Optimisation of a fermentation process for butanol production by particle swarm optimisation (PSO)

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Reactive power planning under conditional-value-at-risk assessment using chance-constrained optimisation

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Neurons derived from P19 embryonic carcinoma cells as a platform for biosensor applications - optimisation and characterisation

P19 is a mouse-derived embryonal carcinoma cell line capable of differentiation toward ectodermal, mesodermal and endodermal lineages and could thus be differentiated into neurons. Different culture conditions were tested to optimise and increase the efficiency of neuronal differentiation since the population of P19-derived neurons was reported to be heterogeneous with respect to the morphology and neurotransmitters they synthesise. P19-derived neurons were cultured on microelectrode arrays as cell aggregates and as dissociated cells. Improved neuronal maturation was shown by the presence of microtubule associated protein 2, neurofilament and synaptophysin formation when initiation of neuronal differentiation was prolonged. High initial cell density cultures and coating of surfaces with polyethylenimine-laminin further improved neuronal maturation of differentiated P19 cells. Increased spontaneous activities of the P19-derived neurons were correspondingly recorded. Two to three hours recordings were performed between 17 and 25 days when extracellular signals were stabilised. It was found that P19-derived neurons developed network properties as partially synchronised network activities. P19-derived neurons appeared to give inhomogenous response to the 2 major neurotransmitters, -aminobutyric acid (GABA) and glutamate. The P19-derived neuronal networks obtained from optimised protocol in this thesis were predominantly GABAergic. The reproducible long term extracellular recordings performed showed that neurons derived from P19 embryonal carcinoma cells could be applied as a model for cell based biosensor in corporation with microelectrode arrays.

Laser scanning microscopy combined with image restoration to analyse a 3D model of the human epithelial airway barrier

A laser scanning microscope collects information from a thin, focal plane and ignores out of focus information. During the past few years it has become the standard imaging method to characterise cellular morphology and structures in static as well as in living samples. Laser scanning microscopy combined with digital image restoration is an excellent tool for analysing the cellular cytoarchitecture, expression of specific proteins and interactions of various cell types, thus defining valid criteria for the optimisation of cell culture models. We have used this tool to establish and evaluate a three dimensional model of the human epithelial airway wall.

The Choice of Expedient Regulatory Controls on Housing Market Using Short-Term Equilibria Model

Khutoretsky dealt with the problem of maximising a linear utility function (MUF) over the set of short-term equilibria in a housing market by reducing it to a linear programming problem, and suggested a combinatorial algorithm for this problem. Two approaches to the market adjustment were considered: the funding of housing construction and the granting of housing allowances. In both cases, locally optimal regulatory measures can be developed using the corresponding dual prices. The optimal effects (with the regulation expenditures restricted by an amount K) can be found using specialised models based on MUF: a model M1 for choice of the optimum structure of investment in housing construction, and a model M2 for optimum distribution of housing allowances. The linear integer optimisation problems corresponding to these models are initially difficult but can be solved after slight modifications of the parameters. In particular, the necessary modification of K does not exceed the maximum construction cost of one dwelling (for M1) or the maximum size of one housing allowance (for M2). The result is particularly useful since slight modification of K is not essential in practice.

An integral model for natural regeneration of Pinus pinea L. in the Northern Plateau (Spain) = Modelo integral de regeneración natural para Pinus pinea L. en la Meseta Norte (España)

Natural regeneration is an ecological key-process that makes plant persistence possible and, consequently, it constitutes an essential element of sustainable forest management. In this respect, natural regeneration in even-aged stands of Pinus pinea L. located in the Spanish Northern Plateau has not always been successfully achieved despite over a century of pine nut-based management. As a result, natural regeneration has recently become a major concern for forest managers when we are living a moment of rationalization of investment in silviculture. The present dissertation is addressed to provide answers to forest managers on this topic through the development of an integral regeneration multistage model for P. pinea stands in the region. From this model, recommendations for natural regeneration-based silviculture can be derived under present and future climate scenarios. Also, the model structure makes it possible to detect the likely bottlenecks affecting the process. The integral model consists of five submodels corresponding to each of the subprocesses linking the stages involved in natural regeneration (seed production, seed dispersal, seed germination, seed predation and seedling survival). The outputs of the submodels represent the transitional probabilities between these stages as a function of climatic and stand variables, which in turn are representative of the ecological factors driving regeneration. At subprocess level, the findings of this dissertation should be interpreted as follows. The scheduling of the shelterwood system currently conducted over low density stands leads to situations of dispersal limitation since the initial stages of the regeneration period. Concerning predation, predator activity appears to be only limited by the occurrence of severe summer droughts and masting events, the summer resulting in a favourable period for seed survival. Out of this time interval, predators were found to almost totally deplete seed crops. Given that P. pinea dissemination occurs in summer (i.e. the safe period against predation), the likelihood of a seed to not be destroyed is conditional to germination occurrence prior to the intensification of predator activity. However, the optimal conditions for germination seldom take place, restraining emergence to few days during the fall. Thus, the window to reach the seedling stage is narrow. In addition, the seedling survival submodel predicts extremely high seedling mortality rates and therefore only some individuals from large cohorts will be able to persist. These facts, along with the strong climate-mediated masting habit exhibited by P. pinea, reveal that viii the overall probability of establishment is low. Given this background, current management –low final stand densities resulting from intense thinning and strict felling schedules– conditions the occurrence of enough favourable events to achieve natural regeneration during the current rotation time. Stochastic simulation and optimisation computed through the integral model confirm this circumstance, suggesting that more flexible and progressive regeneration fellings should be conducted. From an ecological standpoint, these results inform a reproductive strategy leading to uneven-aged stand structures, in full accordance with the medium shade-tolerant behaviour of the species. As a final remark, stochastic simulations performed under a climate-change scenario show that regeneration in the species will not be strongly hampered in the future. This resilient behaviour highlights the fundamental ecological role played by P. pinea in demanding areas where other tree species fail to persist.

