We would like to thank Dr. Johnathan Williams for his advice on MATLAB data processing.

The method reported was followed in a study that received ethical approval from the Bournemouth University Research Ethics Committee (Reference 15005).
1. Participants
<ol>
	<li>Recruit healthy adults (aged from 23 to 63 years, mean &#177; S.D.; 42.0 &#177; 13.4, body mass 70.4 &#177; 15.3 kg, height 175.5 &#177; 9.8 cm; 15 males, 15 females) to participate in the study. Thirty participants were recruited for this study.</li>
	<li>Ensure that there is no self-reported history of dizziness, balance problems or walking difficulties in the participants.</li>
	<li>Ensure that participants did not suffer from any known neuromuscular injury or condition affecting balance or walking.</li>
</ol>
2. Setup and procedures for gait analysis
<ol>
	<li>Use a gait analysis system (Figure 1) comprised of a dual-belt force plate-instrumented treadmill, a 10-camera motion capture system and a virtual environment that provides optic flow.</li>
	<li>Ensure that the participant is wearing very tight non-reflective clothing such as cycling shorts or leggings.</li>
	<li>Using double sided adhesive tapes attach 25 passive reflective markers and place according to the lower body configuration of the Human Body Model (HBM)7 as detailed in Table 1 and Figure 2. The information in this document is taken from the HBM Reference Manual8.</li>
	<li>Use a joint ruler to take measurements of the required knee and ankle widths for the HBM6.</li>
	<li>Secure participant to a safety harness that is fastened to an overhead frame.</li>
	<li>Start a new session in the database and make sure it is active (highlighted).</li>
	<li>Using the subject tab, create a new participant from the Labeling Skeleton button.</li>
	<li>Browse to the &#39;LowerLimb HBM_N2.vst&#39; file and then enter the name of the participant. The new participant appears in the Subjects pane.</li>
	<li>Go to the Tools pane and open the Subject Preparation tab.</li>
	<li>Zero level the forceplates via the Hardware tab. Make sure that no weight is exerted on the force plates.</li>
	<li>Prepare the participant for the ROM trial by having them ready in the middle of the treadmill.</li>
	<li>To ensure the participant can accustom themselves to the self-paced treadmill, ask them to walk at a comfortable speed for 5 min at the beginning of the session9,10.</li>
	<li>Following the acclimatization and without any delay time, ask the participant to walk for a minimum of 5 min10,11.</li>
	<li>Ensure participants are blinded to the timing of the recordings.</li>
	<li>Ensure to start the treadmill and start data recordings by clicking the Start recording button12. This can be done with integrated software (Table of Materials).</li>
	<li>Stop the recording after acquiring the desired amount of data. It is recommended to collect three sets of 25 cycles.</li>
	<li>Open the processing software (Table of Materials) and remove the high-frequency noise on data, by selecting a low-pass filter to the marker data such as a second order Butterworth filter with a cut-off frequency of 6 Hz.</li>
	<li>Go to File, and then select Export to save as a .csv.</li>
	<li>Determine individual strides from vertical force data and use the foot markers to ascertain gait events13.</li>
	<li>Analyze the gait parameters such as kinematic, kinetic and spatial-temporal data in Matlab R2017a (Supplementary File).</li>
</ol>
3. Setup and procedures for muscle strength test
<ol>
	<li>Use the muscle strength testing equipment (multimodal dynamometer) (Figure 3), to measure participants&#39; muscle strength based on Maximum Voluntary Isometric Contraction (MVIC)14.</li>
	<li>Attach the tool/pad number 701 to the dynamometer exercise head.</li>
	<li>Test participant&#39;s right and left knee isometric muscle strength.</li>
	<li>Test participants in a seated position on a chair with a backrest.</li>
	<li>Using the up/down switch, align the dynamometer axis with the knee joint&#39;s anatomical axis of rotation. Place the pad of the tool centrally at the lower part of the shin of the tibia.</li>
	<li>Keep the knee at 90&#176; flexion, the hip in neutral rotation and abduction, and the foot in plantar flexion.</li>
	<li>Place the participant&#39;s hands on their abdomen and stabilize the trunk, hips, and mid-thigh on the chair with Velcro straps.</li>
	<li>Run a practice trial for participants to get accustomed to the testing maneuver.</li>
	<li>Instruct the participant to extend their knee (exert pressure upwards on the pad) followed by flex (exert pressure downwards on the pad) to exert a maximum contraction on the command Go for 3 s.</li>
	<li>Provide verbal prompts and encouragement (&#34;Push&#34; for upwards and &#34;Pull&#34; for downwards) during the strength testing.</li>
	<li>Ensure that participants are aware they can stop the test immediately if they experience any unusual pain or discomfort.</li>
	<li>Allow participants to rest for 2 min.</li>
	<li>Repeat steps 3.1 - 3.12, three times for the left leg and right leg and record the data in newtons (N).</li>
	<li>Save all the data and export as a report for the analysis.</li>
</ol>

