In situ NIR spectroscopy monitoring of plasmid production processes effect of producing strain, medium composition and the cultivation strategy

BACKGROUNDWhile the pharmaceutical industry keeps an eye on plasmid DNA production for new generation gene therapies, real-time monitoring techniques for plasmid bioproduction are as yet unavailable. This work shows the possibility of in situ monitoring of plasmid production in Escherichia coli cultures using a near infrared (NIR) fiber optic probe. RESULTSPartial least squares (PLS) regression models based on the NIR spectra were developed for predicting bioprocess critical variables such as the concentrations of biomass, plasmid, carbon sources (glucose and glycerol) and acetate. In order to achieve robust models able to predict the performance of plasmid production processes, independently of the composition of the cultivation medium, cultivation strategy (batch versus fed-batch) and E. coli strain used, three strategies were adopted, using: (i) E. coliDH5 cultures conducted under different media compositions and culture strategies (batch and fed-batch); (ii) engineered E. coli strains, MG1655endArecApgi and MG1655endArecA, grown on the same medium and culture strategy; (iii) diverse E. coli strains, over batch and fed-batch cultivations and using different media compositions. PLS models showed high accuracy for predicting all variables in the three groups of cultures. CONCLUSIONNIR spectroscopy combined with PLS modeling provides a fast, inexpensive and contamination-free technique to accurately monitoring plasmid bioprocesses in real time, independently of the medium composition, cultivation strategy and the E. coli strain used.

Monitoring the ex-vivo expansion of human mesenchymal stem/stromal cells in xeno-free microcarrier-based reactor systems by MIR spectroscopy

Human mesenchymal stem/stromal cells (MSCs) have received considerable attention in the field of cell-based therapies due to their high differentiation potential and ability to modulate immune responses. However, since these cells can only be isolated in very low quantities, successful realization of these therapies requires MSCs ex-vivo expansion to achieve relevant cell doses. The metabolic activity is one of the parameters often monitored during MSCs cultivation by using expensive multi-analytical methods, some of them time-consuming. The present work evaluates the use of mid-infrared (MIR) spectroscopy, through rapid and economic high-throughput analyses associated to multivariate data analysis, to monitor three different MSCs cultivation runs conducted in spinner flasks, under xeno-free culture conditions, which differ in the type of microcarriers used and the culture feeding strategy applied. After evaluating diverse spectral preprocessing techniques, the optimized partial least square (PLS) regression models based on the MIR spectra to estimate the glucose, lactate and ammonia concentrations yielded high coefficients of determination (R2 ≥ 0.98, ≥0.98, and ≥0.94, respectively) and low prediction errors (RMSECV ≤ 4.7%, ≤4.4% and ≤5.7%, respectively). Besides PLS models valid for specific expansion protocols, a robust model simultaneously valid for the three processes was also built for predicting glucose, lactate and ammonia, yielding a R2 of 0.95, 0.97 and 0.86, and a RMSECV of 0.33, 0.57, and 0.09 mM, respectively. Therefore, MIR spectroscopy combined with multivariate data analysis represents a promising tool for both optimization and control of MSCs expansion processes.

Comparing wind generation profiles: A circular data approach

The importance of wind power energy for energy and environmental policies has been growing in past recent years. However, because of its random nature over time, the wind generation cannot be reliable dispatched and perfectly forecasted, becoming a challenge when integrating this production in power systems. In addition the wind energy has to cope with the diversity of production resulting from alternative wind power profiles located in different regions. In 2012, Portugal presented a cumulative installed capacity distributed over 223 wind farms [1]. In this work the circular data statistical methods are used to analyze and compare alternative spatial wind generation profiles. Variables indicating extreme situations are analyzed. The hour (s) of the day where the farm production attains its maximum daily production is considered. This variable was converted into circular variable, and the use of circular statistics enables to identify the daily hour distribution for different wind production profiles. This methodology was applied to a real case, considering data from the Portuguese power system regarding the year 2012 with a 15-minutes interval. Six geographical locations were considered, representing different wind generation profiles in the Portuguese system.In this work the circular data statistical methods are used to analyze and compare alternative spatial wind generation profiles. Variables indicating extreme situations are analyzed. The hour (s) of the day where the farm production attains its maximum daily production is considered. This variable was converted into circular variable, and the use of circular statistics enables to identify the daily hour distribution for different wind production profiles. This methodology was applied to a real case, considering data from the Portuguese power system regarding the year 2012 with a 15-minutes interval. Six geographical locations were considered, representing different wind generation profiles in the Portuguese system.

