A practical step length algorithm using lower limb angular velocities

The use of Inertial Measurement Units (IMUs) for spatial gait analysis has opened the door to unconstrained measurements within the home and community. Bandwidth, cost limitations, and ease of use has historically restricted the number and location of sensors worn on the body. In this paper, we describe a four-sensor configuration of IMUs placed on the shanks and thighs that is sufficient to provide an accurate measure of temporal gait parameters, spatial gait parameters, and joint angle dynamics during ambulation. Estimating spatial gait parameters solely from gyroscope data is preferred because gyroscopes are less susceptible to sensor noise and a system comprised of only gyroscopes uses decreased bandwidth compared to a typical 9 degree-of-freedom IMU. The purpose of this study was to determine the validity of a novel method of step length estimation using gyroscopes attached to the shanks and thighs. An Inverted Pendulum Model algorithm (IPM) was proposed to calculate step length, stride length, and gait speed. The algorithm incorporates heel-strike events and average forward velocity per step to make these assessments. IMU algorithm accuracy was determined via concurrent validity with an instrumented walkway and results explained via the collision model of gait. The IPM produced accurate estimates of step length, stride length, and gait speed with a mean difference of 3 cm and an RMSE of 6.6 cm for step length, thus establishing a new approach for spatial gait parameter calculation. The lack of numerical integration in IPM makes it well suited for use in continuous monitoring applications where sensor sampling rates are restricted.     

The discriminant capabilities of stability measures, trunk kinematics, and step kinematics in classifying successful and failed compensatory stepping responses by young adults

This study evaluated the discriminant capability of stability measures, trunk kinematics, and step kinematics to classify successful and failed compensatory stepping responses. In addition, the shared variance between stability measures, step kinematics, and trunk kinematics is reported. The stability measures included the anteroposterior distance (d) between the body center of mass and the stepping limb toe, the margin of stability (MOS), as well as time-to-boundary considering velocity (TTB(v)), velocity and acceleration (TTB(a)), and MOS (TTB(MOS)). Kinematic measures included trunk flexion angle and angular velocity, step length, and the time after disturbance onset of recovery step completion. Fourteen young adults stood on a treadmill that delivered surface accelerations necessitating multiple forward compensatory steps. Thirteen subjects fell from an initial disturbance, but recovered from a second, identical disturbance. Trunk flexion velocity at completion of the first recovery step and trunk flexion angle at completion of the second step had the greatest overall classification of all measures (92.3%). TTB(v) and TTB(a) at completion of both steps had the greatest classification accuracy of all stability measures (80.8%). The length of the first recovery step (r ≤ 0.70) and trunk flexion angle at completion of the second recovery step (r ≤ -0.54) had the largest correlations with stability measures. Although TTB(v) and TTB(a) demonstrated somewhat smaller discriminant capabilities than trunk kinematics, the small correlations between these stability measures and trunk kinematics (|r| ≤ 0.52) suggest that they reflect two important, yet different, aspects of a compensatory stepping response.     

Estimating lipid and lean body mass in small passerine birds using TOBEC,external morphology and subcutaneous fat‐scoring

To assess regression models for lipid and lean body mass in small birds, we recorded live body mass ±0.1 g, total body electrical conductivity (TOBEC; from “third generation” TOBEC machine EM‐SCAN® SA‐3000) or E‐Value, visual fat score (VisFat), and seven body measurements for 52 migratory passerine birds of 13 species (5–40 g). We determined lipid and lean mass of each bird after petroleum‐ether extraction of lipids. We obtained “net”E‐Value (NEV) for each scanned bird by subtracting the E‐Value of the empty bird‐restraining tube, because these showed an inverse temperature dependence (P<0.005). Leave‐one‐out cross validation was used to assess model selection and construct 95% confidence intervals. Although precision of TOBEC increased with bird size (CV of NEV vs. live mass: r=−0.276, P=0.002) and it explained an increasing proportion of variation in lean mass moving from small‐ to medium‐ to large‐bird classes of our data, it did no better than head length in single‐variable prediction of lean or lipid mass and was included in five of the 14 multivariate models we developed. The best multiple regression to predict lean mass included live weight, VisFat, bill length, tarsus and lnNEV (adjusted R²=99.0%); however, the same model lacking only lnNEV yielded aR²=98.9%. A parallel to the above pair of models, but predicting lipid mass, yielded aR²=90.3% and 90.0%, respectively. Subdividing the data by three size classes and three taxa (American redstart Setophaga ruticilla, ovenbird Seiurus aurocapilla, warblers), best‐subset multiple‐regression models predicted lean mass with aR² from 94.7 to 99.6% and lipid mass with aR² from 85.4 to 98.3%. Best models for the size‐ and species‐groups included VisFat and zero to five body measurements, and most included live weight. lnNEV was included only in the models for ovenbird (lipid), warblers (lipid), all birds (both), and large birds (both). Actual lipid mass of all birds was more highly correlated with multiple‐regression‐predicted lipid mass (r=0.955) than with visual subcutaneous fat‐scoring (r=0.683). These multiple‐regression models predicting lipid content using live‐bird measurements and visual fat score as independent variables represent more accurate and precise estimates of actual lipid content in small passerines than any previously published. They are particularly accurate for placing birds into percentage body‐fat classes.     

