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Euroconference on the Biology of Type IV Secretion Processes: bacterial gates into the outer world.

Type IV secretion systems (T4SSs) mediate both protein and ssDNA secretion from a wide range of bacteria into virtually any cell type or into the milieu. It is this versatility that confers on them the ability to participate in many processes of bacterial life that imply communication with their environment. Type IV secretion systems are involved in horizontal DNA transfer to other bacteria and to plant cells, in DNA uptake from the milieu, in toxin secretion into the milieu, and in the injection of virulence factors into the eukaryotic host cell in a number of mammalian and plant pathogens. Recently, a EuroConference addressed the different aspects of the biology of these transmembrane multiprotein complexes, from the crystal structure of the individual components to the modification that the secreted substrates induce in the recipient cell. Significant progress has been made in the understanding of the molecular architecture and mechanism of secretion. The analysis of protein-protein interactions confirms the role of coupling proteins as substrate recruiters for the transporter. The VirB10 component of the complex has come up as a strong candidate for signal transducer. The wide range of effects on the recipient suggests that many effector proteins are secreted. New effector proteins are being identified for both plant and animal pathogens, as are their targets within the host cells. New T4SS members are being identified that perform novel roles, beyond DNA transfer and virulence, such as establishment of symbiotic processes. Our current knowledge of the Biology of Type IV Secretion Processes increases our ability to exploit them as biotechnological tools or to use them as new targets for inhibitors that could constitute a new generation of antimicrobials in the near future.

Animals↗

Evolution of surgical skills training.

Surgical training is changing: one hundred years of tradition is being challenged by legal and ethical concerns for patient safety, work hours restrictions, the cost of operating room time, and complications. Surgical simulation and skills training offers an opportunity to teach and practice advanced skills outside of the operating room environment before attempting them on living patients. Simulation training can be as straight forward as using real instruments and video equipment to manipulate simulated "tissue" in a box trainer. More advanced, virtual reality simulators are now available and ready for widespread use. Early systems have demonstrated their effectiveness and discriminative ability. Newer systems enable the development of comprehensive curricula and full procedural simulations. The Accreditation Council of Graduate Medical Education's (ACGME) has mandated the development of novel methods of training and evaluation. Surgical organizations are calling for methods to ensure the maintenance of skills, advance surgical training, and to credential surgeons as technically competent. Simulators in their current form have been demonstrated to improve the operating room performance of surgical residents. Development of standardized training curricula remains an urgent and important agenda, particularly for minimal invasive surgery. An innovative and progressive approach, borrowing experiences from the field of aviation, can provide the foundation for the next century of surgical training, ensuring the quality of the product. As the technology develops, the way we practice will continue to evolve, to the benefit of physicians and patients.

Clinical Competence↗

Recent trends in robot-assisted therapy environments to improve real-life functional performance after stroke.

Upper and lower limb robotic tools for neuro-rehabilitation are effective in reducing motor impairment but they are limited in their ability to improve real world function. There is a need to improve functional outcomes after robot-assisted therapy. Improvements in the effectiveness of these environments may be achieved by incorporating into their design and control strategies important elements key to inducing motor learning and cerebral plasticity such as mass-practice, feedback, task-engagement, and complex problem solving. This special issue presents nine articles. Novel strategies covered in this issue encourage more natural movements through the use of virtual reality and real objects and faster motor learning through the use of error feedback to guide acquisition of natural movements that are salient to real activities. In addition, several articles describe novel systems and techniques that use of custom and commercial games combined with new low-cost robot systems and a humanoid robot to embody the " supervisory presence" of the therapy as possible solutions to exercise compliance in under-supervised environments such as the home.

Editorial↗

The genetic data environment an expandable GUI for multiple sequence analysis.

An X-Windows-based graphic user interface is presented which allows the seamless integration of numerous existing biomolecular programs into a single analysis environment. This environment is based on a core multiple sequence editor that is linked to external programs by a user-expandable menu system and is supported on Sun and DEC workstations. There is no limitation to the number of external functions that can be linked to the interface. The length and number of sequences that can be handled are limited only by the size of virtual memory present on the workstation. The sequence data itself is used as the reference point from which analysis is done, and scalable graphic views are supported. It is suggested that future software development utilizing this expandable, user-defined menu system and the I/O linkage of external programs will allow biologists to easily integrate expertise from disparate fields into a single environment.

