Name: design, fabrication, and administration of the hand active sensation test (haste)
Uploaded: 2015-09-08T15:00:00
Description: Watch this Scientific Journal Video about Design, Fabrication, and Administration of the Hand Active Sensation Test HASTe at JoVE.com

This work was supported in part by OSU&#8217;s CCTS Program through TL1TR001069 award to Dr. Borstad.&#160; We would like to acknowledge Amelia Siles, DPT, NCS for her valuable measurement insights and Sarah Alexander for her assistance with editing the final draft of this manuscript.

1. Design and Fabrication of Visual Screen and HASTe (See &#34;Table of Materials&#34;&#160;for All Required Equipment and Materials):
<ol>
	<li>Assemble Visual Screen (Figure 1):
	<ol>
		<li>Cut four pieces of 0.5-inch PVC pipe (material a) to 10 inches (these form the base of the screen). Cut three pieces of 0.5-inch PVC pipe to 20 inches (these form the upright portion of the screen).</li>
		<li>Insert two pieces of 10-inch pipe into both PVC Tee&#8217;s (parallel openings) (material b). Insert two ...

<ol>
	<li>Nakayama, H., Jgtgensen, H., Stig, K., Raaschou, H. O., Olsen, T. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Recovery+of+upper+extremity+function+in+stroke+patients:+the+Copenhagen+Stroke+Study.">Recovery of upper extremity function in stroke patients: the Copenhagen Stroke Study.</a> Age (SD). 74, 12 (1994).</li><li>Carey, L. M., Matyas, T. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Frequency+of+discriminative+sensory+loss+in+the+hand+after+stroke+in+a+rehabilitation+setting.">Frequency of discriminative sensory loss in the hand after stroke in a rehabilitation setting.</a> Journal of Rehabilitation Medicine. 43, 257-263 (2011).</li><li>Winward, C. E., Halligan, P. W., Wade, D. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Somatosensory+recovery:+A+longitudinal+study+of+the+first+6+months+after+unilateral+stroke.">Somatosensory recovery: A longitudinal study of the first 6 months after unilateral stroke.</a> Disability &amp; Rehabilitation. 29, 293-299 (2007).</li><li>Sullivan, J. E., Hedman, L. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Sensory+Dysfunction+Following+Stroke:+Incidence,+Significance,+Examination,+and+Intervention.">Sensory Dysfunction Following Stroke: Incidence, Significance, Examination, and Intervention.</a> Topics in Stroke Rehabilitation. 15, 200-217 (2008).</li><li>Connell, L. A., Lincoln, N. B., Radford, K. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Somatosensory+impairment+after+stroke:+frequency+of+different+deficits+and+their+recovery.">Somatosensory impairment after stroke: frequency of different deficits and their recovery.</a> Clinical Rehabilitation. 22, 758 (2008).</li><li>Winward, C. E., Halligan, P. W., Wade, D. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Current+practice+and+clinical+relevance+of+somatosensory+assessment+after+stroke.">Current practice and clinical relevance of somatosensory assessment after stroke.</a> Clinical rehabilitation. 13, 48-55 (1999).</li><li>Torre, 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=Somatosensory-related+limitations+for+bimanual+coordination+after+stroke.">Somatosensory-related limitations for bimanual coordination after stroke.</a> Neurorehabilitation and neural repair. , (2013).</li><li>Nichols-Larsen, D. S., Clark, P. C., Zeringue, A., Greenspan, A., Blanton, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Factors+influencing+stroke+survivors'+quality+of+life+during+subacute+recovery.">Factors influencing stroke survivors' quality of life during subacute recovery.</a> Stroke. 36, 1480-1484 (2005).</li><li>Borstad, A. L., Nichols-Larsen, D. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Assessing+and+treating+Higher-level+Somatosensory+Impairments+Post+Stroke.">Assessing and treating Higher-level Somatosensory Impairments Post Stroke.