Studies on the outer membrane of Pseudomonas aeruginosa and associated resistance properties

The aim of this thesis was to investigate antibacterial agents for use in disinfectant formulation in conjunction with benzalkonium chloride (BKC), and if possible, to synthesise novel agents based upon successful structures. Development of resistance to antibacterial agents following long-term exposure of P. aeruginosa to BKC was also investigated, examining cross-resistance to clinically relevant antibiotics and determining mechanisms of resistance. In this study over 50 compounds were examined for antibacterial action against P. aeruginosa, both alone and in conjunction with BKC. Successful compounds were used to design novel agents, based upon the acridine ring structure, some of which showed synergy with BKC. In 15 of the 16 strains exposed to increasing concentrations of BKC, resistance to the disinfectant arose. Strains PAO1 and OO14 were examined further, each showing stable BKC resistance and a slightly varying profile of cross-resistance. In strain PAO1 alterations in the fatty acids of the cytoplasmic membrane, increase in expression of OprG, decrease in susceptibility to EDTA as an outer membrane permeabilising agent and an increase in negativity of the cell surface charge were observed as cells became more resistant to BKC. In strain OO14 a decrease in whole cell phosphatidylcholine content, a decrease in binding/uptake of BKC and an increase in cell surface hydrophobicity were observed as cells became more resistant to BKC. Resistance to tobramycin in strain OO14 was initially high, but fell as cells were adapted to BKC, this coincided with a quantitative reduction of plasmid DNA in the cells.

Studies on the mechanism of action on antitumour imidazotetrazinones

The imidazotetrazinones are clinically active antitumour agents, temozolomide currently proving successful in the treatment of melanomas and gliomas. The exact nature of the biological processes underlying response are as yet unclear.This thesis attempts to identify the cellular targets important to the cytotoxicity of imidazotetrazinones, to elucidate the pathways by which this damage leads to cell death, and to identify mechanisms by which tumour cells may circumvent this action. The levels of the DNA repair enzymes O6-alkylguanine-DNA-alkyltransferase (O6-AGAT) and 3-methyladenine-DNA-glycosylase (3MAG) have been examined in a range of murine and human cell lines with differential sensitivity to temozolomide. All the cell lines were proficient in 3MAG despite there being 40-fold difference in sensitivity to temozolomide. This suggests that while 3-methyladenine is a major product of temozolomide alkylation of DNA it is unlikely to be a cytotoxic lesion. In contrast, there was a 20-fold variation in O6-AGAT levels and the concentration of this repair enzyme correlated with variations in cytotoxicity. Furthermore, depletion of this enzyme in a resistant, O6-AGAT proficient cell line (Raji), by pre-treatment with the free base O6-methylguanine resulted in 54% sensitisation to the effects of temozolomide. These observations have been extended to 3 glioma cell lines; results that support the view that the cytotoxicity of temozolomide is related to alkylation at the O6-position of guanine and that resistance to this drug is determined by efficient repair of this lesion. It is clear, however, the other factors may influence tumour response since temozolomide showed little differential activity towards 3 established solid murine tumours in vivo, despite different tumour O6-AGAT levels. Unlike mitozolomide, temozolomide is incapable of cross-linking DNA and a mechanism by which O6-methylguanine may exert lethality is unclear. The cytotoxicity of the methyl group may be due to its disruption of DNA-protein interactions, or alternatively cell death may not be a direct result of the alkyl group itself, but manifested by DNA single-strand breaks. Enhanced alkaline elution rates were found for the DNA of Raji cells treated with temozolomide following alkyltransferase depletion, suggesting a relationship between O6-methylguanine and the induction single-strand breaks. Such breaks can activate poly(ADP-ribose) synthetase (ADPRT) an enzyme capable of rapid and lethal depletion of cellular NAD levels. However, at concentrations of temozolomlde relevant in vivo little change in adenine nucleotides was detected in cell lines, although this enzyme would appear important in modulating DNA repair since inhibition of ADPRT potentiated temozolomide cytotoxicity in Raji cells but not O6-AGAT deficient GM892A cells. Cell lines have been reported that are O6-AGAT deficient yet resistant to methylating agents. Thus, resistance to temozolomide may arise not only by removal of the methyl group from the O6-position of guanine, but also from another mechanism involving caffeine-sensitive post-replication repair or mismatch repair activity. A modification of the standard Maxam Gilbert sequencing technique was used to determine the sequence specificity of guanine-N7 alkylation. Temozolomide preferentially alkylated runs of guanines with the intensity of reaction increasing with the number of adjacent guanines in the DNA sequence. Comparable results were obtained with a polymerase-stop assay, although neither technique elucidates the sequence specificity of O6-guanine alkylation. The importance of such specificity to cytotoxicity is uncertain, although guanine-rich sequences are common to the promoter regions of oncogenes. Expression of a plasmid reporter gene under the control of the Ha-ras proto~oncogene promoter was inhibited by alkylation with temozolomide when transfected into cancer cell lines, However, this inhibition did not appear to be related to O6~guanine alkylation and therefore would seem unimportant to the chemotherapeutic activity of temozolomide.

