Enhancing composite Digital Documents Using XML-based Standoff Markup

Document representations can rapidly become unwieldy if they try to encapsulate all possible document properties, ranging from abstract structure to detailed rendering and layout. We present a composite document approach wherein an XMLbased document representation is linked via a shadow tree of bi-directional pointers to a PDF representation of the same document. Using a two-window viewer any material selected in the PDF can be related back to the corresponding material in the XML, and vice versa. In this way the treatment of specialist material such as mathematics, music or chemistry (e.g. via read aloud or play aloud ) can be activated via standard tools working within the XML representation, rather than requiring that application-specific structures be embedded in the PDF itself. The problems of textual recognition and tree pattern matching between the two representations are discussed in detail. Comparisons are drawn between our use of a shadow tree of pointers to map between document representations and the use of a code-replacement shadow tree in technologies such as XBL.

DESIGN OF EFFICIENT IN-NETWORK DATA PROCESSING AND DISSEMINATION FOR VANETS

By providing vehicle-to-vehicle and vehicle-to-infrastructure wireless communications, vehicular ad hoc networks (VANETs), also known as the “networks on wheels”, can greatly enhance traffic safety, traffic efficiency and driving experience for intelligent transportation system (ITS). However, the unique features of VANETs, such as high mobility and uneven distribution of vehicular nodes, impose critical challenges of high efficiency and reliability for the implementation of VANETs. This dissertation is motivated by the great application potentials of VANETs in the design of efficient in-network data processing and dissemination. Considering the significance of message aggregation, data dissemination and data collection, this dissertation research targets at enhancing the traffic safety and traffic efficiency, as well as developing novel commercial applications, based on VANETs, following four aspects: 1) accurate and efficient message aggregation to detect on-road safety relevant events, 2) reliable data dissemination to reliably notify remote vehicles, 3) efficient and reliable spatial data collection from vehicular sensors, and 4) novel promising applications to exploit the commercial potentials of VANETs. Specifically, to enable cooperative detection of safety relevant events on the roads, the structure-less message aggregation (SLMA) scheme is proposed to improve communication efficiency and message accuracy. The scheme of relative position based message dissemination (RPB-MD) is proposed to reliably and efficiently disseminate messages to all intended vehicles in the zone-of-relevance in varying traffic density. Due to numerous vehicular sensor data available based on VANETs, the scheme of compressive sampling based data collection (CS-DC) is proposed to efficiently collect the spatial relevance data in a large scale, especially in the dense traffic. In addition, with novel and efficient solutions proposed for the application specific issues of data dissemination and data collection, several appealing value-added applications for VANETs are developed to exploit the commercial potentials of VANETs, namely general purpose automatic survey (GPAS), VANET-based ambient ad dissemination (VAAD) and VANET based vehicle performance monitoring and analysis (VehicleView). Thus, by improving the efficiency and reliability in in-network data processing and dissemination, including message aggregation, data dissemination and data collection, together with the development of novel promising applications, this dissertation will help push VANETs further to the stage of massive deployment.

Calibration of the detector flight models for the HERMES and SPIRIT nanosatellite missions

The High Energy Rapid Modular Ensemble of Satellites (HERMES) is a new mission concept involving the development of a constellation of six CubeSats in low Earth orbit with new miniaturized instruments that host a hybrid Silicon Drift Detector/GAGG:Ce based system for X-ray and γ-ray detection, aiming to monitor high-energy cosmic transients, such as Gamma Ray Bursts and the electromagnetic counterparts of gravitational wave events. The HERMES constellation will also operate together with the Australian-Italian SpIRIT mission, which will house a HERMES-like detector. The HERMES pathfinder mini-constellation, consisting of six satellites plus SpIRIT, is likely to be launched in 2023. The HERMES detectors are based on the heritage of the Italian ReDSoX collaboration, with joint design and production by INFN-Trieste and Fondazione Bruno Kessler, and the involvement of several Italian research institutes and universities. An application-specific, low-noise, low-power integrated circuit (ASIC) called LYRA was conceived and designed for the HERMES readout electronics. My thesis project focuses on the ground calibrations of the first HERMES and SpIRIT flight detectors, with a performance assessment and characterization of the detectors. The first part of this work addresses measurements and experimental tests on laboratory prototypes of the HERMES detectors and their front-end electronics, while the second part is based on the design of the experimental setup for flight detector calibrations and related functional tests for data acquisition, as well as the development of the calibration software. In more detail, the calibration parameters (such as the gain of each detector channel) are determined using measurements with radioactive sources, performed at different operating temperatures between -20°C and +20°C by placing the detector in a suitable climate chamber. The final part of the thesis involves the analysis of the calibration data and a discussion of the results.

