Isolation of human fungi from soil and identification of two endemic areas of Cryptococcus neoformans and Coccidioides immitis

The present study was carried out in two different areas of Province of Cordoba, Argentina, where there was a suspicious of endemic mycosis. The previous data were the presence of a clinical case of pulmonary cryptococcosis in one area (Alta Gracia) and the previous findings of a high incidence of coccidioidin and cryptococcin reactors in the population of the second one (Villa Dolores). In both areas soil samples for fungi were studied and Cryptococcus neoformans was found in 2/25 samples from Alta Gracia. In Villa Dolores Coccidioides immitis was isolated in 2/40 samples, and C. neoformans in 1/40 samples. Delayed hypersensitivity test with cryptococcin was determined in the population from Alta Gracia and it was found to be 5.3%. Positive cutaneous tests with coccidioidin (33.8%) and cryptococcin (31.9%) in Villa Dolores were obtained. With these findings two endemic areas of systemic mycoses in Cordoba, Argentina were delimited.

Optimization of multi-stage amplifiers in deep-submicron CMOS using a distributed/parallel genetic algorithm

IEEE International Symposium on Circuits and Systems, pp. 724 – 727, Seattle, EUA

Multi-bit sigma-delta modulators with enhanced dynamic-range using non-linear DAC for hearing aids

15th IEEE International Conference on Electronics, Circuits and Systems, Malta

Functional domains of Bacillus subtilis transcription factor AraR and identification of aminoacids important for nucleoprotein complex assembly and effector-binding.

Journal of Bacteriology (Apr 2006) 3024-3036

Framework para aplicações colaborativas multi-plataforma

Dissertação de Mestrado em Engenharia Informática

Simultaneous identification of Trypanosoma cruzi surface and internal antigens reactive to different immunoglobulin classes (radio-immunoblotting)

A radioactive Western-blotting technique was developed by which the reactivity of Immunoglobulins (Igs) from different classes to both membrane radiolabelled and internal parasite antigens is simultaneously identified. The method includes radioiodination of parasites, polypeptide fractionation by SDS-PAGE, Western-blot transfer and autoradiography of the immunoblots developed with anti-Igs conjugates labelled with enzymes. The analysis is then performed by the comparison of common bands on the autoradiograms and the respective substrate stained nitrocellulose blots. This technique was used to analyse T. cruzi trypomastigote surface labelled antigens reactive to IgM, IgA and IgG specific antibodies. A different pattern of reactivity with acute Chagas' disease patients sera was thus obtained.

Multi-agent system for personalization of location-based services

The current ubiquitous network access and increase in network bandwidth are driving the sales of mobile location-aware user devices and, consequently, the development of context-aware applications, namely location-based services. The goal of this project is to provide consumers of location-based services with a richer end-user experience by means of service composition, personalization, device adaptation and continuity of service. Our approach relies on a multi-agent system composed of proxy agents that act as mediators and providers of personalization meta-services, device adaptation and continuity of service for consumers of pre-existing location-based services. These proxy agents, which have Web services interfaces to ensure a high level of interoperability, perform service composition and take in consideration the preferences of the users, the limitations of the user devices, making the usage of different types of devices seamless for the end-user. To validate and evaluate the performance of this approach, use cases were defined, tests were conducted and results gathered which demonstrated that the initial goals were successfully fulfilled.

Argumentation as Distributed Belief Revision: Conflict Resolution in Decentralised Multi-agent Systems

Decentralised co-operative multi-agent systems are computational systems where conflicts are frequent due to the nature of the represented knowledge. Negotiation methodologies, in this case argumentation based negotiation methodologies, were developed and applied to solve unforeseeable and, therefore, unavoidable conflicts. The supporting computational model is a distributed belief revision system where argumentation plays the decisive role of revision. The distributed belief revision system detects, isolates and solves, whenever possible, the identified conflicts. The detection and isolation of the conflicts is automatically performed by the distributed consistency mechanism and the resolution of the conflict, or belief revision, is achieved via argumentation. We propose and describe two argumentation protocols intended to solve different types of identified information conflicts: context dependent and context independent conflicts. While the protocol for context dependent conflicts generates new consensual alternatives, the latter chooses to adopt the soundest, strongest argument presented. The paper shows the suitability of using argumentation as a distributed decentralised belief revision protocol to solve unavoidable conflicts.