</a> Topics in Stroke Rehabilitation. 21, 290-295 (2014).</li><li>Byl, N., Leano, J., Cheney, L. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+Byl-Cheney-Boczai+Sensory+Discriminator:+reliability,+validity,+and+responsiveness+for+testing+stereognosis.">The Byl-Cheney-Boczai Sensory Discriminator: reliability, validity, and responsiveness for testing stereognosis.</a> Journal of Hand Therapy. 15, 315-330 (2002).</li><li>Lincoln, N. B., Jackson, J. M., Adams, S. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Reliability+and+revision+of+the+Nottingham+Sensory+Assessment+for+stroke+patients.">Reliability and revision of the Nottingham Sensory Assessment for stroke patients.</a> Physiotherapy. 84, 358-365 (1998).</li><li>Carey, L. M., Nankervis, , 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=.">.</a> , (2006).</li><li>Ayres, A. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Sensory integration and praxis test (SIPT). , (1989).</li><li>Kalisch, T., Tegenthoff, M., Dinse, H. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Improvement+of+sensorimotor+functions+in+old+age+by+passive+sensory+stimulation.">Improvement of sensorimotor functions in old age by passive sensory stimulation.</a> Clinical Interventions in Aging. 3, 673 (2008).</li><li>Meyer, S., Kattunen, A. H., Thijs, V., Feys, H., Verheyden, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=How+do+somatosensory+deficits+in+the+arm+and+hand+relate+to+upper+limb+impairment,+activity,+and+participation+problems+after+stroke?+A+systematic+Review.">How do somatosensory deficits in the arm and hand relate to upper limb impairment, activity, and participation problems after stroke? A systematic Review.</a> Physical Therapy. 94, (2014).</li><li>Williams, P. S., Basso, D. M., Case-Smith, J., Nichols-Larsen, D. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Development+of+the+Hand+Active+Sensation+Test:+reliability+and+validity.">Development of the Hand Active Sensation Test: reliability and validity.</a> Arch. Phys. Med. Rehabil. 87, 1471-1477 (2006).</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> International Classification of Functioning, Disability and Health (ICF). , (2001).</li><li>McDonnell, M. N., Hillier, S. L., Miles, T. S., Thompson, P. D., Ridding, M. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Influence+of+combined+afferent+stimulation+and+task-specific+training+following+stroke:+a+pilot+randomized+controlled+trial.">Influence of combined afferent stimulation and task-specific training following stroke: a pilot randomized controlled trial.</a> Neurorehabilitation and neural repair. 21, 435-443 (2007).</li><li>Byl, N. N., Pitsch, E. A., Abrams, G. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Functional+outcomes+can+vary+by+dose:+learning-based+sensorimotor+training+for+patients+stable+poststroke.">Functional outcomes can vary by dose: learning-based sensorimotor training for patients stable poststroke.</a> Neurorehabilitation and neural repair. 22, 494 (2008).</li><li>Carey, L., Macdonell, R., Matyas, T. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=SENSe:+Study+of+the+Effectiveness+of+Neurorehabilitation+on+Sensation+A+Randomized+Controlled+Trial.">SENSe: Study of the Effectiveness of Neurorehabilitation on Sensation A Randomized Controlled Trial.</a> Neurorehabilitation and neural repair. 25, 304-313 (2011).</li></ol>

Evidence suggests that similar to motor recovery following stroke, somatosensory recovery requires task specific training. 18,19,20 Therefore, if we aim to improve the hands performance as a haptic receptor, haptic impairments must be identified. 15 The opportunity to quantify haptic ability in the clinic and laboratory has been limited, to some extent, based on the availability of valid and reliable outcome measures. &#160;The large number of stroke survivors with somatosensory impairmen...

