Phosphatidylinositol 4,5-bisphosphate clusters act as molecular beacons for vesicle recruitment.

Synaptic-vesicle exocytosis is mediated by the vesicular Ca(2+) sensor synaptotagmin-1. Synaptotagmin-1 interacts with the SNARE protein syntaxin-1A and acidic phospholipids such as phosphatidylinositol 4,5-bisphosphate (PIP2). However, it is unclear how these interactions contribute to triggering membrane fusion. Using PC12 cells from Rattus norvegicus and artificial supported bilayers, we show that synaptotagmin-1 interacts with the polybasic linker region of syntaxin-1A independent of Ca(2+) through PIP2. This interaction allows both Ca(2+)-binding sites of synaptotagmin-1 to bind to phosphatidylserine in the vesicle membrane upon Ca(2+) triggering. We determined the crystal structure of the C2B domain of synaptotagmin-1 bound to phosphoserine, allowing development of a high-resolution model of synaptotagmin bridging two different membranes. Our results suggest that PIP2 clusters organized by syntaxin-1 act as molecular beacons for vesicle docking, with the subsequent Ca(2+) influx bringing the vesicle membrane close enough for membrane fusion.

Molecular beacons with a homo-DNA stem: improving target selectivity

Molecular beacons (MBs) are stem-loop DNA probes used for identifying and reporting the presence and localization of nucleic acid targets in vitro and in vivo via target-dependent dequenching of fluorescence. A drawback of conventional MB design is present in the stem sequence that is necessary to keep the MBs in a closed conformation in the absence of a target, but that can participate in target binding in the open (target-on) conformation, giving rise to the possibility of false-positive results. In order to circumvent these problems, we designed MBs in which the stem was replaced by an orthogonal DNA analog that does not cross-pair with natural nucleic acids. Homo-DNA seemed to be specially suited, as it forms stable adenine-adenine base pairs of the reversed Hoogsteen type, potentially reducing the number of necessary building blocks for stem design to one. We found that MBs in which the stem part was replaced by homo-adenylate residues can easily be synthesized using conventional automated DNA synthesis. As conventional MBs, such hybrid MBs show cooperative hairpin to coil transitions in the absence of a DNA target, indicating stable homo-DNA base pair formation in the closed conformation. Furthermore, our results show that the homo-adenylate stem is excluded from DNA target binding, which leads to a significant increase in target binding selectivity

Analysis of Mycobacterium tuberculosis in the state of Texas for rifampin resistance using molecular beacons

Mycobacterium tuberculosis, a bacillus known to cause disease in humans since ancient times, is the etiological agent of tuberculosis (TB). The infection is primarily pulmonary, although other organs may also be affected. The prevalence of pulmonary TB disease in the US is highest along the US-Mexico border, and of the four US states bordering Mexico, Texas had the second highest percentage of cases of TB disease among Mexico-born individuals in 1999 (CDC, 2001). Between the years of 1993 and 1998, the prevalence of drug-resistant (DR) TB was 9.1% among Mexican-born individuals and 4.4% among US-born individuals (CDC, 2001). In the same time period, the prevalence of multi-drug resistant (MDR) TB was 1.4% among Mexican-born individuals and 0.6% among US-born individuals (CDC, 2001). There is a renewed urgency in the quest for faster and more effective screening, diagnosis, and treatment methods for TB due to the resurgence of tuberculosis in the US during the mid-1980s and early 1990s (CDC, 2007a), and the emergence of drug-resistant, multidrug-resistant, and extremely drug-resistant tuberculosis worldwide. Failure to identify DR and MDR-TB quickly leads to poorer treatment outcomes (CDC, 2007b). The recent rise in TB/HIV comorbidity further complicates TB control efforts. The gold standard for identification of DR-TB requires mycobacterial growth in culture, a technique taking up to three weeks, during which time DR/MDR-TB individuals harboring resistant organisms may be receiving inappropriate treatment. The goal of this study was to determine the sensitivity and specificity of real-time quantitative polymerase chain reaction (qPCR) using molecular beacons in the Texas population. qPCR using molecular beacons is a novel approach to detect mycobacterial mutations conferring drug resistance. This technique is time-efficient and has been shown to have high sensitivity and specificity in several populations worldwide. Rifampin (RIF) susceptibility was chosen as the test parameter because strains of M. tuberculosis which are resistant to RIF are likely to also be MDR. Due to its status as a point of entry for many immigrants into the US, control efforts against TB and drug-resistant TB in Texas is a vital component of prevention efforts in the US as a whole. We show that qPCR using molecular beacons has high sensitivity and specificity when compared with culture (94% and 87%, respectively) and DNA sequencing (90% and 96%, respectively). We also used receiver operator curve analysis to calculate cutoff values for the objective determination of results obtained by qPCR using molecular beacons. ^