Environmental Decision Support: a Multi-Agent Approach

The environmental management domain is vast and encompasses many identifiable activities: impact assessment, planning, project evaluation, etc. In particular, this paper focusses on the modelling of the project evaluation activity. The environmental decision support system under development aims to provide assistance to project developers in the selection of adequate locations, guaranteeing the compliance with the applicable regulations and the existing development plans as well as satisfying the specified project requirements. The inherent multidisciplinarity features of this activity lead to the adoption of the Multi-Agent paradigm, and, in particular, to the modelling of the involved agencies as a community of cooperative autonomous agents, where each agency contributes with its share of problem solving to the final system’s recommendation. To achieve this behaviour the many conclusions of the individual agencies have to be justifiably accommodated: not only they may differ, but can be interdependent, complementary, irreconcilable, or simply, independent. We propose different solutions (involving both local and global consistency) to support the adequate merge of the distinct perspectives that inevitably arise during this type of decision making.

Belief Revision in Multi-Agent Systems

The ability to respond sensibly to changing and conflicting beliefs is an integral part of intelligent agency. To this end, we outline the design and implementation of a Distributed Assumption-based Truth Maintenance System (DATMS) appropriate for controlling cooperative problem solving in a dynamic real world multi-agent community. Our DATMS works on the principle of local coherence which means that different agents can have different perspectives on the same fact provided that these stances are appropriately justified. The belief revision algorithm is presented, the meta-level code needed to ensure that all system-wide queries can be uniquely answered is described, and the DATMS’ implementation in a general purpose multi-agent shell is discussed.

Consistency and Context Management in a Multi Agent Belief Revision Testbed

Multi-agent architectures are well suited for complex inherently distributed problem solving domains. From the many challenging aspects that arise within this framework, a crucial one emerges: how to incorporate dynamic and conflicting agent beliefs? While the belief revision activity in a single agent scenario is concentrated on incorporating new information while preserving consistency, in a multi-agent system it also has to deal with possible conflicts between the agents perspectives. To provide an adequate framework, each agent, built as a combination of an assumption based belief revision system and a cooperation layer, was enriched with additional features: a distributed search control mechanism allowing dynamic context management, and a set of different distributed consistency methodologies. As a result, a Distributed Belief Revision Testbed (DiBeRT) was developed. This paper is a preliminary report presenting some of DiBeRT contributions: a concise representation of external beliefs; a simple and innovative methodology to achieve distributed context management; and a reduced inter-agent data exchange format.

Beliefs and Conflicts in a Real World Multi-Agent System

In a real world multiagent system, where the agents are faced with partial, incomplete and intrinsically dynamic knowledge, conflicts are inevitable. Frequently, different agents have goals or beliefs that cannot hold simultaneously. Conflict resolution methodologies have to be adopted to overcome such undesirable occurrences. In this paper we investigate the application of distributed belief revision techniques as the support for conflict resolution in the analysis of the validity of the candidate beams to be produced in the CERN particle accelerators. This CERN multiagent system contains a higher hierarchy agent, the Specialist agent, which makes use of meta-knowledge (on how the con- flicting beliefs have been produced by the other agents) in order to detect which beliefs should be abandoned. Upon solving a conflict, the Specialist instructs the involved agents to revise their beliefs accordingly. Conflicts in the problem domain are mapped into conflicting beliefs of the distributed belief revision system, where they can be handled by proven formal methods. This technique builds on well established concepts and combines them in a new way to solve important problems. We find this approach generally applicable in several domains.

Multi-agent system for distributed manufacturing scheduling with genetic algorithms and tabu search

Computerized scheduling methods and computerized scheduling systems according to exemplary embodiments. A computerized scheduling method may be stored in a memory and executed on one or more processors. The method may include defining a main multi-machine scheduling problem as a plurality of single machine scheduling problems; independently solving the plurality of single machine scheduling problems thereby calculating a plurality of near optimal single machine scheduling problem solutions; integrating the plurality of near optimal single machine scheduling problem solutions into a main multi-machine scheduling problem solution; and outputting the main multi-machine scheduling problem solution.

Multi-elemental analysis of ready-to-eat “baby leaf” vegetables using microwave digestion and high-resolution continuum source atomic absorption spectrometry

The mineral content (phosphorous (P), potassium (K), sodium (Na), calcium (Ca), magnesium (Mg), iron (Fe), manganese (Mn), zinc (Zn) and copper (Cu)) of eight ready-to-eat baby leaf vegetables was determined. The samples were subjected to microwave-assisted digestion and the minerals were quantified by High-Resolution Continuum Source Atomic Absorption Spectrometry (HR-CS-AAS) with flame and electrothermal atomisation. The methods were optimised and validated producing low LOQs, good repeatability and linearity, and recoveries, ranging from 91% to 110% for the minerals analysed. Phosphorous was determined by a standard colorimetric method. The accuracy of the method was checked by analysing a certified reference material; results were in agreement with the quantified value. The samples had a high content of potassium and calcium, but the principal mineral was iron. The mineral content was stable during storage and baby leaf vegetables could represent a good source of minerals in a balanced diet. A linear discriminant analysis was performed to compare the mineral profile obtained and showed, as expected, that the mineral content was similar between samples from the same family. The Linear Discriminant Analysis was able to discriminate different samples based on their mineral profile.

