It is known that cell density influences the maturation process of in vitro neuronal networks. Neuronal cultures plated with different cell densities differ in number of synapses per neuron and thus in single neuron synaptic transmission, which results in a density-dependent neuronal network activity. Although many authors provided detailed information about the effects of cell density on neuronal culture activity, a dedicated report of density and age influence on neuronal hippocampal culture activity has not yet been reported. Therefore, this work aims at providing reference data to researchers that set up an experimental study on hippocampal neuronal cultures, helping in planning and decoding the experiments. In this work, we analysed the effects of both neuronal density and culture age on functional attributes of maturing hippocampal cultures. We characterized the electrophysiological activity of neuronal cultures seeded at three different cell densities, recording their spontaneous electrical activity over maturation by means of MicroElectrode Arrays (MEAs). We had gather data from 86 independent hippocampal cultures to achieve solid statistic results, considering the high culture-to-culture variability. Network activity was evaluated in terms of simple spiking, burst and network burst features. We observed that electrical descriptors were characterized by a functional peak during maturation, followed by a stable phase (for sparse and medium density cultures) or by a decrease phase (for high dense neuronal cultures). Moreover, 900 cells/mm2 cultures showed characteristics suitable for long lasting experiments (e.g. chronic effect of drug treatments) while 1800 cells/mm2 cultures should be preferred for experiments that require intense electrical activity (e.g. to evaluate the effect of inhibitory molecules). Finally, cell cultures at 3600 cells/mm2 are more appropriate for experiments in which time saving is relevant (e.g. drug screenings). These results are intended to be a reference for the planning of in vitro neurophysiological and neuropharmacological experiments with MEAs.
Breathing frequency (fB) is an important vital sign that—if appropriately monitored—may help to predict clinical adverse events. Inertial sensors open the door to the development of low-cost, wearable, and easy-to-use breathing-monitoring systems. The present paper proposes a new posture-independent processing algorithm for breath-by-breath extraction of breathing temporal parameters from chest-wall inclination change signals measured using inertial measurement units. An important step of the processing algorithm is dimension reduction (DR) that allows the extraction of a single respiratory signal starting from 4-component quaternion data. Three different DR methods are proposed and compared in terms of accuracy of breathing temporal parameter estimation, in a group of healthy subjects, considering different breathing patterns and different postures; optoelectronic plethysmography was used as reference system. In this study, we found that the method based on PCA-fusion of the four quaternion components provided the best fB estimation performance in terms of mean absolute errors (<2 breaths/min), correlation (r > 0.963) and Bland–Altman Analysis, outperforming the other two methods, based on the selection of a single quaternion component, identified on the basis of spectral analysis; particularly, in supine position, results provided by PCA-based method were even better than those obtained with the ideal quaternion component, determined a posteriori as the one providing the minimum estimation error. The proposed algorithm and system were able to successfully reconstruct the respiration-induced movement, and to accurately determine the respiratory rate in an automatic, position-independent manner.
Immersive virtual reality (IVR) offers new possibilities to perform treatments in an ecological and interactive environment with multimodal online feedbacks. Sixteen school-aged children (mean age 11 ± 2.4 years) with Bilateral CP-diplegia, attending mainstream schools were recruited for a pilot study in a pre-post treatment experimental design. The intervention was focused on walking competences and endurance and performed by the Gait Real-time Analysis Interactive Lab (GRAIL), an innovative treadmill platform based on IVR. The participants underwent eighteen therapy sessions in 4 weeks. Functional evaluations, instrumental measures including GAIT analysis and parental questionnaire were utilized to assess the treatment effects. Walking pattern (stride length left and right side, respectively p = 0.001 and 0.003; walking speed p = 0.001), endurance (6MWT, p = 0.026), gross motor abilities (GMFM-88, p = 0.041) and most kinematic and kinetic parameters significantly improved after the intervention. The changes were mainly predicted by age and cognitive abilities. The effect could have been due to the possibility of IVR to foster integration of motor/perceptual competences beyond the training of the walking ability, giving a chance of improvement also to older and already treated children.
Single-treatment approaches seem to be more effective than mixed approaches, independently from the duration (4 or 10 weeks). RAGT seems to have similar effect with respect to the traditional TOP, at least over 10 weeks.
