Conditional and inducible transgene expression in endothelial and hematopoietic cells using Cre/loxP and tetracycline-off systems

In the present study, the tetracycline-off and Cre/loxP systems were combined to gain temporal and spatial control of transgene expression. Mice were generated that carried three transgenes: Tie2-tTA, tet-O-Cre and either the ZEG or ZAP reporter. Tie2-tTA directs expression of tetracycline-controlled transactivator (tTA) in endothelial and hematopoietic cells under the control of the Tie2 promoter. Tet-O-Cre produces Cre recombinase from a minimal promoter containing the tet-operator (tetO). ZEG or ZAP contains a strong promoter and a loxP-flanked stop sequence, followed by an enhanced green fluorescence protein (EGFP) or human placental alkaline phosphatase (hPLAP) reporter. In the presence of tetracycline, the tTA transactivator produced by Tie-2-tTA is disabled and Cre is not expressed. In the absence of tetracycline, the tTA binds tet-O-Cre to drive the expression of Cre, which recombines the loxP sites of the ZEG or ZAP transgene and results in reporter gene expression. In the present study, the expression of the ZEG or ZAP reporter genes in embryos and adult animals with and without tetracycline treatment was examined. In the presence of tetracycline, no reporter gene expression was observed. When tetracycline was withdrawn, Cre excision was activated and the reporter genes were detected in endothelial and hematopoietic cells. These results demonstrate that this system may be used to bypass embryonic lethality and access adult phenotypes.

Biomechanical evaluation of different angle-stable locking plate systems for mandibular surgery

PURPOSE To compare the initial stability and stability after fatigue of three different locking systems (Synthes(®), Stryker(®) and Medartis(®)) for mandibular fixation and reconstruction. METHOD Standard mandible locking plates with identical profile height (1,5 mm), comparable length and screws with identical diameter (2,0 mm) were used. Plates were fixed with six screws according a preparation protocol. Four point bending tests were then performed using artificial bone material to compare their initial stability and failure limit under realistic loading conditions. Loading of the plates was performed using of a servo hydraulic driven testing machine. The stiffness of the implant/bone construct was calculated using a linear regression on the experimental data included in a range of applied moment between 2 Nm and 6 Nm. RESULTS No statistical difference in the elastic stiffness was visible between the three types of plate. However, differences were observed between the systems concerning the maximal load supported. The Stryker and Synthes systems were able to support a significantly higher moment. CONCLUSION For clinical application all systems show good and reliable results. Practical aspects such as handling, possible angulation of screw fixation, possibility of screw/plate removal, etc. may favour one or the other plating system.

Computer simulations of sample preconcentration in carrier-free systems and isoelectric focusing in microchannels using simple ampholytes.

In this work, electrophoretic preconcentration of protein and peptide samples in microchannels was studied theoretically using the 1D dynamic simulator GENTRANS, and experimentally combined with MS. In all configurations studied, the sample was uniformly distributed throughout the channel before power application, and driving electrodes were used as microchannel ends. In the first part, previously obtained experimental results from carrier-free systems are compared to simulation results, and the effects of atmospheric carbon dioxide and impurities in the sample solution are examined. Simulation provided insight into the dynamics of the transport of all components under the applied electric field and revealed the formation of a pure water zone in the channel center. In the second part, the use of an IEF procedure with simple well defined amphoteric carrier components, i.e. amino acids, for concentration and fractionation of peptides was investigated. By performing simulations a qualitative description of the analyte behavior in this system was obtained. Neurotensin and [Glu1]-Fibrinopeptide B were separated by IEF in microchannels featuring a liquid lid for simple sample handling and placement of the driving electrodes. Component distributions in the channel were detected using MALDI- and nano-ESI-MS and data were in agreement with those obtained by simulation. Dynamic simulations are demonstrated to represent an effective tool to investigate the electrophoretic behavior of all components in the microchannel.