The role of the efflux mechanisms in multidrug resistance in Mycobacterium tuberculosis

Abstract The emergence of multi and extensively drug resistant tuberculosis (MDRTB and XDRTB) has increased the concern of public health authorities around the world. The World Health Organization has defined MDRTB as tuberculosis (TB) caused by organisms resistant to at least isoniazid and rifampicin, the main first-line drugs used in TB therapy, whereas XDRTB refers to TB resistant not only to isoniazid and rifampicin, but also to a fluoroquinolone and to at least one of the three injectable second-line drugs, kanamycin, amikacin and capreomycin. Resistance in Mycobacterium tuberculosis is mainly due to the occurrence of spontaneous mutations and followed by selection of mutants by subsequent treatment. However, some resistant clinical isolates do not present mutations in any genes associated with resistance to a given antibiotic, which suggests that other mechanism(s) are involved in the development of drug resistance, namely the presence of efflux pump systems that extrude the drug to the exterior of the cell, preventing access to its target. Increased efflux activity can occur in response to prolonged exposure to subinhibitory concentrations of anti-TB drugs, a situation that may result from inadequate TB therapy. The inhibition of efflux activity with a non-antibiotic inhibitor may restore activity of an antibiotic subject to efflux and thus provide a way to enhance the activity of current anti-TB drugs. The work described in this thesis foccus on the study of efflux mechanisms in the development of multidrug resistance in M. tuberculosis and how phenotypic resistance, mediated by efflux pumps, correlates with genetic resistance. In order to accomplish this goal, several experimental protocols were developed using biological models such as Escherichia coli, the fast growing mycobacteria Mycobacterium smegmatis, and Mycobacterium avium, before their application to M. tuberculosis. This approach allowed the study of the mechanisms that result in the physiological adaptation of E. coli to subinhibitory concentrations of tetracycline (Chapter II), the development of a fluorometric method that allows the detection and quantification of efflux of ethidium bromide (Chapter III), the characterization of the ethidium bromide transport in M. smegmatis (Chapter IV) and the contribution of efflux activity to macrolide resistance in Mycobacterium avium complex (Chapter V). Finally, the methods developed allowed the study of the role of efflux pumps in M. tuberculosis strains induced to isoniazid resistance (Chapter VI). By this manner, in Chapter II it was possible to observe that the physiological adaptation of E. coli to tetracycline results from an interplay between events at the genetic level and protein folding that decrease permeability of the cell envelope and increase efflux pump activity. Furthermore, Chapter III describes the development of a semi-automated fluorometric method that allowed the correlation of this efflux activity with the transport kinetics of ethidium bromide (a known efflux pump substrate) in E. coli and the identification of efflux inhibitors. Concerning M. smegmatis, we have compared the wild-type M. smegmatis mc2155 with knockout mutants for LfrA and MspA for their ability to transport ethidium bromide. The results presented in Chapter IV showed that MspA, the major porin in M. smegmatis, plays an important role in the entrance of ethidium bromide and antibiotics into the cell and that efflux via the LfrA pump is involved in low-level resistance to these compounds in M. smegmatis. Chapter V describes the study of the contribution of efflux pumps to macrolide resistance in clinical M. avium complex isolates. It was demonstrated that resistance to clarithromycin was significantly reduced in the presence of efflux inhibitors such as thioridazine, chlorpromazine and verapamil. These same inhibitors decreased efflux of ethidium bromide and increased the retention of [14C]-erythromycin in these isolates. Finaly, the methods developed with the experimental models mentioned above allowed the study of the role of efflux pumps on M. tuberculosis strains induced to isoniazid resistance. This is described in Chapter VI of this Thesis, where it is demonstrated that induced resistance to isoniazid does not involve mutations in any of the genes known to be associated with isoniazid resistance, but an efflux system that is sensitive to efflux inhibitors. These inhibitors decreased the efflux of ethidium bromide and also reduced the minimum inhibitory concentration of isoniazid in these strains. Moreover, expression analysis showed overexpression of genes that code for efflux pumps in the induced strains relatively to the non-induced parental strains. In conclusion, the work described in this thesis demonstrates that efflux pumps play an important role in the development of drug resistance, namely in mycobacteria. A strategy to overcome efflux-mediated resistance may consist on the use of compounds that inhibit efflux activity, restoring the activity of antimicrobials that are efflux pump substrates, a useful approach particularly in TB where the most effective treatment regimens are becoming uneffective due to the increase of MDRTB/XDRTB.