Simulação de um processo de pirólise rápida de biomassa

Mestrado em Engenharia Química - Ramo Optimização Energética na Indústria Química

O papel dos sistemas de Non-Intrusive Load Monitoring para a eficiência energética do setor residencial

A sustentabilidade energética do planeta é uma preocupação corrente e, neste sentido, a eficiência energética afigura-se como sendo essencial para a redução do consumo em todos os setores de atividade. No que diz respeito ao setor residencial, o indevido comportamento dos utilizadores aliado ao desconhecimento do consumo dos diversos aparelhos, são factores impeditivos para a redução do consumo energético. Uma ferramenta importante, neste sentido, é a monitorização de consumos nomeadamente a monitorização não intrusiva, que apresenta vantagens económicas relativamente à monitorização intrusiva, embora levante alguns desafios na desagregação de cargas. Abordou-se então, neste documento, a temática da monitorização não intrusiva onde se desenvolveu uma ferramenta de desagregação de cargas residenciais, sobretudo de aparelhos que apresentavam elevados consumos. Para isso, monitorizaram-se os consumos agregados de energia elétrica, água e gás de seis habitações do município de Vila Nova de Gaia. Através da incorporação dos vetores de água e gás, a acrescentar ao da energia elétrica, provou-se que a performance do algoritmo de desagregação de aparelhos poderá aumentar, no caso de aparelhos que utilizem simultaneamente energia elétrica e água ou energia elétrica e gás. A eficiência energética é também parte constituinte deste trabalho e, para tal, implementaram-se medidas de eficiência energética para uma das habitações em estudo, de forma a concluir as que exibiam maior potencial de poupança, assim como rápidos períodos de retorno de investimento. De um modo geral, os objetivos propostos foram alcançados e espera-se que num futuro próximo, a monitorização de consumos não intrusiva se apresente como uma solução de referência no que respeita à sustentabilidade energética do setor residencial.

Gestão Inteligente de uma Habitação considerando o Controlo de Cargas, de Microgeração e Veículos Elétricos em Função do seu Perfil de Utilização

Os consumidores finais são vistos, no novo paradigma da operação das redes elétricas, como intervenientes ativos com capacidade para gerir os seus recursos energéticos, nomeadamente as cargas, as unidades de produção, os veículos elétricos e a participação em eventos de Demand Response. Tem sido evidente um aumento do consumo de energia, sendo que o setor residencial representa uma importante parte do consumo global dos países desenvolvidos. Para que a participação ativa dos consumidores seja possível, várias abordagens têm vindo a ser propostas, com ênfase nas Smart Grids e nas Microgrids. Diversos sistemas têm sido propostos e desenvolvidos com o intuito de tornar a operação dos sistemas elétricos mais flexível. Neste contexto, os sistemas de gestão de instalações domésticas apresentam-se como um elemento fulcral para a participação ativa dos consumidores na gestão energética, permitindo aos operadores de sistema coordenarem a produção mas também a procura. No entanto, é importante identificar as vantagens da implementação e uso de sistemas de gestão de energia elétrica para os consumidores finais. Nesta dissertação são propostas metodologias de apoio ao consumidor doméstico na gestão dos recursos energéticos existentes e a implementação das mesmas na plataforma de simulação de um sistema de gestão de energia desenvolvido para consumidores domésticos, o SCADA House Intelligent Management (SHIM). Para tal, foi desenvolvida uma interface que permite a simulação em laboratório do sistema de gestão desenvolvido. Adicionalmente, o SHIM foi incluído no simulador Multi-Agent Smart Grid Simulation Plataform (MASGriP) permitindo a simulação de cenários considerando diferentes agentes. Ao nível das metodologias desenvolvidas são propostos diferentes algoritmos de gestão dos recursos energéticos existentes numa habitação, considerando utilizadores com diferentes tipos de recursos (cargas; cargas e veículos elétricos; cargas, veículos elétricos e microgeração). Adicionalmente é proposto um método de gestão dinâmica das cargas para eventos de Demand Response de longa duração, considerando as características técnicas dos equipamentos. Nesta dissertação são apresentados cinco casos de estudos em que cada um deles tem diferentes cenários de simulação. Estes casos de estudos são importantes para verificar a viabilidade da implementação das metodologias propostas para o SHIM. Adicionalmente são apresentados na dissertação perfis reais dos vários recursos energéticos e de consumidores domésticos que são, posteriormente, utilizados para o desenvolvimento dos casos de estudo e aplicação das metodologias.

Defining Electricity Tariffs Using the Knowledge About the Consumers Profiles in ELECON Project

The increasing importance of the integration of distributed generation and demand response in the power systems operation and planning, namely at lower voltage levels of distribution networks and in the competitive environment of electricity markets, leads us to the concept of smart grids. In both traditional and smart grid operation, non-technical losses are a great economic concern, which can be addressed. In this context, the ELECON project addresses the use of demand response contributions to the identification of non-technical losses. The present paper proposes a methodology to be used by Virtual Power Players (VPPs), which are entities able to aggregate distributed small-size resources, aiming to define the best electricity tariffs for several, clusters of consumers. A case study based on real consumption data demonstrates the application of the proposed methodology.