Algorithms↗

Mixing realities? An application of augmented reality for the treatment of cockroach phobia.

Augmented reality (AR) refers to the introduction of virtual elements in the real world. That is, the person is seeing an image composed of a visualization of the real world, and a series of virtual elements that, at that same moment, are super-imposed on the real world. The most important aspect of AR is that the virtual elements supply to the person relevant and useful information that is not contained in the real world. AR has notable potential, and has already been used in diverse fields, such as medicine, the army, coaching, engineering, design, and robotics. Until now, AR has never been used in the scope of psychological treatment. Nevertheless, AR presents various advantages. Just like in the classical systems of virtual reality, it is possible to have total control over the virtual elements that are super-imposed on the real world, and how one interacts with those elements. AR could involve additional advantages; on one side it could be less expensive since it also uses the real world (this does not need to be modeled), and it could facilitate the feeling of presence (the sensation of being there), and reality judgment (the fact of judging the experience as real) of the person since the environment he or she is in, and what he or she is seeing is, in fact the "reality." In this paper, we present the data of the first case study in which AR has been used for the treatment of a specific phobia, cockroaches phobia. It addresses a system of AR that permits exposure to virtual cockroaches super-imposed on the real world. In order to carry out the exposure, the guidelines of Ost with respect to "one-session treatment" were followed. The results are promising. The participant demonstrated notable fear and avoidance in the behavioral avoidance test before the treatment, and not only was an important decrease in the scores of fear and avoidance observed after the treatment, but also the participant was capable of approaching, interacting, and killing live cockroaches immediately following the treatment. The results are maintained in a follow-up conducted 1 month after the termination of the treatment.

Adult↗

Nucleosomes: a solution to a crowded intracellular environment?

The emergence of eukaryotes was accompanied by two major events that concern their genome and are of crucial significance when considered in terms of macromolecular crowding: (i) a substantial increase in the amount of DNA, and (ii) its confinement within a defined space. The resulting highly crowded environment would have strongly promoted DNA self-assembly processes, leading to extremely condensed and thermodynamically stable DNA aggregates. Such structural transitions have indeed been observed in vitro, as well as in virtually all cellular systems in which a nucleosomal assembly is absent. In this appear we raise the hypothesis that upon transition from prokaryotic systems to eukaryotes, the nucleosomes were rendered essential in order to negate extensive DNA condensation processes that would have resulted from excluded volume effects. By suppressing such processes, the nucleosomes act to maintain and regulate the conformational space of the DNA, thus enabling conformational flexibility and reversible structural modulations.

Animals↗

Coronavirus IBV-induced membrane fusion occurs at near-neutral pH.

The lysosomotropic agent NH4Cl caused a reduction of 80-95% in the number of chick kidney (CK) cells and Vero cells infected by infectious bronchitis virus (IBV) strain Beaudette, as determined by immunofluorescence at the end of the first replication cycle. Inhibition only occurred when NH4Cl was present during the first 2 h after infection. Syncytium formation was studied during replication of IBV-Beaudette in Vero cells. Some cell-cell fusion occurred at pH 7.0 and pH 6.5 but it was optimal at pH 6.7. IBV strain UK/123/82 did not replicate in Vero cells and was studied in CK cells in which it grew well but without forming syncytia. In contrast to IBV-Beaudette, NH4Cl had virtually no effect on the replication of UK/123/82. The results show that the IBV spike glycoprotein induces membrane fusion at near neutral pH although some IBV strains may require a mildly acidic environment for the efficient uncoating of the virion RNA.

Ammonium Chloride↗

L(+)-lactate does not affect twitch and tetanic responses in mechanically skinned mammalian muscle fibres.

