Volcano-Shaped Scanning Probe Microscopy Probe for Combined Force-Electrogram Recordings from Excitable Cells.

Atomic force microscopy based approaches have led to remarkable advances in the field of mechanobiology. However, linking the mechanical cues to biological responses requires complementary techniques capable of recording these physiological characteristics. In this study, we present an instrument for combined optical, force, and electrical measurements based on a novel type of scanning probe microscopy cantilever composed of a protruding volcano-shaped nanopatterned microelectrode (nanovolcano probe) at the tip of a suspended microcantilever. This probe enables simultaneous force and electrical recordings from single cells. Successful impedance measurements on mechanically stimulated neonatal rat cardiomyocytes in situ were achieved using these nanovolcano probes. Furthermore, proof of concept experiments demonstrated that extracellular field potentials (electrogram) together with contraction displacement curves could simultaneously be recorded. These features render the nanovolcano probe especially suited for mechanobiological studies aiming at linking mechanical stimuli to electrophysiological responses of single cells.

Intracellular Recording of Cardiomyocyte Action Potentials with Nanopatterned Volcano-Shaped Microelectrode Arrays.

Micronanotechnology-based multielectrode arrays have led to remarkable progress in the field of transmembrane voltage recording of excitable cells. However, providing long-term optoporation- or electroporation-free intracellular access remains a considerable challenge. In this study, a novel type of nanopatterned volcano-shaped microelectrode (nanovolcano) is described that spontaneously fuses with the cell membrane and permits stable intracellular access. The complex nanostructure was manufactured following a simple and scalable fabrication process based on ion beam etching redeposition. The resulting ring-shaped structure provided passive intracellular access to neonatal rat cardiomyocytes. Intracellular action potentials were successfully recorded in vitro from different devices, and continuous recording for more than 1 h was achieved. By reporting transmembrane action potentials at potentially high spatial resolution without the need to apply physical triggers, the nanovolcanoes show distinct advantages over multielectrode arrays for the assessment of electrophysiological characteristics of cardiomyocyte networks at the transmembrane voltage level over time.

Assessing blue wildebeests' vigilance, grouping and foraging responses to perceived predation risk using playback experiments.

Two aspects of reactive antipredator behaviour are still unclear for ungulates. First, when there is a direct predation threat, how do prey balance antipredator and social vigilance to learn a predator's location and assess the risk? Second, how do an individual's group and environment affect its responses? We tested the responses of adult females in 101 groups of wildebeest to playbacks of lion roars or car noises in Etosha National Park, Namibia. We analysed how the times they spent in different types categories of vigilance, and their within-group density, were affected by the playbacks and how a range of social and environmental variables affected those responses. Females increased their antipredator vigilance but not their social vigilance, after lion roars but not car noises, suggesting that they mostly relied on their own vigilance rather than social information to try to find the source of the lion roars. Females' antipredator vigilance increased more when they were further from cover and with other prey species, suggesting that both circumstances increased their perception of risk. They 'bunched' more after lion roars than car noises and their bite rates decreased as they bunched. Animals' use of social information about threats is likely to be context-dependent.

Ion beam etching redeposition for 3D multimaterial nanostructure manufacturing.

A novel fabrication method based on the local sputtering of photoresist sidewalls during ion beam etching is presented. This method allows for the manufacture of three-dimensional multimaterial nanostructures at the wafer scale in only four process steps. Features of various shapes and profiles can be fabricated at sub-100-nm dimensions with unprecedented freedom in material choice. Complex nanostructures such as nanochannels, multimaterial nanowalls, and suspended networks were successfully fabricated using only standard microprocessing tools. This provides an alternative to traditional nanofabrication techniques, as well as new opportunities for biosensing, nanofluidics, nanophotonics, and nanoelectronics.

Separation of blood microsamples by exploiting sedimentation at the microscale.

Microsample analysis is highly beneficial in blood-based testing where cutting-edge bioanalytical technologies enable the analysis of volumes down to a few tens of microliters. Despite the availability of analytical methods, the difficulty in obtaining high-quality and standardized microsamples at the point of collection remains a major limitation of the process. Here, we detail and model a blood separation principle which exploits discrete viscosity differences caused by blood particle sedimentation in a laminar flow. Based on this phenomenon, we developed a portable capillary-driven microfluidic device that separates blood microsamples collected from finger-pricks and delivers 2 µL of metered serum for bench-top analysis. Flow cytometric analysis demonstrated the high purity of generated microsamples. Proteomic and metabolomic analyses of the microsamples of 283 proteins and 1351 metabolite features was consistent with samples generated via a conventional centrifugation method. These results were confirmed by a clinical study scrutinising 8 blood markers in obese patients.