Mechanisms of adaptation from a multiple to a single step recovery strategy following repeated exposure to forward loss of balance in older adults

When released from an initial, static, forward lean angle and instructed to recover with a single step, some older adults are able to meet the task requirements, whereas others either stumble or fall. The purpose of the present study was to use the concept of margin of stability (MoS) to investigate balance recovery responses in the anterior-posterior direction exhibited by older single steppers, multiple steppers and those that are able to adapt from multiple to single steps following exposure to repeated forward loss of balance. One hundred and fifty-one healthy, community dwelling, older adults, aged 65-80 years, participated in the study. Participants performed four trials of the balance recovery task from each of three initial lean angles. Balance recovery responses in the anterior-posterior direction were quantified at three events; cable release (CR), toe-off (TO) and foot contact (FC), for trials performed at the intermediate lean angle. MoS was computed as the anterior-posterior distance between the forward boundary of the Base of Support (BoS) and the vertical projection of the velocity adjusted centre of mass position (XCoM). Approximately one-third of participants adapted from a multiple to a single step recovery strategy following repeated exposure to the task. MoS at FC for the single and multiple step trials in the adaptation group were intermediate between the exclusively single step group and the exclusively multiple step group, with the single step trials having a significant, 3.7 times higher MoS at FC than the multiple step trials. Consistent with differences between single and multiple steppers, adaptation from multiple to single steps was attributed to an increased BoS at FC, a reduced XCoM at FC and an increased rate of BoS displacement from TO to FC. Adaptations occurred within a single test session and suggest older adults that are close to the threshold of successful recovery can rapidly improve dynamic stability following repeated exposure to a forward loss of balance.     

Minimal step length necessary for recovery of forward balance loss with a single step

Although the boundary conditions necessary to trigger a step in reaction to a forward balance loss have been predicted in previous research, the relationship between minimal step length needed for balance recovery with this single step and the center of mass (COM) motion state (i.e., its position and velocity) remains unknown. The purpose of this paper was to present a theoretical framework within which the minimal step length needed for balance recovery can be estimated. We therefore developed a simplified four-segment sagittal model of human body stepping for balance recovery. The work-energy principle of the Newtonian mechanics was employed in the simulation to determine the amount of excess mechanical energy that can be absorbed as a function of step length and the corresponding eccentric joint work that can be generated in a single step. We found that an increase in initial forward velocity and a greater forward shift of the COM require a corresponding increase in the minimal step length needed for balance recovery. Furthermore, the minimal step length is also a function of the muscle strength at the ankle: the lower the muscle strength, the greater the minimal step length required. Our theoretical framework reduces the complexity associated with previous studies relying on forward dynamics and iterative optimization processes. This method may also be applied to study aspects of balance control such as the prevention of balance loss in the posterior or mediolateral direction.     

