Determination of chitin content in fungal cell wall: an alternative flow cytometric method

The conventional methods used to evaluate chitin content in fungi, such as biochemical assessment of glucosamine release after acid hydrolysis or epifluorescence microscopy, are low throughput, laborious, time-consuming, and cannot evaluate a large number of cells. We developed a flow cytometric assay, efficient, and fast, based on Calcofluor White staining to measure chitin content in yeast cells. A staining index was defined, its value was directly related to chitin amount and taking into consideration the different levels of autofluorecence. Twenty-two Candida spp. and four Cryptococcus neoformans clinical isolates with distinct susceptibility profiles to caspofungin were evaluated. Candida albicans clinical isolate SC5314, and isogenic strains with deletions in chitin synthase 3 (chs3Δ/chs3Δ) and genes encoding predicted Glycosyl Phosphatidyl Inositol (GPI)-anchored proteins (pga31Δ/Δ and pga62Δ/Δ), were used as controls. As expected, the wild-type strain displayed a significant higher chitin content (P < 0.001) than chs3Δ/chs3Δ and pga31Δ/Δ especially in the presence of caspofungin. Ca. parapsilosis, Ca. tropicalis, and Ca. albicans showed higher cell wall chitin content. Although no relationship between chitin content and antifungal drug susceptibility phenotype was found, an association was established between the paradoxical growth effect in the presence of high caspofungin concentrations and the chitin content. This novel flow cytometry protocol revealed to be a simple and reliable assay to estimate cell wall chitin content of fungi.

A novel flow cytometric protocol for assessment of yeast cell adhesion

Microbial adhesion is a field of recognized relevance and, as such, an impressive array of tools has been developed to understand its molecular mechanisms and ultimately for its quantification. Some of the major limitations found within these methodologies concern the incubation time, the small number of cells analyzed, and the operator's subjectivity. To overcome these aspects, we have developed a quantitative method to measure yeast cells' adhesion through flow cytometry. In this methodology, a suspension of yeast cells is mixed with green fluorescent polystyrene microspheres (uncoated or coated with host proteins). Within 2 h, an adhesion profile is obtained based on two parameters: percentage and cells-microsphere population's distribution pattern. This flow cytometry protocol represents a useful tool to quantify yeast adhesion to different substrata in a large scale, providing manifold data in a speedy and informative manner.

Multi-apprentice learning for meta-heuristics parameter tuning in a multi agent scheduling system

The scheduling problem is considered in complexity theory as a NP-hard combinatorial optimization problem. Meta-heuristics proved to be very useful in the resolution of this class of problems. However, these techniques require parameter tuning which is a very hard task to perform. A Case-based Reasoning module is proposed in order to solve the parameter tuning problem in a Multi-Agent Scheduling System. A computational study is performed in order to evaluate the proposed CBR module performance.

Case-based reasoning for meta-heuristics self-parameterization in a multi-agent scheduling system

A novel agent-based approach to Meta-Heuristics self-configuration is proposed in this work. Meta-heuristics are examples of algorithms where parameters need to be set up as efficient as possible in order to unsure its performance. This paper presents a learning module for self-parameterization of Meta-heuristics (MHs) in a Multi-Agent System (MAS) for resolution of scheduling problems. The learning is based on Case-based Reasoning (CBR) and two different integration approaches are proposed. A computational study is made for comparing the two CBR integration perspectives. In the end, some conclusions are reached and future work outlined.

Cooperation mechanism for team-work based multi-agent system in dynamic scheduling through meta-heuristics

This paper describes a Multi-agent Scheduling System that assumes the existence of several Machines Agents (which are decision-making entities) distributed inside the Manufacturing System that interact and cooperate with other agents in order to obtain optimal or near-optimal global performances. Agents have to manage their internal behaviors and their relationships with other agents via cooperative negotiation in accordance with business policies defined by the user manager. Some Multi Agent Systems (MAS) organizational aspects are considered. An original Cooperation Mechanism for a Team-work based Architecture is proposed to address dynamic scheduling using Meta-Heuristics.