αvβ8 Integrin Interacts with RhoGDI1 to Regulate Rac1 and Cdc42 Activation and Drive Glioblastoma Cell Invasion

As an interface between the circulatory and central nervous systems, the neurovascular unit is vital to the development and survival of tumors. The malignant brain cancer glioblastoma multiforme (GBM) displays invasive growth behaviors that are major impediments to surgical resection and targeted therapies. Adhesion and signaling pathways that drive GBM cell invasion remain largely uncharacterized. Here we have utilized human GBM cell lines, primary patient samples, and pre-clinical mouse models to demonstrate that integrin αvβ8 is a major driver of GBM cell invasion. β8 integrin is overexpressed in many human GBM cells, with higher integrin expression correlating with increased invasion and diminished patient survival. Silencing β8 integrin in human GBM cells leads to impaired tumor cell invasion due to hyperactivation of the Rho GTPases Rac1 and Cdc42. β8 integrin associates with Rho GDP Dissociation Inhibitor 1 (RhoGDI1), an intracellular signaling effector that sequesters Rho GTPases in their inactive GDP-bound states. Silencing RhoGDI1 expression or uncoupling αvβ8 integrin-RhoGDI1 protein interactions blocks GBM cell invasion due to Rho GTPase hyperactivation. These data reveal for the first time that αvβ8 integrin, via interactions with RhoGDI1, suppresses activation of Rho proteins to promote GBM cell invasiveness. Hence, targeting the αvβ8 integrin-RhoGDI1 signaling axis may be an effective strategy for blocking GBM cell invasion.

Magnetic susceptibility and electric conductivity of marine surficial sediments by benthic electromagnetic profiling

Distribution, accumulation and diagenesis of surficial sediments in coastal and continental shelf systems follow complex chains of localized processes and form deposits of great spatial variability. Given the environmental and economic relevance of ocean margins, there is growing need for innovative geophysical exploration methods to characterize seafloor sediments by more than acoustic properties. A newly conceptualized benthic profiling and data processing approach based on controlled source electromagnetic (CSEM) imaging permits to coevally quantify the magnetic susceptibility and the electric conductivity of shallow marine deposits. The two physical properties differ fundamentally insofar as magnetic susceptibility mostly assesses solid particle characteristics such as terrigenous or iron mineral content, redox state and contamination level, while electric conductivity primarily relates to the fluid-filled pore space and detects salinity, porosity and grain-size variations. We develop and validate a layered half-space inversion algorithm for submarine multifrequency CSEM with concentric sensor configuration. Guided by results of modeling, we modified a commercial land CSEM sensor for submarine application, which was mounted into a nonconductive and nonmagnetic bottom-towed sled. This benthic EM profiler Neridis II achieves 25 soundings/second at 3-4 knots over continuous profiles of up to hundred kilometers. Magnetic susceptibility is determined from the 75 Hz in-phase response (90% signal originates from the top 50 cm), while electric conductivity is derived from the 5 kHz out-of-phase (quadrature) component (90% signal from the top 92 cm). Exemplary survey data from the north-west Iberian margin underline the excellent sensitivity, functionality and robustness of the system in littoral (~0-50 m) and neritic (~50-300 m) environments. Susceptibility vs. porosity cross-plots successfully identify known lithofacies units and their transitions. All presently available data indicate an eminent potential of CSEM profiling for assessing the complex distribution of shallow marine surficial sediments and for revealing climatic, hydrodynamic, diagenetic and anthropogenic factors governing their formation.

Methods and Tools for Objective Assessment of Psychomotor Skills in Laparoscopic Surgery

Training and assessment paradigms for laparoscopic surgical skills are evolving from traditional mentor–trainee tutorship towards structured, more objective and safer programs. Accreditation of surgeons requires reaching a consensus on metrics and tasks used to assess surgeons’ psychomotor skills. Ongoing development of tracking systems and software solutions has allowed for the expansion of novel training and assessment means in laparoscopy. The current challenge is to adapt and include these systems within training programs, and to exploit their possibilities for evaluation purposes. This paper describes the state of the art in research on measuring and assessing psychomotor laparoscopic skills. It gives an overview on tracking systems as well as on metrics and advanced statistical and machine learning techniques employed for evaluation purposes. The later ones have a potential to be used as an aid in deciding on the surgical competence level, which is an important aspect when accreditation of the surgeons in particular, and patient safety in general, are considered. The prospective of these methods and tools make them complementary means for surgical assessment of motor skills, especially in the early stages of training. Successful examples such as the Fundamentals of Laparoscopic Surgery should help drive a paradigm change to structured curricula based on objective parameters. These may improve the accreditation of new surgeons, as well as optimize their already overloaded training schedules.

