The formulation of artificial reference standards for use within the ELISPOT assay

Whether to assess the functionality of equipment or as a determinate for the accuracy of assays, reference standards are essential for the purposes of standardisation and validation. The ELISPOT assay, developed over thirty years ago, has emerged as a leading immunological assay in the development of novel vaccines for the assessment of efficacy. However, with its widespread use, there is a growing demand for a greater level of standardisation across different laboratories. One of the major difficulties in achieving this goal has been the lack of definitive reference standards. This is partly due to the ex vivo nature of the assay, which relies on cells being placed directly into the wells. Thus, the aim of this thesis was to produce an artificial reference standard using liposomes, for use within the assay. Liposomes are spherical bilayer vesicles with an enclosed aqueous compartment and therefore are models for biological membranes. Initial work examined pre-design considerations in order to produce an optimal formulation that would closely mimic the action of the cells ordinarily placed on the assay. Recognition of the structural differences between liposomes and cells led to the formulation of liposomes with increased density. This was achieved by using a synthesised cholesterol analogue. By incorporating this cholesterol analogue in liposomes, increased sedimentation rates were observed within the first few hours. The optimal liposome formulation from these studies was composed of 2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC), cholesterol (Chol) and brominated cholesterol (Brchol) at a 16:4:12 µMol ratio, based on a significantly higher (p<0.01) sedimentation (as determined by a percentage transmission of 59 ± 5.9 % compared to the control formulation at 29 ± 12 % after four hours). By considering a range of liposome formulations ‘proof of principle’ for using liposomes as ELISPOT reference standards was shown; recombinant IFN? cytokine was successfully entrapped within vesicles of different lipid compositions, which were able to promote spot formation within the ELISPOT assay. Using optimised liposome formulations composed of phosphatidylcholine with or without cholesterol (16 µMol total lipid) further development was undertaken to produce an optimised, scalable protocol for the production of liposomes as reference standards. A linear increase in spot number by the manipulation of cytokine concentration and/or lipid concentrations was not possible, potentially due to the saturation that occurred within the base of wells. Investigations into storage of the formulations demonstrated the feasibility of freezing and lyophilisation with disaccharide cryoprotectants, but also highlighted the need for further protocol optimisation to achieve a robust reference standard upon storage. Finally, the transfer of small-scale production to a medium lab-scale batch (40 mL) demonstrated this was feasible within the laboratory using the optimised protocol.

A multivariate approach to bidding strategies

A multivariate approach to bidding strategy is presented in comparison with previous standard approaches. An optimal formulation is derived and a method of parameter estimation proposed. A case study illustrates the derivation of optimal and other strategic mark up values against a single bidder. Concluding remarks concern extensions to multiple competitors differing levels of information, and sensitivity analysis.

Evaluating Alternative Nutrient Sources in Subsistence-Level Aquaponic Systems

Many food production methods are both economically and environmentally unsustainable. Our project investigated aquaponics, an alternative method of agriculture that could address these issues. Aquaponics combines fish and plant crop production in a symbiotic, closed-loop system. We aimed to reduce the initial and operating costs of current aquaponic systems by utilizing alternative feeds. These improvements may allow for sustainable implementation of the system in rural or developing regions. We conducted a multi-phase process to determine the most affordable and effective feed alternatives for use in an aquaponic system. At the end of two preliminary phases, soybean meal was identified as the most effective potential feed supplement. In our final phase, we constructed and tested six full-scale aquaponic systems of our own design. Data showed that soybean meal can be used to reduce operating costs and reliance on fishmeal. However, a more targeted investigation is needed to identify the optimal formulation of alternative feed blends.

Evaluation of a Water-soluble Bioadhesive Patch for Photodynamic Therapy of Vulval Lesions

An innovative bioadhesive patch intended primarily as a vulval drug delivery system and, specifically, as a means to deliver photosensitisers, or their prodrugs, for photodynamic purposes is described. The patch was formulated with a copolymer of methyl vinyl ether and maleic anhydride (PMVE/MA) as a bioadhesive matrix and poly(vinyl chloride) as a drug-impervious backing layer. Adhesive strength to neonate porcine skin, as a model substrate, was evaluated using peel and tensile testing measurements. Acceptabilities of non-drug loaded patches were appraised using human volunteers and visual-analogue scoring devices. An optimal formulation, with water uptake and peel strengths appropriate for vulval drug delivery, was cast from a 20% (w/w) PMVE/MA solution and adhered with a strength of approximately 1.7 N cm-2. Patient evaluation demonstrated comfort and firm attachment for up to 4 h in mobile patients. Aminolevulinic acid, a commonly used photosensitiser, was formulated into the candidate formulation and applied to vulval intraepithelial neoplastic lesions. Fluorescence under ultraviolet illumination revealed protoporphyrin synthesis. The patch achieves the extended application times obligatory in topical photodynamic therapy of vulval lesions, thereby contributing to potential methods for the eradication of neoplastic lesions in the lower female reproductive tract.

