Effect of position on the mechanical interaction between the rib cage and abdomen in preterm infants.

To determine the influence of body position on chest wall and pulmonary function, we studied the ventilatory, pulmonary mechanics, and thoracoabdominal motion profiles in 20 preterm infants recovering from respiratory disease who were positioned in both the supine and prone position. Thoracoabdominal motion was assessed from measurements of relative rib cage and abdominal movement and the calculated phase angle (an index of thoracoabdominal synchrony) of the rib and abdomen Lissajous figures. The ventilatory and pulmonary function profiles were assessed from simultaneous measurements of transpulmonary pressure, airflow, and tidal volume. The infants were studied in quiet sleep, and the order of positioning was randomized across patients. The results demonstrated no significant difference in ventilatory and pulmonary function measurements as a function of position. In contrast, there was a significant reduction (-49%) in the phase angle of the Lissajous figures and an increase (+66%) in rib cage motion in prone compared with the supine position. In addition, the degree of improvement in phase angle in the prone position was correlated to the severity of asynchrony in the supine position. We speculate that the improvement in thoracoabdominal synchrony in the prone position is related to alterations of chest wall mechanics and respiratory muscle tone mediated by a posturally related shift in the area of apposition of the diaphragm to the anterior inner rib cage wall and increase in passive tension of the muscles of the rib cage. This study suggests that the mechanical advantage associated with prone positioning may confer a useful alternative breathing pattern to the preterm infant in whom elevated respiratory work loads and respiratory musculoskeletal immaturity may predispose to respiratory failure.     

Charge movement in gating-locked HCN channels reveals weak coupling of voltage sensors and gate

HCN (hyperpolarization-activated cyclic nucleotide gated) pacemaker channels have an architecture similar to that of voltage-gated K⁺ channels, but they open with the opposite voltage dependence. HCN channels use essentially the same positively charged voltage sensors and intracellular activation gates as K⁺ channels, but apparently these two components are coupled differently. In this study, we examine the energetics of coupling between the voltage sensor and the pore by using cysteine mutant channels for which low concentrations of Cd²⁺ ions freeze the open–closed gating machinery but still allow the sensors to move. We were able to lock mutant channels either into open or into closed states by the application of Cd²⁺ and measure the effect on voltage sensor movement. Cd²⁺ did not immobilize the gating charge, as expected for strict coupling, but rather it produced shifts in the voltage dependence of voltage sensor charge movement, consistent with its effect of confining transitions to either closed or open states. From the magnitude of the Cd²⁺-induced shifts, we estimate that each voltage sensor produces a roughly three- to sevenfold effect on the open–closed equilibrium, corresponding to a coupling energy of ∼1.3–2 kT per sensor. Such coupling is not only opposite in sign to the coupling in K⁺ channels, but also much weaker.     

COMPUTER PROGRAM FOR THREE-DIMENSIONAL RECONSTRUCTIONS AND NUMERICAL ANALYSES OF PLANT ORGANS FROM SERIAL SECTIONS

A computer program (SECTION) is presented which allows for the three-dimensional reconstruction of serial cross sections through biological materials. The program provides a numerical analysis of perimeter length and transverse area of each anatomical feature designated in a cross section, as well as surface area and volume computations for features that pass from one section to another. In addition to rotating and tilting the 3-D reconstruction in any desired orientation, the program has editing capabilities which allow different combinations of anatomical features to be shown in neutral gray outline or interconnected by colored lines.     

