The efficiency of encapsulation within surface rehydrated polymersomes

The key to the use of polymersomes as effective molecular delivery systems is in the ability to design processing routes that can efficiently encapsulate the molecular payload. We have evaluated various surface rehydration mechanisms for encapsulation, in each case characterizing the morphologies formed using DLS and confocal microscopy as well as determining the encapsulation efficiency for the hydrophilic dye Rhodamine B. In contrast to bulk methods, where the encapsulation efficiencies are low, we find that higher efficiencies can be obtained by the rehydration of thin films. We relate these results to the non-equilibrium mechanisms that underlie vesicle formation and discuss how an understanding of these mechanisms can help optimize encapsulation efficiencies. Our conclusion is that, even considering the good encapsulation efficiency, surface methods are still unsuitable for the massive scale-up needed when applied to commercial mass market molecular delivery scenarios. However, targeting more specialized applications for high value ingredients (like pharmaceuticals) might be more feasible.

The Hydration chemistry of blended portland blastfurnace slag cements for radiactive waste encapsulation

Blended Portland-blastfumace slag cements provide a suitable matrix for the encapsulation of low and intermediate level waste due to their inherantly low connective porosity and provide a highly alkaline and strongly reduced chemical environment. The hydration mechanism of these materials is complex and involves several competing chemical reactions. This thesis investigates three main areas: 1) The developing chemical shrinkage of the system shows that the underlying kinetics are dominantly linear and estimates of the activation energy of the slag made by this method and by conduction calorimetry show it to be c.53 kJ/mol. 2) Examination of the soUd phase reveals that caldum hydroxide is initially precipitated and subsequently consumed during hydration. The absolute rate of slag hydration is investigated by chemical and thermal methods and an estimation of the average silicate chain length (3 silicate units) by NMR is presented. 3) The developing pore solution chemistry shows that the system becomes rapidly alkaline (pH 13 - 13.5) and subsequently strongly reduced. Ion chromatography shows the presence of reduced sulphur species which are associated with the onset of reducing conditions. In the above studies, close control of the hydration temperature was maintained and the operation of a temperature controlled pore fluid extration press is reported.

The application of robotics to the turbine blade encapsulation process

A survey of the existing state-of-the-art of turbine blade manufacture highlights two operations that have not been automated namely that of loading of a turbine blade into an encapsulation die, and that of removing a machined blade from the encapsulation block. The automation of blade decapsulation has not been pursued. In order to develop a system to automate the loading of an encapsulation die a prototype mechanical handling robot has been designed together with a computer controlled encapsulation die. The robot has been designed as a mechanical handling robot of cylindrical geometry, suitable for use in a circular work cell. It is the prototype for a production model to be called `The Cybermate'. The prototype robot is mechanically complete but due to unforeseen circumstances the robot control system is not available (the development of the control system did not form a part of this project), hence it has not been possible to fully test and assess the robot mechanical design. Robot loading of the encapsulation die has thus been simulated. The research work with regard to the encapsulation die has focused on the development of computer controlled, hydraulically actuated, location pins. Such pins compensate for the inherent positional inaccuracy of the loading robot and reproduce the dexterity of the human operator. Each pin comprises a miniature hydraulic cylinder, controlled by a standard bidirectional flow control valve. The precision positional control is obtained through pulsing of the valves under software control, with positional feedback from an 8-bit transducer. A test-rig comprising one hydraulic location pin together with an opposing spring loaded pin has demonstrated that such a pin arrangement can be controlled with a repeatability of +/-.00045'. In addition this test-rig has demonstrated that such a pin arrangement can be used to gauge and compensate for the dimensional error of the component held between the pins, by offsetting the pin datum positions to allow for the component error. A gauging repeatability of +/- 0.00015' was demonstrated. This work has led to the design and manufacture of an encapsulation die comprising ten such pins and the associated computer software. All aspects of the control software except blade gauging and positional data storage have been demonstrated. Work is now required to achieve the accuracy of control demonstrated by the single pin test-rig, with each of the ten pins in the encapsulation die. This would allow trials of the complete loading cycle to take place.