Additive manufacturing of hierarchical injectable scaffolds for tissue engineering.

We present a 3D-printing technology allowing free-form fabrication of centimetre-scale injectable structures for minimally invasive delivery. They result from the combination of 3D printing onto a cryogenic substrate and optimisation of carboxymethylcellulose-based cryogel inks. The resulting highly porous and elastic cryogels are biocompatible, and allow for protection of cell viability during compression for injection. Implanted into the murine subcutaneous space, they are colonized with a loose fibrovascular tissue with minimal signs of inflammation and remain encapsulation-free at three months. Finally, we vary local pore size through control of the substrate temperature during cryogenic printing. This enables control over local cell seeding density in vitro and over vascularization density in cell-free scaffolds in vivo. In sum, we address the need for 3D-bioprinting of large, yet injectable and highly biocompatible scaffolds and show modulation of the local response through control over local pore size. STATEMENT OF SIGNIFICANCE: This work combines the power of 3D additive manufacturing with clinically advantageous minimally invasive delivery. We obtain porous, highly compressible and mechanically rugged structures by optimizing a cryogenic 3D printing process. Only a basic commercial 3D printer and elementary control over reaction rate and freezing are required. The porous hydrogels obtained are capable of withstanding delivery through capillaries up to 50 times smaller than their largest linear dimension, an as yet unprecedented compression ratio. Cells seeded onto the hydrogels are protected during compression. The hydrogel structures further exhibit excellent biocompatibility 3 months after subcutaneous injection into mice. We finally demonstrate that local modulation of pore size grants control over vascularization density in vivo. This provides proof-of-principle that meaningful biological information can be encoded during the 3D printing process, deploying its effect after minimally invasive implantation.

Impedance-based real-time position sensor for lab-on-a-chip devices.

This paper presents the theoretical and experimental development of an integrated position sensor for lab-on-a-chip devices. The interest for single cell analysis is growing. However, this requires monitoring and controlling cell displacements in real time during their journey in the chip. Due to the high number of cells that must be monitored at the same time, classical vision-based sensors are not suitable. This paper aims to present an alternative based on impedance measurement. The position of the cells is obtained from the variation of impedance measured between two electrodes. This technique presents several advantages: the sensor is integrated into the chip, the measurement electrodes are compatible with the fabrication process of actuation electrodes for dielectrophoresis, the sampling time of the sensor is high and the position of the cells can be obtained in real time. This article highlights the concept of position-sensitive impedance sensing. The design of the chip, and in particular of the electrodes, is discussed to improve the sensitivity and repeatability of the measurement. The issue of real-time detection in a noisy environment is solved by using an extended Kalman filter. As a first proof of concept, this article presents experimental validation on a 1D case to determine the longitudinal position of 8.7 µm diameter beads in a channel.

A reproducible method for µm precision alignment of PDMS microchannels with on-chip electrodes using a mask aligner.

This paper describes a reproducible method for µm precision alignment of polydimethylsiloxane (PDMS) microchannels with coplanar electrodes using a conventional mask aligner for lab-on-a-chip applications. It is based on the use of a silicon mold in combination with a PMMA sarcophagus for precise control of the parallelism between the top and bottom surfaces of molded PDMS. The alignment of the fabricated PDMS slab with electrodes patterned on a glass chip is then performed using a conventional mask aligner with a custom-made steel chuck and magnets. This technique allows to bond and align chips with a resolution of less than 2 µm.

Design and fabrication of a nanofibrous polycaprolactone tubular nerve guide for peripheral nerve tissue engineering using a two-pole electrospinning system.

Nerve guidance conduits are considered to be the new generation of scaffolds designed for nerve disorders. A tubular construct with a highly aligned fibrous structure, mimicking the endoneurium layer surrounding inner axons of a nerve fascicle, is a suitable candidate for a nerve guide. In this paper a new approach for the fabrication of 3D tubular nerve guides is introduced using simulation of a two-pole electrospinning system and describing its mechanism. The structure of this scaffold is then optimized using the Taguchi statistical method and after morphological studies by scanning electron microscopy, the crystallinity, tensile strength and protein adsorption of these highly aligned fibres are investigated, comparing them with semi-aligned and random fibres produced via conventional mandrel electrospinning. Cell attachment, proliferation and migration of PC12 neuronal like cells are studied on highly aligned, semi aligned and random structures, and morphological change and elongation are observed in PC12 cells. The results of these studies suggest that conduits fabricated using two-pole electrospinning are a suitable and promising scaffold for peripheral and even spinal nerve regeneration. This nerve guide has a great potential for further advanced modifications and regeneration in higher levels.