Optimization of amaranth flour films plasticized with glycerol and sorbitol by multi-response analysis

The optimal formulation for the preparation of amaranth flour films plasticized with glycerol and sorbitol was obtained by a multi-response analysis. The optimization aimed to achieve films with higher resistance to break, moderate elongation and lower solubility in water. The influence of plasticizer concentration (Cg, glycerol or Cs, sorbitol) and process temperature (Tp) on the mechanical properties and solubility of the amaranth flour films was initially studied by response surface methodology (RSM). The optimized conditions obtained were Cg 20.02 g glycerol/100 g flour and Tp 75 degrees C, and Cs 29.6 g sorbitol/100 g flour and Tp 75 degrees C. Characterization of the films prepared with these formulations revealed that the optimization methodology employed in this work was satisfactory. Sorbitol was the most suitable plasticizer. It furnished amaranth flour films that were more resistant to break and less permeable to oxygen, due to its greater miscibility with the biopolymers present in the flour and its lower affinity for water. (C) 2011 Elsevier Ltd. All rights reserved.

Optimization of the surimi production from Mechanically Recovered Fish Meat (MRFM) using response surface methodology

The by-products generated from industrial filleting of tilapia surimi can be used for the manufacture of surimi. The surimi production uses large amounts of water, which generates a wastewater rich in organic compounds (lipids, soluble proteins and blood). Optimizing the number of washing cycles will contribute to a more sustainable production. A mathematical model of mechanically recovered tilapia meat (Oreochromis niloticus) for the processing of surimi (minced fish washing cycles and tapioca starch addition) based on two quality parameters (texture and moisture) was constructed by applying the response surface methodology (RSM). Each factor had an important effect on the moisture and texture of surimi. This study found that the optimal formulation for producing the best surimi using the by-products of tilapia filleting in manufacturing fish burger were the addition of 10% tapioca starch and three minced fish washing cycles. A microstructural evaluation supported the findings of the mathematical model. Practical Applications: The use of mechanically recovered fish meat (MRFM) for the production of surimi enables the utilization of the by-products of filleting fish. However, the inferior quality of the surimi produced from MRFM in relation to that produced with fillets necessitates the addition of starch; secondly, surimi production consumes a large volume of water. RSM provides a valuable means for optimizing the number of washing cycles and starch amounts utilized in fish burger production. Tapioca starch, widely produced in Brazil, has desirable characteristics (surface sheen, smooth texture, neutral taste and clarity in solution) for use in MRFM-produced surimi. © 2013 Wiley Periodicals, Inc.

Modelos equivalentes de parques eólicos usando algoritmos genéticos

Esta dissertação apresenta uma metodologia baseada em algoritmo genético (AG) para determinar modelos dinâmicos equivalentes de parques eólicos com geradores de indução em gaiola de esquilo ( GIGE) e geradores de indução duplamente alimentados ( GIDA), apresentando parâmetros elétricos e mecânicos distintos. A técnica se baseia em uma formulação multiobjetiva solucionada por um AG para minimizar os erros quadráticos das potências ativa e reativa entre modelo de um único gerador equivalente e o modelo do parque eólico investigado. A influência do modelo equivalente do parque eólico no comportamento dinâmico dos geradores síncronos é também investigada por meio do método proposto. A abordagem é testada em um parque eólico de 10MW composto por quatro turbinas eólicas ( 2x2MW e 2x3MW), consistindo alternadamente de geradores GIGE e GIDA interligados a uma barra infinita e posteriormente a rede elétrica do IEEE 14 barras. Os resultados obtidos pelo uso do modelo dinâmico detalhado para a representação do parque eólico são comparados aos do modelo equivalente proposto para avaliar a precisão e o custo computacional do modelo proposto.

CIMBNC- Combination and interactions of three kinds of nanoclays structural modifiers, for the colored bio-nanocomposites optimization

Biopolymers do not have competitive prices, which has prevented their industrial exploitation on a global scale so far. In this context, Using nanoclays, improvements in certain biopolymer properties, mainly mechanical and thermal, have been achieved. However, research has been much less focused on changing optical properties through the incorporation of nanoclays. At the same time, current research has focused on obtaining nanopigments, by organic dyes adsoptions into different nanoclays in order to achieve sustainable colouring and high performance materials. By combining advances in these lines of research, biodegradable composites with optimal mechanical and optical properties can be obtained. The aim of this work is to find the optimal formulation of naturally sourced nanopigments, incorporate them into a biological origin epoxy resin, and obtain a significant improvement in their mechanical, and optical properties. We combine three structural modifiers in the nanopigment synthesis: surfactant, silane and mordant salt. The latter was selected in order to replicate the mordant textile dyeing with natural dyes. Using a Taguchi’s desing L8, we look for the effect of the presence of the modifiers, the pH acidification, and the interactions effect between the synthesis factors. Three natural dyes were selected: chlorophyll, beta-carotene, and beetroot extract. Furthermore we use two kinds of laminar nanoclays, differentiated by the ion exchange charge: montmorillonite, and hydrotalcite. Then the thermal, mechanical and colorimetric characterization of the bionanocomposite materials was carried out. The optimal conditions to obtain the best bionanocomposite materials are using acid pH, and modifying the nanoclays with mordant and surfactant.