Domain-Specific Multi-Modeling of Security Concerns in Service-Oriented Architectures

As a common reference for many in-development standards and execution frameworks, special attention is being paid to Service-Oriented Architectures. SOAs modeling, however, is an area in which a consensus has not being achieved. Currently, standardization organizations are defining proposals to offer a solution to this problem. Nevertheless, until very recently, non-functional aspects of services have not been considered for standardization processes. In particular, there exists a lack of a design solution that permits an independent development of the functional and non-functional concerns of SOAs, allowing that each concern be addressed in a convenient manner in early stages of the development, in a way that could guarantee the quality of this type of systems. This paper, leveraging on previous work, presents an approach to integrate security-related non-functional aspects (such as confidentiality, integrity, and access control) in the development of services.

Multi Domain-Specific Modeling of the Security Concerns of Service-Oriented Architectures

Service-Oriented Architectures (SOA), and Web Services (WS), the technology generally used to implement them, achieve the integration of heterogeneous technologies, providing interoperability, and yielding the reutilization of pre-existent systems. Model-driven development methodologies provide inherent benefits such as increased productivity, greater reuse, and better maintainability, to name a few. Efforts on achieving model-driven development of SOAs already exist, but there is currently no standard solution that addresses non-functional aspects of these services as well. This paper presents an approach to integrate these non-functional aspects in the development of web services, with an emphasis on security.

Application of a species-specific PCR-RFLP to identify Ancylostoma eggs directly from canine faeces

Apart from their veterinary importance, the hookworms Ancylostoma caninum, Ancylostoma braziliense and Ancylostoma caninum are also capable of causing zoonotic disease in humans. A highly sensitive and species-specific PCR-RFLP technique was utilised to detect and differentiate the various canine Ancylostoma spp directly from eggs in faeces. This technique was utilised to screen 101 canine faecal samples from parasite endemic tea growing communities in Assam, India, as part as an ongoing epidemiological investigation into canine parasitic zoonoses. The prevalence of hookworms in dogs was found to be 98% using a combination of PCR and conventional microscopy. Overall, 36% of dogs were found positive for single hookworm infections with A. caninum, 24% positive for single infections with A. braziliense and 38% had mixed infections with both A. caninum and A. braziliense. No dogs were found positive for A. ceylanicum in the community under study. The high prevalence of A. caninum and A. braziliense in dogs in this community may account for the high incidence of cutaneous larva migrans (CLM) observed among the human population residing at the tea estates. The PCR-RFLP technique described herein allows epidemiological screening of canine hookworms to be conducted rapidly, with ease and accuracy, and has the potential to be applied to a number of different clinical, pharmacological and epidemiological situations. (C) 2004 Elsevier B.V. All rights reserved.

Application of recombinant antibody library for screening specific antigens in a bovine sperm cell subpopulation

Antibody phage display libraries are a useful tool in proteomic analyses. This study evaluated an antibody recombinant library for identification of sex-specific proteins on the sperm cell surface. The Griffin.1 library was used to produce phage antibodies capable of recognizing membrane proteins from Nelore sperm cells. After producing soluble monoclonal scFv, clones were screened on Simental sperm cells by flow cytometry and those that bound to 40-60% of cells were selected. These clones were re-analyzed using Nelore sperm cells and all clones bound to 40-60% of cells. Positive clones were submitted to a binding assay against male and female bovine leukocytes by flow cytometry and one clone preferentially bound to male cells. The results indicate that phage display antibodies are an alternative method for identification of molecules markers on sperm cells. (C) 2007 Elsevier B.V. All rights reserved.

A sensitive and specific assay for glutathione with potential application to glutathione disulphide, using high-performance liquid chromatography-tandem mass spectrometry

We have utilised the combination of sensitivity and specificity afforded by coupling high-performance liquid chromatography (HPLC) to a tandem mass spectrometer (MS-MS) to produce an assay which is suitable for assaying glutathione (GSH) concentrations in liver tissue. The sensitivity suggests it may also be suitable for extrahepatic tissues, The method has been validated for GSH using mouse liver samples and also allows the assay of GSSG. The stability of GSH under conditions relevant to the assay has been determined. A 20-mul amount of a diluted methanol extract of tissue is injected with detection limits of 0.2 pmol for GSH and 2 pmol for GSSG. The HPLC uses an Altima C-18 (150X4.6 mm, 5 mum) column at 35 degreesC. Chromatography utilises a linear gradient from 0 to 10% methanol in 0.1% formic acid over 5 min, with a final isocratic stage holding at 10% methanol for 5 min. Total flow rate is 0.8 ml/min. The transition from the M+H ion (308.1 m/z for GSH, and 613.3 m/z for GSSG) to the 162.0 m/z (GSH) and 355.3 m/z (GSSG) fragments are monitored. (C) 2001 Elsevier Science B.V. All rights reserved.