<ol>
	<li>Lu, T. W., Chang, C. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Biomechanics+of+human+movement+and+its+clinical+applications.">Biomechanics of human movement and its clinical applications.</a> The Kaohsiung Journal of Medical Sciences. 28 (2 Suppl), S13-S25 (2012).</li><li>Kaufman, K., An, K., Firestein, G. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Kelley and Firestein's Textbook of Rheumatology (Tenth Edition). , 78-89 (2017).</li><li>Sloot, L. H., van der Krogt, M. M., Harlaar, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Self-paced+versus+fixed+speed+treadmill+walking.">Self-paced versus fixed speed treadmill walking.</a> Gait &amp; Posture. 39 (1), 478-484 (2014).</li><li>Beaton, D. E., O'Driscoll, S. W., Richards, R. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Grip+strength+testing+using+the+BTE+work+simulator+and+the+jamar+dynamometer:+A+comparative+study.">Grip strength testing using the BTE work simulator and the jamar dynamometer: A comparative study.</a> The Journal of Hand Surgery. 20 (2), 293-298 (1995).</li><li>Jindal, P., Narayan, A., Ganesan, S., MacDermid, J. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Muscle+strength+differences+in+healthy+young+adults+with+and+without+generalized+joint+hypermobility:+a+cross-sectional+study.">Muscle strength differences in healthy young adults with and without generalized joint hypermobility: a cross-sectional study.</a> BMC Sports Science, Medicine &amp; Rehabilitation. 8, 12 (2016).</li><li>Muehlbauer, T., Granacher, U., Borde, R., Hortob&#225;gyi, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Non-Discriminant+Relationships+between+Leg+Muscle+Strength,+Mass+and+Gait+Performance+in+Healthy+Young+and+Old+Adults.">Non-Discriminant Relationships between Leg Muscle Strength, Mass and Gait Performance in Healthy Young and Old Adults.</a> Gerontology. 64 (1), 11-18 (2018).</li><li>van den Bogert, A. J., Geijtenbeek, T., Even-Zohar, O., Steenbrink, F., Hardin, E. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+real-time+system+for+biomechanical+analysis+of+human+movement+and+muscle+function.">A real-time system for biomechanical analysis of human movement and muscle function.</a> Medical &amp; Biological Engineering &amp; Computing. 51 (10), 1069-1077 (2013).</li><li>. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> HBM2 Reference Manual. , 9-11 (2017).</li><li>Sloot, L. H., van der Krogt, M. M., Harlaar, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effects+of+adding+a+virtual+reality+environment+to+different+modes+of+treadmill+walking.">Effects of adding a virtual reality environment to different modes of treadmill walking.</a> Gait Posture. 39 (3), 939-945 (2014).</li><li>Liu, W. Y., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Reproducibility+and+Validity+of+the+6-Minute+Walk+Test+Using+the+Gait+Real-Time+Analysis+Interactive+Lab+in+Patients+with+COPD+and+Healthy+Elderly.">Reproducibility and Validity of the 6-Minute Walk Test Using the Gait Real-Time Analysis Interactive Lab in Patients with COPD and Healthy Elderly.</a> PLoS One. 11 (9), e0162444 (2016).</li><li>Herman, T., Mirelman, A., Giladi, N., Schweiger, A., Hausdorff, J. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Executive+Control+Deficits+as+a+Prodrome+to+Falls+in+Healthy+Older+Adults:+A+Prospective+Study+Linking+Thinking,+Walking,+and+Falling.">Executive Control Deficits as a Prodrome to Falls in Healthy Older Adults: A Prospective Study Linking Thinking, Walking, and Falling.</a> The Journals of Gerontology: Series A. 65 (10), 1086-1092 (2010).</li><li>Geijtenbeek, T., Steenbrink, F., Otten, B., Even-Zohar, O. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Proceedings+of+the+10th+International+Conference+on+Virtual+Reality+Continuum+and+Its+Applications+in+Industry.">Proceedings of the 10th International Conference on Virtual Reality Continuum and Its Applications in Industry.</a> , 201-208 (2011).</li><li>Zeni, J. A., Higginson, J. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Gait+parameters+and+stride-to-stride+variability+during+familiarization+to+walking+on+a+split-belt+treadmill.">Gait parameters and stride-to-stride variability during familiarization to walking on a split-belt treadmill.</a> Clinical Biomechanics (Bristol, Avon). 25 (4), 383-386 (2010).</li><li>Meldrum, D., Cahalane, E., Conroy, R., Fitzgerald, D., Hardiman, O. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Maximum+voluntary+isometric+contraction:+reference+values+and+clinical+application.">Maximum voluntary isometric contraction: reference values and clinical application.</a> Amyotroph Lateral Sclerosis. 8 (1), 47-55 (2007).