A novel method to determine lean body water using localized skin biopsies: correlation between lean skin water and lean body water in an overhydration model

To determine the relationship between total body water (TBW) fraction and local water content measured in the skin (SW) this study assessed eight anesthetized piglets in an overhydration model. TBW was assessed by deuterium oxide dilution and body mass measurements taken throughout the experiments, and by whole body carcass analysis at the end of each experiment. Additionally, extracellular water and plasma volume were assessed using bromide dilution and Evan's blue dilution, respectively. SW was assessed by tissue biopsies taken at 60-min intervals throughout the experiment. Lean body water (LBW) fraction and lean skin water (LSW) fraction were assessed by extracting the fat from the carcass and biopsy samples. A correlation does exist between TBW fraction and SW fraction with r2=0.58 (P<0.05); however, the strongest correlation occurred between the LBW fraction and LSW fraction with r2=0.87 (P<0.05) and an SE of prediction of 0.77%. These data demonstrate that LSW gives an accurate and precise estimate of LBW and could therefore be used to determine the hydration index in appropriate research settings.     

The effect of cane length and step height on muscle strength and body balance of elderly people in a stairway environment

Background

It has been reported that 75% of stairway accidents occur while descending stairs. Using a cane can help to prevent older people and those with limited mobility from falling. However, studies have shown that two-thirds of older cane users use a cane that is longer than the recommended length, which may cause unnecessary muscular loads. This study aims to assess balance and muscular load in older people descending different height steps with different cane lengths.

Methods

Nine participants (5 males and 4 females) aged over 65 years participated in this study. Cane length and stair height were independent variables. Electromyography signals were recorded from the biceps brachii of the arm that usually held the cane and from both gastrocnemius muscles. In addition, the center of pressure (CoP) was assessed as an indicator of balance in older people descending a step.

Results

Descending from higher steps resulted in the use of greater arm and leg strength at the time of first foot contact. However, cane length did not affect any of the root mean square values. In addition, the CoP Stabilometric Parameters showed that mean distance, antero-posterior mean distance, total excursions, antero-posterior total excursions, mean velocity, and antero-posterior mean velocity were significantly affected by step height, but not by cane length.

Conclusions

If cane length is within the currently suggested range, then it has little effect on the force load on the arm and legs when descending a step. Step height has a greater effect than cane length on the strategies used by older people to maintain stability.     

Photogrammetry: a useful tool for three‐dimensional morphometric analysis of small mammals

Recently, the improvement of methods for shape analysis has revolutionized the field of morphometrics. While three‐dimensional (3D) imaging technology is increasingly available, many studies of 3D structures still use two‐dimensional (2D) data, even when this may result in the loss of important information. This is particularly conspicuous in the study of small mammals, as devices precise enough for 3D digitization of small objects are the most expensive. Thus, the development of low‐cost methods aimed to recover 3D shape from small mammals would be of wide interest. Photogrammetry allows for obtaining 3D data with a lower cost than other 3D techniques, but it has not been previously applied to the study of small mammals. Accordingly, here we test the suitability of photogrammetric techniques to obtain 3D landmarks on mouse skulls as a model for small mammals. Shape and size of 3D models obtained with photogrammetric techniques were consistent among replicates, even when different sets of photographs were used. The linear measurements obtained from 3D models produced here were highly correlated with measurements obtained with callipers on actual crania, and differences among both sets of measures were smaller than those among individuals in most of the tested measures. These results show for the first time that photogrammetry is a precise technique for 3D shape analysis of small mammals. Photogrammetry also proved to be accurate for obtaining linear measurements between 3D landmarks; however, further studies are needed to demonstrate that this technique is also accurate to recreate 3D shapes.     

Simplified low-cost methodology to establish,histologically process and analyze three-dimensional cancer cell spheroid arrays

Differently of two-dimensional cell culture, three-dimensional (3D) multicellular spheroid model allows cells to establish cell-cell/cell-matrix interactions over the entire cell surface, more closely mimicking tumor microenvironments and cellular subpopulations with specific standards of morphology, differentiation and gene expression. Thenceforth several methodologies involving or the 3D cell aggregates generation or its histological processing and analysis have emerged, but in general they are laborious, expensive and complex to set up as a routine technique. Thus, we developed a complete methodology, detailing a simple, accessible and low-cost step by step, including 1) the 3D cell aggregate generation using hanging drop technique; 2) providing a simple way to assess morphological parameters of generated spheroids; followed by 3) a multiple and organized histological processing, keeping several individual spheroids inside an agarose apparatus, maintaining a known order and position of each ones, similar to tissue microarray principle; 4) until the last step, where it is allowed a simultaneous histological composition analysis of several spheroid slices, organized side by side, in a same block section, through conventional stainings or 5) immunostaining against different molecular markers. Therefore, the present methodology aims to popularize 3D cell culture, allowing to make this a regular technique in basic cell biology research, once all steps are performed without using onerous reagents, materials or equipment. In addition to bring the agarose apparatus as a simple low cost novelty, allowing high-throughput analysis of several spheroids simultaneously in an organized manner.     