Synthesis and degradation of biodegradable polyanhydride microspheres for encapsulation of proteins.

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Encapsulation of phase change materials using rice-husk-char

This paper explored a new approach to prepare phase change microcapsules using carbon-based particles via Pickering emulsions for energy storage applications. Rice-husk-char, a by-product in biofuel production, containing 53.58 wt% of carbon was used as a model carbon-based material to encapsulate hexadecane. As a model phase change material, hexadecane was emulsified in aqueous suspensions of rice-husk-char nanoparticles. Water soluble polymers poly(diallyldimethyl-ammonium chloride) and poly(sodium styrene sulfonate) were used to fix the rice-husk-char nanoparticles on the emulsion droplets through layer-by-layer assembly to enhance the structural stability of the microcapsules. The microcapsules formed are composed of a thin shell encompassing a large core consisting of hexadecane. Thermal gravimetrical and differential scanning calorimeter analyses showed the phase change enthalpy of 80.9 kJ kg−1 or 120.0 MJ m−3. Design criteria of phase change microcapsules and preparation considerations were discussed in terms of desired applications. This work demonstrated possible utilisations of biomass-originated carbon-based material for thermal energy recovery and storage applications, which can be a new route of carbon capture and utilisation.

True real-time change data capture with web service database encapsulation

This research is investigating the claim that Change Data Capture (CDC) technologies capture data changes in real-time. Based on theory, our hypothesis states that real-time CDC is not achievable with traditional approaches (log scanning, triggers and timestamps). Traditional approaches to CDC require a resource to be polled, which prevents true real-time CDC. We propose an approach to CDC that encapsulates the data source with a set of web services. These web services will propagate the changes to the targets and eliminate the need for polling. Additionally we propose a framework for CDC technologies that allow changes to flow from source to target. This paper discusses current CDC technologies and presents the theory about why they are unable to deliver changes in real-time. Following, we discuss our web service approach to CDC and accompanying framework, explaining how they can produce real-time CDC. The paper concludes with a discussion on the research required to investigate the real-time capabilities of CDC technologies. © 2010 IEEE.

What nominal phrases are all about: the atypical case of indefinite pronouns with a determiner

This paper presents novel data that challenge the traditional categorial understanding of the nominal phrase. The established use of an indefinite pronoun with a determiner in French (ce quelqu'un, du n'importe quoi, un je ne sais quoi) contravenes assumptions both about pronouns, which should not be embedded, and nominal phrases, which should be headed by a noun. Analysed here for the first time, the embedding of a pronoun under a determiner is shown to find its justification in the semantic import of the construction. The anaphoric role guaranteeing referential continuity is promoted by a strong determiner; weak determiners typically contribute to constructing a designative use of the pronoun when a more precise characterisation cannot or will not be provided. How this construction would be analysed in the Minimalist Programme is presented to suggest that the phrase satisfies semantic requirements, which resolves the paradoxes of its traditional definition

Effect of vesicle size on tissue localization and immunogenicity of liposomal DNA vaccines

The formulation of plasmid DNA (pDNA) in cationic liposomes is a promising strategy to improve the potency of DNA vaccines. In this respect, physicochemical parameters such as liposome size may be important for their efficacy. The aim of the current study was to investigate the effect of vesicle size on the in vivo performance of liposomal pDNA vaccines after subcutaneous vaccination in mice. The tissue distribution of cationic liposomes of two sizes, 500 nm (PDI 0.6) and 140 nm (PDI 0.15), composed of egg PC, DOPE and DOTAP, with encapsulated OVA-encoding pDNA, was studied by using dual radiolabeled pDNA-liposomes. Their potency to elicit cellular and humoral immune responses was investigated upon application in a homologous and heterologous vaccination schedule with 3 week intervals. It was shown that encapsulation of pDNA into cationic lipsomes resulted in deposition at the site of injection, and strongest retention was observed at large vesicle size. The vaccination studies demonstrated a more robust induction of OVA-specific, functional CD8+ T-cells and higher antibody levels upon vaccination with small monodisperse pDNA-liposomes, as compared to large heterodisperse liposomes or naked pDNA. The introduction of a PEG-coating on the small cationic liposomes resulted in enhanced lymphatic drainage, but immune responses were not improved when compared to non-PEGylated liposomes. In conclusion, it was shown that the physicochemical properties of the liposomes are of crucial importance for their performance as pDNA vaccine carrier, and cationic charge and small size are favorable properties for subcutaneous DNA vaccination.