Imagine the feel of a loved one&#8217;s hand. It is calloused or smooth? Strong or delicate? Warm or cool? Do you hold it in a handshake or with fingers intertwined? If you have evoked a memory, it is likely grounded in your ability to use movement of your hands to solicit somatosensory information, which is called haptic touch, or active sensation. Characteristics that can be determined with haptic touch include size, shape, weight, texture, surface compliance, and temperature. &#160;Haptic receptors include proprioceptors, which are found in the skin, muscles, tendons, and joints, as well as cutaneous receptors, both of which are activated during manual exploration of objects. Different hand actions aid in the determination of object characteristics. For example, repeatedly performing a lateral rubbing motion across an object may expose texture properties, or lifting the object in the hand may reveal object weight. Therefore, both manual dexterity and somatosensation are vital to haptic performance and the human experience.
Following stroke, both somatosensory and motor impairments contribute to diminished haptic performance. 1 Poststroke motor impairments are common and well defined, with approximately 70% of stroke survivors experiencing some level of paresis. 1 Somatosensory impairments after stroke are also common, occurring in 47-89% 2-4 of individuals. Researchers agree that both cutaneous and proprioceptive impairments are common after stroke. For a thorough characterization of the consequences of stroke on somatosensory domains the reader is referred to the works by of Carey et al., 2 Connell et al., 5 and Sullivan. 4
Somatosensory impairments contribute to diminished rehabilitation outcomes, 6 upper limb coordination, 7 function and quality of life.8 However, quantification of somatosensory impairments, especially active sensation, is lacking in clinical practice in part due to the fact that somatosensory loss is less apparent and more difficult to quantify than motor impairments.9 Measures which have been developed to quantify active sensation include the Byl-Chyney-Boczai Sensory Discriminator (BCB), 10 the stereognosis component of the Revised Nottingham Sensory Assessment, 11 the Functional Tactile Object Recognition Tests (fTORT), 12 &#160;the Manual Form Perception Test (a subset of the Sensory Information and Praxis Test), 13 and the Haptic Object Recognition Test (HORT). 14 Although these measures are available, a recent systematic review, which sought to describe how somatosensory impairments in the arm and hand relate to upper limb problems following stroke, concluded that clinicians and researchers currently lack valid and reliable tests of somatosensation. 15 Therefore, addressing the availability of clinically useful and parametrically sound measures of haptic performance is essential.
The Hand Active Sensation Test (HASTe) is an 18-item match-to-sample test of weight and texture discrimination, originally published by Williams and colleagues in 2006. 16 The HASTe is a measure of haptic touch and is sensitive to haptic impairments in individuals with stroke (indicated by fewer than 13 correct matches). As the HASTe seeks to measure haptic perception, as minimum criteria, individuals need the abilities to grasp and lift with the hand and arm and follow the test instructions. In the International Classification of Functioning, Disability and Health (ICF) model, 17 the HASTe is considered an activity level measurement. The HASTe takes between 15-30 min to administer per hand tested. Advantages of the HASTe include that it is inexpensive, total of material prices for 2015 estimated at $100, and easy to construct and that it&#8217;s 18-point scale provides greater resolution regarding performance than more common dichotomous characterization of &#8220;intact&#8221; or &#8220;impaired&#8221;.
The purpose of this paper is to describe the design, fabrication and administration of the HASTe. While it is possible to infer the test set-up from the original HASTe publication, this paper provides detailed methods for fabricating a visual screen and a HASTe test kit, both to be used during testing. The equipment required for assembly, as well as a detailed list of all required materials is listed in the Table of&#160;Materials. A single sheet with instructions for administering and scoring the HASTe is also provided as the following:
APPENDIX 1: HASTe&#160; Administration Instructions and Score Sheet &#160;&#160;&#160; &#160;&#160;&#160; Participant ID#_______&#160; Date_______
Set Up:&#160; Seat the test participant at a table (table ~29 inches high and chair seat ~18 inches high) with, initially, the dominant or less impaired upper extremity resting on the table and placed under the assembled visual screen. During the test, keep all objects in numerical order to maintain organization.
Demonstration Trials:&#160; Administer two demonstration trials prior to scoring. Provide the participant with objects A and B, and instruct him/her to compare the objects&#8217; weights. Then, provide the participant with objects A and C, and instruct him/her to compare the objects&#8217; textures. Offer feedback only for the demonstration trials.
Scored Trials:&#160; There will be 18 trials scored per extremity, with a maximum of 5 min allowed for each. Instruct the participant to manually explore the specified test object (identified &#8220;test&#8221; in the tables below) with the dominant or less impaired hand, then explore each of the three possible object matches (numbered otherwise within each trial box below) with this same hand. The possible matches will vary by either weight or texture, but never by both within one trial.&#160; Instruct the participant to find the match of the original object and tell him/her they can touch each object as many times as needed to determine an answer.&#160; Do not inform the participant as to which object property (texture or weight) he or she is matching within a trial, and do not assist the participant with manual exploration of the objects. Indicate the participant&#8217;s final answer for each trial in the tables below by circling the number of that object. Test the more impaired upper extremity after the participant completes the test with the less impaired upper extremity. Using the tables below, score the test by determining the number of correct matches from a total of 18 trials per hand (one box per trial below).
Verbal Instructions to Participant: &#8220;Use one hand to manually explore the test object first. Then, explore each of the three possible matches, which vary by either weight or texture but never by both within one trial.&#160; Find the match. You can touch each object as many times as you need to determine your answer. There will be 18 trials. You have a maximum of 5 min for each trial.&#8221;
<img alt="Score card" src="/files/ftp_upload/53178/53178scorecard.jpg" />