This study investigated whether a high intracellular concentration of L(+)-lactate (30 mM) affects normal excitation-contraction coupling in skeletal muscle. Electrical stimulation was used to elicit action potentials in the (sealed) transverse-tubular system of mechanically skinned muscle fibres, giving rise to twitch and tetanic force responses. As the sarcolemma was absent, lactate could be applied to the cytoplasmic environment via the bathing solution (at a constant pH of 7.1) and its effect examined independently of other metabolic changes that occur during muscle fatigue. The presence of 30 mM lactate had virtually no effect on direct activation of the contractile apparatus by Ca2+. Lactate also had no significant effect on either the rate of rise or the peak of the twitch response, with the only detectable effect being a slight (13%) slowing in its relaxation rate. As the amplitude of the twitch response (approximately 60% of maximum force) may be regarded as a sensitive indicator of the amount of Ca2+ released by an action potential, there was evidently to change in Ca2+ release in the presence of lactate. Lactate also had no significant effect on the rate of rise and peak force of the tetanic response or on its subsequent relaxation. Additional experiments, in which the sarcoplasmic reticulum was emptied of Ca2+ (in a caffeine solution) and reloaded repeatedly, showed no significant effect of 30 mM lactate on Ca2+ uptake. This study shows that the presence of L(+)-lactate does not inhibit excitation-contraction coupling in mechanically skinned fibres.

Animals↗

Astrocyte reactivity influences the number of presynaptic terminals apposed to spinal motoneurons after axotomy.

Although synaptic plasticity is a widespread phenomenon, the underlying mechanisms leading to its occurrence are virtually unknown. In this sense, glial cells, especially astrocytes, may have a role in network changes of the nervous system, influencing the retraction of boutons as well as providing a proper perisynaptic environment, thereby affecting the replacement of inputs. Interestingly, the glial reaction does vary between strains of rats and mice. In this sense, we present evidence that C57BL/6J and A/J isogenic mice present different astrocyte reactivity after a peripheral lesion in vivo as well as in vitro, by analyzing primary cell cultures. Such a difference in the glial reaction has a direct influence on in vivo number of pre-synaptic terminals and on in vitro synaptogenesis.

Animals↗

Clinical evaluation of the GlucoWatch biographer: a continual, non-invasive glucose monitor for patients with diabetes.

A device providing frequent, automatic, and non-invasive glucose measurements for persons with diabetes has been developed: the GlucoWatch biographer. This device extracts glucose through intact skin via reverse iontophoresis where it is detected by an amperometric biosensor. The biographer can provide glucose readings every 20 min for 12 h. The performance of this device was evaluated in two large clinical studies in a controlled clinical environment (n=231), and the home environment (n=124). Accuracy of the biographer was evaluated by comparing the automatic biographer readings to serial finger-stick blood glucose (BG) measurements. Biographer performance was comparable in both environments. Mean difference between biographer and finger-stick measurements was -0.01 and 0.26 mmol l(-1) for the clinical and home environments, respectively. The mean absolute value of the relative difference was 1.06 and 1.18 mmol l(-1) for the same studies. Correlation coefficient (r) between biographer and finger-stick measurements was 0.85 and 0.80 for the two studies. In both studies, over 94% of the biographer readings were in the clinically acceptable A+B region of the Clarke Error Grid. A slight positive bias is observed for the biographer readings at low BG levels. Biographer accuracy is relatively constant over all rates of BG changes, except when BG decreases more than 10 mmol l(-1) h(-1), which occurred for only 0.2% of points in the home environment study. Biographer precision, as measured by CV%, is approx. 10%. Skin irritation, characterized by erythema and edema, was either non-existent or mild in >90% of subjects and resolved in virtually all subjects without treatment in several days.

Blood Glucose Self-Monitoring↗

Autophosphorylation dependent destabilization of the insulin receptor kinase domain: tryptophan-1175 reports changes in the catalytic cleft.