Development and evaluation of a novel intranasal spray for the delivery of amantadine

The aim of this study was to develop and characterize an intranasal delivery system for amantadine hydrochloride (AMT). Optimal formulations consisted of a thermosensitive polymer Pluronic® 127 and either carboxymethyl cellulose or chitosan which demonstrated gel transition at nasal cavity temperatures (34 ± 1°C). Rheologically, the loss tangent (Tan δ) confirmed a 3-stage gelation phenomena at 34 ± 1°C and non-Newtonian behavior. Storage of optimized formulation carboxymethyl cellulose and optimal formulation chitosan at 4°C for 8 weeks resulted in repeatable release profiles at 34°C when sampled, with a Fickian mechanism earlier on but moving toward anomalous transport by week 8. Polymers (Pluronic® 127, carboxymethyl cellulose, and chitosan) demonstrated no significant cellular toxicity to human nasal epithelial cells up to 4 mg/mL and up to 1 mM for AMT (IC50: 4.5 ± 0.05 mM). Optimized formulation carboxymethyl cellulose and optimal formulation chitosan demonstrated slower release across an in vitro human nasal airway model (43%-44% vs 79 ± 4.58% for AMT). Using a human nasal cast model, deposition into the olfactory regions (potential nose-to-brain) was demonstrated on nozzle insertion (5 mm), whereas tilting of the head forward (15°) resulted in greater deposition in the bulk of the nasal cavity.

Tail gas catalyzed N2O decomposition over Fe-beta zeolite. On the promoting role of framework connected AlO6 sites in the vicinity of Fe by controlled dealumination during exchange

A novel route to prepare highly active and stable N2O decomposition catalysts is presented, based on Fe-exchanged beta zeolite. The procedure consists of liquid phase Fe(III) exchange at low pH. By varying the pH systematically from 3.5 to 0, using nitric acid during each Fe(III)-exchange procedure, the degree of dealumination was controlled, verified by ICP and NMR. Dealumination changes the presence of neighbouring octahedral Al sites of the Fe sites, improving the performance for this reaction. The so-obtained catalysts exhibit a remarkable enhancement in activity, for an optimal pH of 1. Further optimization by increasing the Fe content is possible. The optimal formulation showed good conversion levels, comparable to a benchmark Fe-ferrierite catalyst. The catalyst stability under tail gas conditions containing NO, O2 and H2O was excellent, without any appreciable activity decay during 70 h time on stream. Based on characterisation and data analysis from ICP, single pulse excitation NMR, MQ MAS NMR, N2 physisorption, TPR(H2) analysis and apparent activation energies, the improved catalytic performance is attributed to an increased concentration of active sites. Temperature programmed reduction experiments reveal significant changes in the Fe(III) reducibility pattern with the presence of two reduction peaks; tentatively attributed to the interaction of the Fe-oxo species with electron withdrawing extraframework AlO6 species, causing a delayed reduction. A low-temperature peak is attributed to Fe-species exchanged on zeolitic AlO4 sites, which are partially charged by the presence of the neighbouring extraframework AlO6 sites. Improved mass transport phenomena due to acid leaching is ruled out. The increased activity is rationalized by an active site model, whose concentration increases by selectively washing out the distorted extraframework AlO6 species under acidic (optimal) conditions, liberating active Fe species.

Optimal hydrothermal load flow: Formulation and a successive approximation solution for fixed head systems

This paper deals with the optimal load flow problem in a fixed-head hydrothermal electric power system. Equality constraints on the volume of water available for active power generation at the hydro plants as well as inequality constraints on the reactive power generation at the voltage controlled buses are imposed. Conditions for optimal load flow are derived and a successive approximation algorithm for solving the optimal generation schedule is developed. Computer implementation of the algorithm is discussed, and the results obtained from the computer solution of test systems are presented.

