Efficacy and toxicity of biocides used in handling sterile pharmaceuticals

This thesis has sought to investigate disinfection agents and procedures which may provide sanitisation against bacterial spores. A hard-surface disinfection test method was designed to ascertain which combinations of biocide and application method were most effective against bacterial spores. A combination of spraying and wiping was the most effective method of disinfection against Bacillus spores, with wiping found to play a key role in spore removal. The most efficacious of the biocides investigated was the 6% hydrogen peroxide. Vaporised Hydrogen Peroxide (VHP) gassing was more effective than traditional disinfection. In addition to efficacy, the toxic potential of the biocides to human airway epithelial cells in vitro was evaluated. Toxicity against human bronchial and nasal epithelial cells was assessed by determining cell viability, inflammatory status, protein oxidation and epithelial cell layer integrity. In addition the cell death mechanism following biocide exposure was investigated. There was a decrease in viable cells following exposure to all biocides when applied at practical concentrations. Almost all of the biocides tested elicited a pro-inflammatory response from the cells as measured by IL-8 production. All biocides increased protein oxidation as measured by thiol and carbonyl levels. Measurement of transepithelial electrical resistance and paracellular permeability indicated biocide-dependent decrease in epithelial cell barrier function. The cellular response was biased towards necrotic rather than apoptotic death. The use of biocides, although efficacious to some effects against Bacillus spores, will require careful monitoring for adverse health effects on personnel.

A quantitative analysis of optic nerve axons in elderly control subjects and patients with Alzheimer's disease

Objective: To study the density and cross-sectional area of axons in the optic nerve in elderly control subjects and in cases of Alzheimer's disease (AD) using an image analysis system. Methods: Sections of optic nerves from control and AD patients were stained with toluidine blue to reveal axon profiles. Results: The density of axons was reduced in both the center and peripheral portions of the optic nerve in AD compared with control patients. Analysis of axons with different cross-sectional areas suggested a specific loss of the smaller sized axons in AD, i.e., those with areas less that 1.99 μm2. An analysis of axons >11 μm2 in cross-sectional area suggested no specific loss of the larger axons in this group of patients. Conclusions: The data suggest that image analysis provides an accurate and reproducible method of quantifying axons in the optic nerve. In addition, the data suggest that axons are lost throughout the optic nerve with a specific loss of the smaller-sized axons. Loss of the smaller axons may explain the deficits in color vision observed in a significant proportion of patients with AD.

Measuring business growth:high-growth firms and their contribution to employment in the UK

The Measuring Business Growth report is a comprehensive look at UK business growth over the past decade. It makes a powerful case that a small number of high-growth businesses are responsible for the lion's share of job creation and prosperity. It is the counterpart to Business Growth and Innovation, which considers the wider benefits of growth businesses, their socio-economic impact, and the relationship between growth and innovation. This has significant implications for the direction of economic policy. It suggests that focusing attention on growing businesses and promoting excellence, far from being an elitist policy, gives rise to widespread job creation and prosperity.

Synthesis and characterization of dual-functionalized core-shell fluorescent microspheres for bioconjugation and cellular delivery

The efficient transport of micron-sized beads into cells, via a non-endocytosis mediated mechanism, has only recently been described. As such there is considerable scope for optimization and exploitation of this procedure to enable imaging and sensing applications to be realized. Herein, we report the design, synthesis and characterization of fluorescent microsphere-based cellular delivery agents that can also carry biological cargoes. These core-shell polymer microspheres possess two distinct chemical environments; the core is hydrophobic and can be labeled with fluorescent dye, to permit visual tracking of the microsphere during and after cellular delivery, whilst the outer shell renders the external surfaces of the microspheres hydrophilic, thus facilitating both bioconjugation and cellular compatibility. Cross-linked core particles were prepared in a dispersion polymerization reaction employing styrene, divinylbenzene and a thiol-functionalized co-monomer. These core particles were then shelled in a seeded emulsion polymerization reaction, employing styrene, divinylbenzene and methacrylic acid, to generate orthogonally functionalized core-shell microspheres which were internally labeled via the core thiol moieties through reaction with a thiol reactive dye (DY630-maleimide). Following internal labeling, bioconjugation of green fluorescent protein (GFP) to their carboxyl-functionalized surfaces was successfully accomplished using standard coupling protocols. The resultant dual-labeled microspheres were visualized by both of the fully resolvable fluorescence emissions of their cores (DY630) and shells (GFP). In vitro cellular uptake of these microspheres by HeLa cells was demonstrated conventionally by fluorescence-based flow cytometry, whilst MTT assays demonstrated that 92% of HeLa cells remained viable after uptake. Due to their size and surface functionalities, these far-red-labeled microspheres are ideal candidates for in vitro, cellular delivery of proteins, as described in the accompanying paper.

A method of fundamental solutions for transient heat conduction

In this paper we investigate an application of the method of fundamental solutions (MFS) to transient heat conduction. In almost all of the previously proposed MFS for time-dependent heat conduction the fictitious sources are located outside the time-interval of interest. In our case, however, these sources are instead placed outside the space domain of interest in the same manner as is done for stationary heat conduction. A denseness result for this method is discussed and the method is numerically tested showing that accurate numerical results can be obtained. Furthermore, a test example with boundary singularities shows that it is advisable to remove such singularities before applying the MFS.

Radiolabelling of antigen and liposomes for vaccine biodistribution studies

A relatively simple and effective method to follow the movement of pharmaceutical preparations such as vaccines in biodistribution studies is to radiolabel the components. Whilst single radiolabelling is common practice, in vaccine systems containing adjuvants the ability to follow both the adjuvant and the antigen is favourable. To this end, we have devised a dual-radiolabelling method whereby the adjuvant (liposomes) is labelled with 3H and the antigen (a subunit protein) with 125I. This model is stable and reproducible; we have shown release of the radiolabels to be negligible over periods of up to 1 week in foetal calf serum at 37° C. In this paper we describe the techniques which enable the radiolabelling of various components, assessing stability and processing of samples which all for their application in biodistribution studies. Furthermore we provide examples derived from our studies using this model in tuberculosis vaccine biodistribution studies. © 2010 by the authors.

Mere equities and third parties in leasehold law

Examines the concept of a "mere equity" in the context of the Land Registration Act 2002 s.116(b). Considers, by reference to case law, the nature and status of a mere equity and equities coming within the category of equitable rights binding third parties, including a landlord's right to rectification of a lease, the right to set aside a lease and a tenant's right to relief against forfeiture of a lease. Comments on the extent to which s.116(b) requires a mere equity to be more than just procedural and to be an equitable proprietary right capable of binding successors in title.

Joint tenancies and English commorientes:a question of survivorship or severance?

The aim of this article is to provide a critical appraisal of the English law in relation to the doctrine of commorientes with particular reference to its implications in respect of property held on a joint tenancy. The article suggests a measure of reform which would produce a fairer dsitribution of joint property in circusmtances where all joint tenants have died in a common disaster and it cannot be ascertained which joint tenant died first.

Co-owners and severance - determining beneficial entitlement after Stack

The article seeks to examine the effect of a severance of a joint beneficial title following the House of Lords' ruling in Stack v Dowden [2007] 2 AC 432. If, under Stack, the parties' beneficial interests fall to be treated as identical to their legal interests (in effect, giving rise to a joint tenancy both at law and in equity), how does the mechanism of severance of a joint beneficial tenancy actually operate so as to give rise to separate and unequal shares where the parties' common intention has changed by virtue of post-acquisition events? The question is examined by reference to an ambulatory and new constructive trust, as well as where the parties have entered into an express declaration of trust of their beneficial interests.