Multiplex detection of four pathogenic retroviruses using molecular beacons

We describe a multiplex nucleic acid assay that identifies and determines the abundance of four different pathogenic retroviruses (HIV-1, HIV-2, and human T-lymphotrophic virus types I and II). Retroviral DNA sequences are amplified in a single, sealed tube by simultaneous PCR assays, and the resulting amplicons are detected in real time by the hybridization of four differently colored, amplicon-specific molecular beacons. The color of the fluorescence generated in the course of amplification identifies which retroviruses are present, and the number of thermal cycles required for the intensity of each color to rise significantly above background provides an accurate measure of the number of copies of each retroviral sequence that were present originally in the sample. Fewer than 10 retroviral genomes can be detected. Moreover, 10 copies of a rare retrovirus can be detected in the presence of 100,000 copies of an abundant retrovirus. Ninety-six samples can be analyzed in 3 hr on a single plate, and the use of a closed-tube format eliminates crossover contamination. Utilizing previously well characterized clinical samples, we demonstrate that each of the pathogenic retroviruses can be identified correctly and no false positives occur. This assay enables the rapid and reliable screening of donated blood and transplantable tissues.

Development of guidelines for installation of intersection control beacons. Final report.

Federal Highway Administration, Washington, D.C.

Bonfires to beacons : Federal civil aviation policy under the Air commerce act, 1926-1938 /

Mode of access: Internet.

The American coast pilot : containing the courses and distances between the principal harbours, capes, and headlands, on the coast of North and South America; with directions for sailing into the same; describing the soundings, bearings of the light-houses and beacons from the rocks, shoals, ledges, &c. With the prevailing winds, settings of the currents, &c. and the latitudes and longitudes of the principal harbours and capes. Together with a tide table / By Edmund M. Blunt. To which is annexed, the Shipmaster's assistant; containing disbursements and other ship accounts, &c. /

Issued without the Shipmaster's assistant which was published separately in 1822 under the title: The merchant's and shipmaster's assistant.

Exploring the transformative potential of Bluetooth beacons in higher education

The growing ubiquity of smartphones and tablet devices integrated into personal, social and professional life, facilitated by expansive communication networks globally, has the potential to disrupt higher education. Academics and students are considering the future possibilities of exploiting these tools and utilising networks to consolidate and expand knowledge, enhancing learning gain. Bluetooth beacon technology has been developed by both Apple and Google as a way to situate digital information within physical spaces, and this paper reflects on a beacon intervention in a contemporary art school in higher education conducted by the authors intended to develop a situated community of practice in Art & Design. The paper describes the project, including relevant theoretical foundations and background to the beacon technology, with regards to the potential of using these devices to create a connected learning community by enhancing learning and facilitating knowledge creation in a borderless learning space.

Os faróis da sociedade de informação: uma análise crítica sobre a situação da competência em informação no Brasil

This paper presents and reviews the recommendations done by experts during the specialists meeting held in the city of Alexandria, Egypt, in the end of 2005 and, according to this information, the Brazilian situation is analyzed. Internationalization and institutionalization of information literacy and lifelong learning as essential factors to the development of the nations are also explored. Beacons of the Information Society translate the vision and concepts involved. In Brazil, the actions around information literacy are not a consensus. The challenges to be faced include: to discover forms to foster and to appropriately disseminate national and local knowledge, to advance discussions and deepen the subject, to discover adequate alternatives for disseminating information practices that encompass distinct professional groups and populations, to overcome structural development gaps. As a matter that permeates each and every process of learning, research, development, problem-solving and decision-making, information literacy went beyond the boundaries of librarianship and transformed itself into a world transdiciplinary movement, even under the aegis of different denominations and emphasis.