Preparation, characterisation and entrapment of a non-glycosidic threitol ceramide into liposomes for presentation to invariant natural killer T cells

Dendritic cells (DCs) are able to present glycolipids to invariant natural killer T (iNKT) cells in vivo. Very few compounds have been found to stimulate iNKT cells, and of these, the best characterised is the glycolipid a-galactosylceramide, which stimulates the production of large quantities of interferon-gamma (IFN-?) and interleukin-4 (IL-4). However, aGalCer leads to overstimulation of iNKT cells. It has been demonstrated that the aGalCer analogue, threitol ceramide (ThrCer 2), successfully activates iNKT cells and overcomes the problematic iNKT cell activation-induced anergy. In this study, ThrCer 2 has been inserted into the bilayers of liposomes composed of a neutral lipid, 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC), or dimethyldioctadecylammonium bromide (DDA), a cationic lipid. Incorporation efficiencies of ThrCer within the liposomes was 96% for DSPC liposomes and 80% for DDA liposomes, with the vesicle size (large multilamellar vs. small unilamellar vesicles) making no significant difference. Langmuir-Blodgett studies suggest that both DSPC and DDA stack within the monolayer co-operatively with the ThrCer molecules with no condensing effect. In terms of cellular responses, IFN-? secretion was higher for cells treated with small DDA liposomes compared with the other liposome formulations, suggesting that ThrCer encapsulation in this liposome formulation resulted in a higher uptake by DCs.

Liposomes:Formulation and characterisation as contrast agents and as vaccine delivery systems

Liposome systems are well reported for their activity as vaccine adjuvants; however novel lipid-based microbubbles have also been reported to enhance the targeting of antigens into dendritic cells (DCs) in cancer immunotherapy (Suzuki et al 2009). This research initially focused on the formulation of gas-filled lipid coated microbubbles and their potential activation of macrophages using in vitro models. Further studies in the thesis concentrated on aqueous-filled liposomes as vaccine delivery systems. Initial work involved formulating and characterising four different methods of producing lipid-coated microbubbles (sometimes referred to as gas-filled liposomes), by homogenisation, sonication, a gas-releasing chemical reaction and agitation/pressurisation in terms of stability and physico-chemical characteristics. Two of the preparations were tested as pressure probes in MRI studies. The first preparation composed of a standard phospholipid (DSPC) filled with air or nitrogen (N2), whilst in the second method the microbubbles were composed of a fluorinated phospholipid (F-GPC) filled with a fluorocarbon saturated gas. The studies showed that whilst maintaining high sensitivity, a novel contrast agent which allows stable MRI measurements of fluid pressure over time, could be produced using lipid-coated microbubbles. The F-GPC microbubbles were found to withstand pressures up to 2.6 bar with minimal damage as opposed to the DSPC microbubbles, which were damaged at above 1.3 bar. However, it was also found that DSPC-filled with N2 microbubbles were also extremely robust to pressure and their performance was similar to that of F-GPC based microbubbles. Following on from the MRI studies, the DSPC-air and N2 filled lipid-based microbubbles were assessed for their potential activation of macrophages using in vitro models and compared to equivalent aqueous-filled liposomes. The microbubble formulations did not stimulate macrophage uptake, so studies thereafter focused on aqueous-filled liposomes. Further studies concentrated on formulating and characterising, both physico-chemically and immunologically, cationic liposomes based on the potent adjuvant dimethyldioctadecylammonium (DDA) and immunomodulatory trehalose dibehenate (TDB) with the addition of polyethylene glycol (PEG). One of the proposed hypotheses for the mechanism behind the immunostimulatory effect obtained with DDA:TDB is the ‘depot effect’ in which the liposomal carrier helps to retain the antigen at the injection site thereby increasing the time of vaccine exposure to the immune cells. The depot effect has been suggested to be primarily due to their cationic nature. Results reported within this thesis demonstrate that higher levels of PEG i.e. 25 % were able to significantly inhibit the formation of a liposome depot at the injection site and also severely limit the retention of antigen at the site. This therefore resulted in a faster drainage of the liposomes from the site of injection. The versatility of cationic liposomes based on DDA:TDB in combination with different immunostimulatory ligands including, polyinosinic-polycytidylic acid (poly (I:C), TLR 3 ligand), and CpG (TLR 9 ligand) either entrapped within the vesicles or adsorbed onto the liposome surface was investigated for immunogenic capacity as vaccine adjuvants. Small unilamellar (SUV) DDA:TDB vesicles (20-100 nm native size) with protein antigen adsorbed to the vesicle surface were the most potent in inducing both T cell (7-fold increase) and antibody (up to 2 log increase) antigen specific responses. The addition of TLR agonists poly(I:C) and CpG to SUV liposomes had small or no effect on their adjuvanticity. Finally, threitol ceramide (ThrCer), a new mmunostimulatory agent, was incorporated into the bilayers of liposomes composed of DDA or DSPC to investigate the uptake of ThrCer, by dendritic cells (DCs), and presentation on CD1d molecules to invariant natural killer T cells. These systems were prepared both as multilamellar vesicles (MLV) and Small unilamellar (SUV). It was demonstrated that the IFN-g secretion was higher for DDA SUV liposome formulation (p<0.05), suggesting that ThrCer encapsulation in this liposome formulation resulted in a higher uptake by DCs.