Domain-Specific Language for Coordination patterns

The integration and composition of software systems requires a good architectural design phase to speed up communications between (remote) components. However, during implementation phase, the code to coordinate such components often ends up mixed in the main business code. This leads to maintenance problems, raising the need for, on the one hand, separating the coordination code from the business code, and on the other hand, providing mechanisms for analysis and comprehension of the architectural decisions once made. In this context our aim is at developing a domain-specific language, CoordL, to describe typical coordination patterns. From our point of view, coordination patterns are abstractions, in a graph form, over the composition of coordination statements from the system code. These patterns would allow us to identify, by means of pattern-based graph search strategies, the code responsible for the coordination of the several components in a system. The recovering and separation of the architectural decisions for a better comprehension of the software is the main purpose of this pattern language

Optical processing devices based on a-SiCH multilayer architectures

Red, green and blue optical signals were directed to an a-SiC:H multilayered device, each one with a specific transmission rate. The combined optical signal was analyzed by reading out, under different applied voltages, the generated photocurrent. Results show that when a chromatic time dependent wavelength combination with different transmission rates irradiates the multilayered structure, the device operates as a tunable wavelength filter and can be used in wavelength division multiplexing systems for short range communications. An application to fluorescent proteins detection is presented. (C) 2010 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim

Oriented Tailoring of Plastic Antibodies for Prostate Specific Antigen and Application of the Imprinted Material as Ionophore in Potentiometric Detection

Prostate Specific Antigen (PSA) is the biomarker of choice for screening prostate cancer throughout the population, with PSA values above 10 ng/mL pointing out a high probability of associated cancer1. According to the most recent World Health Organization (WHO) data, prostate cancer is the commonest form of cancer in men in Europe2. Early detection of prostate cancer is thus very important and is currently made by screening PSA in men over 45 years old, combined with other alterations in serum and urine parameters. PSA is a glycoprotein with a molecular mass of approximately 32 kDa consisting of one polypeptide chain, which is produced by the secretory epithelium of human prostate. Currently, the standard methods available for PSA screening are immunoassays like Enzyme-Linked Immunoabsorbent Assay (ELISA). These methods are highly sensitive and specific for the detection of PSA, but they require expensive laboratory facilities and high qualify personal resources. Other highly sensitive and specific methods for the detection of PSA have also become available and are in its majority immunobiosensors1,3-5, relying on antibodies. Less expensive methods producing quicker responses are thus needed, which may be achieved by synthesizing artificial antibodies by means of molecular imprinting techniques. These should also be coupled to simple and low cost devices, such as those of the potentiometric kind, one approach that has been proven successful6. Potentiometric sensors offer the advantage of selectivity and portability for use in point-of-care and have been widely recognized as potential analytical tools in this field. The inherent method is simple, precise, accurate and inexpensive regarding reagent consumption and equipment involved. Thus, this work proposes a new plastic antibody for PSA, designed over the surface of graphene layers extracted from graphite. Charged monomers were used to enable an oriented tailoring of the PSA rebinding sites. Uncharged monomers were used as control. These materials were used as ionophores in conventional solid-contact graphite electrodes. The obtained results showed that the imprinted materials displayed a selective response to PSA. The electrodes with charged monomers showed a more stable and sensitive response, with an average slope of -44.2 mV/decade and a detection limit of 5.8X10-11 mol/L (2 ng/mL). The corresponding non-imprinted sensors showed smaller sensitivity, with average slopes of -24.8 mV/decade. The best sensors were successfully applied to the analysis of serum samples, with percentage recoveries of 106.5% and relatives errors of 6.5%.

Development of a new chlamydiales-specific real-time PCR and its application to respiratory clinical samples.