The Road from Community Ideas to Organisational Innovation: A Life Cycle Survey of Idea Management Systems

This paper introduces a new emerging software component, the idea management system, which helps to gather, organise, select and manage the innovative ideas provided by the communities gathered around organisations or enterprises. We define the notion of the idea life cycle, which provides a framework for characterising tools and techniques that drive the evolution of community submitted data inside idea management systems. Furthermore, we show the dependencies between the community-created information and the enterprise processes that are a result of using idea management systems and point out the possible benefits.

Development of a grid connected micro wind generator. A practical activity for the course on electric generation with wind energy

This paper describes a practical activity, part of the renewable energy course where the students have to build their own complete wind generation system, including blades, PM-generator, power electronics and control. After connecting the system to the electric grid the system has been tested during real wind scenarios. The paper will describe the electric part of the work surface-mounted permanent magnet machine design criteria as well as the power electronics part for the power control and the grid connection. A Kalman filter is used for the voltage phase estimation and current commands obtained in order to control active and reactive power. The connection to the grid has been done and active and reactive power has been measured in the system.

Performance optimization in switched reluctance motor drives

In this paper, switched reluctance motors (SRM) are proposed as an alternative for electric power assisted steering (EPAS) applications. A prototype machine has been developed as very attractive design for a steering electric motor, both from a cost and size perspective. A fourphase 8/6 SRM drive is designed for a rack type EPAS which should provide a maximum force of 10 kN. Two-dimension finite element analysis is used to validate the design.

A documentation generator for (C)LP systems

We describe lpdoc, a tool which generates documentation manuals automatically from one or more logic program source files, written in Ciao, ISO-Prolog, and other (C)LP languages. It is particularly useful for documenting library modules, for which it automatically generates a rich description of the module interface. However, it can also be used quite successfully to document full applications. A fundamental advantage of using lpdoc is that it helps maintaining a true correspondence between the program and its documentation, and also identifying precisely to what versión of the program a given printed manual corresponds. The quality of the documentation generated can be greatly enhanced by including within the program text assertions (declarations with types, modes, etc. ...) for the predicates in the program, and machine-readable comments. One of the main novelties of lpdoc is that these assertions and comments are written using the Ciao system asseriion language, which is also the language of communication between the compiler and the user and between the components of the compiler. This allows a significant synergy among specification, debugging, documentation, optimization, etc. A simple compatibility library allows conventional (C)LP systems to ignore these assertions and comments and treat normally programs documented in this way. The documentation can be generated interactively from emacs or from the command line, in many formats including texinfo, dvi, ps, pdf, info, ascii, html/css, Unix nroff/man, Windows help, etc., and can include bibliographic citations and images, lpdoc can also genérate "man" pages (Unix man page format), nicely formatted plain ASCII "readme" files, installation scripts useful when the manuals are included in software distributions, brief descriptions in html/css or info formats suitable for inclusión in on-line Índices of manuals, and even complete WWW and info sites containing on-line catalogs of documents and software distributions. The lpdoc manual, all other Ciao system manuals, and parts of this paper are generated by lpdoc.

An abstract machine for restricted and-parallel execution of logic programs

Although the sequential execution speed of logic programs has been greatly improved by the concepts introduced in the Warren Abstract Machine (WAM), parallel execution represents the only way to increase this speed beyond the natural limits of sequential systems. However, most proposed parallel logic programming execution models lack the performance optimizations and storage efficiency of sequential systems. This paper presents a parallel abstract machine which is an extension of the WAM and is thus capable of supporting ANDParallelism without giving up the optimizations present in sequential implementations. A suitable instruction set, which can be used as a target by a variety of logic programming languages, is also included. Special instructions are provided to support a generalized version of "Restricted AND-Parallelism" (RAP), a technique which reduces the overhead traditionally associated with the run-time management of variable binding conflicts to a series of simple run-time checks, which select one out of a series of compiled execution graphs.

