The effect of pH on PAMAM dendrimer-siRNA complexation:endosomal considerations as determined by molecular dynamics simulation

Intracellular degradation of genes, most notably within the endo-lysosomal compartment is considered a significant barrier to (non-viral) gene delivery in vivo. Previous reports based on in vitro studies claim that carriers possessing a mixture of primary, secondary and tertiary amines are able to buffer the acidic environment within the endosome, allowing for timely release of their contents, leading to higher transfection rates. In this report, we adopt an atomistic molecular dynamics (MD) simulation approach, comparing the complexation of 21-bp siRNA with low-generation polyamidoamine (PAMAM) dendrimers (G0 and G1) at both neutral and acidic pHs, the latter of which mimics the degradative environment within maturing 'late-endosomes'. Our simulations reveal that the time taken for the dendrimer-gene complex (dendriplex) to reach equilibrium is appreciably longer at low pH and this is accompanied by more compact packaging of the dendriplex, as compared to simulations performed at neutral pH. We also note larger absolute values of calculated binding free energies of the dendriplex at low pH, indicating a higher dendrimer-nucleic acid affinity in comparison with neutral pH. These novel simulations provide a more detailed understanding of low molecular-weight polymer-siRNA behavior, mimicking the endosomal environment and provide input of direct relevance to the "proton sponge theory", thereby advancing the rational design of non-viral gene delivery systems.

Semantic and organization considerations in database conceptual modelling : the Semantic Conceptual Organizational Model (SECOM)

This thesis presents a new approach to designing large organizational databases. The approach emphasizes the need for a holistic approach to the design process. The development of the proposed approach was based on a comprehensive examination of the issues of relevance to the design and utilization of databases. Such issues include conceptual modelling, organization theory, and semantic theory. The conceptual modelling approach presented in this thesis is developed over three design stages, or model perspectives. In the semantic perspective, concept definitions were developed based on established semantic principles. Such definitions rely on meaning - provided by intension and extension - to determine intrinsic conceptual definitions. A tool, called meaning-based classification (MBC), is devised to classify concepts based on meaning. Concept classes are then integrated using concept definitions and a set of semantic relations which rely on concept content and form. In the application perspective, relationships are semantically defined according to the application environment. Relationship definitions include explicit relationship properties and constraints. The organization perspective introduces a new set of relations specifically developed to maintain conformity of conceptual abstractions with the nature of information abstractions implied by user requirements throughout the organization. Such relations are based on the stratification of work hierarchies, defined elsewhere in the thesis. Finally, an example of an application of the proposed approach is presented to illustrate the applicability and practicality of the modelling approach.

Teaching and learning German language and culture in higher education in Britain:problems, challenges and didactical implications. A case study

The aim of this thesis is to explore key aspects and problems of the institutionalised teaching and learning of German language and culture in the context of German Studies in British Higher Education (HE). This investigation focuses on teaching and learning experiences in one department of German Studies in the UK, which is the micro-context of the present study, in order to provide an in-depth insight into real-life problems, strengths and weaknesses as they occur in the practice of teaching and learning German. Following Lamb (2004) and Holliday (1994), the present study acts on the assumption that each micro-context does not exist in vacuo but is always embedded in a wider socio-political and education environment, namely the macro-context, which largely determines how and what is taught. The macro-analysis of the present study surveys the socio-political developments that have recently affected the sector of modern languages and specifically the discipline of German Studies in the UK. It demonstrates the impact they have had on teaching and learning German at the undergraduate level in Britain. This context is interesting inasmuch as the situation in Britain is to a large extent a paradigmatic example of the developments in German Studies in English-speaking countries. Subsequently, the present study explores learning experiences of a group of thirty-five first year students. It focuses on their previous experiences in learning German, exposure to the target language, motivation, learning strategies and difficulties encountered, when learning German at the tertiary level. Then, on the basis of interviews with five lecturers of German, teaching experience in the context under study is explored, problems and successful teaching strategies discussed.

Simultaneous measurement of temperature and strain: cross sensitivity considerations

Interferometric sensors using optical fibers as a transduction medium have been shown to be sensitive to a variety of physical measurands. A result of this is that the resolution of a system designed to sense strain, for example, may be compromised by fluctuations in the temperature of the environment. The possibility of simultaneously determining the strain and temperature applied to the same piece of highly birefringent fiber is discussed. Second-order effects are shown to be important for long sensing lengths or in the presence of high strains or temperature changes. The results of experiments carried out to verify the theoretical predictions are also described.

Formulation design considerations for oral vaccines

Whilst oral vaccination is a potentially preferred route in terms of patient adherence and mass vaccination, the ability to formulate effective oral vaccines remains a challenge. The primary barrier to oral vaccination is effective delivery of the vaccine through the GI tract owing to the many obstacles it presents, including low pH, enzyme degradation and bile-salt solubilization, which can result in breakdown/deactivation of a vaccine. For effective immune responses after oral administration, particulates need to be taken up bythe M cells however, these are few in number. To enhance M-cell uptake, particle characteristics can be optimized with particle size, surface charge, targeting groups and bioadhesive properties all being considerations. Yet improved uptake may not translate into enhanced immune responses and formulating particulates with inherent adjuvant properties can offer advantages. Within this article, we establish the options available for consideration when building effective oral particulate vaccines.

Performance impact of asymmetric filtered 40Gb/s RZ DPSK system:tight optical filtering considerations

We present the impact of Asymmetric filtering of strong (e.g. 35 GHz) optical filters on the performance of 42.7 Gb/s 50% RZ-DPSK systems. The performance is evaluated when offsetting the filter by substantial amounts and it is found that with an offset of almost half the bit rate there is a significant improvement in the calculated Q.

Characterization of asymmetric filtered 40 Gb/s RZ-DPSK system-strong filtering considerations

We present the impact of frequency offsetting of strong (e.g. 35 GHz) optical filters on the performance of 42.7 Gb/s 50% RZ-DPSK systems. The performance is evaluated when offsetting the filter by substantial amounts and it is found that with an offset of almost half the bit rate there is a significant improvement in the calculated 'Q' (> 1 dB). We deployed balanced, constructive single ended and destructive single ended detection, so that we could investigate the physical origins of the penalty reduction of asymmetric filtering of 42.7 Gb/s 50% RZ-DPSK system.

Performance improvement of asymmetrical filtered 40GB/s RZ-DPSK receiver design-strong filtering considerations

We present a novel differential phase shift keying receiver design under strong optical filtering. The receiver design is based on asymmetrical filtering at the destructive port of the Mach Zehnder Interferometer. The asymmetrical filtered receiver design can significantly increase performance by 2 to 4.7dB in calculated "Q".

Country attractiveness for offshoring and offshore outsourcing:additional considerations

In this article, we review aspects relating to the attractiveness of India for information technology offshore – outsourcing. Our starting point is that, indeed, India will remain competitive in the short-medium term. However, more importantly, we move on to argue that country attractiveness is becoming a less important issue. We consider an alternative approach to analyze country attractive in which the client's strategic intent behind going offshore and the vendor's global dispersedness and its local knowledge define the attractiveness of the firm's offshoring strategy.