A Bias Compensated Cross-Relation approach to Thermocouple Characterisation

The measurement of fast changing temperature fluctuations is a challenging problem due to the inherent limited bandwidth of temperature sensors. This results in a measured signal that is a lagged and attenuated version of the input. Compensation can be performed provided an accurate, parameterised sensor model is available. However, to account for the in influence of the measurement environment and changing conditions such as gas velocity, the model must be estimated in-situ. The cross-relation method of blind deconvolution is one approach for in-situ characterisation of sensors. However, a drawback with the method is that it becomes positively biased and unstable at high noise levels. In this paper, the cross-relation method is cast in the discrete-time domain and a bias compensation approach is developed. It is shown that the proposed compensation scheme is robust and yields unbiased estimates with lower estimation variance than the uncompensated version. All results are verified using Monte-Carlo simulations.

Noise resilient discrete-time cross-relation sensor characterisation

In-situ characterisation of thermocouple sensors is a challenging problem. Recently the authors presented a blind characterisation technique based on the cross-relation method of blind identification. The method allows in-situ identification of two thermocouple probes, each with a different dynamic response, using only sampled sensor measurement data. While the technique offers certain advantages over alternative methods, including low estimation variance and the ability to compensate for noise induced bias, the robustness of the method is limited by the multimodal nature of the cost function. In this paper, a normalisation term is proposed which improves the convexity of
the cost function. Further, a normalisation and bias compensation hybrid approach is presented that exploits the advantages of both normalisation and bias compensation. It is found that the optimum of the hybrid cost function is less biased and more stable than when only normalisation is applied. All results were verified by simulation.

Characterisation of polyethylene powders for rotational moulding and effect of powder size and shape on densification behaviour

The particle size, shape and distribution of a range of rotational moulding polyethylenes (PEs) ground to powder was investigated using a novel visual data acquisition and analysis system (TP Picture®), developed by Total Petrochemicals. Differences in the individual particle shape factors of the powder samples were observed and correlations with the grinding conditions were determined. When heated, the bubble dissolution behaviour of the same powders was investigated and the shape factor correlated with densification rate, bubble size and bubble distribution.

A Bias Compensated Cross-Relation approach to Thermocouple Characterisation

The measurement of fast changing temperature fluctuations is a challenging problem due to the inherent limited bandwidth of temperature sensors. This results in a measured signal that is a lagged and attenuated version of the input. Compensation can be performed provided an accurate, parameterised sensor model is available. However, to account for the influence of the measurement environment and changing conditions such as gas velocity, the model must be estimated in-situ. The cross-relation method of blind deconvolution is one approach for in-situ characterisation of sensors. However, a drawback with the method is that it becomes positively biased and unstable at high noise levels. In this paper, the cross-relation method is cast in the discrete-time domain and a bias compensation approach is developed. It is shown that the proposed compensation scheme is robust and yields unbiased estimates with lower estimation variance than the uncompensated version. All results are verified using Monte-Carlo simulations.

Characterisation of a pleistocene debris-avalanche deposit in the Tenteniguada Basin, Gran Canaria Island, Spain

[EN] We studied a large debris-avalanche deposit of Pleistocene age in the Tenteniguada Basin, Gran Canaria Island, Spain. This deposit, which is well preserved because it is mostly covered by basanite lava flows, has distinctive matrix and block facies, hummocky topography and internal structures typical of debris avalanches. However, neither syneruptive lavas nor some characteristic features of volcanic debris-avalanche deposits, such as a stratovolcano edifice or a horseshoe-shaped crater, are present. The occurrence of internal features characteristic of volcanic avalanche deposits could be attributed to the volcanic materials involved in the movement rather than to the triggering of the avalanche during a volcanic eruption.