[Self-regulation and virtual reality in forensic psychiatry: An emphasis on theoretical underpinnings]. / Applications de la réalité virtuelle en psychiatrie légale : la perspective de l'autorégulation comme cadre théorique.

INTRODUCTION: Forensic psychiatry is the field whose expertise is the assessment and treatment of offending behaviours, in particular when offenses are related to mental illness. An underlying question for all etiological models concerns the manner in which an individual's behaviours are organized. Specifically, it becomes crucial to understand how certain individuals come to display maladaptive behaviours in a given environment, especially when considering issues such as offenders' responsibility and their ability to change their behaviours. VIRTUAL REALITY: Thanks to its ability to generate specific environments, associated with a high experimental control on generated simulations, virtual reality is gaining recognition in forensic psychiatry. Virtual reality has generated promising research data and may turn out to be a remarkable clinical tool in the near future. While research has increased, a conceptual work about its theoretical underpinnings is still lacking. However, no important benefit should be expected from the introduction of a new tool (as innovative as virtual reality) without an explicit and heuristic theoretical framework capable of clarifying its benefits in forensic psychiatry. OBJECTIVES: Our paper introduces self-regulation perspective as the most suitable theoretical framework for virtual reality in forensic psychiatry. It will be argued that virtual reality does not solely help to increase ecological validity. However, it does allow one to grant access to an improved understanding of violent offending behaviours by probing into the underlying mechanisms involved in the self-regulation of behaviours in a dynamical environment. Illustrations are given as well as a discussion regarding perspectives in the use of virtual reality in forensic psychiatry.

Seasonal variation in accumulation of persistent organic pollutants in an Arctic marine benthic food web.

The aim of the present study was to investigate seasonal variation in persistent organic pollutant (POP) concentrations, as well as food-web biomagnification, in an Arctic, benthic marine community. Macrozoobenthos, demersal fish and common eiders were collected both inside and outside of Kongsfjorden, Svalbard, during May, July and October 2007. The samples were analysed for a selection of legacy chlorinated POPs. Overall, low levels of POPs were measured in all samples. Although POP levels and accumulation patterns showed some seasonal variation, the magnitude and direction of change was not consistent among species. Overall, seasonality in bioaccumulation in benthic biota was less pronounced than in the pelagic system in Kongsfjorden. In addition, the results indicate that δ(15)N is not a good predictor for POP-levels in benthic food chains. Other factors, such as feeding strategy (omnivory, necrophagy versus herbivory), degree of contact with the sediment, and a high dependence on particulate organic matter (POM), with low POP-levels and high δ(15)N-values (due to bacterial isotope enrichment), seem to govern the uptake of the different POPs and result in loads deviating from what would be expected consulting the trophic position alone.

Environmental consequences of uranium atmospheric releases from fuel cycle facility: II. The atmospheric deposition of uranium and thorium on plants.

Uranium and thorium isotopes were measured in cypress leaves, wheat grains and lettuce taken in the surroundings of the uranium conversion facility of Malvési (South of France). The comparison of activity levels and activity ratios (namely (238)U/(232)Th and (230)Th/(232)Th) in plants with those in aerosols taken at this site and plants taken far from it shows that aerosols emitted by the nuclear site (uranium releases in the atmosphere by stacks and (230)Th-rich particles emitted from artificial ponds collecting radioactive waste mud) accounts for the high activities recorded in the plant samples close to the site. The atmospheric deposition process onto the plants appears to be the dominant process in plant contamination. Dry deposition velocities of airborne uranium and thorium were measured as 4.6 × 10(-3) and 5.0 × 10(-3) m s(-1), respectively.

[Sleep and academic performance in young elite athletes]. / Sommeil et résultats scolaires chez les jeunes sportifs de haut niveau.

RATIONALE: In French law (Code du Sport), the status of elite athlete is allowed for young athletes beginning at the age of 12 years. For these young athletes, the aim is to reach the highest level of performance in their sport without compromising academic performance. Training time is therefore often substantial and sleep patterns appear to play a key role in performance recovery. The aim of this study was to assess sleep patterns and their effects on academic performance in young elite athletes. MATERIAL AND METHODS: Sleep patterns were assessed using questionnaires completed during a specific information-based intervention on sports medicine topics. The academic performance of young elite athletes was assessed by collecting their grades (transmitted by their teachers). RESULTS: Sleep patterns were assessed for 137 young elite athletes (64 females, 73 males; mean age, 15.7 years) and academic performance for 109 of them. Daily sleep duration during school periods (8h22 ± 38 min) were shorter compared to holidays and week-ends (10h02 ± 1h16, P<0.0001). Fifty-six athletes (41 %) subjectively estimated their sleep quality as poor or just sufficient. Poor sleep quality was correlated with poor academic performance in this specific athlete population. DISCUSSION: Sleep is the most important period for recovery from daily activity, but little information is available regarding the specific population of young elite athletes. The results reported herein suggest insufficiency (quantitatively and qualitatively) of sleep patterns in some of the young athletes, possibly leading to detrimental effects on athletic performance. Moreover, disturbed sleep patterns may also impact academic performance in young elite athletes. CONCLUSION: Teachers, athletic trainers, physicians, and any other professionals working with young elite athletes should pay particular attention to this specific population regarding the possible negative repercussions of poor sleep patterns on academic and athletic performance.