Quantification of metabolically active biomass using Methylene Blue dye Reduction Test (MBRT): measurement of CFU in about 200 s

Quantification of viable cells is a critical step in almost all biological experiments. Despite its importance, the methods developed so far to differentiate between viable and non-viable cells suffer from major limitations such as being time intensive, inaccurate and expensive. Here, we present a method to quantify viable cells based on reduction of methylene blue dye in cell cultures. Although the methylene blue reduction method is well known to check the bacterial load in milk, its application in the quantification of viable cells has not been reported. We have developed and standardized this method by monitoring the dye reduction rate at each time point for growth of Escherichia coli. The standard growth curve was monitored using this technique. The Methylene Blue dye Reduction Test (MBRT) correlates very well with Colony Forming Units (CFU) up to a 800 live cells as established by plating. The test developed is simple, accurate and fast (200 s) as compared to available techniques. We demonstrate the utility of the developed assay to monitor CFU rapidly and accurately for E. coli, Bacillus subtilis and a mixed culture of E. coli and B. subtilis. This assay, thus, has a wide applicability to all types of aerobic organisms.     

Tension responses of frog skeletal muscle to ramp and step length changes

Tension responses to ramp shortening of varying speed in whole muscle or single fibres from the plateau of an isometric tetanus, revealed at least two distinct phases. There was a fast initial drop in tension followed by a change of slope and a definite inflexion on the tension record. As the velocity of the imposed length change was increased, the inflexion point appeared at a lower tension. Similar inflexions were not observed during ramp releases to an elastic band or a segment of semitendinosus tendon. The tension records obtained with moderately fast ramp length changes to contracting muscle reflect the T1 and T2 phases of the tension transients.     

Computer-aided 2D and 3D quantification of human stem cell fate from in vitro samples using Volocity high performance image analysis software

Accurate automated cell fate analysis of immunostained human stem cells from 2- and 3-dimensional (2D-3D) images would improve efficiency in the field of stem cell research. Development of an accurate and precise tool that reduces variability and the time needed for human stem cell fate analysis will improve productivity and interpretability of the data across research groups. In this study, we have created protocols for high performance image analysis software Volocity? to classify and quantify cytoplasmic and nuclear cell fate markers from 2D-3D images of human neural stem cells after in vitro differentiation. To enhance 3D image capture efficiency, we optimized the image acquisition settings of an Olympus FV10i? confocal laser scanning microscope to match our quantification protocols and improve cell fate classification. The methods developed in this study will allow for a more time efficient and accurate software based, operator validated, stem cell fate classification and quantification from 2D and 3D images, and yield the highest ≥94.4% correspondence with human recognized objects.     

Increasing step width reduces the requirements for subtalar joint moments and powers

The subtalar joint (STJ) contributes to the absorption and generation of mechanical energy (and power) during walking to maintain frontal plane stability. Previous observational studies have suggested that there may be a relationship between step width and STJ supination moment. This study directly tests the hypothesis that walking with a step width greater than preferred would reduce STJ moments, energy absorption, and power generation requirements, while increasing energy absorption at the hip during initial contact. Participants (n = 12, 7 females) were asked to walk on an instrumented treadmill at a constant velocity and cadence at a range of fixed step widths ranging from 0.1 to 0.4 times leg length (L). Walking at step widths greater than preferred (0.149 ± 0.04 L) reduced peak STJ moments at initial contact and propulsion which subsequently reduced the negative and positive work performed at the STJ. There was a 43% reduction in energy absorption (negative work) and approximately 30% decrease in positive work at the STJ as step width increased from 0.1 L to 0.4 L. An increase in energy absorption at the knee and hip was evident with an increase in step width during initial contact, although minimal mechanical changes were observed at the proximal joints during propulsion. These results suggest an increase in step width reduces the forces generated by muscles at the STJ across stance and is therefore likely to be beneficial in the prevention and treatment of their injuries. In terms of rehabilitation, the increase in mechanical costs occurring due to an increase in energy absorption by the hip and knee is of minimal concern.     