Assessing the impact of adaptive accessibility on the optimal transport policy implementation by using an integrated land-use/transport model for Madrid

Accessibility is an essential concept widely used to evaluate the impact of land-use and transport strategies in transport and urban planning. Accessibility is typically evaluated by using a transport model or a land-use model independently or successively without a feedback loop, thus neglecting the interaction effects between the two systems and the induced competition effects among opportunities due to accessibility improvements. More than a mere methodological curiosity, failure to account for land- use/transport interactions and the competition effect may result in large underestimation of the policy effects. With the recent development of land-use and transport interaction (LUTI) models, there is a growing interest in using these models to adequately measure accessibility and evaluate its impact. The current study joins this research stream by embedding an accessibility measure in a LUTI model with two main aims. The first aim is to account for adaptive accessibility, namely the adjustment of the potential accessibility due to the effect of competition among opportunities (e.g., workplaces) as a result of improved accessibility. LUTI models are particularly suitable for assessing adaptive accessibility because the competition factor is a function of the number of jobs, which is related to land-use attractiveness and the number of workers which is related, among other factors, to the transport demand. The second aim is to identify the optimal implementation scenario of policy measures on the basis of the potential and adaptive accessibility and analyse the results in terms of social welfare and accessibility. The metropolitan area of Madrid is used as a case-study and two transport policy instruments, namely a cordon toll and bus frequency increase, have been chosen for the simulation study in order to present the usefulness of the approach to urban planners and policy makers. The MARS model (Metropolitan Activity Relocation Simulator) calibrated for Madrid was employed as the analysis tool. The impact of accessibility is embedded in the model through a social welfare function that includes not only costs and benefits to both road users and transport operators, but also costs and benefits for the government and society in general (external costs). An optimisation procedure is performed by the MARS model for maximizing the value of objective function in order to find the best (optimal) policy imp lementations intensity (i.e., price, frequency). Last, the two policy strategies are evaluated in terms of their accessibility. Results show that the accessibility with competition factor influences the optimal policy implementation level and also generates different results in terms of social welfare. In addition, mapping the difference between the potential and the adaptive accessibility indicators shows that the main changes occur in areas where there is a strong competition among land-use opportunities.