Protein kinases are regulated by conformational or chemical changes which facilitate access of substrates to the active site and promote correct orientations of catalytically essential residues and water molecules. The switch between basal and activated states of the insulin receptor's kinase domain (IRKD) results from autophosphorylation. We investigated the effects of IRKD autophosphorylation on the conformational stability by guanidine hydrochloride (GdnHCl) dependent denaturation and by iodide quenching of intrinsic fluorescence. Tryptophan residues of the recombinant soluble IRKD (residues R953-S1355) were excited at a lambdaex of 295 nm, and emission spectra were analyzed for centroid (a characteristic of average polarity of the indole rings' environments) and integrated fluorescence intensity over the lambdaem range of 310-420 nm. Denaturation profiles of both apo- and phospho-IRKD forms are complex with at least three distinct unfolding transitions. The first and last transitions were reversible and cooperative and had midpoints at 0.4 or 0.7 M GdnHCl and 2.4 or 2.7 M GdnHCl, respectively; transitions of phospho-IRKD occurred at lower GdnHCl concentrations. Calculations of free energy of unfolding suggested a loss of approximately 2.3 kcal/mol of stabilization for the first transition and approximately 1.5 kcal/mol for the third transition. Circular dichroism showed subtle changes in secondary structure over the first transition and global unfolding over the last transition. The first transition reports changes primarily in the local environment of W1175, which is near the catalytic loop and is conserved among protein tyrosine kinases. W1175 is also the dominant fluorophore of the native emission spectrum. Iodide quenching of W1175 was virtually undetectable in the apo-IRKD but significant in the phospho-IRKD, suggesting that W1175 exposure to small solutes is strongly dependent on the conformation of the activation loop. These studies indicate that autophosphorylation, while exposing the catalytic center, also produces a conformer less stable than the apoenzyme.

Amino Acid Substitution↗

Purification of glycoproteins by selective transport using concanavalin-mediated reverse micellar extraction.

A novel methodology for coupling liquid-liquid extraction with affinity interaction has been developed to selectively and efficiently purify and separate glycoproteins. The basis for the separation is the selective extraction of glycoproteins from an aqueous solution into a reverse micellar organic phase by using concanavalin A (a sugar-binding lectin) as a facilitative carrier. Specifically, horseradish peroxidase (a common glycoprotein) can be bound to concanavalin A in an aqueous phase and then extracted into an AOT-isooctane organic phase with negligible loss in enzyme activity. Virtually no extraction of peroxidase occurs in the absence of concanavalin A. Electron spin resonance studies have shown that the large lectin-glycoprotein complex (96,000 daltons) resides in a nonaqueous environment within the reverse micelle, perhaps at the surfactant, water-pool interface; hence, extraction of the large complex is feasible. The facilitative extraction has been extended to selective transport of peroxidase from a mixture of peroxidase and alkaline phosphatase (a nonglycosylated protein). This results in an efficient separation strategy with a separation factor of 16.

Chemistry Techniques, Analytical↗

Experimental practices in economics: a methodological challenge for psychologists?

This target article is concerned with the implications of the surprisingly different experimental practices in economics and in areas of psychology relevant to both economists and psychologists, such as behavioral decision making. We consider four features of experimentation in economics, namely, script enactment, repeated trials, performance-based monetary payments, and the proscription against deception, and compare them to experimental practices in psychology, primarily in the area of behavioral decision making. Whereas economists bring a precisely defined "script" to experiments for participants to enact, psychologists often do not provide such a script, leaving participants to infer what choices the situation affords. By often using repeated experimental trials, economists allow participants to learn about the task and the environment; psychologists typically do not. Economists generally pay participants on the basis of clearly defined performance criteria; psychologists usually pay a flat fee or grant a fixed amount of course credit. Economists virtually never deceive participants; psychologists, especially in some areas of inquiry, often do. We argue that experimental standards in economics are regulatory in that they allow for little variation between the experimental practices of individual researchers. The experimental standards in psychology, by contrast, are comparatively laissez-faire. We believe that the wider range of experimental practices in psychology reflects a lack of procedural regularity that may contribute to the variability of empirical findings in the research fields under consideration. We conclude with a call for more research on the consequences of methodological preferences, such as the use on monetary payments, and propose a "do-it-both-ways" rule regarding the enactment of scripts, repetition of trials, and performance-based monetary payments. We also argue, on pragmatic grounds, that the default practice should be not to deceive participants.

Deception↗

Innovation in health care: developments in NHS trusts.