<table border="0" cellpadding="0" cellspacing="0">
	<tbody>
		<tr>
			<td>Equipment Needed for Assembly</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Tape measure</td>
			<td></td>
			<td></td>
			<td>To measure lengths of materials</td>
		</tr>
		<tr>
			<td>PVC saw</td>
			<td></td>
			<td></td>
			<td>To cut PVC pieces to appropriate lengths given below</td>
		</tr>
		<tr>
			<td>Scissors</td>
			<td></td>
			<td></td>
			<td>To cut paper, cork and laminating material</td>
		</tr>
		<tr>
			<td>Scale accurate to 0.1 ounce</td>
			<td></td>
			<td></td>
			<td>To determine exact weight of test objects</td>
		</tr>
		<tr>
			<td>Sharpie Permanent Marker</td>
			<td></td>
			<td></td>
			<td>To label test item number and weight</td>
		</tr>
		<tr>
			<td>Visual Screen Materials</td>
			<td>Company</td>
			<td>Catalog Number</td>
			<td></td>
		</tr>
		<tr>
			<td>0.5-inch PVC Pipe (Sch. 40 Plain-End Pipe)</td>
			<td>Home Depot</td>
			<td>530048</td>
			<td>104 inches total, will be cut into four 10-inch and three 20-inch pieces</td>
		</tr>
		<tr>
			<td>0.5-inch PVC Tee (Sch. 40 SxSxS Tee)</td>
			<td>Home Depot</td>
			<td>406005RMC</td>
			<td>Two pieces</td>
		</tr>
		<tr>
			<td>PVC Elbow (Lasco 0.5-inch Dia 90 degree PVC Sch 40 Side Outlet Elbow)</td>
			<td>Home Depot</td>
			<td>413005RMC</td>
			<td>Two pieces</td>
		</tr>
		<tr>
			<td>Pillowcase</td>
			<td></td>
			<td></td>
			<td>One to be hung from the PVC frame as a visual shield</td>
		</tr>
		<tr>
			<td>ACCO Binder Clips, Medium</td>
			<td>amazon.com</td>
			<td>72050</td>
			<td>Two to hold the pillowcase to the PVC frame</td>
		</tr>
		<tr>
			<td>Testing Kit Materials</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>1.5-inch PVC Sch. 40 DWV Plain End Pipe</td>
			<td>Home Depot</td>
			<td>531111</td>
			<td>85 inches total, will be cut into 21 4-inch pieces</td>
		</tr>
		<tr>
			<td>Quartet Cork Roll, 1/16-inch thick</td>
			<td>amazon.com</td>
			<td>NA</td>
			<td>1 roll, 24x48 inches, will be cut into 42 4x7-inch pieces to cover all test and example items</td>
		</tr>
		<tr>
			<td>Oatey all purpose cement for CPVC and PVC</td>
			<td>Home Depot</td>
			<td>308213</td>
			<td>8 ounce can, to use to adhere cork, paper and end caps to PVC test items</td>
		</tr>
		<tr>
			<td>Avery Self-Adhesive Laminating Roll</td>
			<td>amazon.com</td>
			<td>73610</td>
			<td>One 24x600-inch roll, will be cut into six 4x7-inches pieces to cover cork on six test objects</td>
		</tr>
		<tr>
			<td>Brown Builder&#39;s Paper</td>
			<td>Home Depot</td>
			<td>35140</td>
			<td>One roll, will be cut into six 4x7-inch pieces to cover cork on six test objects</td>
		</tr>
		<tr>
			<td>3M Pro Grade 9 Sandpaper</td>
			<td>Home Depot</td>
			<td>25060P-G</td>
			<td>One piece 4x7 inches, to use to cover one example item</td>
		</tr>
		<tr>
			<td>Ranger Glossy Cardstock</td>
			<td>amazon.com</td>
			<td>NA</td>
			<td>One 8.5x11-inch, 10-Pack, white, will be cut into two 4x7-inch pieces to cover example items</td>
		</tr>
		<tr>
			<td>Marblex-durable self modeling clay in moist form</td>
			<td>amaco.com/shop/</td>
			<td>X-242</td>
			<td>One 5-pound package, used to achieve correct weights of test objects</td>
		</tr>
		<tr>
			<td>Medium density polyethylene packing foam</td>
			<td>amazon.com</td>
			<td>NA</td>
			<td>One foam sheet, 220 poly, charcoal, 2x24x18 inches, to fill the remaining space in test objects after clay has been inserted</td>
		</tr>
		<tr>
			<td>Knock Out Plug for 1.5-inch PVC</td>
			<td>Home Depot</td>
			<td>85000</td>
			<td>42 caps to seal the ends of the 21 test items</td>
		</tr>
		<tr>
			<td>Sterilite 6-quart plastic storage box</td>
			<td>Home Depot</td>
			<td>16428960</td>
			<td>One to store/transport test objects</td>
		</tr>
	</tbody>
</table>