Register Allocation and Optimal Spill code Scheduling in Software Pipelined Loops using 0-1 Integer Linear Programming Formulation

In achieving higher instruction level parallelism, software pipelining increases the register pressure in the loop. The usefulness of the generated schedule may be restricted to cases where the register pressure is less than the available number of registers. Spill instructions need to be introduced otherwise. But scheduling these spill instructions in the compact schedule is a difficult task. Several heuristics have been proposed to schedule spill code. These heuristics may generate more spill code than necessary, and scheduling them may necessitate increasing the initiation interval. We model the problem of register allocation with spill code generation and scheduling in software pipelined loops as a 0-1 integer linear program. The formulation minimizes the increase in initiation interval (II) by optimally placing spill code and simultaneously minimizes the amount of spill code produced. To the best of our knowledge, this is the first integrated formulation for register allocation, optimal spill code generation and scheduling for software pipelined loops. The proposed formulation performs better than the existing heuristics by preventing an increase in II in 11.11% of the loops and generating 18.48% less spill code on average among the loops extracted from Perfect Club and SPEC benchmarks with a moderate increase in compilation time.

A code for designing optimal networks by implicit enumeration using the all-interconnection inequality formulation : (a programming manual for ILLODIE-AIF) /

Thesis (M.S.)--University of Illinois at Urbana-Champaign.

A study of certain factors affecting computation times in designing optimal NOR networks by the implicit enumeration method using the feed-forward formulation /

Bibliography: p. 27-29.

Addressing issues in sparseness, ecological bias and formulation of the adjacency matrix in Bayesian spatio-temporal analysis of disease counts

The main objective of this PhD was to further develop Bayesian spatio-temporal models (specifically the Conditional Autoregressive (CAR) class of models), for the analysis of sparse disease outcomes such as birth defects. The motivation for the thesis arose from problems encountered when analyzing a large birth defect registry in New South Wales. The specific components and related research objectives of the thesis were developed from gaps in the literature on current formulations of the CAR model, and health service planning requirements. Data from a large probabilistically-linked database from 1990 to 2004, consisting of fields from two separate registries: the Birth Defect Registry (BDR) and Midwives Data Collection (MDC) were used in the analyses in this thesis. The main objective was split into smaller goals. The first goal was to determine how the specification of the neighbourhood weight matrix will affect the smoothing properties of the CAR model, and this is the focus of chapter 6. Secondly, I hoped to evaluate the usefulness of incorporating a zero-inflated Poisson (ZIP) component as well as a shared-component model in terms of modeling a sparse outcome, and this is carried out in chapter 7. The third goal was to identify optimal sampling and sample size schemes designed to select individual level data for a hybrid ecological spatial model, and this is done in chapter 8. Finally, I wanted to put together the earlier improvements to the CAR model, and along with demographic projections, provide forecasts for birth defects at the SLA level. Chapter 9 describes how this is done. For the first objective, I examined a series of neighbourhood weight matrices, and showed how smoothing the relative risk estimates according to similarity by an important covariate (i.e. maternal age) helped improve the model’s ability to recover the underlying risk, as compared to the traditional adjacency (specifically the Queen) method of applying weights. Next, to address the sparseness and excess zeros commonly encountered in the analysis of rare outcomes such as birth defects, I compared a few models, including an extension of the usual Poisson model to encompass excess zeros in the data. This was achieved via a mixture model, which also encompassed the shared component model to improve on the estimation of sparse counts through borrowing strength across a shared component (e.g. latent risk factor/s) with the referent outcome (caesarean section was used in this example). Using the Deviance Information Criteria (DIC), I showed how the proposed model performed better than the usual models, but only when both outcomes shared a strong spatial correlation. The next objective involved identifying the optimal sampling and sample size strategy for incorporating individual-level data with areal covariates in a hybrid study design. I performed extensive simulation studies, evaluating thirteen different sampling schemes along with variations in sample size. This was done in the context of an ecological regression model that incorporated spatial correlation in the outcomes, as well as accommodating both individual and areal measures of covariates. Using the Average Mean Squared Error (AMSE), I showed how a simple random sample of 20% of the SLAs, followed by selecting all cases in the SLAs chosen, along with an equal number of controls, provided the lowest AMSE. The final objective involved combining the improved spatio-temporal CAR model with population (i.e. women) forecasts, to provide 30-year annual estimates of birth defects at the Statistical Local Area (SLA) level in New South Wales, Australia. The projections were illustrated using sixteen different SLAs, representing the various areal measures of socio-economic status and remoteness. A sensitivity analysis of the assumptions used in the projection was also undertaken. By the end of the thesis, I will show how challenges in the spatial analysis of rare diseases such as birth defects can be addressed, by specifically formulating the neighbourhood weight matrix to smooth according to a key covariate (i.e. maternal age), incorporating a ZIP component to model excess zeros in outcomes and borrowing strength from a referent outcome (i.e. caesarean counts). An efficient strategy to sample individual-level data and sample size considerations for rare disease will also be presented. Finally, projections in birth defect categories at the SLA level will be made.