778 resultados para Isolation protocol


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The pathogenesis-related (PR) protein superfamily is widely distributed in the animal, plant, and fungal kingdoms and is implicated in human brain tumor growth and plant pathogenesis. The precise biological activity of PR proteins, however, has remained elusive. Here we report the characterization, cloning and structural homology modeling of Tex31 from the venom duct of Conus textile. Tex31 was isolated to >95% purity by activity-guided fractionation using a para-nitroanilide substrate based on the putative cleavage site residues found in the propeptide precursor of conotoxin TxVIA. Tex31 requires four residues including a leucine N-terminal of the cleavage site for efficient substrate processing. The sequence of Tex31 was determined using two degenerate PCR primers designed from N-terminal and tryptic digest Edman sequences. A BLAST search revealed that Tex31 was a member of the PR protein superfamily and most closely related to the CRISP family of mammalian proteins that have a cysteine-rich C-terminal tail. A homology model constructed from two PR proteins revealed that the likely catalytic residues in Tex31 fall within a structurally conserved domain found in PR proteins. Thus, it is possible that other PR proteins may also be substrate-specific proteases.

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Recent progresses in the software development world has assisted a change in hardware from heavy mainframes and desktop machines to unimaginable small devices leading to the prophetic "third computing paradigm", Ubiquitous Computing. Still, this novel unnoticeable devices lack in various capabilities, like computing power, storage capacity and human interface. Connectivity associated to this devices is also considered an handicap which comes generally associated expensive and limited protocols like GSM and UMTS. Considering this scenario as background, this paper presents a minimal communication protocol introducing better interfaces for limited devices. Special attention has been paid to the limitations of connectivity, storage capacity and scalability of the developed software applications. Illustrating this new protocol, a case-study is presented addressing car sensors communicating with a central

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With the number of elderly people increasing tremendously worldwide, comes the need for effective methods to maintain or improve older adults' cognitive performance. Using continuous neurofeedback, through the use of EEG techniques, people can learn how to train and alter their brain electrical activity. A software platform that puts together the proposed rehabilitation methodology has been developed: a digital game protocol that supports neurofeedback training of alpha and theta rhythms, by reading the EEG activity and presenting it back to the subject, interleaved with neurocognitive tasks such as n-Back and Corsi Block-Tapping. This tool will be used as a potential rehabilitative platform for age-related memory impairments.

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OBJECTIVE: To evaluate the microbiological quality of pasteurized milk commercialized in Rio de Janeiro, Brazil, and determine serologically enteropathogenic Escherichia coli (EPEC) strains in E. coli isolates obtained from milk samples. METHODS: Ninety samples of pasteurized milk -- types B and C -- of three different commercial brands, purchased in supermarkets and bakeries in Rio de Janeiro, were examined. The amount of total and fecal coliform bacteria was estimated using the Most Probable Number technique. Mesophilic, psychrotrophic, and thermoduric microorganism counts were determined by the Standard Plate Count technique. Isolation and identification of E. coli were carried out using conventional physiological tests. Commercial antisera were used for serological characterization of EPEC. RESULTS: The three milk brands analyzed revealed bacterial counts above the regulated values of the Brazilian government. It was found that among 208 strains of E. coli isolated, 46 (22.1%) were serologically classified as EPEC. The most common EPEC serogroup was O55 (15.2%). CONCLUSIONS: Though recent studies on virulence factors indicate that not all strains serologically classified as EPEC are able to attaching/effacing lesion, it is believed that the isolation of EPEC serogroups from pasteurized milk represent a potential risk for children, as well as an indicative of the presence of other enteropathogens.

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Screening programs, particularly the inclusion of specific orthoptic tests to detect visual abnormalities, varies among countries. This study aims to: 1) describes expert perception of issues related with children visual screening; 2) identify specific orthoptic tests to detect visual abnormalities in children visual screening.

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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.

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Mestrado em Engenharia Electrotécnica e de Computadores

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Wireless Sensor Networks (WSNs) have been attracting increasing interests in the development of a new generation of embedded systems with great potential for many applications such as surveillance, environment monitoring, emergency medical response and home automation. However, the communication paradigms in Wireless Sensor Networks differ from the ones attributed to traditional wireless networks, triggering the need for new communication protocols and mechanisms. In this Technical Report, we present a survey on communication protocols for WSNs with a particular emphasis on the lower protocol layers. We give a particular focus to the MAC (Medium Access Control) sub-layer, since it has a prominent influence on some relevant requirements that must be satisfied by WSN protocols, such as energy consumption, time performance and scalability. We overview some relevant MAC protocol solutions and discuss how they tackle the trade-off between the referred requirements.

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The overall goal of the REMPLI project is to design and implement a communication infrastructure for distributed data acquisition and remote control operations using the power grid as the communication medium. The primary target application is remote meter reading with high time resolution, where the meters can be energy, heat, gas, or water meters. The users of the system (e.g. utility companies) will benefit from the REMPLI system by gaining more detailed information about how energy is consumed by the end-users. In this context, the power-line communication (PLC) is deployed to cover the distance between utility company’s Private Network and the end user. This document specifies a protocol for real-time PLC, in the framework of the REMPLI project. It mainly comprises the Network Layer and Data Link Layer. The protocol was designed having into consideration the specific aspects of the network: different network typologies (star, tree, ring, multiple paths), dynamic changes in network topology (due to network maintenance, hazards, etc.), communication lines strongly affected by noise.