Adaptive Learning in Games: Defining Profiles of Competitor Players

Artificial Intelligence has been applied to dynamic games for many years. The ultimate goal is creating responses in virtual entities that display human-like reasoning in the definition of their behaviors. However, virtual entities that can be mistaken for real persons are yet very far from being fully achieved. This paper presents an adaptive learning based methodology for the definition of players’ profiles, with the purpose of supporting decisions of virtual entities. The proposed methodology is based on reinforcement learning algorithms, which are responsible for choosing, along the time, with the gathering of experience, the most appropriate from a set of different learning approaches. These learning approaches have very distinct natures, from mathematical to artificial intelligence and data analysis methodologies, so that the methodology is prepared for very distinct situations. This way it is equipped with a variety of tools that individually can be useful for each encountered situation. The proposed methodology is tested firstly on two simpler computer versus human player games: the rock-paper-scissors game, and a penalty-shootout simulation. Finally, the methodology is applied to the definition of action profiles of electricity market players; players that compete in a dynamic game-wise environment, in which the main goal is the achievement of the highest possible profits in the market.

Epidemiology and antimicrobial resistance of B. fragilis group organisms isolated from clinical specimen and human intestinal microbiota

Epidemiological aspects and the antimicrobial susceptibility profile of the Bacteroides fragilis group isolated from clinical and human intestinal specimens were examined in this study. B. fragilis group strains were isolated from 46 (37%) of 124 clinical specimens and the source of the samples was: Blood culture (3), intraabdominal infection (27), brain abscess (2), soft tissue infection (17), respiratory sinus (3), pleural aspirate (9), breast abscess (3), surgical infected wound (22), pelvic inflammatory disease (22), chronic otitis media (9) and miscellaneous (7). Intraabdominal and soft tissue infections were responsible for more than half of the clinical isolates. Susceptibility to penicillin, cefoxitin, tetracycline, metronidazole, chloramphenicol and clindamycin was examined. All isolates were susceptible to metronidazole and chloramphenicol. For clindamycin and cefoxitin the resistance rates observed were 21.7% and 10.9% respectively. Susceptibility profiles varied among the different species tested. A total of 37 species of B. fragilis group isolated from intestinal microbiota of individuals who had no antimicrobial therapy for at least 1 month before the sampling was also examined. All strains were also susceptible to chloramphenicol and motronidazole and the resistance rates to clindamycin and cefoxitin were 19.4% and 5.4% respectively. A few institutions, in Brazil, have monitored the antimicrobial susceptibility of B. fragilis group strains isolated from anaerobic infections. The resistance rates to cefoxitin and clindamycin and the variation in susceptibility patterns among the species isolated in this study emphasize the need for monitoring of susceptibility patterns of B. fragilis group organisms isolated, especially at our University Hospitals.

Using Typical Consumption Profiles to Stablish the Consumption Level for Short Time Periods

The current practices in the consumption metering by electricity utilities is currently largely based on monthly consumption reading. The consumption metering device is always calculating the cumulative consumption. Then, it is possible to calculate the difference between the actual and the previous consumption evaluation in order to estimate the monthly consumption. The power systems planning needs in many aspects to handle consumption data obtained for shorter periods, namely in the Demand Response programs planning. The work presented in this paper is based on the application of typical consumption profiles that are previously defined for a certain power system area. Such profiles are then used in order to estimate the 15 minutes consumption for a certain consumer or consumer type.

Cerebrospinal fluid profiles in acquired immunodeficiency syndrome with and without neurocryptococcosis

Cryptococcosis is one of the most common fungal infections of the central nervous system (CNS) in AIDS patients and meningoencephalitis or meningitis is a frequently observed manifestation. However, systematic studies of cerebrospinal fluid (CSF) composition from AIDS patients with CNS cryptococcosis have been few. CSF samples from 114 HIV seropositive patients whose clinical complaint suggested CNS involvement, were analyzed; 32 samples from patients diagnosed as having neurocryptococcosis (Group 1) and 82 samples from patients with no identified neurological disfunction (Group 2). Based on cytological and biochemical results, two distinct profiles were observed: Normal (Group 1 = 31%, Group 2 = 39%); Abnormal (Group 1 = 69%, Group 2 = 61%). Lymphocytes were the most frequent cells in both groups. Our CSF cytological and biochemical findings showed that in AIDS patients liquoric abnormalities are quite frequent, non-specific and difficult to interpret. In these circumstances a systematic search to identify the etiologic agent using microbiological and/or immunological assays must be routinely performed