Estimation of discretization errors in contact pressure measurements

Contact pressure measurements in total knee replacements are often made using a discrete sensor such as the Tekscan K-Scan sensor. However, no method currently exists for predicting the magnitude of sensor discretization errors in contact force, peak pressure, average pressure, and contact area, making it difficult to evaluate the accuracy of such measurements. This study identifies a non-dimensional area variable, defined as the ratio of the number of perimeter elements to the total number of elements with pressure, which can be used to predict these errors. The variable was evaluated by simulating discrete pressure sensors subjected to Hertzian and uniform pressure distributions with two different calibration procedures. The simulations systematically varied the size of the sensor elements, the contact ellipse aspect ratio, and the ellipse's location on the sensor grid. In addition, contact pressure measurements made with a K-Scan sensor on four different total knee designs were used to evaluate the magnitude of discretization errors under practical conditions. The simulations predicted a strong power law relationship (r(2)>0.89) between worst-case discretization errors and the proposed non-dimensional area variable. In the total knee experiments, predicted discretization errors were on the order of 1-4% for contact force and peak pressure and 3-9% for average pressure and contact area. These errors are comparable to those arising from inserting a sensor into the joint space or truncating pressures with pressure sensitive film. The reported power law regression coefficients provide a simple way to estimate the accuracy of experimental measurements made with discrete pressure sensors when the contact patch is approximately elliptical.     

Thoracoabdominal asynchrony failed to grade airway obstructions in foals.

Respiratory inductive plethysmography (RIP) can be used to obtain a valid measure of tidal volume in humans. This device also compares the contributions to ventilation of the thorax and abdomen. Although thoracoabdominal asynchrony is a prominent clinical feature for patients with airway obstruction, the accuracy of the RIP device to assess the severity of obstruction is unclear. This study analyzes how well RIP variables reflect the degree of a fixed external inspiratory plus expiratory resistive load in foals. Foals were employed because the species and age group are commonly afflicted with respiratory disease. Eight conscious, sedated (xylazine 1.25 mg/kg body wt) foals were subjected to randomly ordered resistive loads at the airway opening and, on a separate day, to histamine aerosol challenge. During resistive loading, phase angle changed significantly, as did phase relation (P < or = 0.05). However, no significant correlation was found between the degree of change in resistive load and the degree to which phase angle or relation was altered (r(s) = 0.41 and 0.25, respectively). In addition, neither phase angle nor relation changed significantly with histamine challenge. We conclude that, although RIP variables changed markedly with fixed upper airway resistive loading, the degree to which they changed was erratic and therefore not useful for grading these obstructions. Furthermore, RIP variables were insensitive measures of histamine-induced bronchoconstriction.     

Mode shift of the voltage sensors in Shaker K+ channels is caused by energetic coupling to the pore domain

The voltage sensors of voltage-gated ion channels undergo a conformational change upon depolarization of the membrane that leads to pore opening. This conformational change can be measured as gating currents and is thought to be transferred to the pore domain via an annealing of the covalent link between voltage sensor and pore (S4-S5 linker) and the C terminus of the pore domain (S6). Upon prolonged depolarizations, the voltage dependence of the charge movement shifts to more hyperpolarized potentials. This mode shift had been linked to C-type inactivation but has recently been suggested to be caused by a relaxation of the voltage sensor itself. In this study, we identified two ShakerIR mutations in the S4-S5 linker (I384N) and S6 (F484G) that, when mutated, completely uncouple voltage sensor movement from pore opening. Using these mutants, we show that the pore transfers energy onto the voltage sensor and that uncoupling the pore from the voltage sensor leads the voltage sensors to be activated at more negative potentials. This uncoupling also eliminates the mode shift occurring during prolonged depolarizations, indicating that the pore influences entry into the mode shift. Using voltage-clamp fluorometry, we identified that the slow conformational change of the S4 previously correlated with the mode shift disappears when uncoupling the pore. The effects can be explained by a mechanical load that is imposed upon the voltage sensors by the pore domain and allosterically modulates its conformation. Mode shift is caused by the stabilization of the open state but leads to a conformational change in the voltage sensor.     

