Video analysis of chemotactic locomotion of stored human polymorphonuclear leukocytes

Previous studies of the storage of polymorphonuclear leukocytes (PMNs) have used an empirical approach to define "optimal" conditions. To date, no storage conditions have been described which satisfactorily preserve the chemotactic function of PMNs beyond 24 h. In an effort to define the precise nature of the storage lesion, we studied the chemotactic locomotion of freshly isolated PMNs and PMNs which had been suspended in citrate-phosphate-dextrose-adenine (CPD-A1) plasma and stored in PVC bags, at 20-22 degrees C for 24 h. We used time-lapse video recording and computer image analysis to quantitate the motion of PMNs migrating under agarose. The positions of individual motile cells were traced at 1-min intervals for 5 min. The following parameters were used to quantitate migration: speed (distance/min), persistence of locomotion index (velocity/speed), orientation angle (the angle of the vector describing the next displacement of a cell relative to a direct line toward the chemoattractant), and chemotropic index (cosine of the orientation angle). After 24 h of storage, the following changes were observed: fewer cells migrated, the speed of migrating cells was reduced by 25%, the persistence of locomotion index decreased by 7%, which indicates that migrating cells made slightly more/wider turns, and the chemotropic index was decreased by 30%, which indicates that migrating cells were less accurate in their orientation toward the chemoattractant. Apparently, the storage of PMNs selectively impairs the ability of some cells to orient accurately in a chemotactic gradient and changes the distribution of these locomotor parameters within the population.     

Dynamic motion planning of 3D human locomotion using gradient-based optimization

Since humans can walk with an infinite variety of postures and limb movements, there is no unique solution to the modeling problem to predict human gait motions. Accordingly, we test herein the hypothesis that the redundancy of human walking mechanisms makes solving for human joint profiles and force time histories an indeterminate problem best solved by inverse dynamics and optimization methods. A new optimization-based human-modeling framework is thus described for predicting three-dimensional human gait motions on level and inclined planes. The basic unknowns in the framework are the joint motion time histories of a 25-degree-of-freedom human model and its six global degrees of freedom. The joint motion histories are calculated by minimizing an objective function such as deviation of the trunk from upright posture that relates to the human model's performance. A variety of important constraints are imposed on the optimization problem, including (1) satisfaction of dynamic equilibrium equations by requiring the model's zero moment point (ZMP) to lie within the instantaneous geometrical base of support, (2) foot collision avoidance, (3) limits on ground-foot friction, and (4) vanishing yawing moment. Analytical forms of objective and constraint functions are presented and discussed for the proposed human-modeling framework in which the resulting optimization problems are solved using gradient-based mathematical programming techniques. When the framework is applied to the modeling of bipedal locomotion on level and inclined planes, acyclic human walking motions that are smooth and realistic as opposed to less natural robotic motions are obtained. The aspects of the modeling framework requiring further investigation and refinement, as well as potential applications of the framework in biomechanics, are discussed.     

Experimental and analytical tools for studying the human microbiome

The human microbiome substantially affects many aspects of human physiology, including metabolism, drug interactions and numerous diseases. This realization, coupled with ever-improving nucleotide sequencing technology, has precipitated the collection of diverse data sets that profile the microbiome. In the past 2 years, studies have begun to include sufficient numbers of subjects to provide the power to associate these microbiome features with clinical states using advanced algorithms, increasing the use of microbiome studies both individually and collectively. Here we discuss tools and strategies for microbiome studies, from primer selection to bioinformatics analysis.     

An analytical estimation of the energy cost for legged locomotion

Legged locomotion requires the determination of a number of parameters such as stride period, stride length, order of leg movements, leg trajectory, etc. How are these parameters determined? It has been reported that the locomotor patterns of many legged animals exhibit common characteristics, which suggests that there exists a basic strategy for legged locomotion. In this study we derive an equation to estimate the cost of transport for legged locomotion and examine a criterion of the minimization of the transport cost as a candidate of the strategy. The obtained optimal locomotor pattern that minimizes the cost suitably represents many characteristics of the pattern observed in legged animals. This suggests that the locomotor pattern of legged animals is well optimized with regard to the energetic cost. The result also suggests that the existence of specific gait patterns and the phase transition between them could be the result due to optimization; they are induced by the change in the distribution of ground reaction forces for each leg during locomotion.     