Essential parts of the R&D in virtually all UK biomedical innovations are executed within teaching hospitals, which are undergoing major organizational changes as part of the ongoing NHS reforms. Examines the impact of the changing structural and regulatory environment on the process of biomedical innovation management of two groups of technologically novel projects with the NHS. Finds that regulatory changes were tending to centralize and formalize innovation management, but that end users were playing an active role in directing the course of new treatment development. Problems of allocation of R&D resources within teaching hospitals were still unresolved, although some evidence from the London teaching hospitals suggested that hospital managers were able to deal with conflicting demands where they perceived that research excellence could be an asset rather than a cost.

Decision Making, Organizational↗

Elucidation of the inhibitory factors of yogurt against Salmonella typhimurium.

The inhibitory nature of yogurt against contaminating microorganisms has been studied extensively. Nevertheless, the factors responsible for the death of Salmonella typhimurium in yogurt have not been elucidated. An understanding of these factors is important for the determination of yogurt's safety to consumer health. Yogurt fermented for 18 h at 42 C had a stable environment with the following conditions: pH 3.85, oxidation-reduction potential -80 mV, lactic acid concentration 158 mM, and acetic acid concentration 3.7 mM. Under these conditions, lactic acid was responsible for virtually all of yogurt's bactericidal activity against S. typhimurium at 37 C. Die-off rates were observed when these conditions were reproduced artificially in milk (artificial milk yogurt) and when lactic acid was added back to 18-h yogurt from which acids were removed by passage of the whey through a Dowex 1 (Cl-) anion exchange column (cationic yogurt). Factors that augmented lactic acid inhibition of S. typhimurium were low pH and low oxidation-reduction potential. The die-off rate of S. typhimurium was more rapid in yogurt whey (yogurt minus the casein fraction) than in whole yogurt, indicating that the casein fraction was partially able to protect Salmonella.

Acetates↗

Comparative assessment of two interfaces for delivering a multimedia medical course in the French-speaking Virtual Medical University (UMVF).

The UMVF aims at helping medical students during their normal curriculum via the facilities provided by Internet based techniques. This paper describes a comparative assessment of two interfaces delivering a multimedia course: a conventional web server (WS) and an integrated e-learning platform in the form of a Virtual Campus (VC). Eleven students were arbitrarily divided into two groups. We used a qualitative method for comparing their acceptance of the on line course provided by the two different interfaces. The two groups were globally satisfied. However, a decrease in satisfaction was noted at the end of the experimentation in the VC group. This may be explained by a more complex Graphical User Interface (GUI) of the VC and some constraints which do not exist with the WS. The current e-learning platforms are probably not optimised for working conditions where presential and virtual activities are mixed. We think that a new type of "light" platforms should be developed for these specific working conditions. Students of the two groups also had limitations about the multimedia environment. They may change their opinion if they get more accustomed with the multimedia environment and if their teachers make a more adequate use of the multimedia techniques.

Academic Medical Centers↗

Telebaby videostreaming of newborns over Internet.

New information technologies are used to fill gaps in our need for communication. Hospitals are now starting to discover the possibilities of multimedia communication for their patients. Telebaby is a project of the University Medical Centre Utrecht that allows parents to view their newborn on the intensive care, high care or medium care unit at home. Standard Internet technology for distributing multimedia allows parents to virtually "visit" their baby more often and give parents the feeling of control. A non-complex technical system like Telebaby is easily adopted by the parents, implementing it in a closed environment like a hospital is the real challenge.

Academic Medical Centers↗

Interactive stereoscopic rendering of volumetric environments.

We present an efficient stereoscopic rendering algorithm supporting interactive navigation through large-scale 3D voxel-based environments. In this algorithm, most of the pixel values of the right image are derived from the left image by a fast 3D warping based on a specific stereoscopic projection geometry. An accelerated volumetric ray casting then fills the remaining gaps in the warped right image. Our algorithm has been parallelized on a multiprocessor by employing effective task partitioning schemes and achieved a high cache coherency and load balancing. We also extend our stereoscopic rendering to include view-dependent shading and transparency effects. We have applied our algorithm in two virtual navigation systems, flythrough over terrain and virtual colonoscopy, and reached interactive stereoscopic rendering rates of more than 10 frames per second on a 16-processor SGI Challenge.

Algorithms↗