A Distributed System for the Dissemination of DGPS Data through the Internet

In this paper we describe a low cost distributed system intended to increase the positioning accuracy of outdoor navigation systems based on the Global Positioning System (GPS). Since the accuracy of absolute GPS positioning is insufficient for many outdoor navigation tasks, another GPS based methodology – the Differential GPS (DGPS) – was developed in the nineties. The differential or relative positioning approach is based on the calculation and dissemination of the range errors of the received GPS satellites. GPS/DGPS receivers correlate the broadcasted GPS data with the DGPS corrections, granting users increased accuracy. DGPS data can be disseminated using terrestrial radio beacons, satellites and, more recently, the Internet. Our goal is to provide mobile platforms within our campus with DGPS data for precise outdoor navigation. To achieve this objective, we designed and implemented a three-tier client/server distributed system that, first, establishes Internet links with remote DGPS sources and, then, performs campus-wide dissemination of the obtained data. The Internet links are established between data servers connected to remote DGPS sources and the client, which is the data input module of the campus-wide DGPS data provider. The campus DGPS data provider allows the establishment of both Intranet and wireless links within the campus. This distributed system is expected to provide adequate support for accurate outdoor navigation tasks.

Real Time Internet DGPS Service

The accuracy of the Navigation Satellite Timing and Ranging (NAVSTAR) Global Positioning System (GPS) measurements is insufficient for many outdoor navigation tasks. As a result, in the late nineties, a new methodology – the Differential GPS (DGPS) – was developed. The differential approach is based on the calculation and dissemination of the range errors of the GPS satellites received. GPS/DGPS receivers correlate the broadcasted GPS data with the DGPS corrections, granting users increased accuracy. DGPS data can be disseminated using terrestrial radio beacons, satellites and, more recently, the Internet. Our goal is to provide mobile platforms within our campus with DGPS data for precise outdoor navigation. To achieve this objective, we designed and implemented a three-tier client/server distributed system that establishes Internet links with remote DGPS sources and performs campus-wide dissemination of the obtained data. The Internet links are established between data servers connected to remote DGPS sources and the client, which is the data input module of the campus-wide DGPS data provider. The campus DGPS data provider allows the establishment of both Intranet and wireless links within the campus. This distributed system is expected to provide adequate support for accurate (submetric) outdoor navigation tasks.

Implementation of a Campus wide DGPS Data Server

Although the Navigation Satellite Timing and Ranging (NAVSTAR) Global Positioning System (GPS) is, de facto, the standard positioning system used in outdoor navigation, it does not provide, per se, all the features required to perform many outdoor navigational tasks. The accuracy of the GPS measurements is the most critical issue. The quest for higher position readings accuracy led to the development, in the late nineties, of the Differential Global Positioning System (DGPS). The differential GPS method detects the range errors of the GPS satellites received and broadcasts them. The DGPS/GPS receivers correlate the DGPS data with the GPS satellite data they are receiving, granting users increased accuracy. DGPS data is broadcasted using terrestrial radio beacons, satellites and, more recently, the Internet. Our goal is to have access, within the ISEP campus, to DGPS correction data. To achieve this objective we designed and implemented a distributed system composed of two main modules which are interconnected: a distributed application responsible for the establishment of the data link over the Internet between the remote DGPS stations and the campus, and the campus-wide DGPS data server application. The DGPS data Internet link is provided by a two-tier client/server distributed application where the server-side is connected to the DGPS station and the client-side is located at the campus. The second unit, the campus DGPS data server application, diffuses DGPS data received at the campus via the Intranet and via a wireless data link. The wireless broadcast is intended for DGPS/GPS portable receivers equipped with an air interface and the Intranet link is provided for DGPS/GPS receivers with just a RS232 DGPS data interface. While the DGPS data Internet link servers receive the DGPS data from the DGPS base stations and forward it to the DGPS data Internet link client, the DGPS data Internet link client outputs the received DGPS data to the campus DGPS data server application. The distributed system is expected to provide adequate support for accurate (sub-metric) outdoor campus navigation tasks. This paper describes in detail the overall distributed application.