Customisable e-training programmes based on trainees profiles

Dissertation presented at Faculdade de Ciências e Tecnologia of Universidade Nova de Lisboa to obtain the Master degree in Electrical and Computer Engineering

Lettuce (Lactuca sativa L.) leaf-proteome profiles after exposure to cylindrospermopsin and a microcystin-LR/cylindrospermopsin mixture: A concentration-dependent response

The intensification of agricultural productivity is an important challenge worldwide. However, environmental stressors can provide challenges to this intensification. The progressive occurrence of the cyanotoxins cylindrospermopsin (CYN) and microcystin-LR (MC-LR) as a potential consequence of eutrophication and climate change is of increasing concern in the agricultural sector because it has been reported that these cyanotoxins exert harmful effects in crop plants. A proteomic-based approach has been shown to be a suitable tool for the detection and identification of the primary responses of organisms exposed to cyanotoxins. The aim of this study was to compare the leaf-proteome profiles of lettuce plants exposed to environmentally relevant concentrations of CYN and a MC-LR/CYN mixture. Lettuce plants were exposed to 1, 10, and 100 lg/l CYN and a MC-LR/CYN mixture for five days. The proteins of lettuce leaves were separated by twodimensional electrophoresis (2-DE), and those that were differentially abundant were then identified by matrix-assisted laser desorption/ionization time of flight-mass spectrometry (MALDI-TOF/TOF MS). The biological functions of the proteins that were most represented in both experiments were photosynthesis and carbon metabolism and stress/defense response. Proteins involved in protein synthesis and signal transduction were also highly observed in the MC-LR/CYN experiment. Although distinct protein abundance patterns were observed in both experiments, the effects appear to be concentration-dependent, and the effects of the mixture were clearly stronger than those of CYN alone. The obtained results highlight the putative tolerance of lettuce to CYN at concentrations up to 100 lg/l. Furthermore, the combination of CYN with MC-LR at low concentrations (1 lg/l) stimulated a significant increase in the fresh weight (fr. wt) of lettuce leaves and at the proteomic level resulted in the increase in abundance of a high number of proteins. In contrast, many proteins exhibited a decrease in abundance or were absent in the gels of the simultaneous exposure to 10 and 100 lg/l MC-LR/CYN. In the latter, also a significant decrease in the fr. wt of lettuce leaves was obtained. These findings provide important insights into the molecular mechanisms of the lettuce response to CYN and MC-LR/CYN and may contribute to the identification of potential protein markers of exposure and proteins that may confer tolerance to CYN and MC-LR/CYN. Furthermore, because lettuce is an important crop worldwide, this study may improve our understanding of the potential impact of these cyanotoxins on its quality traits (e.g., presence of allergenic proteins).

ANTI M. leprae IgM ANTIBODY DETERMINATION BY ULTRAMICROIMMUNOENZYMATIC (UMELISA HANSEN) FOR THE DIAGNOSIS AND MONITORING LEPROSY

The relationship between the IgM antibody response, antigenic load as well as the clinical improvement after chemotherapy was studied in order to obtain useful data for the early diagnosis and monitoring leprosy. A level of 82% (94/115) agreement was obtained between IgM UMELISA HANSEN and slitskin smear examination. Discrepant results were observed in 16 patients who showed positive IgM response despite negative by the skin smear examination. In these patients, the IgM response was seen to be associated to the early signal for bacilli recurrence in the skin. In one of these patients the presence of bacilli was demonstrated in the skin, two months after IgM antibodies being detected by UMELISA HANSEN. Also in one of the treated patients positive by both diagnostic techniques, a remarkable decrease in the IgM antibody levels was seen, correlating with a significant clinical improvement. Moreover it was found a direct relationship between the IgM antibody response and bacterial antigenic load, regardless the time elapsed in the disease's evolution.

Criteria for the selection of foster families and monitoring of placements.: Comparative study of the application of the Casey Foster Applicant Inventory-Applicant Version (CFAI-A)

Based on a literature review, this article frames different stages of the foster care process, identifying a set of standardized measures in the American and Portuguese contexts which, if implemented, could contribute towards higher levels of foster success. The article continues with the presentation of a comparative study, based on the application of the Casey Foster Applicant Inventory-Applicant Version (CFAI-A) questionnaire, in the aforementioned contexts. Taking a comparative analyses of CFAI-A's psychometric characteristics in four different samples as a starting point, one discovered that despite the fact that the questionnaire was adapted to Portuguese reality, it kept the quality values presented on the American samples. It specifically shows significant values regarding reliability and validity. This questionnaire, which aims to assess the potential of foster families, also supports the technical staff's decision making process regarding the monitoring and support of foster families, while it also promotes a better decision in the placement process towards the child's integration and development.