Physicochemical characterisation of protein ingredients prepared from milk by ultrafiltration or microfiltration for application in formulated nutritional products

Formulated food systems are becoming more sophisticated as demand grows for the design of structural and nutritional profiles targeted at increasingly specific demographics. Milk protein is an important bio- and techno-functional component of such formulations, which include infant formula, sports supplements, clinical beverages and elderly nutrition products. This thesis outlines research into ingredients that are key to the development of these products, namely milk protein concentrate (MPC), milk protein isolate (MPI), micellar casein concentrate (MCC), β-casein concentrate (BCC) and serum protein concentrate (SPC). MPC powders ranging from 37 to 90% protein (solids basis) were studied for properties of relevance to handling and storage of powders, powder solubilisation and thermal processing of reconstituted MPCs. MPC powders with ≥80% protein were found to have very poor flowability and high compressibility; in addition, these high-protein MPCs exhibited poor wetting and dispersion characteristics during rehydration in water. Heat stability studies on unconcentrated (3.5%, 140°C) and concentrated (8.5%, 120°C) MPC suspensions, showed that suspensions prepared from high-protein MPCs coagulated much more rapidly than lower protein MPCs. β-casein ingredients were developed using membrane processing. Enrichment of β-casein from skim milk was performed at laboratory-scale using ‘cold’ microfiltration (MF) at <4°C with either 1000 kDa molecular weight cut-off or 0.1 µm pore-size membranes. At pilot-scale, a second ‘warm’ MF step at 26°C was incorporated for selective purification of micellised β-casein from whey proteins; using this approach, BCCs with β-casein purity of up to 80% (protein basis) were prepared, with the whey protein purity of the SPC co-product reaching ~90%. The BCC ingredient could prevent supersaturated solutions of calcium phosphate (CaP) from precipitating, although the amorphous CaP formed created large micelles that were less thermo-reversible than those in CaP-free systems. Another co-product of BCC manufacture, MCC powder, was shown to have superior rehydration characteristics compared to traditional MCCs. The findings presented in this thesis constitute a significant advance in the research of milk protein ingredients, in terms of optimising their preparation by membrane filtration, preventing their destabilisation during processing and facilitating their effective incorporation into nutritional formulations designed for consumers of a specific age, lifestyle or health status

Generation and Characterisation of a Canine EGFP-HMGA2 Prostate Cancer In Vitro Model

The architectural transcription factor HMGA2 is abundantly expressed during embryonic development. In several malignant neoplasias including prostate cancer, high re-expression of HMGA2 is correlated with malignancy and poor prognosis. The let-7 miRNA family is described to regulate HMGA2 negatively. The balance of let-7 and HMGA2 is discussed to play a major role in tumour aetiology. To further analyse the role of HMGA2 in prostate cancer a stable and highly reproducible in vitro model system is precondition. Herein we established a canine CT1258-EGFP-HMGA2 prostate cancer cell line stably overexpressing HMGA2 linked to EGFP and in addition the reference cell line CT1258-EGFP expressing solely EGFP to exclude EGFP-induced effects. Both recombinant cell lines were characterised by fluorescence microscopy, flow cytometry and immunocytochemistry. The proliferative effect of ectopically overexpressed HMGA2 was determined via BrdU assays. Comparative karyotyping of the derived and the initial CT1258 cell lines was performed to analyse chromosome consistency. The impact of the ectopic HMGA2 expression on its regulator let-7a was analysed by quantitative real-time PCR. Fluorescence microscopy and immunocytochemistry detected successful expression of the EGFP-HMGA2 fusion protein exclusively accumulating in the nucleus. Gene expression analyses confirmed HMGA2 overexpression in CT1258-EGFP-HMGA2 in comparison to CT1258-EGFP and native cells. Significantly higher let-7a expression levels were found in CT1258-EGFP-HMGA2 and CT1258-EGFP. The BrdU assays detected an increased proliferation of CT1258-HMGA2-EGFP cells compared to CT1258-EGFP and native CT1258. The cytogenetic analyses of CT1258-EGFP and CT1258-EGFP-HMGA2 resulted in a comparable hyperdiploid karyotype as described for native CT1258 cells. To further investigate the impact of recombinant overexpressed HMGA2 on CT1258 cells, other selected targets described to underlie HMGA2 regulation were screened in addition. The new fluorescent CT1258-EGFP-HMGA2 cell line is a stable tool enabling in vitro and in vivo analyses of the HMGA2-mediated effects on cells and the development and pathogenesis of prostate cancer.