Intolerance of ambiguity, gender stereotypes, and attitudes to sexuality

The premise of this thesis is that Western thought is characterised by the need to enforce binary classification in order to structure the world. Classifications of sexuality and gender both embody this tendency, which has been largely influenced by Judeo-Christian tradition. Thus, it is argued that attitudes to sexuality, particularly homosexuality are, in part, a function of the way in which we seek to impose structure on the world. From this view, it is (partly) the ambiguity, inherent in gender and sexual variation, which evokes negative responses. The thesis presents a series of inter-linked studies examining attitudes to various aspects of human sexuality, including the human body, non-procreative sex acts (anal an oral sex) and patterns of sexuality that depart from the hetero-homo dichotomy. The findings support the view that attitudes to sexuality are significantly informed by gender-role stereotypes, with negative attitudes linked to intolerance of ambiguity. Male participants show large differences in their evaluations of male and female bodies, and of male and female sexual actors, than do female participants. Male participants also show a greater negativity to gay male sexual activity than do female participants, but males perceive lesbian sexuality similarly to heterosexuality. Male bodies are rated as being less 'permeable' than female bodies and male actors are more frequently identified as being the instigators of sexual acts. Crucial to the concept of heterosexism is the assumption that 'femininity' is considered inherently inferior to 'masculinity'. Hence, the findings provide an empirical basis for making connections between heterosexism and sexism, and therefore between the psychology of women, and gay and lesbian psychology.

Ultrasonic inspection of austenitic stainless steel weldments

Ultrasonic waves interact in a complex manner with the metallurgical structure of austenitic weldments resulting in ambiguity when interpreting reflections and at times in misinterpretation of defect positions. In this work, current knowledge of the structure of austenitic welds is outlined, and the influence of this structure on the propagation of ultrasonic waves is reviewed. Using an established and highly accurate technique, data on velocity variations as a function of the angle between the direction of soundwave propagation and the axes of preferred grain orientation existing in such welds, are experimentally obtained. These results and existing theory are used to provide quantitative evidence of (i) anisotropy factors in austenitic welds, (ii) beam skewing effects for different wave modes and polarizations, and (iii) the extent of acoustic impedance mismatch between parent and weld metals. The existence of "false" indications is demonstrated, and suggestions are made into their nature. The effectiveness of conventional transverse wave techniques for inspecting artificial and real defects existing in austenitic weldments is experimentally investigated, the limitations are demonstrated, and possible solutions are proposed. The possibilities offered by the use of longitudinal angle probes for ultrasonic inspection of real and artificial defects existing in austenitic weldments are experimentally investigated, and parameters such as probe angle, frequency and scanning position are evaluated. Detailed work has been carried out on the interaction of ultrasound with fatigue and corrosion-fatigue cracks in the weld metal and the heat affected zones (HAZs) of 316 and 347 types of austenitic weldments, together with the influence of elastic compressive stresses, defect topography and defect geometry. Practical applications of all results are discussed, and more effective means of ultrasonic inspection of austenitic weldments are suggested.

The effect of stimulus location on the major components of the visual evoked response

The topographical distribution of the pattern reversal Visual Evoked Response (VER) was recorded from a localised montage of 20 electrodes over the visual cortex. The response was recorded after stimulation with a black and white checkerboard stimulus. The effect of field location on the major components was investigated in 11 subjects (age range (23-55). The major components of the half field response were; a negative around 75ms (N75) followed by a positivity around 80ms (P80), then a positivity around 100ms (P100) followed by another positivity at around 120ms (P120) and a negativity at approximately 145ms (N145). No effect of field size could be demonstrated on either the amplitude or latency of the late negativity, N145. No significant effect of field size or location was shown on the latency of the P100 response. A delay previously shown in the upper half field response was therefore not substantiated. In contrast the amplitude of the major positivity, P100 was significantly affected by the field size and location. The amplitude of both P100 and N145 were significantly reduced following upper field stimulation when compared with the lower field response. No significant amplitude difference between the upper and lower field responses was demonstrated using electroretinography, the amplitude may therefore be reduced as a result of the ventral position of the upper field representation on the visual cortex. The lateral half field VEP was compared with the distribution of the visual evoked magnetic response (VEMR). The distribution of the VEMR supported the proposal that the paradoxical lateralisation of the VEP half field response is the result of the source being directed ipsilaterally. The morphology of the VEP following octant and double octant stimulation suggests that the response is generated in the striate cortex, with a reversal in response distribution following stimulation of the upper vertical and horizontal meridia.

