The serological diagnosis of staphylococcal infective endocarditis

Objectives: Establishing the diagnosis of infective endocarditis (IE) can be difficult when blood cultures remain sterile or echocardiography is inconclusive. Staphylococcus aureus is a common aetiological microorganism in IE and is associated with severe valvular destruction and increased mortality. Early diagnosis using culture and antibiotic independent tests would be preferable to allow prompt antibiotic administration. We have developed and evaluated 2 serological assays for the rapid identification of a staphylococcal aetiology in infective endocarditis. The assays measure IgG against whole cells of S. aureus and IgG against lipid S, a novel extracellular antigen released by Gram-positive microorganisms. Methods: Serum was collected from 130 patients with IE and 94 control patients. IgG against whole cells of S. aureus and against lipid S was measured by enzyme linked immunosorbent assay (ELISA). Results: Anti-lipid S IgG titres were higher in IE caused by Gram-positive microorganisms than in controls (p < 0.0001) and higher in staphylococcal IE than in both controls and IE caused by other microorganisms (p = 0.0003). Anti-whole cell staphylococcal IgG was significantly higher in serum from patients with staphylococcal IE than in IE caused by other microorganisms and control samples (p < 0.0001). Conclusion: High anti-whole cell IgG titres are predictive of a staphylococcal aetiology in IE. Elevated serum anti-lipid S IgG titres are predictive of Gram-positive infection compared to controls, very high titres being associated with staphylococcal IE. © 2005 The British Infection Society.

Biosynthesis and analysis of plant oxylipins

The term oxylipin is applied to the generation of oxygenated products of polyunsaturated fatty acids that can arise either through non-enzymatic or enzymatic processes generating a complex array of products, including alcohols, aldehydes, ketones, acids and hydrocarbon gases. The biosynthetic origin of these products has revealed an array of enzymes involved in their formation and more recently a radical pathway. These include lipoxygenases and α-dioxygenase that insert both oxygen atoms in to the acyl chain to initiate the pathways, to specialised P450 monooxygenases that are responsible for their downstream processing. This latter group include enzymes at the branch points such as allene oxide synthase, leading to jasmonate signalling, hydroperoxide lyase, responsible for generating pathogen/pest defensive volatiles and divinyl ether synthases and peroxygenases involved in the formation of antimicrobial compounds. The complexity of the products generated raises significant challenges for their rapid identification and quantification using metabolic screening methods. Here the current developments in oxylipin metabolism are reviewed together with the emerging technologies required to expand this important field of research that underpins advances in plant-pest/pathogen interactions.

Identification of optically clear regions of ternary polymer blends using a novel rapid screening method

The application of a rapid screening method for the construction of ternary phase diagrams is described for the first time, providing detailed visualization of phase boundaries in solvent-mediated blends. Our new approach rapidly identifies ternary blend compositions that afford optically clear materials, useful for applications where transparent films are necessary. The use of 96-well plates and a scanning plate reader has enabled rapid optical characterization to be carried out by transmission spectrophotometry (450 nm), whilst the nature and extent of crystallinity was examined subsequently by wide angle X-ray scattering (WAXS). The moderating effect of cellulose acetate butyrate can be visualized as driving the position of the phase boundaries in poly(l-lactic acid)/polycaprolactone (PLLA/PCL) blends. More surprisingly, the boundaries are critically dependent on the molecular weight of the crystallizable PLLA and PCL, with higher molecular weight polymers leading to blends with reduced phase separation. On the other hand, the propensity to crystallize was more evident in shorter chains. WAXS provides a convenient way of characterizing the contribution of the individual blend components to the crystalline regions across the range of blend compositions. © 2013 Society of Chemical Industry.

Identification and molecular mechanisms of the rapid tonicity-induced relocalization of the aquaporin 4 channel

The aquaporin family of integral membrane proteins is comprised of channels that mediate cellular water flow. Aquaporin 4 (AQP4) is highly expressed in the glial cells of the central nervous system and facilitates the osmotically-driven pathological brain swelling associated with stroke and traumatic brain injury. Here we show that AQP4 cell surface expression can be rapidly and reversibly regulated in response to changes of tonicity in primary cortical rat astrocytes and in transfected HEK293 cells. The translocation mechanism involves protein kinase A (PKA) activation, influx of extracellular calcium and activation of calmodulin. We identify five putative PKA phosphorylation sites and use site-directed mutagenesis to show that only phosphorylation at one of these sites, serine- 276, is necessary for the translocation response. We discuss our findings in the context of the identification of new therapeutic approaches to treating brain oedema.

A specific and rapid neural signature for parental instinct

Darwin originally pointed out that there is something about infants which prompts adults to respond to and care for them, in order to increase individual fitness, i.e. reproductive success, via increased survivorship of one's own offspring. Lorenz proposed that it is the specific structure of the infant face that serves to elicit these parental responses, but the biological basis for this remains elusive. Here, we investigated whether adults show specific brain responses to unfamiliar infant faces compared to adult faces, where the infant and adult faces had been carefully matched across the two groups for emotional valence and arousal, as well as size and luminosity. The faces also matched closely in terms of attractiveness. Using magnetoencephalography (MEG) in adults, we found that highly specific brain activity occurred within a seventh of a second in response to unfamiliar infant faces but not to adult faces. This activity occurred in the medial orbitofrontal cortex (mOFC), an area implicated in reward behaviour, suggesting for the first time a neural basis for this vital evolutionary process. We found a peak in activity first in mOFC and then in the right fusiform face area (FFA). In mOFC the first significant peak (p<0.001) in differences in power between infant and adult faces was found at around 130 ms in the 10-15 Hz band. These early differences were not found in the FFA. In contrast, differences in power were found later, at around 165 ms, in a different band (20-25 Hz) in the right FFA, suggesting a feedback effect from mOFC. These findings provide evidence in humans of a potential brain basis for the "innate releasing mechanisms" described by Lorenz for affection and nurturing of young infants. This has potentially important clinical applications in relation to postnatal depression, and could provide opportunities for early identification of families at risk.

