists is to observe a protein's structure in the native environment of the cell itself. Until recently, this seemed to be more of a dream than a reality. Advances in nuclear magnetic resonance (NMR) spectroscopy techniques, however, have now made possible the observation of mechanistic events at the molecular level of protein structure. Smith and colleagues, in an exciting contribution, review emerging ‘in-cell NMR’ techniques that demonstrate the potential to monitor biological activities by NMR in real time in native physiological environments. A current drawback of NMR as a structure determination tool derives from size limitations of the molecule under investigation and the structures of large proteins and their complexes are therefore typically intractable by NMR. A solution to this challenge is the use of selective isotope labeling of the target protein, which results in a marked reduction of the complexity of NMR spectra and allows dynamic processes even in very large proteins and even ribosomes to be investigated. Kerfah and co-workers introduce methyl-specific isotopic labeling as a molecular tool-box, and review its applications to the solution NMR analysis of large proteins. Tyagi and Lemke next examine single-molecule FRET and crosslinking following the co-translational incorporation of non-canonical amino acids (ncAAs); the goal here is to move beyond static snap-shots of proteins and their complexes and to observe them as dynamic entities. The encoding of ncAAs through codon-suppression technology allows biomolecules to be investigated with diverse structural biology methods. In their article, Tyagi and Lemke discuss these approaches and speculate on the design of improved host organisms for ‘integrative structural biology research’. Our volume concludes with two contributions that resolve particular bottlenecks in the protein structure determination pipeline. The contribution by Crepin and co-workers introduces the concept of polyproteins in contemporary structural biology. Polyproteins are widespread in nature. They represent long polypeptide chains in which individual smaller proteins with different biological function are covalently linked together. Highly specific proteases then tailor the polyprotein into its constituent proteins. Many viruses use polyproteins as a means of organizing their proteome. The concept of polyproteins has now been exploited successfully to produce hitherto inaccessible recombinant protein complexes. For instance, by means of a self-processing synthetic polyprotein, the influenza polymerase, a high-value drug target that had remained elusive for decades, has been produced, and its high-resolution structure determined. In the contribution by Desmyter and co-workers, a further, often imposing, bottleneck in high-resolution protein structure determination is addressed: The requirement to form stable three-dimensional crystal lattices that diffract incident X-ray radiation to high resolution. Nanobodies have proven to be uniquely useful as crystallization chaperones, to coax challenging targets into suitable crystal lattices. Desmyter and co-workers review the generation of nanobodies by immunization, and highlight the application of this powerful technology to the crystallography of important protein specimens including G protein-coupled receptors (GPCRs). Recombinant protein production has come a long way since Peter Lobban's hypothesis in the late 1960s, with recombinant proteins now a dominant force in structural biology. The contributions in this volume showcase an impressive array of inventive approaches that are being developed and implemented, ever increasing the scope of recombinant technology to facilitate the determination of elusive protein structures. Powerful new methods from synthetic biology are further accelerating progress. Structure determination is now reaching into the living cell with the ultimate goal of observing functional molecular architectures in action in their native physiological environment. We anticipate that even the most challenging protein assemblies will be tackled by recombinant technology in the near future.

Polynomial Expansions for Solutions of Higher-Order Bessel Heat Equation in Quantum Calculus

Mathematics Subject Class.: 33C10,33D60,26D15,33D05,33D15,33D90

Enjoying the third age! Discourse, identity and liminality in extra-care communities

Extra-care housing has been an important and growing element of housing and care for older people in the United Kingdom since the 1990s. Previous studies have examined specific features and programmes within extra-care locations, but few have studied how residents negotiate social life and identity. Those that have, have noted that while extra care brings many health-related and social benefits, extra-care communities can also be difficult affective terrain. Given that many residents are now ‘ageing in place’ in extra care, it is timely to revisit these questions of identity and affect. Here we draw on the qualitative element of a three-year, mixed-method study of 14 extra-care villages and schemes run by the ExtraCare Charitable Trust. We follow Alemàn in regarding residents' ambivalent accounts of life in ExtraCare as important windows on the way in which liminal residents negotiate the dialectics of dependence and independence. However, we suggest that the dialectic of interest here is that of the third and fourth age, as described by Gilleard and Higgs. We set that dialectic within a post-structuralist/Lacanian framework in order to examine the different modes of enjoyment that liminal residents procure in ExtraCare's third age public spaces and ideals, and suggest that their complaints can be read in three ways: as statements about altered material conditions; as inter-subjective bolstering of group identity; and as fantasmatic support for liminal identities. Finally, we examine the implications that this latter psycho-social reading of residents' complaints has for enhancing and supporting residents' wellbeing.

A nanoliter volume nuclear magnetic resonance (NMR) system using tunneling magneto-resistive (TMR) sensors to recognize biomolecules

The need to incorporate advanced engineering tools in biology, biochemistry and medicine is in great demand. Many of the existing instruments and tools are usually expensive and require special facilities.^ With the advent of nanotechnology in the past decade, new approaches to develop devices and tools have been generated by academia and industry. ^ One such technology, NMR spectroscopy, has been used by biochemists for more than 2 decades to study the molecular structure of chemical compounds. However, NMR spectrometers are very expensive and require special laboratory rooms for their proper operation. High magnetic fields with strengths in the order of several Tesla make these instruments unaffordable to most research groups.^ This doctoral research proposes a new technology to develop NMR spectrometers that can operate at field strengths of less than 0.5 Tesla using an inexpensive permanent magnet and spin dependent nanoscale magnetic devices. This portable NMR system is intended to analyze samples as small as a few nanoliters.^ The main problem to resolve when downscaling the variables is to obtain an NMR signal with high Signal-To-Noise-Ratio (SNR). A special Tunneling Magneto-Resistive (TMR) sensor design was developed to achieve this goal. The minimum specifications for each component of the proposed NMR system were established. A complete NMR system was designed based on these minimum requirements. The goat was always to find cost effective realistic components. The novel design of the NMR system uses technologies such as Direct Digital Synthesis (DDS), Digital Signal Processing (DSP) and a special Backpropagation Neural Network that finds the best match of the NMR spectrum. The system was designed, calculated and simulated with excellent results.^ In addition, a general method to design TMR Sensors was developed. The technique was automated and a computer program was written to help the designer perform this task interactively.^