design, fabrication, and administration of the hand active sensation test (haste)

The concept of personalizing neurologic rehabilitation, based on individual impairments, has experienced a recent surge. In parallel, the number of outcome measures of upper extremity motor performance has grown. However, clinicians and researchers lack practical, quantitative measures of the hand&#8217;s natural role as a receptor of the environment. The Hand Active Sensation Test (HASTe), developed by Williams and colleagues in 2006, is a valid and reliable measure of haptic performance. Though not available commercially, the HASTe can be fabricated from inexpensive materials, and it has been used successfully to identify impairments in haptic touch in individuals with stroke. (Williams, 2006). This paper presents the methods of design and fabrication of the HASTe testing kit, as well as a visual screen to be used during administration, and instructions for the tests administration and scoring.

Williams and colleagues published the original study, which established the reliability and validity of the HASTe, in 2006. Based on that study, individuals scoring fewer than 13 correct matches on the HASTe are considered to have impaired haptic performance, while those scoring between 13 and 18 are considered to have unimpaired haptic performance. 16 When an individual scores 6 or fewer correct matches, it is considered a chance score, based on the odds in an 18 trial/3 item match to sample design.
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Watch this Scientific Journal Video about Design, Fabrication, and Administration of the Hand Active Sensation Test HASTe at JoVE.com

Design, Fabrication, and Administration of the Hand Active Sensation Test HASTe

The Hand Active Sensation Test (HASTe) is a valid and reliable measure of haptic performance, which has been used successfully to identify impaired haptic touch in individuals with stroke. The purpose of this paper is to describe the design, fabrication and administration of the HASTe.

Design, Fabrication, and Administration of the Hand Active Sensation Test (HASTe)

The concept of personalizing neurologic rehabilitation, based on individual impairments, has experienced a recent surge. In parallel, the number of ...