Automatic detection of gait events: a case study using inductive learning techniques

One of the problems which occurs in the development of a control system for functional electrical stimulation of the lower limbs is to detect accurately specific events within the gait cycle. We present a method for the classification of phases of the gait cycle using the artificial intelligence technique of inductive learning. Both the terminology of inductive learning and the algorithm used for the analyses are fully explained. Given a set of examples of sensor data from the gait events that are to be delected, the inductive learning algorithm is able to produce a decision tree (or set of rules) which classify the data using a minimum number of sensors. The nature of the redundancy of the sensor set is examined by progressively removing combinations of sensors and noting the effect on both the size of the decision trees produced and their classification accuracy on ‘unseen’ testing data. Since the algorithm is able to calculate which sensors are more important (informative), comparisons with the intuitive appreciation of sensor importance of five researchers in the fields were made, revealing that those sensors which appear intuitively most informative may, in fact, provide the least information. Comparison results with the standard statistical classification technique of linear discriminant analysis are also presented, showing the relative simplicity of the inductively derived rules together with their good classification accuracy. In addition to the control of FES, such techniques are also applicable to automatic gait analysis and the construction of expert systems for diagnosis of gait pathologies.     

Technical note: A stereological analysis of the cross‐sectional variability of the femoral osteon population

Unbiased selection of regions of interest (ROIs) and unbiased definition of histological structures are needed to improve the repeatability of microscopic methods for age at death determination and to reduce operator subjectivity. We present results obtained by selecting ROIs according to stereological principles on a sample of 28 femoral cross sections of Caucasoid males aged 20–89 years. A regular grid was overlaid on the cross section, and the ROIs were selected as close as possible to the periosteum in the anterior, lateral, and medial regions. The areas consisting of all intact secondary osteons plus fragments were outlined and osteon population density, percent osteon population, area, and perimeter were calculated using stereological methods and software. Overall, the analyses of intra‐ and inter‐section variability showed no significant difference between the ROIs, i.e., the location within the cross section of the ROIs does not affect the outcome of the analyses. The individual variability was found to be higher in adults aged 30–55 years than in other age ranges. ranges. Am J Phys Anthropol 2010. © 2010 Wiley‐Liss, Inc.     

Use of a triaxial magnetometer for respiratory measurements

We describe a triaxial magnetometer (Tri-mag) system, which consists of a transmitter, four sensors, a processing unit, and a personal computer (PC). The Tri-mag processing unit outputs the position of each sensor relative to the transmitter in three orthogonal coordinates, and this information is communicated to the PC. First, we demonstrated that within a defined octant of a sphere in which the center is the transmitter, we can measure radial distances with an accuracy of +/- 1 mm over a range extending from 10 to 70 cm from the transmitter. Second, we recorded the three-dimensional movement of sensors on the anterior and posterior surfaces of the chest wall during maximum voluntary ventilation in four normal men; all sensors were placed in the midsagittal plane of the body. Anterior sensors were located on the sternum at the level of the third intercostal space and at 2 cm above the umbilicus, whereas posterior sensors were located on the posterior spine at the same vertical levels as the anterior sensors. In all subjects the following was found. 1) Both anterior sensors moved anterior and cephalad during inspiration. The anterior thoracic sensor showed greater vertical than anteroposterior (A-P) movement, whereas the anterior abdominal sensor showed greater A-P than vertical movement. 2) Inspiration was associated with spinal extension, whereas expiration was associated with spinal flexion. Third, we used Tri-mag information to 1) measure tidal volume (VT) over a range extending from 500 ml to inspiratory capacity and 2) measure the change in end-expiratory lung volume (EELV) over a range extending from FRC to FRC plus a minimum of 1.5 liters. Our results indicate that greater than 96% of the changes in VT and greater than 82% of the changes in EELV can be accounted for by changes in A-P, vertical, and lateral dimensions of the chest wall.     