Microencapsulation studies with P(HB-HV) polymers

Microencapsulation processes, based upon the concept of solvent evaporation, have been employed within these studies to prepare microparticles from poly--hydroxybutyrate homopolymers and copolymers thereof with 3-hydroxyvalerate [P(HB-HV) polymers]. Variations in the preparative technique have facilitated the manufacture of two structurally distinct forms of microparticle. Thus, monolithic microspheres and reservoir-type microcapsules have been respectively fabricated by single and double emulsion-solvent evaporation processes. The objective of the studies reported in chapter three is to asses how a range of preparative variables affect the yield, shape and surface morphology of P(HB-HV) microcapsules. The following chapter then describes how microcapsule morphology in general, and microcapsule porosity in particular, can be regulated by blending the fabricating P(HB-HV) polymer with poly--caprolactone [PCL]. One revelation of these studies is the ability to generate uniformly microporous microcapsules from blends of various high molecular weight P(HB-HV) polymers with a low molecular weight form of PCL. These microcapsules are of particular interest because they may have the potential to facilitate the release of an encapsulated macromolecule via an aqueous diffusion mechanism which is not reliant on polymer degradation. In order to investigate this possibility, one such formulation is used in chapter five to encapsulate a wide range of different macromolecules, whose in vitro release behaviour is subsequently evaluated. The studies reported in chapter six centre on the preparation and characterization of hydrocortisone-loaded microspheres, prepared from a range of P(HB-HV) polymers, using a single emulsion-solvent evaporation process. In this chapter, the influence of the organic phase viscosity on the efficiency of drug encapsulation is the focus of initial investigations. Thereafter, it is shown how the strategies previously adopted for the regulation of microcapsule morphology can also be applied to single emulsion systems, with profound implications for the rate of drug release.