The role of experts in policy making

As public policy issues increasingly have a technical aspect to them an interactive relationship has developed between science and policy. The aim of this thesis is to investigate the two aspects of this relationship: the influence of science on policy and the influence of policy implications on science. Most existing studies in this area treat only one or other of these aspects. Furthermore, they tend to provide interesting case study material but very little theoretical analysis. This thesis attempts to overcome these problems by dealing with both aspects of the interaction between science and policy and by providing theoretical models of this relationship. The thesis combines the theoretical development of these models with the analysis of three empirical case studies: the controversy in Britain over smoking and health; the application of educational psychology to the development of education policy in Britain; the controversy over the health effect of lead in the environment. The theoretical models are developed in Part 1. In Part 2 the empirical case studies are presented and in Part 3 the theoretical material is assessed in the light of these case studies. The main thesis of this study is that there is a fundamental mismatch between science and policy-making. Criticism is always essential in science. However, when science is involved in the policy process, either scientific claims are not subjected to a significant level of criticism or they are scrutinized so closely that no view achieves general consensus and conflicting advice results. In this situation, contrary to the traditional view, science can generate uncertainty. The role which science plays in the policy process is influenced by this level of criticism, by the context of political power and by the progress of an issue through the various stages of the policy process.

Evaluating productive efficiency:comparative study of commercial banks in Gulf countries

Financial institutes are an integral part of any modern economy. In the 1970s and 1980s, Gulf Cooperation Council (GCC) countries made significant progress in financial deepening and in building a modern financial infrastructure. This study aims to evaluate the performance (efficiency) of financial institutes (banking sector) in GCC countries. Since, the selected variables include negative data for some banks and positive for others, and the available evaluation methods are not helpful in this case, so we developed a Semi Oriented Radial Model to perform this evaluation. Furthermore, since the SORM evaluation result provides a limited information for any decision maker (bankers, investors, etc...), we proposed a second stage analysis using classification and regression (C&R) method to get further results combining SORM results with other environmental data (Financial, economical and political) to set rules for the efficient banks, hence, the results will be useful for bankers in order to improve their bank performance and to the investors, maximize their returns. Mainly there are two approaches to evaluate the performance of Decision Making Units (DMUs), under each of them there are different methods with different assumptions. Parametric approach is based on the econometric regression theory and nonparametric approach is based on a mathematical linear programming theory. Under the nonparametric approaches, there are two methods: Data Envelopment Analysis (DEA) and Free Disposal Hull (FDH). While there are three methods under the parametric approach: Stochastic Frontier Analysis (SFA); Thick Frontier Analysis (TFA) and Distribution-Free Analysis (DFA). The result shows that DEA and SFA are the most applicable methods in banking sector, but DEA is seem to be most popular between researchers. However DEA as SFA still facing many challenges, one of these challenges is how to deal with negative data, since it requires the assumption that all the input and output values are non-negative, while in many applications negative outputs could appear e.g. losses in contrast with profit. Although there are few developed Models under DEA to deal with negative data but we believe that each of them has it is own limitations, therefore we developed a Semi-Oriented-Radial-Model (SORM) that could handle the negativity issue in DEA. The application result using SORM shows that the overall performance of GCC banking is relatively high (85.6%). Although, the efficiency score is fluctuated over the study period (1998-2007) due to the second Gulf War and to the international financial crisis, but still higher than the efficiency score of their counterpart in other countries. Banks operating in Saudi Arabia seem to be the highest efficient banks followed by UAE, Omani and Bahraini banks, while banks operating in Qatar and Kuwait seem to be the lowest efficient banks; this is because these two countries are the most affected country in the second Gulf War. Also, the result shows that there is no statistical relationship between the operating style (Islamic or Conventional) and bank efficiency. Even though there is no statistical differences due to the operational style, but Islamic bank seem to be more efficient than the Conventional bank, since on average their efficiency score is 86.33% compare to 85.38% for Conventional banks. Furthermore, the Islamic banks seem to be more affected by the political crisis (second Gulf War), whereas Conventional banks seem to be more affected by the financial crisis.