Fast and slow voltage sensor movements in HERG potassium channels

HERG encodes an inwardly-rectifying potassium channel that plays an important role in repolarization of the cardiac action potential. Inward rectification of HERG channels results from rapid and voltage-dependent inactivation gating, combined with very slow activation gating. We asked whether the voltage sensor is implicated in the unusual properties of HERG gating: does the voltage sensor move slowly to account for slow activation and deactivation, or could the voltage sensor move rapidly to account for the rapid kinetics and intrinsic voltage dependence of inactivation? To probe voltage sensor movement, we used a fluorescence technique to examine conformational changes near the positively charged S4 region. Fluorescent probes attached to three different residues on the NH₂-terminal end of the S4 region (E518C, E519C, and L520C) reported both fast and slow voltage-dependent changes in fluorescence. The slow changes in fluorescence correlated strongly with activation gating, suggesting that the slow activation gating of HERG results from slow voltage sensor movement. The fast changes in fluorescence showed voltage dependence and kinetics similar to inactivation gating, though these fluorescence signals were not affected by external tetraethylammonium blockade or mutations that alter inactivation. A working model with two types of voltage sensor movement is proposed as a framework for understanding HERG channel gating and the fluorescence signals.     

A chemiluminescence sensor array based on nanomaterials for discrimination of teas

In recent years, electronic tongue and nose devices have been developed that consist of an array of cross‐responsive sensors. In this study, we report a chemiluminescence (CL) sensor array based on oxidation at twelve different catalytic nanomaterial locations for the discrimination of eight teas. CL response patterns or “fingerprints” were obtained for a given compound on the sensor array and then discriminated through linear discriminant analysis. The experiments demonstrate that the sensor array had excellent differentiability and reversibility. Copyright © 2012 John Wiley & Sons, Ltd.     

The relationship between Q gamma and Ca release from the sarcoplasmic reticulum in skeletal muscle

Asymmetric membrane currents and fluxes of Ca2+ release were determined in skeletal muscle fibers voltage clamped in a Vaseline-gap chamber. The conditioning pulse protocol 1 for suppressing Ca2+ release and the "hump" component of charge movement current (I gamma), described in the first paper of this series, was applied at different test pulse voltages. The amplitude of the current suppressed during the ON transient reached a maximum at slightly suprathreshold test voltages (-50 to -40 mV) and decayed at higher voltages. The component of charge movement current suppressed by 20 microM tetracaine also went through a maximum at low pulse voltages. This anomalous voltage dependence is thus a property of I gamma, defined by either the conditioning protocol or the tetracaine effect. A negative (inward-going) phase was often observed in the asymmetric current during the ON of depolarizing pulses. This inward phase was shown to be an intramembranous charge movement based on (a) its presence in the records of total membrane current, (b) its voltage dependence, with a maximum at slightly suprathreshold voltages, (c) its association with a "hump" in the asymmetric current, (d) its inhibition by interventions that reduce the "hump", (e) equality of ON and OFF areas in the records of asymmetric current presenting this inward phase, and (f) its kinetic relationship with the time derivative of Ca release flux. The nonmonotonic voltage dependence of the amplitude of the hump and the possibility of an inward phase of intramembranous charge movement are used as the main criteria in the quantitative testing of a specific model. According to this model, released Ca2+ binds to negatively charged sites on the myoplasmic face of the voltage sensor and increases the local transmembrane potential, thus driving additional charge movement (the hump). This model successfully predicts the anomalous voltage dependence and all the kinetic properties of I gamma described in the previous papers. It also accounts for the inward phase in total asymmetric current and in the current suppressed by protocol 1. According to this model, I gamma accompanies activating transitions at the same set of voltage sensors as I beta. Therefore it should open additional release channels, which in turn should cause more I gamma, providing a positive feedback mechanism in the regulation of calcium release.     