</li><li>Ancillao, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Modern Functional Evaluation Methods for Muscle Strength and Gait Analysis. , 133 (2018).</li><li>Mun, J. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+method+for+the+reduction+of+skin+marker+artifacts+during+walking+:+Application+to+the+knee.">A method for the reduction of skin marker artifacts during walking : Application to the knee.</a> KSME International Journal. 17 (6), 825-835 (2003).</li><li>Liu, P. C., Liu, J. F., Chen, L. Y., Xia, K., Wu, X. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Intermittent+pneumatic+compression+devices+combined+with+anticoagulants+for+prevention+of+symptomatic+deep+vein+thrombosis+after+total+knee+arthroplasty:+a+pilot+study.">Intermittent pneumatic compression devices combined with anticoagulants for prevention of symptomatic deep vein thrombosis after total knee arthroplasty: a pilot study.</a> Therapeutics and Clinical Risk Management. 13, 179-183 (2017).</li><li>Al-Amri, M., Al Balushi, H., Mashabi, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Intra-rater+repeatability+of+gait+parameters+in+healthy+adults+during+self-paced+treadmill-based+virtual+reality+walking.">Intra-rater repeatability of gait parameters in healthy adults during self-paced treadmill-based virtual reality walking.</a> Computer Methods in Biomechanics and Biomedical Engineering. 20 (16), 1669-1677 (2017).</li><li>Zeni, J., Richards, J., Higginson, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Two+simple+methods+for+determining+gait+events+during+treadmill+and+overground+walking+using+kinematic+data.">Two simple methods for determining gait events during treadmill and overground walking using kinematic data.</a> Gait &amp; Posture. 27 (4), 710-714 (2008).</li><li>Tsaopoulos, D. E., Baltzopoulos, V., Richards, P. J., Maganaris, C. N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mechanical+correction+of+dynamometer+moment+for+the+effects+of+segment+motion+during+isometric+knee-extension+tests.">Mechanical correction of dynamometer moment for the effects of segment motion during isometric knee-extension tests.</a> Journal of Applied Physiology. 111 (1), 68-74 (2011).</li><li>Abernethy, P., Wilson, G., Logan, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Strength+and+power+assessment.+Issues,+controversies+and+challenges.">Strength and power assessment. Issues, controversies and challenges.</a> Sports Medicine. 19 (6), 401-417 (1995).</li><li>Kroll, W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Reliability+of+a+Selected+Measure+of+Human+Strength.">Reliability of a Selected Measure of Human Strength.</a> Research Quarterly, American Association for Health, Physical Education and Recreation. 33 (3), 410-417 (1962).</li><li>Anzak, A., Tan, H., Pogosyan, A., Brown, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Doing+better+than+your+best:+loud+auditory+stimulation+yields+improvements+in+maximal+voluntary+force.">Doing better than your best: loud auditory stimulation yields improvements in maximal voluntary force.</a> Experimental Brain Research. 208 (2), 237-243 (2011).</li><li>Belkhiria, C., De Marco, G., Driss, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effects+of+verbal+encouragement+on+force+and+electromyographic+activations+during+exercise.">Effects of verbal encouragement on force and electromyographic activations during exercise.</a> Journal of Sports Medicine and Physical Fitness. 58 (5), 750-757 (2018).</li><li>Bickers, M. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Does+verbal+encouragement+work?+The+effect+of+verbal+encouragement+on+a+muscular+endurance+task.">Does verbal encouragement work? The effect of verbal encouragement on a muscular endurance task.</a> Clinical Rehabilitation. 7 (3), 196-200 (1993).</li><li>Karaba-Jakovljevic, D., Popadic-Gacesa, J., Grujic, N., Barak, O., Drapsin, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Motivation+and+motoric+tests+in+sports.">Motivation and motoric tests in sports.</a> Medicinki Pregled. 60 (5-6), 231-236 (2007).</li><li>Andreacci, J. L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+effects+of+frequency+of+encouragement+on+performance+during+maximal+exercise+testing.">The effects of frequency of encouragement on performance during maximal exercise testing.</a> Journal of Sports Science. 20 (4), 345-352 (2002).</li><li>Rendos, N. K., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Variations+in+Verbal+Encouragement+Modify+Isokinetic+Performance.">Variations in Verbal Encouragement Modify Isokinetic Performance.</a> Journal of Strength and Conditioning Research. 33 (3), 708-716 (2019).</li></ol>