Contractile equilibration of single cells to step changes in extracellular stiffness

Extracellular stiffness has been shown to alter long timescale cell behaviors such as growth and differentiation, but the cellular response to changes in stiffness on short timescales is poorly understood. By studying the contractile response of cells to dynamic stiffness conditions using an atomic force microscope, we observe a seconds-timescale response to a step change in extracellular stiffness. Specifically, we observe acceleration in contraction velocity (μm/min) and force rate (nN/min) upon a step decrease in stiffness and deceleration upon a step increase in stiffness. Interestingly, this seconds-timescale response to a change in extracellular stiffness is not altered by inhibiting focal adhesion signaling or stretch-activated ion channels and is independent of cell height and contraction force. Rather, the response timescale is altered only by disrupting cytoskeletal mechanics and is well described by a simple mechanical model of a constant velocity actuator pulling against an internal cellular viscoelastic network. Consistent with the predictions of this model, we find that an osmotically expanding hydrogel responds to step changes in extracellular stiffness in a similar manner to cells. We therefore propose that an initial event in stiffness sensing is establishment of a mechanical equilibrium that balances contraction of the viscoelastic cytoskeleton with deformation of the extracellular matrix.     

Creation of accurate 3D models of harbor porpoises (Phocoena phocoena) using 3D photogrammetry

Creating accurate 3D models of marine mammals is valuable for assessment of body condition, computational fluids dynamics models of locomotion, and for education. However, the methods for creating 3D models are not well-developed. We used photography and video to create 3D photogrammetry models of harbor porpoises (Phocoena phocoena). We accessed one live adult female (155.5 cm total length), and two dead animals, one juvenile (110 cm total length) and one calf (88 cm total length). We accessed the two dead individuals through a stranding network in Germany, and the live individual through the Fjord and Baelt research center in Denmark. For all porpoises, we used still photographs from hand-held cameras, drone video, and synchronized GoPro videos to create 3D photogrammetric models. We used Blender software, and other 3D reconstruction software, to recreate the 3D body meshes, and confirmed the accuracy of each of the 3D body meshes by comparing digital measures on the 3D models to original measures taken on the specimens. We also provide a colored, animated version of the live harbor porpoise for educational purposes. These open-access 3D models can be used to develop methods to study body morphometrics and condition, and to study bioenergetics and locomotion costs.     

Walking speed and spatiotemporal step mean measures are reliable during feedback-controlled treadmill walking; however,spatiotemporal step variability is not reliable

The purpose of the study was to compare the effects of a feedback-controlled treadmill (FeedbackTM) to a traditional fixed-speed treadmill (FixedTM) on spatiotemporal gait means, variability, and dynamics. The study also examined inter-session reliability when using the FeedbackTM. Ten young adults walked on the FeedbackTM for a 5-minute familiarization followed by a 16-minute experimental trial. They returned within one week and completed a 5-minute familiarization followed by a 16-minute experimental trial each for FeedbackTM and FixedTM conditions. Mean walking speed and step time, length, width, and speed means and coefficient of variation were calculated from all experimental conditions. Step time, length, width, and speed gait dynamics were analyzed using detrended fluctuation analysis. Mean differences between experimental trials were determined using ANOVAs and reliability between FeedbackTM sessions was determined by intraclass correlation coefficient. No difference was found in mean walking speed nor spatiotemporal variables, with the exception of step width, between the experimental trials. All mean spatiotemporal variables demonstrated good to excellent reliability between sessions, while coefficient of variation was not reliable. Gait dynamics of step time, length, width, and speed were significantly more persistent during the FeedbackTM condition compared to FixedTM, especially step speed. However, gait dynamics demonstrated fair to poor reliability between FeedbackTM sessions. When walking on the FeedbackTM, users maintain a consistent set point, yet the gait dynamics around the mean are different when compared to walking on a FixedTM. In addition, spatiotemporal gait dynamics and variability may not be consistent across separate days when using the FeedbackTM.     