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Real-time Digital Imaging of Leukocyte-endothelial Interaction in Ischemia-reperfusion Injury (IRI) of the Rat Cremaster Muscle

Ischemia-reperfusion injury (IRI) has been implicated in a large array of pathological conditions such as cerebral stroke, myocardial infarction, intestinal ischemia as well as following transplant and cardiovascular surgery.1 Reperfusion of previously ischemic tissue, while essential for the prevention of irreversible tissue injury, elicits excessive inflammation of the affected tissue. Adjacent to the production of reactive oxygen species, activation of the complement system and increased microvascular permeability, the activation of leukocytes is one of the principle actors in the pathological cascade of inflammatory tissue damage during reperfusion.2, 3 Leukocyte activation is a multistep process consisting of rolling, firm adhesion and transmigration and is mediated by a complex interaction between adhesion molecules in response to chemoattractants such as complement factors, chemokines, or platelet-activating factor.4
While leukocyte rolling in postcapillary venules is predominantly mediated by the interaction of selectins5 with their counter ligands, firm adhesion of leukocytes to the endothelium is selectin-controlled via binding to intercellular adhesion molecules (ICAM) and vascular cellular adhesion molecules (VCAM).6, 7 
Gold standard for the in vivo observation of leukocyte-endothelial interaction is the technique of intravital microscopy, first described in 1968.8
Though various models of IRI (ischemia-reperfusion injury) have been described for various organs, 9-12 only few are suitable for direct visualization of leukocyte recruitment in the microvascular bed on a high level of image quality.8 
We here promote the digital intravital epifluorescence microscopy of the postcapillary venule in the cremasteric microcirculation of the rat 13 as a convenient method to qualitatively and quantitatively analyze leukocyte recruitment for IRI-research in striated muscle tissue and provide a detailed manual for accomplishing the technique. We further illustrate common pitfalls and provide useful tips which should enable the reader to truly appreciate, and safely perform the method.
In a step by step protocol we depict how to get started with respiration controlled anesthesia under sufficient monitoring to keep the animal firmly anesthetized for longer periods of time. We then describe the cremasteric preparation as a thin flat sheet for outstanding optical resolution and provide a protocol for leukocyte imaging in IRI that has been well established in our laboratories.

Real-time Digital Imaging of Leukocyte-endothelial Interaction in Ischemia-reperfusion Injury IRI of the Rat Cremaster Muscle

Digital intravital epifluorescence microscopy of postcapillary venules in the cremasteric microcirculation is a convenient method to gain insights into leukocyte-endothelial interaction in vivo in ischemia-reperfusion injury (IRI) of striated muscle tissue. We here provide a detailed protocol to safely perform the technique and discuss its applications and limitations.

Ischemia-reperfusion injury (IRI) has been implicated in a large array of pathological conditions such as cerebral stroke, myocardial infarction, ...

Utility of Dissociated Intrinsic Hand Muscle Atrophy in the Diagnosis of Amyotrophic Lateral Sclerosis

The split hand phenomenon refers to predominant wasting of thenar muscles and is an early and specific feature of amyotrophic lateral sclerosis (ALS). A novel split hand index (SI) was developed to quantify the split hand phenomenon, and its diagnostic utility was assessed in ALS patients. The split hand index was derived by dividing the product of the compound muscle action potential (CMAP) amplitude recorded over the abductor pollicis brevis and first dorsal interosseous muscles by the CMAP amplitude recorded over the abductor digiti minimi muscle. In order to assess the diagnostic utility of the split hand index, ALS patients were prospectively assessed and their results were compared to neuromuscular disorder patients. The split hand index was significantly reduced in ALS when compared to neuromuscular disorder patients (P&#60;0.0001). Limb-onset ALS patients exhibited the greatest reduction in the split&#160;hand index, and a value of 5.2 or less reliably differentiated ALS from other neuromuscular disorders. Consequently, the split&#160;hand index appears to be a novel diagnostic biomarker for ALS, perhaps facilitating an earlier diagnosis.

Dissociated atrophy of intrinsic hand muscles, termed the split hand, appears to be a specific feature of amyotrophic lateral sclerosis (ALS).&#160;Consequently, a novel neurodiagnostic test, termed the split hand&#160;index, was developed to quantify the clinical phenomenon of the split hand, which differentiated ALS from mimic disorders.

The split hand phenomenon refers to predominant wasting of thenar muscles and is an early and specific feature of amyotrophic lateral sclerosis (ALS). A ...