Experimental analysis and planning using the Salmonella/microsomes test

A possible statistical treatment of the results obtained in Salmonella typhimurium TA1950, TA1535, TA1537, TA1538, TA98 and TA100 using the Salmonella/microsomes test was investigated. An analysis of independent repeated experiments pointed to the divergence of the data obtained. Consequently, it has been recommended to carry out the statistical treatment of experimental data within the limits of dispersion analysis with the subsequent use of the Scheffe's method for multiple comparisons. To stabilize the dispersion of experimental data, it is suggested to express the number of revertant colonies as lnX. A minimal volume of the data sample needed for resolving experimental tasks has been calculated. In standard experiments aimed at revealing the mutagenic activity of the compounds used, three Petri dishes should be used in each variant of experiments.     

Exotendons for assistance of human locomotion

Background  

Powered robotic exoskeletons for assistance of human locomotion are currently under development for military and medical applications. The energy requirements for such devices are excessive, and this has become a major obstacle for practical applications. Legged locomotion in many animals, however, is very energy efficient. We propose that poly-articular elastic mechanisms are a major contributor to the economy of locomotion in such specialized animals. Consequently, it should be possible to design unpowered assistive devices that make effective use of similar mechanisms.     

Adiabatic transformability hypothesis of human locomotion

It is hypothesized that metabolic and mechanical changes in human locomotion associated with changes in speed v are constrained by two attractive strategies: $Q_{{\text{metab}}} = 1{\text{ and }}\Delta Q_{{\text{metab}}} /\Delta v = {\text{a}}$ positive definite constant. $Q_{{\text{metab}}} = \Delta {\rm E}_{\text{k}} {\text{s}}^{{\text{ - 1}}} /{\text{ml O}}_{\text{2}} {\text{s}}^{{\text{ - 1}}} $ where ΔEs^?1 is the summed increments and decrements per unit time in the translational and rotational kinetic energies of the body's segments and ml O₂s^?1 is the rate at which chemical energy is dissipated. The expected constancy of ΔQ _metab/Δv _metab was derived from an extension of Ehrenfest's adiabatic hypothesis by which transformations (increases, decreases) in locomotion v can be considered as adiabatic, even though the biological conditions are nonconservative and non-rate-limited. The expected significance of Q _metab=1 was derived from stability considerations of the symmetry per stride of stored and dissipated energy. An experimental evaluation was provided by collecting metabolic and mechanical measures on walking (10 subjects) and running (9 subjects) at progressively greater treadmill speeds but within the aerobic limit. Results revealed that walking was restricted to ometab ? 1 with a nonlinear trajectory in v×Q _metab coordinates shaped by Q _metab=1 (primarily) and the constancy of ΔQ _metab/Δv. Running satisfied Q _metab > 1, with a linear trajectory in v×Q _metab coordinates conforming to ΔQ _metab/_Δv=a constant, with the constant predicted from invariants in the mechanical space v×ΔE _ks^?1. Results also suggested that the metabolic costs of running might be predictable from measures made in the v×ΔE _ks^?1 space.     

Assessment of human locomotion by using an insole measurement system and artificial neural networks

A new method for measuring and characterizing free-living human locomotion is presented. A portable device was developed to objectively record and measure foot-ground contact information in every step for up to 24h. An artificial neural network (ANN) was developed to identify the type and intensity of locomotion. Forty subjects participated in the study. The subjects performed level walking, running, ascending and descending stairs at slow, normal and fast speeds determined by each subject, respectively. The device correctly identified walking, running, ascending and descending stairs (accuracy 98.78%, 98.33%, 97.33%, and 97.29% respectively) among different types of activities. It was also able to determine the speed of walking and running. The correlation between actual speed and estimated speed is 0.98, p< 0.0001. The average error of walking and running speed estimation is -0.050+/-0.747 km/h (mean +/- standard deviation). The study has shown the measurement of duration, frequency, type, and intensity of locomotion highly accurate using the new device and an ANN. It provides an alternative tool to the use of a gait lab to quantitatively study locomotion with high accuracy via a small, light and portable device, and to do so under free-living conditions for the clinical applications.     