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A recent trend in distributed computer-controlled systems (DCCS) is to interconnect the distributed computing elements by means of multi-point broadcast networks. Since the network medium is shared between a number of network nodes, access contention exists and must be solved by a medium access control (MAC) protocol. Usually, DCCS impose real-time constraints. In essence, by real-time constraints we mean that traffic must be sent and received within a bounded interval, otherwise a timing fault is said to occur. This motivates the use of communication networks with a MAC protocol that guarantees bounded access and response times to message requests. PROFIBUS is a communication network in which the MAC protocol is based on a simplified version of the timed-token protocol. In this paper we address the cycle time properties of the PROFIBUS MAC protocol, since the knowledge of these properties is of paramount importance for guaranteeing the real-time behaviour of a distributed computer-controlled system which is supported by this type of network.

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The marriage of emerging information technologies with control technologies is a major driving force that, in the context of the factory-floor, is creating an enormous eagerness for extending the capabilities of currently available fieldbus networks to cover functionalities not considered up to a recent past. Providing wireless capabilities to such type of communication networks is a big share of that effort. The RFieldbus European project is just one example, where PROFIBUS was provided with suitable extensions for implementing hybrid wired/wireless communication systems. In RFieldbus, interoperability between wired and wireless components is achieved by the use specific intermediate networking systems operating as repeaters, thus creating a single logical ring (SLR) network. The main advantage of the SLR approach is that the effort for protocol extensions is not significant. However, a multiple logical ring (MLR) approach provides traffic and error isolation between different network segments. This concept was introduced in, where an approach for a bridge-based architecture was briefly outlined. This paper will focus on the details of the inter-Domain Protocol (IDP), which is responsible for handling transactions between different network domains (wired or wireless) running the PROFIBUS protocol.

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Future industrial control/multimedia applications will increasingly impose or benefit from wireless and mobile communications. Therefore, there is an enormous eagerness for extending currently available industrial communications networks with wireless and mobility capabilities. The RFieldbus European project is just one example, where a PROFIBUS-based hybrid (wired/wireless) architecture was specified and implemented. In the RFieldbus architecture, interoperability between wired and wireless components is achieved by the use specific intermediate networking systems operating at the physical layer level, i.e. operating as repeaters. Instead, in this paper we will focus on a bridge-based approach, which presents several advantages. This concept was introduced in (Ferreira, et al., 2002), where a bridge-based approach was briefly outlined. Then, a specific Inter-Domain Protocol (IDP) was proposed to handle the Inter-Domain transactions in such a bridge-based approach (Ferreira, et al., 2003a). The major contribution of this paper is in extending these previous works by describing the protocol extensions to support inter-cell mobility in such a bridge-based hybrid wired/wireless PROFIBUS networks.

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Coronary artery disease (CAD) is currently one of the most prevalent diseases in the world population and calcium deposits in coronary arteries are one direct risk factor. These can be assessed by the calcium score (CS) application, available via a computed tomography (CT) scan, which gives an accurate indication of the development of the disease. However, the ionising radiation applied to patients is high. This study aimed to optimise the protocol acquisition in order to reduce the radiation dose and explain the flow of procedures to quantify CAD. The main differences in the clinical results, when automated or semiautomated post-processing is used, will be shown, and the epidemiology, imaging, risk factors and prognosis of the disease described. The software steps and the values that allow the risk of developingCADto be predicted will be presented. A64-row multidetector CT scan with dual source and two phantoms (pig hearts) were used to demonstrate the advantages and disadvantages of the Agatston method. The tube energy was balanced. Two measurements were obtained in each of the three experimental protocols (64, 128, 256 mAs). Considerable changes appeared between the values of CS relating to the protocol variation. The predefined standard protocol provided the lowest dose of radiation (0.43 mGy). This study found that the variation in the radiation dose between protocols, taking into consideration the dose control systems attached to the CT equipment and image quality, was not sufficient to justify changing the default protocol provided by the manufacturer.

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This paper addresses sensor network applications which need to obtain an accurate image of physical phenomena and do so with a high sampling rate in both time and space. We present a fast and scalable approach for obtaining an approximate representation of all sensor readings at high sampling rate for quickly reacting to critical events in a physical environment. This approach is an improvement on previous work in that after the new approach has undergone a startup phase then the new approach can use a very small sampling period.

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We consider reliable communications in Body Area Networks (BAN), where a set of nodes placed on human body are connected using wireless links. In order to keep the Specific Absorption Rate (SAR) as low as possible for health safety reasons, these networks operate in low transmit power regime, which however, is known to be error prone. It has been observed that the fluctuations of the Received Signal Strength (RSS) at the nodes of a BAN on a moving person show certain regularities and that the magnitude of these fluctuations are significant (5 - 20 dB). In this paper, we present BANMAC, a MAC protocol that monitors and predicts the channel fluctuations and schedules transmissions opportunistically when the RSS is likely to be higher. The MAC protocol is capable of providing differentiated service and resolves co-channel interference in the event of multiple co-located BANs in a vicinity. We report the design and implementation details of BANMAC integrated with the IEEE 802.15.4 protocol stack. We present experimental data which show that the packet loss rate (PLR) of BANMAC is significantly lower as compared to that of the IEEE 802.15.4 MAC. For comparable PLR, the power consumption of BANMAC is also significantly lower than that of the IEEE 802.15.4. For co-located networks, the convergence time to find a conflict-free channel allocation was approximately 1 s for the centralized coordination mechanism and was approximately 4 s for the distributed coordination mechanism.