The pharmacology of adrenomedullin 2/intermedin

Adrenomedullin 2 (AM2) or intermedin is a member of the calcitonin gene-related peptide (CGRP)/calcitonin family of peptides and was discovered in 2004. Unlike other members of this family, no unique receptor has yet been identified for it. It is extensively distributed throughout the body. It causes hypotension when given peripherally, but when given into the CNS, it increases blood pressure and causes sympathetic activation. It also increases prolactin release, is anti-diuretic and natriuretic and reduces food intake. Whilst its effects resemble those of AM, it is frequently more potent. Some characterization of AM2 has been done on molecularly defined receptors; the existing data suggest that it preferentially activates the AM receptor formed from calcitonin receptor-like receptor and receptor activity modifying protein 3. On this complex, its potency is generally equivalent to that of AM. There is no known receptor-activity where it is more potent than AM. In tissues and in animals it is frequently antagonised by CGRP and AM antagonists; however, situations exist in which an AM2 response is maintained even in the presence of supramaximal concentrations of these antagonists. Thus, there is a partial mismatch between the pharmacology seen in tissues and that on cloned receptors. The only AM2 antagonists are peptide fragments, and these have limited selectivity. It remains unclear as to whether novel AM2 receptors exist or whether the mismatch in pharmacology can be explained by factors such as metabolism. © 2011 The British Pharmacological Society.

Aspirin and alterations in DNA repair proteins in the SW480 colorectal cancer cell line

Regular aspirin intake is associated with a reduction in the incidence of colorectal cancer. Aspirin has been shown to be cytotoxic to colorectal cancer cells in vitro. The molecular basis for this cytotoxicity is controversial, with a number of competing hypotheses in circulation. One suggestion is that the protective effect is related to the induction of expression of the DNA mismatch repair (MMR) proteins hMLH1, hMSH2, hMSH6 and hPMS2 in DNA MMR proficient cells. We report that treatment of the DNA MMR competent/p53 mutant colorectal cancer cell line SW480 with 1 mM aspirin for 48 h caused changes in mRNA expression of several key genes involved in DNA damage signalling pathways, including a significant down-regulation in transcription of the genes ATR, BRCA1 and MAPK12. Increases in the transcription of XRCC3 and GADD45alpha genes are also reported. Regulation of these genes could potentially have profound effects on colorectal cancer cells and may play a role in the observed chemo-protective effect of aspirin in vivo. Although a correlation was not seen between transcript and protein levels of ATR, BRCA1 and GADD45alpha, an increase in XRCC3 encoded protein expression upon aspirin treatment in SW480 cells was observed by immunoblotting, immunofluorescence and immunohistochemical analysis. This is the first report of XRCC3 gene transcription and encoded protein expression being susceptible to exposure to the non-steroidal anti-inflammatory drug, aspirin. Furthermore, this study indicates that alterations in gene transcription seen in microarray studies must be verified at the protein level.

An accurate Markov model for slotted CSMA/CA algorithm in IEEE 802.15.4 networks

In this letter we propose an Markov model for slotted CSMA/CA algorithm working in a non-acknowledgement mode, specified in IEEE 802.15.4 standard. Both saturation throughput and energy consumption are modeled as functions of backoff window size, number of contending devices and frame length. Simulations show that the proposed model can achieve a very high accuracy (less than 1% mismatch) if compared to all existing models (bigger than 10% mismatch).

Stochastically driven instability in rotating shear flows

Origin of hydrodynamic turbulence in rotating shear flows is investigated. The particular emphasis is on flows whose angular velocities decrease but specific angular momenta increase with increasing radial coordinate. Such flows are Rayleigh stable, but must be turbulent in order to explain observed data. Such a mismatch between the linear theory and observations/experiments is more severe when any hydromagnetic/magnetohydrodynamic instability and the corresponding turbulence therein is ruled out. The present work explores the effect of stochastic noise on such hydrodynamic flows. We focus on a small section of such a flow which is essentially a plane shear flow supplemented by the Coriolis effect. This also mimics a small section of an astrophysical accretion disk. It is found that such stochastically driven flows exhibit large temporal and spatial correlations of perturbation velocities, and hence large energy dissipations, that presumably generate instability. A range of angular velocity profiles (for the steady flow), starting with the constant angular momentum to that of the constant circular velocity are explored. It is shown that the growth and roughness exponents calculated from the contour (envelope) of the perturbed flows are all identical, revealing a unique universality class for the stochastically forced hydrodynamics of rotating shear flows. This work, to the best of our knowledge, is the first attempt to understand origin of instability and turbulence in the three-dimensional Rayleigh stable rotating shear flows by introducing additive stochastic noise to the underlying linearized governing equations. This has important implications in resolving the turbulence problem in astrophysical hydrodynamic flows such as accretion disks.