The authors have nothing to disclose.

The contribution of this study is to accurately and comprehensively describe within one protocol the techniques for combined gait analysis and muscle strength testing that have not previously been described together.
In order to achieve accurate results for gait analysis, there are two areas that require maximum attention: 1) marker placements and 2) acclimatization time. The accuracy of the measured data is heavily dependent on the accuracy of the model used. The other key factors that affect...

The discipline of biomechanics primarily involves the study of stress, strain, loads and motion of biological systems - solid and fluid alike. It also involves the modelling of mechanical effects on the structure, size, shape and movement of the body1. For many years, developments in this field have improved our understanding of normal and pathologic gait, mechanics of neuromuscular control, and mechanics of growth and form2.
The main objective of this article is to present a comprehensive methodology on two of the latest technologies available to measure lower limb biomechanics of individuals. The gait analysis system measures and quantifies gait biomechanics by using a self-paced (SP) treadmill in combination with an augmented reality environment, which integrates a SP algorithm to regulate the treadmill&#39;s speed, as described by Sloot et al3. The muscle strength testing equipment is used as an assessment and a treatment tool for upper extremity rehabilitation4. This device can objectively assess a variety of physiological patterns of movement or job simulation tasks in isometric and isotonic modes. It is currently recognized as the gold standard for upper limb strength measurement5 but the evidence related specifically to the lower limb remains unclear. This paper explains the detailed protocol for completing an assessment of gait and isometric strength for the lower extremity.
Within biomechanical analysis, it is useful to combine assessments of functional performance (such as gait analysis) with specific tests of muscular performance. This is because whilst it may be assumed that increased muscle strength improves functional performance, this may not always be apparent6. This understanding is required for the improved future design of rehabilitation protocols and research strategies to assess these approaches.

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>701 Small lever</td>
			<td>Baltimore Therapeutic Equipment Company (BTE)</td>
			<td>Not Available - Online link provided in description</td>
			<td>The unique attachment designed for the Primus RS to measure Knee Extension/Flexion - <a href="https://store.btetech.com/collections/primus/products/701-small-lever" target="_blank">https://store.btetech.com/collections/primus/products/701-small-lever</a></td>
		</tr>
		<tr>
			<td>D-Flow Software - Vresion 3.26</td>
			<td>Motekforce Link</td>
			<td>Not Available - Online link provided in description</td>
			<td>Software used to control GRAIL system - <a href="https://summitmedsci.co.uk/products/motek-dflow-hbm-software/" target="_blank">https://summitmedsci.co.uk/products/motek-dflow-hbm-software/</a></td>
		</tr>
		<tr>
			<td>Gait Offline Analysis (GOAT) - Version 2.3</td>
			<td>Motekforce Link</td>
			<td>Not Available - Online link provided in description</td>
			<td>Software used for the analysis of the gait parameters - <a href="https://www.motekmedical.com/product/grail/" target="_blank">https://www.motekmedical.com/product/grail/</a></td>
		</tr>
		<tr>
			<td>Gait Real-time Analysis Interactive Lab (GRAIL)</td>
			<td>Motekforce Link</td>
			<td>Not Available - Online link provided in description</td>
			<td>GRAIL system measures and quantifies gait biomechanics by using a virtual reality based self-paced (SP) treadmill - <a href="https://www.motekmedical.com/product/grail/" target="_blank">https://www.motekmedical.com/product/grail/</a></td>
		</tr>
		<tr>
			<td>Leg Pad for 701</td>
			<td>Baltimore Therapeutic Equipment Company (BTE)</td>
			<td>Not Available - Online link provided in description</td>
			<td>The unique attachment designed for the Primus RS to measure Knee Extension/Flexion - <a href="https://store.btetech.com/collections/primus/products/701-802-leg-pad" target="_blank">https://store.btetech.com/collections/primus/products/701-802-leg-pad</a></td>
		</tr>
		<tr>
			<td>Positioning Chair</td>
			<td>Baltimore Therapeutic Equipment Company (BTE)</td>
			<td>Not Available - Online link provided in description</td>
			<td>Participant Positioning Chair is designed for assessment and treatment of the lower exteremeties. The chair is designed for multiple positions. <a href="https://www.btetech.com/product/primus/" target="_blank">https://www.btetech.com/product/primus/</a></td>
		</tr>
		<tr>
			<td>Primus RS</td>
			<td>Baltimore Therapeutic Equipment Company (BTE)</td>
			<td>Not Available - Online link provided in description</td>
			<td>Primus RS equipment captures and reports real time objective data in Isotonic, Isometric, and Isokinetic resistance modes - <a href="https://www.btetech.com/wp-content/uploads/BTE-Rehabilitation-Equipment-PrimusRS-Brochure-1.pdf" target="_blank">https://www.btetech.com/wp-content/uploads/BTE-Rehabilitation-Equipment-PrimusRS-Brochure-1.pdf</a></td>
		</tr>
	</tbody>
</table>