Nuclear magnetic resonance spectroscopic and computational investigations of chloroperoxidase catalyzed regio- and enantio-selective transformations

Chloroperoxidase (CPO) is the most versatile heme-containing enzyme that catalyzes a broad spectrum of reactions. The remarkable feature of this enzyme is the high regio- and enantio-selectivity exhibited in CPO-catalyzed oxidation reactions. The aim of this dissertation is to elucidate the structural basis for regio- and enantio-selective transformations and investigate the application of CPO in biodegradation of synthetic dyes. ^ To unravel the mechanism of CPO-catalyzed regioselective oxidation of indole, the dissertation explored the structure of CPO-indole complex using paramagnetic relaxation and molecular modeling. The distances between the protons of indole and the heme iron revealed that the pyrrole ring of indole is oriented toward the heme with its 2-H pointing directly at the heme iron. This provides the first experimental and theoretical explanation for the "unexpected" regioselectivity of CPO-catalyzed indole oxidation. Furthermore, the residues including Leu 70, Phe 103, Ile 179, Val 182, Glu 183, and Phe 186 were found essential to the substrate binding to CPO. These results will serve as a lighthouse in guiding the design of CPO mutants with tailor-made activities for biotechnological applications. ^ To understand the origin of the enantioselectivity of CPO-catalyzed oxidation reactions, the interactions of CPO with substrates such as 2-(methylthio)thiophene were investigated by nuclear magnetic resonance spectroscopy (NMR) and computational techniques. In particular, the enantioselectivity is partly explained by the binding orientation of substrates. In third facet of this dissertation, a green and efficient system for degradation of synthetic dyes was developed. Several commercial dyes such as orange G were tested in the CPO-H2O 2-Cl- system, where degradation of these dyes was found very efficient. The presence of halide ions and acidic pH were found necessary to the decomposition of dyes. Significantly, the results revealed that this degradation of azo dyes involves a ferric hypochlorite intermediate of CPO (Fe-OCl), compound X.^

Effects of sex, third grade reading achievement and motivation as predictors of fourth grade reading achievement of Hispanic students: A path analysis

This study explored the topic of motivation for intermediate students combining both an objective criterion measure (i.e., standardized test scores) and the self-report of students on self-concept and value of reading. The purpose of this study was to examine how third grade reading achievement correlated with the motivation of fourth grade boys and girls, and, in turn, how motivation related to fourth grade reading achievement. The participants were fourth grade students (n=207) attending two public, elementary schools in Miami-Dade County who were of primarily Hispanic origin or descent. Data were collected using the Reading Survey portion of the Motivation to Read Profile (1996) which measures self-concept and value of reading in order to measure motivation and the Third and Fourth Grade Reading Florida Comprehensive Assessment Tests 2.0 (FCAT 2.0) to assess achievement. First, a one way Analysis of Variance (ANOVA) was conducted to determine whether motivation differed significantly between fourth grade boys and girls. Second, a path analysis was used to determine whether motivation mediated or moderated the association between FCAT 2.0 third and fourth grade scores. Results of the ANOVA indicated that motivation, as measured by the Motivation to Read Profile did not differ significantly by sex. Results from the path analysis indicated that the model was significant and that third grade FCAT 2.0 scores accounted for a significant amount of the variance in fourth grade FCAT 2.0 scores once motivation was entered. Results of the study demonstrated that motivation partially mediates, but does not moderate the relationship between FCAT 2.0 third and fourth grade scores. In conclusion, it can be determined that past student achievement for fourth grade students plays a role in current student achievement when motivation is also considered. It is therefore important in order to improve the quality of fourth grade student's current performance to take into account a student's motivation and past achievement. An effort must be made to address students' motivational needs whether through school wide programs or at the classroom level in addition or in conjunction with cognition. Future research on the effect of self-concept in reading achievement is recommended.

Whole-brain patterns of 1H-magnetic resonance spectroscopy imaging in Alzheimer's disease and dementia with Lewy bodies

Acknowledgments We thank Craig Lambert for his help in processing the MRS data. The study was funded by the Sir Jules Thorn Charitable Trust (grant ref: 05/JTA) and was supported by the National Institute for Health Research (NIHR) Newcastle Biomedical Research Centre and the Biomedical Research Unit in Lewy Body Dementia based at Newcastle upon Tyne Hospitals National Health Service (NHS) Foundation Trust and Newcastle University and the NIHR Biomedical Research Centre and Biomedical Research Unit in Dementia based at Cambridge University Hospitals NHS Foundation Trust and the University of Cambridge. The views expressed are those of the authors and not necessarily those of the NHS, the NIHR or the Department of Health.

2nd order Bragg rasonance generated in a 45° tilted fiber grating and its application in a fiber laser

We report the generation of a 13dB 2nd order Bragg resonance in a conventionally UV inscribed 45° tilted fiber grating, showing strong polarization dependency and its application for singe polarization output of a fiber laser. © 2010 Optical Society of America.