ABS4GD: a multi-agent system that simulates group decision processes considering emotional and argumentative aspects

Emotion although being an important factor in our every day life it is many times forgotten in the development of systems to be used by persons. In this work we present an architecture for a ubiquitous group decision support system able to support persons in group decision processes. The system considers the emotional factors of the intervenient participants, as well as the argumentation between them. Particular attention will be taken to one of components of this system: the multi-agent simulator, modeling the human participants, considering emotional characteristics, and allowing the exchanges of hypothetic arguments among the participants.

ISEM: a multi-agent system that simulates competitive electronic marketplaces

With the increasing importance of large commerce across the Internet it is becoming increasingly evident that in a few years the Iternet will host a large number of interacting software agents. a vast number of them will be economically motivated, and will negociate a variety of goods and services. It is therefore important to consider the economic incentives and behaviours of economic software agents, and to use all available means to anticipate their collective interactions. This papers addresses this concern by presenting a multi-agent market simulator designed for analysing agent market strategies based on a complete understanding of buyer and seller behaviours, preference models and pricing algorithms, consideting risk preferences. The system includes agents that are capable of increasing their performance with their own experience, by adapting to the market conditions. The results of the negotiations between agents are analysed by data minig algorithms in order to extract rules that give agents feedback to imprive their strategies.

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Mestrado em Engenharia Informática

Sensing Cloud Optimization to Solve ED of Units with Valve-Point Effects and Multi-fuels

In this paper a solution to an highly constrained and non-convex economical dispatch (ED) problem with a meta-heuristic technique named Sensing Cloud Optimization (SCO) is presented. The proposed meta-heuristic is based on a cloud of particles whose central point represents the objective function value and the remaining particles act as sensors "to fill" the search space and "guide" the central particle so it moves into the best direction. To demonstrate its performance, a case study with multi-fuel units and valve- point effects is presented.

Assessment of saccharide fractionation by ultrafiltration and nanofiltration

This paper addresses the investigation of the fractionation of saccharide mixtures and saccharide mixtures with calcium using ultrafiltration (UF) and nanofiltration (NF). A set of cellulose acetate membranes covered a wide range of molecular weight cut-off (MWCO) ranging from 250 to 46,000 Da and the total feed concentration of saccharides mixtures varied from 1550 to 4700 ppm with the ratio of the two saccharides-solutes (glucose to raffinose) being kept constant at the value of 1.8. The evolution pattern of the saccharide concentration ratio in the UF/NF permeate streams displayed a dependence on the membrane MWCO, on the total sugar concentration and on the presence of calcium ions. For the highest total sugar content, the membranes with MWCO from 2000 to 7000 Da showed saccharide fractionation capability that was enhanced in the presence of calcium. The Steric Pore Flow Model was used to predict individual solute permeation behaviours and to assess the deviations to steric hindered transport of the solutes in multi-component saccharide solutions. (C) 2008 Elsevier B.V. All rights reserved.

Study of the one dimensional and transient bioheat transfer equation: Multi-layer solution development and applications

: In this work we derive an analytical solution given by Bessel series to the transient and one-dimensional (1D) bioheat transfer equation in a multi-layer region with spatially dependent heat sources. Each region represents an independent biological tissue characterized by temperature-invariant physiological parameters and a linearly temperature dependent metabolic heat generation. Moreover, 1D Cartesian, cylindrical or spherical coordinates are used to define the geometry and temperature boundary conditions of first, second and third kinds are assumed at the inner and outer surfaces. We present two examples of clinical applications for the developed solution. In the first one, we investigate two different heat source terms to simulate the heating in a tumor and its surrounding tissue, induced during a magnetic fluid hyperthermia technique used for cancer treatment. To obtain an accurate analytical solution, we determine the error associated with the truncated Bessel series that defines the transient solution. In the second application, we explore the potential of this model to study the effect of different environmental conditions in a multi-layered human head model (brain, bone and scalp). The convective heat transfer effect of a large blood vessel located inside the brain is also investigated. The results are further compared with a numerical solution obtained by the Finite Element Method and computed with COMSOL Multi-physics v4.1 (c). (c) 2013 Elsevier Ltd. All rights reserved.