Ischemic Tissue Injury in the Dorsal Skinfold Chamber of the Mouse: A Skin Flap Model to Investigate Acute Persistent Ischemia

Despite profound expertise and advanced surgical techniques, ischemia-induced complications ranging from wound breakdown to extensive tissue necrosis are still occurring, particularly in reconstructive flap surgery. Multiple experimental flap models have been developed to analyze underlying causes and mechanisms and to investigate treatment strategies to prevent ischemic complications. The limiting factor of most models is the lacking possibility to directly and repetitively visualize microvascular architecture and hemodynamics. The goal of the protocol was to present a well-established mouse model affiliating these before mentioned lacking elements. Harder et al. have developed a model of a musculocutaneous flap with a random perfusion pattern that undergoes acute persistent ischemia and results in ~50% necrosis after 10 days if kept untreated. With the aid of intravital epi-fluorescence microscopy, this chamber model allows repetitive visualization of morphology and hemodynamics in different regions of interest over time. Associated processes such as apoptosis, inflammation, microvascular leakage and angiogenesis can be investigated and correlated to immunohistochemical and molecular protein assays. To date, the model has proven feasibility and reproducibility in several published experimental studies investigating the effect of pre-, peri- and postconditioning of ischemically challenged tissue.

The window of the murine dorsal skinfold chamber presented visualizes a zone of acute persistent ischemia of a musculocutaneous flap. Intravital epi-fluorescence microscopy permits for direct and repetitive assessment of the microvasculature and quantification of hemodynamics. Morphologic and hemodynamic results can further be correlated with histological and molecular analyses.

Despite profound expertise and advanced surgical techniques, ischemia-induced complications ranging from wound breakdown to extensive tissue necrosis are ...

Single-stage Dynamic Reanimation of the Smile in Irreversible Facial Paralysis by Free Functional Muscle Transfer

Unilateral facial paralysis is a common disease that is associated with significant functional, aesthetic and psychological issues. Though idiopathic facial paralysis (Bell&#8217;s palsy) is the most common diagnosis, patients can also present with a history of physical trauma, infectious disease, tumor, or iatrogenic facial paralysis. Early repair within one year of injury can be achieved by direct nerve repair, cross-face nerve grafting or regional nerve transfer. It is due to muscle atrophy that in long lasting facial paralysis complex reconstructive methods have to be applied. Instead of one single procedure, different surgical approaches have to be considered to alleviate the various components of the paralysis.
The reconstruction of a spontaneous dynamic smile with a symmetric resting tone is a crucial factor to overcome the functional deficits and the social handicap that are associated with facial paralysis. Although numerous surgical techniques have been described, a two-stage approach with an initial cross-facial nerve grafting followed by a free functional muscle transfer is most frequently applied. In selected patients however, a single-stage reconstruction using the motor nerve to the masseter as donor nerve is superior to a two-stage repair. The gracilis muscle is most commonly used for reconstruction, as it presents with a constant anatomy, a simple dissection and minimal donor site morbidity.
Here we demonstrate the pre-operative work-up, the post-operative management, and precisely describe the surgical procedure of single-stage microsurgical reconstruction of the smile by free functional gracilis muscle transfer in a step by step protocol. We further illustrate common pitfalls and provide useful tips which should enable the reader to truly comprehend the procedure. We further discuss indications and limitations of the technique and demonstrate representative results.

The use of the masseteric nerve as donor nerve represents a single-stage alternative to the criterion standard two-stage procedure of cross-facial nerve grafting and free muscle transfer in facial paralysis. We provide a detailed description to safely perform this technique with a gracilis muscle transfer and discuss indications and limitations.

Unilateral facial paralysis is a common disease that is associated with significant functional, aesthetic and psychological issues. Though idiopathic ...