Management information:design of a system for change identification

This thesis deals with the problem of Information Systems design for Corporate Management. It shows that the results of applying current approaches to Management Information Systems and Corporate Modelling fully justify a fresh look to the problem. The thesis develops an approach to design based on Cybernetic principles and theories. It looks at Management as an informational process and discusses the relevance of regulation theory to its practice. The work proceeds around the concept of change and its effects on the organization's stability and survival. The idea of looking at organizations as viable systems is discussed and a design to enhance survival capacity is developed. It takes Ashby's theory of adaptation and developments on ultra-stability as a theoretical framework and considering conditions for learning and foresight deduces that a design should include three basic components: A dynamic model of the organization- environment relationships; a method to spot significant changes in the value of the essential variables and in a certain set of parameters; and a Controller able to conceive and change the other two elements and to make choices among alternative policies. Further considerations of the conditions for rapid adaptation in organisms composed of many parts, and the law of Requisite Variety determine that successful adaptive behaviour requires certain functional organization. Beer's model of viable organizations is put in relation to Ashby's theory of adaptation and regulation. The use of the Ultra-stable system as abstract unit of analysis permits developing a rigorous taxonomy of change; it starts distinguishing between change with in behaviour and change of behaviour to complete the classification with organizational change. It relates these changes to the logical categories of learning connecting the topic of Information System design with that of organizational learning.

The development of an expert system for the identification anodic coating process defects as a contribution to the dissemination of anodizing technology

Initially this thesis examines the various mechanisms by which technology is acquired within anodizing plants. In so doing the history of the evolution of anodizing technology is recorded, with particular reference to the growth of major markets and to the contribution of the marketing efforts of the aluminium industry. The business economics of various types of anodizing plants are analyzed. Consideration is also given to the impact of developments in anodizing technology on production economics and market growth. The economic costs associated with work rejected for process defects are considered. Recent changes in the industry have created conditions whereby information technology has a potentially important role to play in retaining existing knowledge. One such contribution is exemplified by the expert system which has been developed for the identification of anodizing process defects. Instead of using a "rule-based" expert system, a commercial neural networks program has been adapted for the task. The advantages of neural networks over 'rule-based' systems is that they are better suited to production problems, since the actual conditions prevailing when the defect was produced are often not known with certainty. In using the expert system, the user first identifies the process stage at which the defect probably occurred and is then directed to a file enabling the actual defects to be identified. After making this identification, the user can consult a database which gives a more detailed description of the defect, advises on remedial action and provides a bibliography of papers relating to the defect. The database uses a proprietary hypertext program, which also provides rapid cross-referencing to similar types of defect. Additionally, a graphics file can be accessed which (where appropriate) will display a graphic of the defect on screen. A total of 117 defects are included, together with 221 literature references, supplemented by 48 cross-reference hyperlinks. The main text of the thesis contains 179 literature references. (DX186565)

Use of narrow mass-window, high-resolution extracted product ion chromatograms for the sensitive and selective identification of protein modifications

Protein modifications, including oxidative modifications, glycosylations, and oxidized lipid-protein adducts, are becoming increasingly important as biomarkers and in understanding disease etiology. There has been a great deal of interest in mapping these on Apo B100 from low density lipoprotein (LDL). We have used extracted ion chromatograms of product ions generated using a very narrow mass window from high-resolution tandem mass spectrometric data collected on a rapid scanning quadrupole time-of-flight (QTOF) instrument, to selectively and sensitively detect modified peptides and identify the site and nature of a number of protein modifications in parallel. We have demonstrated the utility of this method by characterizing for the first time oxidized phospholipid adducts to LDL and human serum albumin and for the detection of glycosylation and kynurenin formation from the oxidation of tryptophan residues in LDL. © 2013 American Chemical Society.

Identification of aspirin analogues that repress NF-κB signalling and demonstrate anti-proliferative activity towards colorectal cancer in vitro and in vivo

Substantial evidence indicates that aspirin and related non-steroidal anti-inflammatory drugs (NSAIDs) have potential as chemopreventative/therapeutic agents. However, these agents cannot be universally recommended for prevention purposes due to their potential side-effect profiles. Here, we compared the growth inhibitory and mechanistic activity of aspirin to two novel analogues, diaspirin (DiA) and fumaryl diaspirin (F-DiA). We found that the aspirin analogues inhibited cell proliferation and induced apoptosis of colorectal cancer cells at significantly lower doses than aspirin. Similar to aspirin, we found that an early response to the analogues was a reduction in levels of cyclin D1 and stimulation of the NF-κB pathway. This stimulation was associated with a significant reduction in basal levels of NF-κB transcriptional activity, in keeping with previous data for aspirin. However, in contrast to aspirin, DiA and F-DiA activity was not associated with nucleolar accumulation of RelA. For all assays, F-DiA had a more rapid and significant effect than DiA, identifying this agent as particularly active against colorectal cancer. Using a syngeneic colorectal tumour model in mice, we found that, while both agents significantly inhibited tumour growth in vivo, this effect was particularly pronounced for F-DiA. These data identify two compounds that are active against colorectal cancer in vitro and in vivo. They also identify a potential mechanism of action of these agents and shed light on the chemical structures that may be important for the antitumour effects of aspirin.