Evaluation of a Quantitative Magnetic Resonance Imaging System for Whole Body Composition Analysis in Rodents

We evaluated the EchoMRI‐900 combination rat and mouse quantitative magnetic resonance (QMR) body composition method in comparison to traditional whole‐body chemical carcass composition analysis (CCA) for measurements of fat and fat‐free mass in rodents. Live and postmortem (PM) QMR fat and lean mass measurements were obtained for lean, obese and outbred strains of rats and mice, and compared with measurements obtained using CCA. A second group of rats was measured before and after 18 h food or water deprivation. Significant positive correlations between QMR and CCA fat and lean mass measurements were shown for rats and mice. Although all live QMR fat and lean measurements were more precise than CCA for rats, values obtained for mice significantly differed from CCA for lean mass only. QMR performed PM slightly overestimated fat and lean values relative to live QMR but did not show lower precision than live QMR. Food deprivation reduced values for both fat and lean mass; water deprivation reduced estimates of lean mass only. In summary, all measurements using this QMR system were comparable to those obtained by CCA, but with higher overall precision, similar to previous reports for the murine QMR system. However, PM QMR measurements slightly overestimated live QMR values, and lean and fat mass measurements in this QMR system are influenced by hydration status and animal size, respectively. Despite these caveats, we conclude that the EchoMRI QMR system offers a fast in vivo method of body composition analysis, well correlated to but with greater overall precision than CCA.     

Control of center of mass motion state through cuing and decoupling of spontaneous gait parameters in level walking

Can the center of mass (COM) motion state, i.e., its position and velocity relative to the base of support (BOS), which dictate gait stability, be predictably controlled by the global gait parameters of step length and gait speed, or by extension, cadence? The precise relationships among step length and gait speed, and the COM motion state are unknown, partially due to the interdependence between step length and gait speed and the difficulty in independent control of both parameters during spontaneous level walking. The purposes of this study were to utilize simultaneous audio-visual cuing to independently manipulate step length and gait speed, and to determine the extent to which the COM position and velocity can be subsequently controlled. Fifty-six young adults were trained at one of the three gait patterns in which both the step length and gait speed were targeted simultaneously. The results showed that the cuing could successfully “decouple” gait speed from step length. Although this approach did yield reliable control of the COM velocity through manipulation of gait speed (R²=0.97), the manipulation of step length yielded less precise control of COM position (R²=0.60). This latter control appears to require manipulation of an additional degree-of-freedom at the local segment level, such that the inclusion of trunk inclination with step length improved the prediction of COM position (R²=0.80).     

Genotype and diet effects in lean and obese Zucker rats fed either safflower or coconut oil diets

Previously we reported that suckling lean heterozygous (FA/fa) Zucker rats had a number of adipose tissue measurements intermediate between those of homozygous lean (FA/FA) and obese (fa/fa) rats. However, in young adult male rats maintained on a low-fat diet, these differences were no longer apparent (i.e., values for the two lean genotypes were similar). In the present study we determined whether the heterozygous effect of the "fa" gene was dependent on the consumption of a high-fat diet. Mother rats were fed high-fat diets containing either safflower (SOD) or coconut (COD) oil throughout mating and lactation. Homozygous lean male and female rats were bred, as well as obese male and lean heterozygous female rats. Suckling rats were studied at 17 days of age. Additional male rats were maintained on the same diet as their mothers until 11-12 weeks of age. Obese suckling rats had higher body weights than lean pups. Inguinal fat pad weights and pad-to-body weight ratios followed the pattern of obese greater than lean (FA/fa) pups that were greater than lean (FA/FA) pups. A similar relationship was found for adipose tissue lipogenic enzyme activities. At 11-12 weeks of age, measurements followed the general pattern of obese rats having greater values than lean rats (i.e., FA/fa = FA/FA). SOD-fa/fa rats had higher hepatic lipogenic enzyme activities than COD-fa/fa rats. In addition, SOD rats had higher fat cell numbers than COD rats. These results suggest that specific fatty acids can alter adipocyte proliferation and/or differentiation in vivo. In addition, there appears to be a defect of fatty acid regulation in livers of genetically obese rats. The heterozygous effect of the "fa" gene in suckling Zucker rats was confirmed. However, high-fat feeding did not result in a heterozygous effect in young adult lean male rats. We will next evaluate the role of sex on this effect.