A documentation generator for logic programming systems

We describe lpdoc, a tool which generates documentation manuals automatically from one or more logic program source files, written in ISO-Prolog, Ciao, and other (C)LP languages. It is particularly useful for documenting library modules, for which it automatically generates a rich description of the module interface. However, it can also be used quite successfully to document full applications. The documentation can be generated in many formats including t e x i n f o, dvi, ps, pdf, inf o, html/css, Unix nrof f/man, Windows help, etc., and can include bibliographic citations and images, lpdoc can also genérate "man" pages (Unix man page format), nicely formatted plain ascii "readme" files, installation scripts useful when the manuals are included in software distributions, brief descriptions in html/css or inf o formats suitable for inclusión in on-line Índices of manuals, and even complete WWW and inf o sites containing on-line catalogs of documents and software distributions. A fundamental advantage of using lpdoc is that it helps maintaining a true correspondence between the program and its documentation, and also identifying precisely to what versión of the program a given printed manual corresponds. The quality of the documentation generated can be greatly enhanced by including within the program text assertions (declarations with types, modes, etc. ...) for the predicates in the program, and machine-readable comments. These assertions and comments are written using the Ciao system assertion language. A simple compatibility library allows conventional (C)LP systems to ignore these assertions and comments and treat normally programs documented in this way. The lpdoc manual, all other Ciao system manuals, and most of this paper, are generated by lpdoc.

An abstract machine based execution model for computer architecture design and efficient implementation of logic programs in parallel.

The term "Logic Programming" refers to a variety of computer languages and execution models which are based on the traditional concept of Symbolic Logic. The expressive power of these languages offers promise to be of great assistance in facing the programming challenges of present and future symbolic processing applications in Artificial Intelligence, Knowledge-based systems, and many other areas of computing. The sequential execution speed of logic programs has been greatly improved since the advent of the first interpreters. However, higher inference speeds are still required in order to meet the demands of applications such as those contemplated for next generation computer systems. The execution of logic programs in parallel is currently considered a promising strategy for attaining such inference speeds. Logic Programming in turn appears as a suitable programming paradigm for parallel architectures because of the many opportunities for parallel execution present in the implementation of logic programs. This dissertation presents an efficient parallel execution model for logic programs. The model is described from the source language level down to an "Abstract Machine" level suitable for direct implementation on existing parallel systems or for the design of special purpose parallel architectures. Few assumptions are made at the source language level and therefore the techniques developed and the general Abstract Machine design are applicable to a variety of logic (and also functional) languages. These techniques offer efficient solutions to several areas of parallel Logic Programming implementation previously considered problematic or a source of considerable overhead, such as the detection and handling of variable binding conflicts in AND-Parallelism, the specification of control and management of the execution tree, the treatment of distributed backtracking, and goal scheduling and memory management issues, etc. A parallel Abstract Machine design is offered, specifying data areas, operation, and a suitable instruction set. This design is based on extending to a parallel environment the techniques introduced by the Warren Abstract Machine, which have already made very fast and space efficient sequential systems a reality. Therefore, the model herein presented is capable of retaining sequential execution speed similar to that of high performance sequential systems, while extracting additional gains in speed by efficiently implementing parallel execution. These claims are supported by simulations of the Abstract Machine on sample programs.

A Satisfaction-based Model for Affect Recognition from Conversational Features in Spoken Dialog Systems

Detecting user affect automatically during real-time conversation is the main challenge towards our greater aim of infusing social intelligence into a natural-language mixed-initiative High-Fidelity (Hi-Fi) audio control spoken dialog agent. In recent years, studies on affect detection from voice have moved on to using realistic, non-acted data, which is subtler. However, it is more challenging to perceive subtler emotions and this is demonstrated in tasks such as labelling and machine prediction. This paper attempts to address part of this challenge by considering the role of user satisfaction ratings and also conversational/dialog features in discriminating contentment and frustration, two types of emotions that are known to be prevalent within spoken human-computer interaction. However, given the laboratory constraints, users might be positively biased when rating the system, indirectly making the reliability of the satisfaction data questionable. Machine learning experiments were conducted on two datasets, users and annotators, which were then compared in order to assess the reliability of these datasets. Our results indicated that standard classifiers were significantly more successful in discriminating the abovementioned emotions and their intensities (reflected by user satisfaction ratings) from annotator data than from user data. These results corroborated that: first, satisfaction data could be used directly as an alternative target variable to model affect, and that they could be predicted exclusively by dialog features. Second, these were only true when trying to predict the abovementioned emotions using annotator?s data, suggesting that user bias does exist in a laboratory-led evaluation.

Benchmarking for stability evaluation of computer systems

In this paper we propose a new benchmark to drive making decisions in maintenance of computer systems. This benchmark is made from load average sample data. The main goal is to improve reliability and performance of a set of devices or components. In particular, the stability of the system is measured in terms of variability of the load. A forecast of the behavior of this stability is also proposal as part of the reporting benchmark. At the final stage, a more stable system is obtained and its global reliability and performance can be then evaluated by means of appropriate specifications.