Model-based identification of motion sensor placement for tracking retraction and elongation of the tongue

Electromagnetic articulography (EMA) is designed to track facial and tongue movements. In practice, the EMA sensors for tracking the movement of the tongue’s surface are placed heuristically. No recommendation exists. Within this paper, a model-based approach providing a mathematical analysis and a computational-based recommendation for the placement of sensors, which is based on the tongue’s envelope of movement, is proposed. For this purpose, an anatomically detailed Finite Element (FE) model of the tongue has been employed to determine the envelope of motion for retraction and elongation using a forward simulation. Two optimality criteria have been proposed to identify a set of optimal sensor locations based on the pre-computed envelope of motion. The first one is based on the assumption that locations exhibiting large displacements contain the most information regarding the tongue’s movement and are less susceptible to measurement errors. The second one selects sensors exhibiting each the largest displacements in the anterior-posterior, superior-inferior, medial-lateral and overall direction. The quality of the two optimality criteria is analysed based on their ability to deduce from the respective sensor locations the corresponding muscle activation parameters of the relevant muscle fibre groups during retraction and elongation by solving the corresponding inverse problem. For this purpose, a statistical analysis has been carried out, in which sensor locations for two different modes of deformation have been subjected to typical measurement errors. Then, for tongue retraction and elongation, the expectation value, the standard deviation, the averaged bias and the averaged coefficient of variation have been computed based on 41 different error-afflicted sensor locations. The results show that the first optimality criteria is superior to the second one and that the averaged bias and averaged coefficient of variation decrease when the number of sensors is increased from 2, 4 to 6 deployable sensors.     

Multiplexed FRET to image multiple signaling events in live cells

We report what to our knowledge is a novel approach for simultaneous imaging of two different Förster resonance energy transfer (FRET) sensors in the same cell with minimal spectral cross talk. Previous methods based on spectral ratiometric imaging of the two FRET sensors have been limited by the availability of suitably bright acceptors for the second FRET pair and the spectral cross talk incurred when measuring in four spectral windows. In contrast to spectral ratiometric imaging, fluorescence lifetime imaging (FLIM) requires measurement of the donor fluorescence only and is independent of emission from the acceptor. By combining FLIM-FRET of the novel red-shifted TagRFP/mPlum FRET pair with spectral ratiometric imaging of an ECFP/Venus pair we were thus able to maximize the spectral separation between our chosen fluorophores while at the same time overcoming the low quantum yield of the far red acceptor mPlum. Using this technique, we could read out a TagRFP/mPlum intermolecular FRET sensor for reporting on small Ras GTP-ase activation in live cells after epidermal growth factor stimulation and an ECFP/Venus Cameleon FRET sensor for monitoring calcium transients within the same cells. The combination of spectral ratiometric imaging of ECFP/Venus and high-speed FLIM-FRET of TagRFP/mPlum can thus increase the spectral bandwidth available and provide robust imaging of multiple FRET sensors within the same cell. Furthermore, since FLIM does not require equal stoichiometries of donor and acceptor, this approach can be used to report on both unimolecular FRET biosensors and protein-protein interactions with the same cell.     

Modelling apical columnar epithelium mechanics from circumferential contractile fibres

Simple columnar epithelia are formed by individual epithelial cells connecting together to form single cell high sheets. They are a main component of many important body tissues and are heavily involved in both normal and cancerous cell activities. Prior experimental observations have identified a series of contractile fibres around the circumference of a cross section located in the upper (apical) region of each cell. While other potential mechanisms have been identified in both the experimental and theoretical literature, these circumferential fibres are considered to be the most likely mechanism controlling movement of this cross section. Here, we investigated the impact of circumferential contractile fibres on movement of the cross section by creating an alternate model where movement is driven from circumferential contractile fibres, without any other potential mechanisms. In this model, we utilised a circumferential contractile fibre representation based on investigations into the movement of contractile fibres as an individual system, treated circumferential fibres as a series of units, and matched our model simulation to experimental geometries. By testing against laser ablation datasets sourced from existing literature, we found that circumferential fibres can reproduce the majority of cross-sectional movements. We also investigated model predictions related to various aspects of cross-sectional movement, providing insights into epithelium mechanics and demonstrating the usefulness of our modelling approach.     