lower limb biomechanical analysis of healthy participants

Biomechanical analysis techniques are useful in the study of human movement. The aim of this study was to introduce a technique for the lower limb biomechanical assessment in healthy participants using commercially available systems. Separate protocols were introduced for the gait analysis and muscle strength testing systems. To ensure maximum accuracy for gait assessment, attention should be given to the marker placements and self-paced treadmill acclimatization time. Similarly, participant positioning, a practice trial, and verbal encouragement are three critical stages in muscle strength testing. The current evidence suggests that the methodology outlined in this article may be effective for the assessment of lower limb biomechanics.

The mean and standard deviation of the spatial-temporal, kinematics, and kinetic gait parameters are given in Table 2. MVIC data for all 30 participants are summarized in Table 3. A typical set of data for the left and right side of one participant showing graphical representation of gait parameters is provided in Figure 4 and Figure 5, respectively.
<p class="jove_content" fo:keep-together.w...

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Lower Limb Biomechanical Analysis of Healthy Participants

This article introduces a comprehensive experimental methodology on two of the latest technologies available to measure the lower limb biomechanics of individuals.

Biomechanical analysis techniques are useful in the study of human movement. The aim of this study was to introduce a technique for the lower limb ...

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8.9K Views.  Bournemouth University. This article introduces a comprehensive experimental methodology on two of the latest technologies available to measure the lower limb biomechanics of individuals.

Video: Lower Limb Biomechanical Analysis of Healthy Participants

TRUE Gene Silencing: Screening of a Heptamer-type Small Guide RNA Library for Potential Cancer Therapeutic Agents

TRUE gene silencing (termed after tRNase ZL-utilizing efficacious gene silencing) is one of the RNA-directed gene silencing technologies, which utilizes an artificial small guide RNA (sgRNA) to guide tRNA 3&#8242; processing endoribonuclease, tRNase ZL, to recognize a target RNA. sgRNAs can be taken up by cells without any transfection reagents and can downregulate their target RNA levels and/or induce apoptosis in human cancer cells. We have screened an sgRNA library containing 156 heptamer-type sgRNAs for the effect on viability of human myeloma and leukemia cells, and found that 20 of them can efficiently induce apoptosis in at least one of the cancer cell lines. Here we present a protocol for screening of a heptamer-type sgRNA library for potential therapeutic drugs against blood cancers. The protocol includes how to construct the sgRNA library, how to assess the effect of each sgRNA on cell viability, and how to further evaluate the effective sgRNAs by flow cytometry. Around 2,000 hits would be expected to be obtained by screening the full-scale sgRNA library composed of 16,384 heptamers.

Here we present a protocol for screening of a heptamer-type sgRNA library for potential therapeutic drugs against blood cancers.

TRUE gene silencing (termed after tRNase ZL-utilizing efficacious gene silencing) is one of the RNA-directed gene silencing technologies, which utilizes ...

Oral Biofilm Sampling for Microbiome Analysis in Healthy Children

Oral biofilm and its molecular analysis provide a basis for investigating various dental research and clinical questions. Knowledge of biofilm composition leads to a better understanding of cariogenic and periopathogenic mechanisms. Microbial changes taking place in the oral cavity during childhood are of interest for several reasons. The evolution of the child oral microbiota and shifts in its composition need to be analyzed further to understand and possibly prevent the onset of disease. At the same time, advanced knowledge of the natural composition of oral biofilm is needed. Early stages of caries-free permanent dentition with healthy gums provide a widely unaffected subgingival habitat that can serve as an in situ baseline for studying features of oral health and disease. Analysis of children's oral biofilm during different stages in life is thus an important theme in the field. Modern molecular analysis methods can provide comprehensive information about the bacterial diversity of such biofilms. To enable microbiota data comparison, it is important to standardize each step in the procedure for molecular data generation. This procedure spans from clinical sampling, Next Generation Sequencing (NGS), bioinformatic data processing, to taxonomic interpretation. One of the most critical factors here is biofilm sampling. Sampling in children is even more challenging in particular due to limited space in subgingival areas. We thus focus on the use of paper points for subgingival sampling. This article provides a detailed protocol for oral biofilm sampling of the subgingival sulcus, the mucosa, and saliva in children.