Non-coding RNAs: multi-tasking molecules in the cell

In the last years it has become increasingly clear that the mammalian transcriptome is highly complex and includes a large number of small non-coding RNAs (sncRNAs) and long noncoding RNAs (lncRNAs). Here we review the biogenesis pathways of the three classes of sncRNAs, namely short interfering RNAs (siRNAs), microRNAs (miRNAs) and PIWI-interacting RNAs (piRNAs). These ncRNAs have been extensively studied and are involved in pathways leading to specific gene silencing and the protection of genomes against virus and transposons, for example. Also, lncRNAs have emerged as pivotal molecules for the transcriptional and post-transcriptional regulation of gene expression which is supported by their tissue-specific expression patterns, subcellular distribution, and developmental regulation. Therefore, we also focus our attention on their role in differentiation and development. SncRNAs and lncRNAs play critical roles in defining DNA methylation patterns, as well as chromatin remodeling thus having a substantial effect in epigenetics. The identification of some overlaps in their biogenesis pathways and functional roles raises the hypothesis that these molecules play concerted functions in vivo, creating complex regulatory networks where cooperation with regulatory proteins is necessary. We also highlighted the implications of biogenesis and gene expression deregulation of sncRNAs and lncRNAs in human diseases like cancer.

Low Profile Multi-Band Antenna for Mobile Communications

In this paper, the design of low profile antennas by using Electromagnetic Band Gap (EBG) structures is introduced. Taking advantage of the fact that they can behave as Perfect Magnetic Conductor (PMC), it is shown that these structures exhibit dual band in-phase reflection at WLAN (Wireless Local Area Network) bands, the 2.4 GHz and 5.2 GHz bands. These structures are applied to PIFA (Planar Inverted-F Antenna) and the results show that it is possible to obtain low profile PIFA's.

Antena multi-banda para comunicações móveis

O desenvolvimento dos sistemas de comunicações móveis tem vindo a ser cada vez maior, fazendo com que os sistemas funcionem em várias bandas de operação. Neste sentido, surge a necessidade de desenvolver antenas que superem aquelas já existentes, ao nível das suas propriedades electromagnéticas, para que os sistemas apresentem uma maior qualidade e possam corresponder às exigências inerentes ao desenvolvimento das sociedades. O objectivo desta dissertação de Mestrado é dimensionar, construir e medir uma antena multi-banda para comunicações móveis, com base em estruturas EBG (Electromagnetic Band-Gap) que melhorem o comportamento electromagnético daquelas já existentes, para a banda de frequências de 2.4 GHz e de 5.2 GHz. Começa-se por fazer-se um estudo acerca do estado da arte de estruturas EBG, muito utilizadas em várias áreas, nomeadamente a área das antenas, área sobre a qual esta dissertação assenta. Posteriormente é feita uma breve introdução às antenas microstrip, particularizando de seguida para antenas PIFA e as suas características. Posteriormente é feito o estudo de uma antena PIFA, com e sem a influência de estruturas EBG, para as bandas de 2.4 GHz e 5.2 GHz. Posteriormente são apresentados e comparados resultados das várias antenas. Da análise desses resultados, verifica-se que é possível obter uma antena de baixo perfil com a utilização de estruturas EBG como plano de massa. Além disso, verifica-se também que é possível diminuir a radiação traseira e aumentar a largura de banda. Finalmente, são apresentadas algumas conclusões e várias propostas de trabalho futuro.

Automatic vehicle detection and classification

This paper presents a proposal for an automatic vehicle detection and classification (AVDC) system. The proposed AVDC should classify vehicles accordingly to the Portuguese legislation (vehicle height over the first axel and number of axels), and should also support profile based classification. The AVDC should also fulfill the needs of the Portuguese motorway operator, Brisa. For the classification based on the profile we propose:he use of Eigenprofiles, a technique based on Principal Components Analysis. The system should also support multi-lane free flow for future integration in this kind of environments.