The functions of WHIRLY1 and REDOX-RESPONSIVE TRANSCRIPTION FACTOR 1 in cross tolerance responses in plants: a hypothesis

Chloroplasts are important sensors of environment change, fulfilling key roles in the regulation of plant growth and development in relation to environmental cues. Photosynthesis produces a repertoire of reductive and oxidative (redox) signals that provide information to the nucleus facilitating appropriate acclimation to a changing light environment. Redox signals are also recognized by the cellular innate immune system allowing activation of non-specific, stress-responsive pathways that underpin cross tolerance to biotic–abiotic stresses. While these pathways have been intensively studied in recent years, little is known about the different components that mediate chloroplast-to-nucleus signalling and facilitate cross tolerance phenomena. Here, we consider the properties of the WHIRLY family of proteins and the REDOX-RESPONSIVE TRANSCRIPTION FACTOR 1 (RRTF1) in relation to chloroplast redox signals that facilitate the synergistic co-activation of gene expression pathways and confer cross tolerance to abiotic and biotic stresses. We propose a new hypothesis for the role of WHIRLY1 as a redox sensor in chloroplast-to-nucleus retrograde signalling leading to cross tolerance, including acclimation and immunity responses. By virtue of its association with chloroplast nucleoids and with nuclear DNA, WHIRLY1 is an attractive candidate coordinator of the expression of photosynthetic genes in the nucleus and chloroplasts. We propose that the redox state of the photosynthetic electron transport chain triggers the movement of WHIRLY1 from the chloroplasts to the nucleus, and draw parallels with the regulation of NONEXPRESSOR OF PATHOGENESIS-RELATED GENES 1 (NPR1).     

Design Optimisation of Plasmonic Metasurfaces for Mid-Infrared High-Sensitivity Chemical Sensing

In this paper, we report on a general method to optimise the optical characteristics of 2D-arrays of plasmonic gold nanoantennas performing as band-pass filter functionalised metasurfaces to be used as high-sensitivity mid-infrared spectroscopic sensors. We demonstrate that it is possible to increase their sensitivity in the detection of chemical and biological substances when the sensors are used in the surface-enhanced infrared absorption (SEIRA) technique. This technique allows revealing the presence of a substance adsorbed on the nanoantennas by measuring its optical absorption under the conditions for which the maximum value of the functionalised metasurface reflectivity occurs at the same wavelength of the substance maximum absorption peak. In particular, numerical simulations based on finite element method of the metasurface detection response demonstrate the possibility to increase the sensor sensitivity of more than four orders of magnitude with respect to that one achievable if the same amount of the substance is deposited on an unstructured planar metal surface. These results can be obtained by acting on the 2D-array periodicity, nanoantenna shape (i.e. rod and cross), size and thickness independently from the wavelength at which the substance absorption occurs. Moreover, in the case of cross-shaped nanoantennas, we report a complete numerical characterisation of the dependence of the metasurface maximum reflectivity and peak wavelength on the variation of the geometrical parameters of both the nanoantennas and the 2D-array.     

Disentangling the effects of genetic and environmental factors on movement behaviour

Individual variability in animal movement behaviour is well documented for many species. However, it remains unclear whether this variability reflects genetic variation, environmental variation or a combination of the two. Here, we conduct a cross‐fostering experiment with the aim of investigating the role of these two components in movement patterns during the post‐fledging dependence period and early natal dispersal of 21 eagle owls Bubo bubo. Our experiment showed that cross‐fostering did not influence any of the movement parameters considered. Movement parameters were, however, affected by the age and sex of the owlets. We therefore suggest that individual variability and family resemblance in movement behaviour during the post‐fledging dependence period and early natal dispersal might not be due to the common genetic origin of siblings, but rather that it originates from factors related to the rearing environment.