Estimating the speed of Drosophila locomotion using an automated behavior detection and analysis system

A fundamental phenotypic trait in Drosophila melanogaster is the speed of movement. Its quantification in response to environmental and experimental factors is highly useful for behavioral and neurological studies. Quantifying this behavioral characteristic in freely moving flies is difficult, and many current systems are limited to evaluating the speed of movement of one fly at a time or rely on expensive, time-consuming methods. Here, we present a novel signal processing method of quantifying the speed of multiple flies using a system with automatic behavior detection and analysis that we previously developed to quantify general activity. By evaluating the shape of the signal wave from recordings of a live and simulated single fly, a metric for speed of movement was found. The feasibility of using this metric to estimate the speed of movement in a population of flies was then confirmed by evaluating recordings taken from populations of flies maintained at two different temperatures. The results were consistent with those reported in the literature. This method provides an automated way of measuring speed of locomotion in a fly population, which will further quantify fly behavioral responses to the environment.     

Extracting phase-dependent human vestibular reflexes during locomotion using both time and frequency correlation approaches

Daily activities, such as walking, may require dynamic modulation of vestibular input onto motoneurons. This dynamic modulation is difficult to identify in humans due to limitations in the delivery and analysis of current vestibular probes, such as galvanic vestibular stimulation. Stochastic vestibular stimulation, however, provides an alternative method to extract human vestibular reflexes. Here, we used time-dependent coherence and time-dependent cross-correlation, coupled with stochastic vestibular stimulation, to investigate the phase dependency of human vestibular reflexes during locomotion. We found that phase-dependent activity from the medial gastrocnemius muscles is correlated with the vestibular signals over the 2- to 20-Hz bandwidth during the stance phase of locomotion. Vestibular-gastrocnemius coherence and time-dependent cross-correlations reached maximums at 21 ± 4 and 23 ± 8% of the step cycle following heel contact and before the period of maximal electromyographic activity (38 ± 5%). These results demonstrate 1) the effectiveness of these techniques in extracting the phase-dependent modulation of vestibulomuscular coupling during a cyclic task; 2) that vestibulomuscular coupling is phasically modulated during locomotion; and 3) that the period of strongest vestibulomuscular coupling does not correspond to the period of maximal electromyographic activity in the gastrocnemius. Therefore, we have shown that stochastic vestibular stimulation, coupled with time-frequency decomposition, provides an effective tool to assess the contribution of vestibular ex-afference to the muscular control during locomotion.     

Preparative size-exclusion chromatography of human serum apolipoproteins using an analytical liquid chromatograph

Apolipoproteins, extracted from human serum high-density lipoproteins, can be resolved and recovered with high yield from a preparative MicroPak TSK Type 3000SW size-exclusion column using Tris-buffered 6 m urea or 6 m guanidinium chloride mobile phases. Adequate resolution of some apolipoprotein pairs is only achieved at low flow velocities and low sample loads, necessitating repetitive injections of small amounts of material for preparative isolation. An analytical high-performance liquid chromatograph equipped with a simplified sample introduction scheme and low-pressure switching valves for fraction collection was used to isolate milligram quantities of HDL apolipoproteins.     

Presenting joint kinematics of human locomotion using phase plane portraits and Poincaré maps

Additional graphical tools are needed to better visualize the joint kinematics of human locomotion. Standard plots in which the joint displacements are plotted against time or percent gait cycle do not provide sufficient information about the dynamics of the system. In this article, a study based on the two graphical tools of nonlinear dynamics to visualize the steady-state kinematics of human gait is presented. An experimental setup was developed to acquire the necessary data for application of the techniques. Twenty young adults, whose medical histories are free of gait pathology, were tested. Computerized electrogoniometers and foot switches were used to measure the kinematic data of the lower extremities and capture four instants of the gait cycle: heel strike, foot flat, heel off, and toe off. Phase plane portraits of each joint were constructed for the sagittal plane by plotting angular velocity against angular displacement. Poincaré maps were obtained by periodically sampling the joint profiles at toe off and plotting the ith iterate against the (i + 1)th one. Phase plane portraits are useful in monitoring the variations of joint velocity and position on the same graph in a more compact form. Poincaré maps are effective in differentiating steady gait from transient locomotion.     