Fluctuation-induced traffic congestion in heterogeneous networks

In studies of complex heterogeneous networks, particularly of the Internet, significant attention was paid to analyzing network failures caused by hardware faults or overload, where the network reaction was modeled as rerouting of traffic away from failed or congested elements. Here we model another type of the network reaction to congestion - a sharp reduction of the input traffic rate through congested routes which occurs on much shorter time scales. We consider the onset of congestion in the Internet where local mismatch between demand and capacity results in traffic losses and show that it can be described as a phase transition characterized by strong non-Gaussian loss fluctuations at a mesoscopic time scale. The fluctuations, caused by noise in input traffic, are exacerbated by the heterogeneous nature of the network manifested in a scale-free load distribution. They result in the network strongly overreacting to the first signs of congestion by significantly reducing input traffic along the communication paths where congestion is utterly negligible. © Copyright EPLA, 2012.

Reduced attentional capacity, but normal processing speed and shifting of attention in developmental dyslexia: evidence from a serial task

We report the performance of a group of adult dyslexics and matched controls in an array-matching task where two strings of either consonants or symbols are presented side by side and have to be judged to be the same or different. The arrays may differ either in the order or identity of two adjacent characters. This task does not require naming – which has been argued to be the cause of dyslexics’ difficulty in processing visual arrays – but, instead, has a strong serial component as demonstrated by the fact that, in both groups, Reaction times (RTs) increase monotonically with position of a mismatch. The dyslexics are clearly impaired in all conditions and performance in the identity conditions predicts performance across orthographic tasks even after age, performance IQ and phonology are partialled out. Moreover, the shapes of serial position curves are revealing of the underlying impairment. In the dyslexics, RTs increase with position at the same rate as in the controls (lines are parallel) ruling out reduced processing speed or difficulties in shifting attention. Instead, error rates show a catastrophic increase for positions which are either searched later or more subject to interference. These results are consistent with a reduction in the attentional capacity needed in a serial task to bind together identity and positional information. This capacity is best seen as a reduction in the number of spotlights into which attention can be split to process information at different locations rather than as a more generic reduction of resources which would also affect processing the details of single objects.

The economics behind the move to 'localism' in England

This paper explores the economic thinking behind the UK Coalition government’s new framework for achieving local growth and the creation of Local Enterprise Partnerships in England. It does so in the light of recent debates about ‘space-neutral’ and ‘place-based’ policymaking. While the British government states its ambition to achieve greater spatial and industrial balance across England (and by implication the UK), we argue that so far at least there is a mismatch between the ‘rhetoric’ and ‘policies’ of local growth and its limitations. These relate to inconsistencies in the way that the different competing economic ideas in circulation within government have been adopted in practice. As a result, the paper highlights six key disconnects and limitations of the economics behind the move in England to local growth.

Cognitive systems associated with the hippocampus of the human brain and their role in behaviour and neurodegenerative disease

Cognitive systems research involves the synthesis of ideas from natural and artificial systems in the analysis, understanding, and design of all intelligent systems. This chapter discusses the cognitive systems associated with the hippocampus (HC) of the human brain and their possible role in behaviour and neurodegenerative disease. The hippocampus (HC) is concerned with the analysis of highly abstract data derived from all sensory systems but its specific role remains controversial. Hence, there have been three major theories concerning its function, viz., the memory theory, the spatial theory, and the behavioral inhibition theory. The memory theory has its origin in the surgical destruction of the HC, which results in severe anterograde and partial retrograde amnesia. The spatial theory has its origin in the observation that neurons in the HC of animals show activity related to their location within the environment. By contrast, the behavioral inhibition theory suggests that the HC acts as a ‘comparator’, i.e., it compares current sensory events with expected or predicted events. If a set of expectations continues to be verified then no alteration of behavior occurs. If, however, a ‘mismatch’ is detected then the HC intervenes by initiating appropriate action by active inhibition of current motor programs and initiation of new data gathering. Understanding the cognitive systems of the hippocampus in humans may aid in the design of intelligent systems involved in spatial mapping, memory, and decision making. In addition, this information may lead to a greater understanding of the course of clinical dementia in the various neurodegenerative diseases in which there is significant damage to the HC.