Query Rewriting Optimisation Techniques for Ontology-Based Data Access

Ontology-Based Data Access (OBDA) permite el acceso a diferentes tipos de fuentes de datos (tradicionalmente bases de datos) usando un modelo más abstracto proporcionado por una ontología. La reescritura de consultas (query rewriting) usa una ontología para reescribir una consulta en una consulta reescrita que puede ser evaluada en la fuente de datos. Las consultas reescritas recuperan las respuestas que están implicadas por la combinación de los datos explicitamente almacenados en la fuente de datos, la consulta original y la ontología. Al trabajar sólo sobre las queries, la reescritura de consultas permite OBDA sobre cualquier fuente de datos que puede ser consultada, independientemente de las posibilidades para modificarla. Sin embargo, producir y evaluar las consultas reescritas son procesos costosos que suelen volverse más complejos conforme la expresividad y tamaño de la ontología y las consultas aumentan. En esta tesis exploramos distintas optimizaciones que peuden ser realizadas tanto en el proceso de reescritura como en las consultas reescritas para mejorar la aplicabilidad de OBDA en contextos realistas. Nuestra contribución técnica principal es un sistema de reescritura de consultas que implementa las optimizaciones presentadas en esta tesis. Estas optimizaciones son las contribuciones principales de la tesis y se pueden agrupar en tres grupos diferentes: -optimizaciones que se pueden aplicar al considerar los predicados en la ontología que no están realmente mapeados con las fuentes de datos. -optimizaciones en ingeniería que se pueden aplicar al manejar el proceso de reescritura de consultas en una forma que permite reducir la carga computacional del proceso de generación de consultas reescritas. -optimizaciones que se pueden aplicar al considerar metainformación adicional acerca de las características de la ABox. En esta tesis proporcionamos demostraciones formales acerca de la corrección y completitud de las optimizaciones propuestas, y una evaluación empírica acerca del impacto de estas optimizaciones. Como contribución adicional, parte de este enfoque empírico, proponemos un banco de pruebas (benchmark) para la evaluación de los sistemas de reescritura de consultas. Adicionalmente, proporcionamos algunas directrices para la creación y expansión de esta clase de bancos de pruebas. ABSTRACT Ontology-Based Data Access (OBDA) allows accessing different kinds of data sources (traditionally databases) using a more abstract model provided by an ontology. Query rewriting uses such ontology to rewrite a query into a rewritten query that can be evaluated on the data source. The rewritten queries retrieve the answers that are entailed by the combination of the data explicitly stored in the data source, the original query and the ontology. However, producing and evaluating the rewritten queries are both costly processes that become generally more complex as the expressiveness and size of the ontology and queries increase. In this thesis we explore several optimisations that can be performed both in the rewriting process and in the rewritten queries to improve the applicability of OBDA in real contexts. Our main technical contribution is a query rewriting system that implements the optimisations presented in this thesis. These optimisations are the core contributions of the thesis and can be grouped into three different groups: -optimisations that can be applied when considering the predicates in the ontology that are actually mapped to the data sources. -engineering optimisations that can be applied by handling the process of query rewriting in a way that permits to reduce the computational load of the query generation process. -optimisations that can be applied when considering additional metainformation about the characteristics of the ABox. In this thesis we provide formal proofs for the correctness of the proposed optimisations, and an empirical evaluation about the impact of the optimisations. As an additional contribution, part of this empirical approach, we propose a benchmark for the evaluation of query rewriting systems. We also provide some guidelines for the creation and expansion of this kind of benchmarks.

Assessing the accessibility impact of transport policy by a land-use and transport interaction model - The case of Madrid

Accessibility is an essential concept widely used to evaluate the impact of transport and land-use strategies in urban planning and policy making. Accessibility is typically evaluated by using separately a transport model or a land-use model. This paper embeds two accessibility indicators (i.e., potential and adaptive accessibility) in a land use and transport interaction (LUTI) model in order to assess transport policies implementation. The first aim is to define the adaptive accessibility, considering the competition factor at territorial level (e.g. workplaces and workers). The second aim is to identify the optimal implementation scenario of policy measures using potential and adaptive accessibility indicators. The analysis of the results in terms of social welfare and accessibility changes closes the paper. Two transport policy measures are applied in Madrid region: a cordon toll and increase bus frequency. They have been simulated through the MARS model (Metropolitan Activity Relocation Simulator, i.e. LUTI model). An optimisation procedure is performed by MARS for maximizing the value of the objective function in order to find the optimal policy implementation (first best). Both policy measures are evaluated in terms of accessibility. Results show that the introduction of the accessibility indicators (potential and adaptive) influence the optimal value of the toll price and bus frequency level, generating different results in terms of social welfare. Mapping the difference between potential and adaptive accessibility indicator shows that the main changes occur in areas where there is a strong competition among different land-use opportunities.

The selection of a model microalgal species as biomaterial for a novel aquatic phytotoxicity assay

A phytotoxicity assay based on the ToxY-PAM dual-channel yield analyser has been developed and successfully incorporated into field assessments for the detection of phytotoxicants in water. As a means of further exploring the scope of the assay application and of selecting a model biomaterial to complement the instrument design, nine algal species were exposed to four chemical substances deemed of priority for water quality monitoring purposes (chlorpyrifos, copper, diuron and nonylphenol ethoxylate). Inter-species differences in sensitivity to the four toxicants varied by a factor of 1.9-100. Measurements of photosystem-II quantum yield using these nine single-celled microalgae as biomaterial corroborated previous studies which have shown that the ToxY-PAM dual-channel yield analyser is a highly sensitive method for the detection of PS-II impacting herbicides. Besides Phaeodactylum tricornutum, the previously applied biomaterial, three other species consistently performed well (Nitzschia closterium, Chlorella vulgaris and Dunaliella tertiolecta) and will be used in further test optimisation experiments. In addition to sensitivity, response time was evaluated and revealed a high degree of variation between species and toxicants. While most species displayed relatively weak and slow responses to copper, C. vulgaris demonstrated an IC10 of 51 μ g L-1, with maximum response measured within 25 minutes and inhibition being accompanied by a large decrease in fluorescence yield. The potential for this C vulgaris-based bioassay to be used for the detection of copper is discussed. There was no evidence that the standard ToxY-PAM protocol, using these unicellular algae species, could be used for the detection of chlorpyrifos or nonylphenol ethoxylate at environmentally relevant levels. © 2005 Elsevier B.V. All rights reserved.