Hazelnut Allergens: Molecular Characterisation, Detection and Clinical Relevance

In the last years, special attention has been devoted to food-induced allergies, from which hazelnut allergy is highlighted. Hazelnut is one of the most commonly consumed tree nuts, being largely used by the food industry in a wide variety of processed foods. It has been regarded as a food with potential health benefits, but also as a source of allergens capable of inducing mild to severe allergic reactions in sensitised individuals. Considering the great number of reports addressing hazelnut allergens, with an estimated increasing trend, this review intends to assemble all the relevant information available so far on the main issues: prevalence of tree nut allergy, clinical threshold levels, molecular characterisation of hazelnut allergens (Cor a 1, Cor a 2, Cor a 8, Cor a 9, Cor a 10, Cor a 11, Cor a 12, Cor a 14 and Cor a TLP) and their clinical relevance, and methodologies for hazelnut allergen detection in foods. A comprehensive overview on the current data about the molecular characterisation of hazelnut allergens is presented, relating biochemical classification and biological function with clinical importance. Recent advances on hazelnut allergen detection methodologies are summarised and compared, including all the novel protein- and DNA-based approaches.

Characterisation of accessions and species of Macadamia to stem infection by Phytophthora cinnamomi

Phytophthora cinnamomi is a major pathogen in most macadamia plantations worldwide. Due to stem lesions, stem cankers and leaf defoliation it results in loss of productivity and tree death. In this study we examined accessions of the four Macadamia species and their hybrids, produced via rooted stem cuttings or germinated seeds, for susceptibility to stem canker and necrotic lesion caused by P. cinnamomi. Plants were wound-inoculated with agar containing P. cinnamomi. The symptoms produced in inoculated plants were used to characterize host susceptibility variation within and among the population. Lesion lengths and severity of stem canker were recorded. The four species and hybrids differed significantly in stem canker severity (P < 0.001) and lesion length (P = 0.04). M. integrifolia and M. tetraphylla hybrids were the most susceptible. M. integrifolia had the greatest stem canker severity and the most extensive lesions above and below the site of inoculation. Restricted lesion sizes were observed in M. ternifolia and M. jansenii. The effects of basal stem diameter and the method of propagation either from cuttings or seed were not significant. The genetic variation in the reactions of macadamia accessions to stem infection by P. cinnamomi is discussed.

Acoustic characterisation of the tonic syllable in portuguese

A particular problem for the automatic prediction of prosody in speech synthesis is the realisation of accented syllables since these are affected by many parameters and are perceptually very salient. For the Portuguese language, in Europe, a set of comprehensive quantitative characterisation data and rules is totally lacking. The present paper is intended to be a quantitative contribution to the solution of this problem. In this paper, a preliminary modelling of duration, intensity and variation of F0 in the tonic syllable will be presented. The dependencies of the model with the syllable position in the word and the word position in the phrase are also presented.

Molecular and biological characterisation of orthobunyaviruses

Orthobunyaviruses are the largest genus within the Bunyaviridae family, with over 170 named viruses classified into 18 serogroups (Elliott and Blakqori, 2001; Plyusnin et al., 2012). Orthobunyaviruses are transmitted by arthropods and have a tripartite negative sense RNA genome, which encodes 4 structural proteins and 2 non-structural proteins. The non-structural protein NSs is the primary virulence factor of orthobunyaviruses and potent antagonist of the type I interferon (IFN) response. However, sequencing studies have identified pathogenic viruses that lack the NSs protein (Mohamed et al., 2009; Gauci et al., 2010). The work presented in this thesis describes the molecular and biological characterisation of divergent orthobunyaviruses. Data on plaque morphology, growth kinetics, protein profiles, sensitivity to IFN and activation of the type I IFN system are presented for viruses in the Anopheles A, Anopheles B, Capim, Gamboa, Guama, Minatitlan, Nyando, Tete and Turlock serogroups. These are complemented with complete genome sequencing and phylogenetic analysis. Low activation of IFN by Tete serogroup viruses, which naturally lack an NSs protein, was also further investigated by the development of a reverse genetics system for Batama virus (BMAV). Recombinant viruses with mutations in the virus nucleocapsid protein amino terminus showed higher activation of type I IFN in vitro and data suggests that low levels of IFN are due to lower activation rather than active antagonism. The anti-orthobunyavirus activity of IFN-stimulated genes IFI44, IFITMs and human and ovine BST2 were also studied, revealing that activity varies not only within the orthobunyavirus genus and virus serogroups but also within virus species. Furthermore, there was evidence of active antagonism of the type I IFN response and ISGs by non-NSs viruses. In summary, the results show that pathogenicity in man and antagonism of the type I IFN response in vitro cannot be predicted by the presence, or absence, of an NSs ORF. They also highlight problems in orthobunyavirus classification with discordance between classical antigen based data and phylogenetic analysis.