In vitro control analysis of an enzyme system: Experimental and analytical developments

In this paper we describe a flow-through system for reconstituting parts of metabolism from purified enzymes. This involves pumping continuously into a reaction chamber, fresh enzymes and reagents so that metabolic reactions occur in the chamber. The waste products leave the chamber via the outflow so that a steady state can be setup. The system we chose consisted of a single enzyme, lactate dehydrogenase. This enzyme was chosen because it consumes NADH in the chamber which could be monitored spectrophotometrically. The aim of the work was to investigate whether a steady state could be achieved in the flow system and whether a metabolic control analysis could be done. We measured two control coefficients, C_LDH and C_pump for the enzyme flux and NADH concentration and confirmed that the summation theorem applied to this system. The advantage of a flow-through system is that the titrations necessary to estimate the control coefficients can be easily and precisely controlled; this means that accurate estimates for the control coefficients can be obtained. In the paper, we discuss some statistical aspects of the data analysis and some possible applications of the technique, including a method to determine the presence of metabolic channelling between two different enzymes.     

Spinal reciprocal inhibition in human locomotion.

The purpose of this paper was to study spinal inhibition during several different motor tasks in healthy human subjects. The short-latency, reciprocal inhibitory pathways from the common peroneal (CP) nerve to the soleus muscle and from the tibial nerve to the tibialis anterior muscle were studied as a depression of ongoing voluntary electromyograph (EMG) activity. First, the effect of stimulus intensity on the amount of inhibition was examined to decide an appropriate stimulation to study the task-dependent modulation of inhibition. Then, the inhibition at one level of stimulation (1.5 x motor threshold) was investigated during standing, walking, and running. The change in slope of inhibition vs. EMG level, which approximates the fraction of ongoing activity that is inhibited, decreased with CP stimulation from 0.52 during standing to 0.30 during fast walking (6 km/h) to 0.17 during running at 9 km/h. Similarly, the slope decreased with tibial nerve stimulation from 0.68 (standing) to 0.42 (fast walking) to 0.35 (running at 9 km/h). All differences, except the last one, were highly significant (P < 0.01, Student's t-test). However, the difference between walking (0.42) and running (0.36) at the same speed (6 km/h) was not significant with tibial nerve stimulation and only significant at P < 0.05 with CP nerve stimulation (0.30, 0.20). Also, the difference between standing (0.52) and slow walking (3 km/h; 0.41) with CP stimulation was not significant, but it was significant (P < 0.01) with tibial nerve stimulation (0.68, 0.49). In conclusion, our findings indicate that spinal reciprocal inhibition decreases substantially with increasing speed and only changes to a lesser extent with task.     

Postural stability of the human mandible during locomotion

Movements of the head and of the mandible relative to the head were measured in human subjects walking and running on a treadmill at various speeds and inclinations. A miniature magnet and piezo-electric accelerometer assembly was mounted on the mandibular incisors, and a Hall-effect sensor along with a second accelerometer mounted on a maxillary incisor along a common vertical axis. Signals from these sensors provided continuous records of vertical head and mandible acceleration, and relative jaw position. Landing on the heel or on the toe in different forms of locomotion was followed by rapid deceleration of the downward movement of the head and slightly less rapid deceleration of the downward movement of the mandible, i.e., the mandible moved downwards relative to the maxilla, then upwards again to near its normal posture within 200 ms. No tooth contact occurred in any forms of gait at any inclination. The movement of the mandible relative to the maxilla depended on the nature and velocity of the locomotion and their effects on head deceleration. The least deceleration and hence mandibular displacement occurred during toe-landing, for example, during "uphill" running. The maximum displacement of the mandible relative to the head was less than 1mm, even at the fastest running speed. The mechanisms that limit the vertical movements of the jaw within such a narrow range are not known, but are likely to include passive soft-tissue visco-elasticity and stretch reflexes in the jaw-closing muscles.