The realisation of JSD specifications in object oriented languages

Jackson System Development (JSD) is an operational software development method which addresses most of the software lifecycle either directly or by providing a framework into which more specialised techniques can fit. The method has two major phases: first an abstract specification is derived that is in principle executable; second the specification is implemented using a variety of transformations. The object oriented paradigm is based on data abstraction and encapsulation coupled to an inheritance architecture that is able to support software reuse. Its claims of improved programmer productivity and easier program maintenance make it an important technology to be considered for building complex software systems. The mapping of JSD specifications into procedural languages typified by Cobol, Ada, etc., involves techniques such as inversion and state vector separation to produce executable systems of acceptable performance. However, at present, no strategy exists to map JSD specifications into object oriented languages. The aim of this research is to investigate the relationship between JSD and the object oriented paradigm, and to identify and implement transformations capable of mapping JSD specifications into an object oriented language typified by Smalltalk-80. The direction which the transformational strategy follows is one whereby the concurrency of a specification is removed. Two approaches implementing inversion - an architectural transformation resulting in a simulated coroutine mechanism being generated - are described in detail. The first approach directly realises inversions by manipulating Smalltalk-80 system contexts. This is possible in Smalltalk-80 because contexts are first class objects and are accessible to the user like any other system object. However, problems associated with this approach are expounded. The second approach realises coroutine-like behaviour in a structure called a `followmap'. A followmap is the results of a transformation on a JSD process in which a collection of followsets is generated. Each followset represents all possible state transitions a process can undergo from the current state of the process. Followsets, together with exploitation of the class/instance mechanism for implementing state vector separation, form the basis for mapping JSD specifications into Smalltalk-80. A tool, which is also built in Smalltalk-80, supports these derived transformations and enables a user to generate Smalltalk-80 prototypes of JSD specifications.

Accessing networked services : a user interface design problem

This research investigates the general user interface problems in using networked services. Some of the problems are: users have to recall machine names and procedures to. invoke networked services; interactions with some of the services are by means of menu-based interfaces which are quite cumbersome to use; inconsistencies exist between the interfaces for different services because they were developed independently. These problems have to be removed so that users can use the services effectively. A prototype system has been developed to help users interact with networked services. This consists of software which gives the user an easy and consistent interface with the various services. The prototype is based on a graphical user interface and it includes the following appJications: Bath Information & Data Services; electronic mail; file editor. The prototype incorporates an online help facility to assist users using the system. The prototype can be divided into two parts: the user interface part that manages interactlon with the user; the communicatIon part that enables the communication with networked services to take place. The implementation is carried out using an object-oriented approach where both the user interface part and communication part are objects. The essential characteristics of object-orientation, - abstraction, encapsulation, inheritance and polymorphism - can all contribute to the better design and implementation of the prototype. The Smalltalk Model-View-Controller (MVC) methodology has been the framework for the construction of the prototype user interface. The purpose of the development was to study the effectiveness of users interaction to networked services. Having completed the prototype, tests users were requested to use the system to evaluate its effectiveness. The evaluation of the prototype is based on observation, i.e. observing the way users use the system and the opinion rating given by the users. Recommendations to improve further the prototype are given based on the results of the evaluation. based on the results of the evah:1ation. . .'. " "', ':::' ,n,<~;'.'

Solubility enhancement of poorly water soluble drugs using liposome technology

The aim of this work is to investigate the various parameters that could control the encapsulation of lipophilic drugs and investigate the influence of the physical properties of poorly water-soluble drugs on bilayer loading. Initial work investigated on the solubilisation of ibuprofen, a model insoluble drug. Drug loading was assessed using HPLC and UV spectrophotometric analysis. Preliminary studies focused on the influence of bilayer composition on drug loading to obtain an optimum cholesterol concentration. This was followed up by studies investigating the effect of longer alkyl chain lipids, unsaturated alkyl chain lipids and charged lipids. The studies also focused on the effects of pH of the hydration medium and addition of the single chain surfactant a-tocopherol. The work was followed up by investigation of a range of insoluble drugs including flurbiprofen, indomethacin, sulindac, mefenamic acid, lignocaine and progesterone to investigate the influence of drugs properties and functional group on liposomal loading. The results show that no defined trend could be obtained linking the drug loading to the different drug properties including molecular weight, log P and other drug specific characteristics. However, the presence of the oppositely charged lipids improved the encapsulation of all the drugs investigated with a similar effect obtained with the substitution of the longer chain lipids. The addition of the single chain surfactant a-tocopherol resulted in enhancement of drug loading and possibly is governed by the log P of the drug candidate. Environmental scanning-electron microscopy (ESEM) was used to dynamically follow the changes in liposome morphology in real time during dehydration thereby providing a alternative assay of liposome formulation and stability. The ESEM analysis clearly demonstrated ibuprofen incorporation enhanced the stability of PC:Chol liposomes.