Identification and characterisation of telomere regulatory and signalling pathways after induction of telomere dysfunction

Telomeres are DNA-protein complexes which cap the ends of eukaryotic linear chromosomes. In normal somatic cells telomeres shorten and become dysfunctional during ageing due to the DNA end replication problem. This leads to activation of signalling pathways that lead to cellular senescence and apoptosis. However, cancer cells typically bypass this barrier to immortalisation in order to proliferate indefinitely. Therefore enhancing our understanding of telomere dysfunction and pathways involved in regulation of the process is essential. However, the pathways involved are highly complex and involve interaction between a wide range of biological processes. Therefore understanding how telomerase dysfunction is regulated is a challenging task and requires a systems biology approach. In this study I have developed a novel methodology for visualisation and analysis of gene lists focusing on the network level rather than individual or small lists of genes. Application of this methodology to an expression data set and a gene methylation data set allowed me to enhance my understanding of the biology underlying a senescence inducing drug and the process of immortalisation respectively. I then used the methodology to compare the effect of genetic background on induction of telomere uncapping. Telomere uncapping was induced in HCT116 WT, p21-/- and p53-/- cells using a viral vector expressing a mutant variant of hTR, the telomerase RNA template. p21-/- cells showed enhanced sensitivity to telomere uncapping. Analysis of a candidate pathway, Mismatch Repair, revealed a role for the process in response to telomere uncapping and that induction of the pathway was p21 dependent. The methodology was then applied to analysis of the telomerase inhibitor GRN163L and synergistic effects of hypoglycaemia with this drug. HCT116 cells were resistant to GRN163L treatment. However, under hypoglycaemic conditions the dose required for ablation of telomerase activity was reduced significantly and telomere shortening was enhanced. Overall this new methodology has allowed our group and collaborators to identify new biology and improve our understanding of processes regulating telomere dysfunction.

Speech and narrative: characterisation techniques in the "Aeneid"

The thesis provides a comprehensive analysis of the characterisation of two of the major figures in the Aeneid, Aeneas and Turnus. Particular attention is paid to their direct speeches, all of which are examined and, where relevant, compared to Homeric models and parallels. To this purpose considerable use is made of the indices in Knauer's Die Aeneis und Homer. A more general comparison is made between the dramatic (direct speech) role of Aeneas and those of Homer's Achilles (Iliad) and Odysseus (Odyssey). An appraisal is made (from the viewpoint of depiction of character) of the relationship between the direct and indirect speeches in the Aeneid. Reasons are given to suggest that it is not mere chance, or for the sake of variety, that certain speeches of Aeneas and Turnus are expressed in oratio obliqua. In addition, the narrative portrayal of Aeneas and Turnus is considered in apposition to that of the speeches. A distinction is drawn between Vergil's direct method of characterisation (direct speeches) and his indirect methods (narrative/oratio obliqua). Inevitably, the analysis involves major consideration of the Roman values which pervade the work. All speeches, thoughts and actions of Aeneas and Turnus are assessed in terms of pietas, impietas, furor, virtus, ratio, clementia, humanitas (etc.). It is shown that individual concepts (such as pietas and impietas) are reflected in Vergil's direct and indirect methods of characterisation. The workings of fate and their relevance to the pietas concept are discussed throughout.