Originally composed of the single family Chlamydiaceae, the Chlamydiales order has extended considerably over the last several decades. Chlamydia-related bacteria were added and classified into six different families and family-level lineages: the Criblamydiaceae, Parachlamydiaceae, Piscichlamydiaceae, Rhabdochlamydiaceae, Simkaniaceae, and Waddliaceae. While several members of the Chlamydiaceae family are known pathogens, recent studies showed diverse associations of Chlamydia-related bacteria with human and animal infections. Some of these latter bacteria might be of medical importance since, given their ability to replicate in free-living amoebae, they may also replicate efficiently in other phagocytic cells, including cells of the innate immune system. Thus, a new Chlamydiales-specific real-time PCR targeting the conserved 16S rRNA gene was developed. This new molecular tool can detect at least five DNA copies and show very high specificity without cross-amplification from other bacterial clade DNA. The new PCR was validated with 128 clinical samples positive or negative for Chlamydia trachomatis or C. pneumoniae. Of 65 positive samples, 61 (93.8%) were found to be positive with the new PCR. The four discordant samples, retested with the original test, were determined to be negative or below detection limits. Then, the new PCR was applied to 422 nasopharyngeal swabs taken from children with or without pneumonia; a total of 48 (11.4%) samples were determined to be positive, and 45 of these were successfully sequenced. The majority of the sequences corresponded to Chlamydia-related bacteria and especially to members of the Parachlamydiaceae family.

Development of a Liver-specific Tet-On Inducible System for AAV Vectors and Its Application in the Treatment of Liver Cancer.

Recombinant adeno-associated virus (rAAV) are effective gene delivery vehicles that can mediate long-lasting transgene expression. However, tight regulation and tissue-specific transgene expression is required for certain therapeutic applications. For regulatable expression from the liver we designed a hepatospecific bidirectional and autoregulatory tetracycline (Tet)-On system (Tet(bidir)Alb) flanked by AAV inverted terminal repeats (ITRs). We characterized the inducible hepatospecific system in comparison with an inducible ubiquitous expression system (Tet(bidir)CMV) using luciferase (luc). Although the ubiquitous system led to luc expression throughout the mouse, luc expression derived from the hepatospecific system was restricted to the liver. Interestingly, the induction rate of the Tet(bidir)Alb was significantly higher than that of Tet(bidir)CMV, whereas leakage of Tet(bidir)Alb was significantly lower. To evaluate the therapeutic potential of this vector, an AAV-Tet(bidir)-Alb-expressing interleukin-12 (IL-12) was tested in a murine model for hepatic colorectal metastasis. The vector induced dose-dependent levels of IL-12 and interferon-γ (IFN-γ), showing no significant toxicity. AAV-Tet(bidir)-Alb-IL-12 was highly efficient in preventing establishment of metastasis in the liver and induced an efficient T-cell memory response to tumor cells. Thus, we have demonstrated persistent, and inducible in vivo expression of a gene from a liver-specific Tet-On inducible construct delivered via an AAV vector and proved to be an efficient tool for treating liver cancer.

Characterization and application of two RANK-specific antibodies with different biological activities.

Antibodies play an important role in therapy and investigative biomedical research. The TNF-family member Receptor Activator of NF-κB (RANK) is known for its role in bone homeostasis and is increasingly recognized as a central player in immune regulation and epithelial cell activation. However, the study of RANK biology has been hampered by missing or insufficient characterization of high affinity tools that recognize RANK. Here, we present a careful description and comparison of two antibodies, RANK-02 obtained by phage display (Newa, 2014 [1]) and R12-31 generated by immunization (Kamijo, 2006 [2]). We found that both antibodies recognized mouse RANK with high affinity, while RANK-02 and R12-31 recognized human RANK with high and lower affinities, respectively. Using a cell apoptosis assay based on stimulation of a RANK:Fas fusion protein, and a cellular NF-κB signaling assay, we showed that R12-31 was agonist for both species. R12-31 interfered little or not at all with the binding of RANKL to RANK, in contrast to RANK-02 that efficiently prevented this interaction. Depending on the assay and species, RANK-02 was either a weak agonist or a partial antagonist of RANK. Both antibodies recognized human Langerhans cells, previously shown to express RANK, while dermal dendritic cells were poorly labeled. In vivo R12-31 agonist activity was demonstrated by its ability to induce the formation of intestinal villous microfold cells in mice. This characterization of two monoclonal antibodies should now allow better evaluation of their application as therapeutic reagents and investigative tools.

Determination of phenobarbital in human plasma by a specific liquid chromatography method: application to a bioequivalence study

A liquid chromatography method was developed and validated for the determination of phenobarbital in human plasma using phenytoin as internal standard. The drugs were extracted from plasma by liquid-liquid extraction and separated isocratically on a C12 analytical column, maintained at 35 ºC, with water:acetonitrile:methanol (58.8:15.2:26, v/v/v) as mobile phase, run at a flow rate of 1.2 mL/min with detection at 205 nm. The method was linear in the range of 0.1-4 μg/mL (r²=0.9999) and demonstrated acceptable results for the precision, accuracy and stability studies. The method was successfully applied for the bioequivalence study of two tablet formulations (test and reference) of phenobarbital 100 mg after single oral dose administration to healthy human volunteers.