Changes in the oral microbiome throughout childhood are of growing interest. Comparison of different microbiome studies reveals a lack of standardized sampling protocols. Limited space makes sampling the sound subgingival sulcus of children challenging. Paper point sampling is presented here in detail as the method of choice for this area.

Oral biofilm and its molecular analysis provide a basis for investigating various dental research and clinical questions. Knowledge of biofilm composition ...

Assessment of the Efficacy of An Osteopathic Treatment in Infants with Biomechanical Impairments to Suckling

Breastfeeding can be challenging for mother-infant dyads experiencing biomechanical suckling difficulties. Although lactation consultants (LCs) all over the world have increased their skills in this field and can provide support to help position the infant at the breast, the impact of their intervention might be limited in the presence of stiff structures in the infants. Here we present a protocol for a randomized controlled trial to assess the efficacy of osteopathic treatment, coupled with lactation consultation, for infants&#39; biomechanical suckling difficulties. It proposes a set-up and a sequence of actions to ensure an optimal context for treatment, as well as a blinding of parents and LCs to the intervention. Data such as the infant&#39;s latch ability measured with the LATCH Assessment Tool, the mother&#39;s nipple pain with a visual analog scale (VAS), and the mother&#39;s perceptions are collected by LCs four times over a 10-day period. Osteopathic lesions are documented by the osteopath, using a standardized assessment grid. Structures of interest are coherent with the anatomical zones involved in latching onto the breast. This protocol also proposes a strategy to document systematically an osteopathic profile of infants with biomechanical suckling difficulties in their first weeks of life. The implementation of this protocol confirms its feasibility for osteopathic assessment and treatment and paves the way for future trials to further explore the efficacy of osteopathic techniques for infants with biomechanical suckling difficulties.

Osteopathy is an emerging field of clinical research. Here we present a protocol to assess the efficacy of an osteopathic intervention coupled with lactation consultation, in infants with biomechanical issues impeding breastfeeding.

Breastfeeding can be challenging for mother-infant dyads experiencing biomechanical suckling difficulties. Although lactation consultants (LCs) all over ...

Preclinical Model of Hind Limb Ischemia in Diabetic Rabbits

Peripheral vascular disease is a widespread clinical problem that affects millions of patients worldwide. A major consequence of peripheral vascular disease is the development of ischemia. In severe cases, patients can develop critical limb ischemia in which they experience constant pain and an increased risk of limb amputation. Current therapies for peripheral ischemia include bypass surgery or percutaneous interventions such as angioplasty with stenting or atherectomy to restore blood flow. However, these treatments often fail to the continued progression of vascular disease or restenosis or are contraindicated due to the overall poor health of the patient. A promising potential approach to treat peripheral ischemia involves the induction of therapeutic neovascularization to allow the patient to develop collateral vasculature. This newly formed network alleviates peripheral ischemia by restoring perfusion to the affected area. The most frequently employed preclinical model for peripheral ischemia utilizes the creation of hind limb ischemia in healthy rabbits through femoral artery ligation. In the past, however, there has been a strong disconnect between the success of preclinical studies and the failure of clinical trials regarding treatments for peripheral ischemia. Healthy animals typically have robust vascular regeneration in response to surgically induced ischemia, in contrast to the reduced vascularity and regeneration in patients with chronic peripheral ischemia. Here, we describe an optimized animal model for peripheral ischemia in rabbits that includes hyperlipidemia and diabetes. This model has reduced collateral formation and blood pressure recovery in comparison to a model with a higher cholesterol diet. Thus, the model may provide better correlation with human patients with compromised angiogenesis from the common co-morbidities that accompany peripheral vascular disease.

We describe a surgical procedure used to induce peripheral ischemia in rabbits with hyperlipidemia and diabetes. This surgery acts as a preclinical model for conditions experienced in peripheral artery disease in patients. Angiography is also described as a means to measure the extent of introduced ischemia and recovery of perfusion.

Peripheral vascular disease is a widespread clinical problem that affects millions of patients worldwide. A major consequence of peripheral vascular ...

The Lower Body Positive Pressure Treadmill for Knee Osteoarthritis Rehabilitation

Here, based on a clinician&rsquo;s point-of-view, we propose a two-model lower body positive pressure (LBPP) protocol (walking and squatting models) in addition to a clinical, functional assessment methodology, including details for further encouragement of the development of non-drug surgical intervention strategies in knee osteoarthritis patients. However, we only present the effect of LBPP training in improvement of pain and knee function in one patient through three-dimensional gait analysis. The exact, long-term effects of this approach should be explored in future studies.