Objective. The minimum clinically important difference (MCID) is a standard way of measuring clinical relevance. The objective of this work was to establish the MCID for the 6-minute walking test (6minWT) and the Gross Motor Function Measure (GMFM-88) in pediatric gait disorders. Methods. A cohort, pretest-posttest study was conducted in a hospitalized care setting. A total of 182 patients with acquired brain injury (ABI) or cerebral palsy (CP) performed 20 robot-assisted gait training sessions complemented with 20 sessions of physical therapy over 4 weeks. Separate MCIDs were calculated using 5 distribution-based approaches, complemented with an anonymized survey completed by clinical professionals. Results. The MCID range for the 6minWT was 20-38 m in the ABI cohort, with subgroup ranges of 20-36 m for GMFCS I-II, 23-46 m for GMFCS III, and 24-46 m for GMFCS IV. MCIDs for the CP population were 6-23 m, with subgroup ranges of 4-28 m for GMFCS I-II, 9-19 m for GMFCS III, and 10-27 m for GMFCS IV. For GMFM-88 total score, MCID values were 1.1%-5.3% for the ABI cohort and 0.1%-3.0% for the CP population. For dimension “D” of the GMFM, MCID ranges were 2.3%-6.5% and 0.8%-5.2% for ABI and CP populations, respectively. For dimension “E,” MCID ranges were 2.8%-6.5% and 0.3%-4.9% for ABI and CP cohorts, respectively. The survey showed a large interquartile range, but the results well mimicked the distribution-based methods. Conclusions. This study identified for the first time MCID ranges for 6minWT and GMFM-88 in pediatric patients with neurological impairments, offering useful insights for clinicians to evaluate the impact of treatments. Distribution-based methods should be used with caution: methods based on pre-post correlation may underestimate MCID when applied to patients with small improvements over the treatment period. Our results should be complemented with estimates obtained using consensus- and anchor-based approaches.
Wearable sensors are becoming increasingly popular for complementing classical clinical assessments of gait deficits. The aim of this review is to examine the existing knowledge by systematically reviewing a large number of papers focusing on the use of wearable inertial sensors for the assessment of gait during the 6-minute walk test (6MWT), a widely recognized, simple, non-invasive, low-cost and reproducible exercise test. After a systematic search on PubMed and Scopus databases, two raters evaluated the quality of 28 full-text articles. Then, the available knowledge was summarized regarding study design, subjects enrolled (number of patients and pathological condition, if any, age, male/female ratio), sensor characteristics (type, number, sampling frequency, range) and body placement, 6MWT protocol and extracted parameters. Results were critically discussed to suggest future directions for the use of inertial sensor devices in the clinics.
Quantitative evaluation of gait has been considered a useful tool with which to identify subtle signs of motor system peculiarities in autism spectrum disorder (ASD). However, there is a paucity of studies reporting gait data in ASD as well as investigating learning processes of locomotor activity. Novel advanced technologies that couple treadmills with virtual reality environments and motion capture systems allows the evaluation of gait patterns on multiple steps and the effects of induced gait perturbations, as well as the ability to manipulate visual and proprioceptive feedbacks. This study aims at describing the gait pattern and motor performance during discrete gait perturbation of drug-naïve, school-aged children with ASD compared to typically developing (TD) peers matched by gender and age. Gait analysis was carried out in an immersive virtual environment using a 3-D motion analysis system with a dual-belt, instrumented treadmill. After 6 min of walking, 20 steps were recorded as baseline. Then, each participant was exposed to 20 trials with a discrete gait perturbation applying a split-belt acceleration to the dominant side at toe-off. Single steps around perturbations were inspected. Finally, 20 steps were recorded during a post-perturbation session. At baseline, children with ASD had reduced ankle flexion moment, greater hip flexion at the initial contact, and greater pelvic anteversion. After the discrete gait perturbation, variations of peak of knee extension significantly differed between groups and correlated with the severity of autistic core symptoms. Throughout perturbation trials, more than 60% of parameters showed reliable adaptation with a decay rate comparable between groups. Overall, these findings depicted gait peculiarities in children with ASD, including both kinetic and kinematic features; a motor adaptation comparable to their TD peers, even though with an atypical pattern; and a motor adaptation rate comparable to TD children but involving different aspects of locomotion. The platform showed its usability with children with ASD and its reliability in the definition of paradigms for the study of motor learning while doing complex tasks, such as gait. The additional possibility to accurately manipulate visual and proprioceptive feedback will allow researchers to systematically investigate motor system features in people with ASD.
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