An impedance-based flow microcytometer for single cell morphology discrimination.

Cell shape is a fundamental biological feature, providing specific information about physiological or pathological cellular conditions. Most of the state-of-the-art microfluidic cytometers, however, only allow simple cell analysis, including viability studies, cell counting and sorting. In this work, we present a non-invasive, label-free device capable of single cell morphology discrimination in continuous flow. The device is based on the principle of liquid electrodes, fabricated in a cross configuration around a sensing zone. This arrangement allows measurement of cell impedance along orthogonal orientations and extraction of an index describing cell shape anisotropy. By adding prior to the sensing volume a series of lateral liquid electrodes, the particle stream was focused toward the channel midline and each cell was oriented in a specific direction before shape sensing. We demonstrate the proof of concept by performing spherical and elongated particle discrimination. As an application, we show that the shape changes experienced during cell division can be monitored and characterized. In particular, budding yeasts at different stages of the mitotic cycle were identified by extracting their anisotropy index.

Smart instrumentation for determination of ligament stiffness and ligament balance in total knee arthroplasty.

Ligament balance is an important and subjective task performed during total knee arthroplasty (TKA) procedure. For this reason, it is desirable to develop instruments to quantitatively assess the soft-tissue balance since excessive imbalance can accelerate prosthesis wear and lead to early surgical revision. The instrumented distractor proposed in this study can assist surgeons on performing ligament balance by measuring the distraction gap and applied load. Also the device allows the determination of the ligament stiffness which can contribute a better understanding of the intrinsic mechanical behavior of the knee joint. Instrumentation of the device involved the use of hall-sensors for measuring the distractor displacement and strain gauges to transduce the force. The sensors were calibrated and tested to demonstrate their suitability for surgical use. Results show the distraction gap can be measured reliably with 0.1mm accuracy and the distractive loads could be assessed with an accuracy in the range of 4N. These characteristics are consistent with those have been proposed, in this work, for a device that could assist on performing ligament balance while permitting surgeons evaluation based on his experience. Preliminary results from in vitro tests were in accordance with expected stiffness values for medial collateral ligament (MCL) and lateral collateral ligament (LCL).

X-ray image enhancement via determinant based feature selection.

Previous work has investigated the feasibility of using Eigenimage-based enhancement tools to highlight abnormalities on chest X-rays (Butler et al in J Med Imaging Radiat Oncol 52:244-253, 2008). While promising, this approach has been limited by computational restrictions of standard clinical workstations, and uncertainty regarding what constitutes an adequate sample size. This paper suggests an alternative mathematical model to the above referenced singular value decomposition method, which can significantly reduce both the required sample size and the time needed to perform analysis. Using this approach images can be efficiently separated into normal and abnormal parts, with the potential for rapid highlighting of pathology.

A custom robot for Transcranial Magnetic Stimulation: first assessment on healthy subjects.

In this paper, a custom robotic system for Transcranial Magnetic Stimulation is assessed in clinical conditions on healthy subjects. A motor cortex mapping is performed using the robotic system with comparison to a manual approach using a neuronavigation system. Stimulation accuracy, repeatability are evaluated as well as the feeling of the system operator and the subject in terms of comfort, tiredness, stress level, ease-of-use. Very encouraging results are obtained on all these aspects, which strengthens the idea of developing robotic assistance for TMS.

Design and test of a MEMS strain-sensing device for monitoring artificial knee implants.

This paper describes the development of a polyimide-based MEMS strain-sensing device. Finite element analysis was used to investigate an artificial knee implant and assist on device design and to optimize sensing characteristics. The sensing element of the device was fabricated using polyimide micromachining with embedded thin-metallic wires and placed into a knee prosthesis. The device was evaluated experimentally in a mechanical knee simulator using static and dynamic axial load conditions similar to those encountered in vivo. Results indicates the sensor is capable of measuring the strain associated to the total axial forces in the range of approximately 4 times body weight with a good sensitivity and accuracy for events happening within 1 s time window.