Here, based on a clinician&#8217;s point-of-view, we propose a two-model lower body positive pressure (LBPP) protocol (walking and squatting models) in addition to a clinical, functional assessment methodology, including details for further encouragement of the development of non-drug surgical intervention strategies in knee osteoarthritis patients. However, we only present the effect of LBPP training in improvement of pain and knee function in one patient through three-dimensional gait analysis. The exact, long-term effects of this approach should be explored in future studies.

Here, based on a clinician&rsquo;s point-of-view, we propose a two-model lower body positive pressure (LBPP) protocol (walking and squatting models) in ...

Virtual Prism Adaptation Therapy: Protocol for Validation in Healthy Adults

Hemispatial neglect is a common impairment after stroke. It is associated with poor functional and social outcomes. Therefore, an adequate intervention is imperative for the successful management of hemispatial neglect. However, the clinical use of various interventions is limited in real clinical practice. Prism adaptation therapy is one of the most evidence-based rehabilitation modalities to treat hemispatial neglect. To overcome any possible shortcoming that may occur with prism therapy, we developed a new system using immersive virtual reality and depth-sensing camera to create a virtual prism adaptation therapy (VPAT). To validate the VPAT system, we designed an experimental protocol investigating the behavioral errors and changes in cortical activation via the VPAT system. Cortical activation was measured by functional near infrared spectroscopy (fNIRS). The experiment consisted of four phases. All four included clicking, pointing or rest applied to right-handed healthy people. Clicking versus pointing was used for investigating the cortical region related with the gross motor task, and pointing with VPAT versus pointing without VPAT was used for investigating the cortical region associated with visuospatial perception. The preliminary results from four healthy participants showed that pointing errors by the VPAT system was similar to the conventional prism adaptation therapy. Further analysis with more participants and fNIRS data, as well as a study in patients with stroke may be required.

This experimental protocol demonstrates the use of virtual prism adaptation therapy (VPAT) in healthy adults and the association between VPAT and functional near infrared spectroscopy to determine the effect of VPAT on cortical activation. Results suggest that VPAT may be feasible and could induce similar behavioral adaptation as conventional prism adaptation therapy.

Hemispatial neglect is a common impairment after stroke. It is associated with poor functional and social outcomes. Therefore, an adequate intervention is ...

On-Site Sampling and Extraction of Brain Tumors for Metabolomics and Lipidomics Analysis

Despite the variety of tools available for cancer diagnosis and classification, methods that enable fast and simple characterization of tumors are still in need. In recent years, mass spectrometry has become a method of choice for untargeted profiling of discriminatory compound as potential biomarkers of a disease. Biofluids are generally considered as preferable matrices given their accessibility and easier sample processing while direct tissue profiling provides more selective information about a given cancer. Preparation of tissues for the analysis via traditional methods is much more complex and time-consuming, and, therefore, not suitable for fast on-site analysis. The current work presents a protocol combining sample preparation and extraction of small molecules on-site, immediately after tumor resection. The sampling device, which is of the size of an acupuncture needle, can be inserted directly into the tissue and then transported to the nearby laboratory for instrumental analysis. The results of metabolomics and lipidomics analyses demonstrate the capability of the approach for the establishment of phenotypes of tumors related to the histological origin of the tumor, malignancy, and genetic mutations, as well as for the selection of discriminating compounds or potential biomarkers. The non-destructive nature of the technique permits subsequent performance of routinely used tests e.g., histological tests, on the same samples used for SPME analysis, thus enabling attainment of more comprehensive information to support personalized diagnostics.

The manuscript presents on-site sampling of human brain tumors with solid phase microextraction followed by their biochemical profiling towards the discovery of biomarkers.

Despite the variety of tools available for cancer diagnosis and classification, methods that enable fast and simple characterization of tumors are still ...

Point-Of-Care Ultrasound Screening for Proximal Lower Extremity Deep Venous Thrombosis

Acute lower extremity deep venous thrombosis (DVT) is a serious vascular disorder that requires accurate and early diagnosis to prevent life-threatening sequelae. While whole leg compression ultrasound with color and spectral Doppler is commonly performed in radiology and vascular labs, point-of-care ultrasound (POCUS) is becoming more common in the acute care setting. Providers appropriately trained in focused POCUS can perform a rapid bedside examination with high sensitivity and specificity in critically ill patients. This paper describes a simplified yet validated approach to POCUS by describing a three-zone protocol for lower extremity DVT POCUS image acquisition. The protocol explains the steps in obtaining vascular images at six compression points in the lower extremity. Beginning at the level of the proximal thigh and moving distally to the popliteal space, the protocol guides the user through each of the compression points in a stepwise manner: from the common femoral vein to the femoral and deep femoral vein bifurcation, and, finally, to the popliteal vein. Further, a visual aid is provided that may assist providers during real-time image acquisition. The goal in presenting this protocol is to help make proximal lower extremity DVT exams more accessible and efficient for POCUS users at the patient&#39;s bedside.