Optimisation of poly-b-hydroxyalkanoate analysis using gas chromatography for enhanced biological phosphorus removal systems

Poly-beta-hydroxyalkanoate (PHA) is a polymer commonly used in carbon and energy storage for many different bacterial cells. Polyphosphate accumulating organisms (PAOs) and glycogen accumulating organisms (GAOs), store PHA anaerobically through metabolism of carbon substrates such as acetate and propionate. Although poly-beta-hydroxybutyrate (PHB)and poly-beta-hydroxyvalerate (PHV) are commonly quantified using a previously developed gas chromatography (GC) method, poly-beta-hydroxy-2-methyl valerate (PH2MV) is seldom quantified despite the fact that it has been shown to be a key PHA fraction produced when PAOs or GAOs metabolise propionate. This paper presents two GC-based methods modified for extraction and quantification of PHB, PHV and PH2MV from enhanced biological phosphorus removal (EBPR) systems. For the extraction Of PHB and PHV from acetate fed PAO and GAO cultures, a 3% sulfuric acid concentration and a 2-20 h digestion time is recommended, while a 10% sulfuric acid solution digested for 20 h is recommended for PHV and PH2MV analysis from propionate fed EBPR systems. (c) 2005 Elsevier B.V. All rights reserved.

A scatterometer neural network sensor model with input noise

The ERS-1 satellite carries a scatterometer which measures the amount of radiation scattered back toward the satellite by the ocean's surface. These measurements can be used to infer wind vectors. The implementation of a neural network based forward model which maps wind vectors to radar backscatter is addressed. Input noise cannot be neglected. To account for this noise, a Bayesian framework is adopted. However, Markov Chain Monte Carlo sampling is too computationally expensive. Instead, gradient information is used with a non-linear optimisation algorithm to find the maximum em a posteriori probability values of the unknown variables. The resulting models are shown to compare well with the current operational model when visualised in the target space.

A scatterometer neural network sensor model with input noise

The ERS-1 satellite carries a scatterometer which measures the amount of radiation scattered back toward the satellite by the ocean's surface. These measurements can be used to infer wind vectors. The implementation of a neural network based forward model which maps wind vectors to radar backscatter is addressed. Input noise cannot be neglected. To account for this noise, a Bayesian framework is adopted. However, Markov Chain Monte Carlo sampling is too computationally expensive. Instead, gradient information is used with a non-linear optimisation algorithm to find the maximum em a posteriori probability values of the unknown variables. The resulting models are shown to compare well with the current operational model when visualised in the target space.

Developing a scalable model of recombinant protein yield from Pichia pastoris:the influence of culture conditions, biomass and induction regime

Background The optimisation and scale-up of process conditions leading to high yields of recombinant proteins is an enduring bottleneck in the post-genomic sciences. Typical experiments rely on varying selected parameters through repeated rounds of trial-and-error optimisation. To rationalise this, several groups have recently adopted the 'design of experiments' (DoE) approach frequently used in industry. Studies have focused on parameters such as medium composition, nutrient feed rates and induction of expression in shake flasks or bioreactors, as well as oxygen transfer rates in micro-well plates. In this study we wanted to generate a predictive model that described small-scale screens and to test its scalability to bioreactors. Results Here we demonstrate how the use of a DoE approach in a multi-well mini-bioreactor permitted the rapid establishment of high yielding production phase conditions that could be transferred to a 7 L bioreactor. Using green fluorescent protein secreted from Pichia pastoris, we derived a predictive model of protein yield as a function of the three most commonly-varied process parameters: temperature, pH and the percentage of dissolved oxygen in the culture medium. Importantly, when yield was normalised to culture volume and density, the model was scalable from mL to L working volumes. By increasing pre-induction biomass accumulation, model-predicted yields were further improved. Yield improvement was most significant, however, on varying the fed-batch induction regime to minimise methanol accumulation so that the productivity of the culture increased throughout the whole induction period. These findings suggest the importance of matching the rate of protein production with the host metabolism. Conclusion We demonstrate how a rational, stepwise approach to recombinant protein production screens can reduce process development time.