Traditionally, lower extremity deep venous thrombosis (DVT) is diagnosed by radiology-performed venous duplex ultrasound. Providers appropriately trained in focused point-of-care ultrasound (POCUS) can perform a rapid bedside examination with high sensitivity and specificity in critically ill patients. We describe the scanning technique for focused POCUS DVT lower extremity examination.

Acute lower extremity deep venous thrombosis (DVT) is a serious vascular disorder that requires accurate and early diagnosis to prevent life-threatening ...

Ultrasound Evaluation of Achilles Tendon Morphology

Measurement of Healthy and Injured Triceps Surae Morphology

Achilles tendon injuries occur throughout the lifespan and can negatively affect quality of life and overall health. Achilles tendinopathy is generally classified as an overuse injury associated with fusiform tendon thickening, neovascularization, and interstitial tendon degeneration. Current literature suggests these structural changes are associated with&#160;symptoms and lower physical activity levels, as well as&#160;symptoms and lower extremity function in the long term. Surgically and non-surgically managed Achilles tendon ruptures result in increased tendon cross-sectional area (CSA) and a lengthened Achilles tendon. Both structural outcomes have clinical implications, as larger CSA positively predicts function, whereas increased tendon lengthening predicts reduced function after Achilles tendon rupture. Given the relationship between structural changes associated with Achilles tendon injuries for both injury severity and injury recovery, it is critical to be able to quantify Achilles tendon structure reliably and accurately. Silbernagel&#39;s group has established a valid and reliable method for efficiently evaluating triceps surae muscle and tendon structure. In this protocol, B-mode musculoskeletal ultrasound imaging is used to measure triceps surae structure, including Achilles tendon thickness and CSA, soleus thickness, and the presence of additional findings (calcifications and bursitis). B-mode extended field-of-view is used to measure Achilles tendon length and gastrocnemius anatomical CSA. Finally, power Doppler is used to identify intratendinous neovascularization. Quantification of triceps surae structure allows for comparison between limbs as well as longitudinal changes in response to exercise and treatment for healthy individuals and those with Achilles tendon injuries. This protocol has been used in many research studies to date and proves valuable in understanding the relationship between tendon structure and injury development, severity, and recovery. As ultrasound devices are becoming more affordable and portable, this protocol proves promising as a clinical tool, given its quick and efficient methods.

Author Spotlight: Exploring the Complexities of Achilles Tendon Injuries &#8212; Research and Future Directions

Ultrasound imaging is becoming more accessible in clinical and research settings, and a consistent protocol will be beneficial for comparison between studies and for clinical interpretations. This protocol for ultrasound evaluation is a valid and reliable method to evaluate Achilles tendon morphology in healthy, tendinopathic, and ruptured tendons.

Achilles tendon injuries occur throughout the lifespan and can negatively affect quality of life and overall health. Achilles tendinopathy is generally ...

Ultrasound in Evaluating Diaphragm Dysfunction and Monitoring Treatment

Measuring Diaphragm Thickness and Function Using Point-of-Care Ultrasound

The diaphragm is the main component of the respiratory muscle pump. Diaphragm dysfunction can cause dyspnea and exercise intolerance, and predisposes affected individuals to respiratory failure. In mechanically ventilated patients, the diaphragm is susceptible to atrophy and dysfunction through disuse and other mechanisms. This contributes to failure to wean and poor long-term clinical outcomes. Point-of-care ultrasound provides a valid and reproducible method for evaluating diaphragm thickness and contractile activity (thickening fraction during inspiration) that can be readily employed by clinicians and researchers alike. This article presents best practices for measuring diaphragm thickness and quantifying diaphragm thickening during tidal breathing or maximal inspiration. Once mastered, this technique can be used to diagnose and prognosticate diaphragm dysfunction, and guide and monitor response to treatment over time in both healthy individuals and acute or chronically ill patients.

Author Spotlight: Unraveling the Impact of Mechanical Ventilation on Diaphragm Function and Patient Outcomes 

Diaphragm thickness and function can be assessed in healthy individuals and critically ill patients using point-of-care ultrasound. This technique offers an accurate, reproducible, feasible, and well-tolerated method for evaluating diaphragm structure and function.

The diaphragm is the main component of the respiratory muscle pump. Diaphragm dysfunction can cause dyspnea and exercise intolerance, and predisposes ...