Step By Step: Microsurgical training method combining two nonliving animal models

The learning of microsurgical techniques and the maintenance of microsurgical skills have been traditionally based on the use of living animals, mainly laboratory rats. This method although extremely valuable can be economically demanding both for the surgeon and the sponsoring institution; it also requires special training facilities that may not always be available or accessible. Furthermore ethical concerns can limit the use of living animals for training purposes. Alternative training methods, such as inert tubes and gloves have not gained popularity among surgeons since they do not offer an experience similar to that of a clinical situation. Non-living animal models include the use of chicken thighs and wings; they offer a practice experience that resembles a clinical situation to a considerable extent. This type of training is relatively cheap and easily available. The microscope and instruments required can be acquired over the internet, and the chicken pieces can be bought at the local supermarket.
This approach allows a motivated trainee to rehearse different types of surgical techniques several times at a reasonable expense, helping to develop or maintain his surgical expertise if more complex facilities are not available. On the current manuscript we describe how to setup a small practice station, how to dissect the specimens, and how to practice both with the chicken thighs and with the chicken wings in a progressive fashion. This approach takes advantage on the versatility of the chicken thigh model and the small size of the chicken wing Brachial artery.

Here we describe how to set up a small microsurgical practice station and a simple and inexpensive method for the training of microsurgery with non-living animal models.

The learning of microsurgical techniques and the maintenance of microsurgical skills have been traditionally based on the use of living animals, mainly ...

A Novel Approach for the Administration of Medications and Fluids in Emergency Scenarios and Settings

The available routes of administration commonly used for medications and fluids in the acute care setting are generally limited to oral, intravenous, or intraosseous routes, but in many patients, particularly in the emergency or critical care settings, these routes are often unavailable or time-consuming to access. A novel device is now available that offers an easy route for administration of medications or fluids via rectal mucosal absorption (also referred to as proctoclysis in the case of fluid administration and subsequent absorption). Although originally intended for the palliative care market, the utility of this device in the emergency setting has recently been described. Specifically, reports of patients being treated for dehydration, alcohol withdrawal, vomiting, fever, myocardial infarction, hyperthyroidism, and cardiac arrest have shown success with administration of a wide variety of medications or fluids (including water, aspirin, lorazepam, ondansetron, acetaminophen, methimazole, and buspirone). Device placement is straightforward, and based on the observation of expected effects from the medication administrations, absorption is rapid. The rapidity of absorption kinetics are further demonstrated in a recent report of the measurement of phenobarbital pharmacokinetics. We describe here the placement and use of this device, and demonstrate methods of pharmacokinetic measurements of medications administered by this method.

This study presents a novel device that offers an easy route to quickly provide medications and fluids in patients with limited or difficult IV access. This small-diameter device is placed in the distal third of the rectum, allowing for ongoing medication and fluids administration.

The available routes of administration commonly used for medications and fluids in the acute care setting are generally limited to oral, intravenous, or ...

Stromal Vascular Fraction-enriched Fat Grafting for the Treatment of Symptomatic End-neuromata

The purpose of this study was to methodically illustrate and highlight the crucial steps of stromal vascular fraction (SVF)-enriched fat grafting as a novel treatment of symptomatic end-neuromata of peripheral sensory nerves, and in this study, specifically of the superficial branch of the radial nerve (SBRN). Despite a multitude of existing treatments, persistent postoperative pain and common pain relapse are still very common, independent of the procedure assessed. The neuroma is microsurgically excised accordingly to standardized protocol. Instead of the relocation of the regenerating nerve stump in neighboring anatomical structures, such as muscle or bone, a fat graft is applied perifocally and acts as a mechanical barrier. In order to reduce the fat resorption rate and boost the regenerative potential of the graft, the highly concentrated SVF is integrated in the grafting. The SVF is isolated from subcutaneous fat by enzymatic and mechanic separation of the lipoaspirate by a specific commercial isolation system. The SVF-enriched fat graft provides both a mechanical barrier and various biological effects at the cellular level, including improving angiogenesis, inflammation, and fibrosis. Both mechanical and biologic effects help to reduce the disorganized axonal outgrowth of the nerve stump during nerve regeneration and hence prevent the recurrence of painful end-neuromata.

The purpose of the study is to illustrate the technical procedure of stromal vascular fraction (SVF)-enriched fat grafting as a novel treatment of symptomatic end-neuromata. This technique provides advantages of both the mechanical barrier and the biological action at the cellular level by the processed and concentrated SVF.

The purpose of this study was to methodically illustrate and highlight the crucial steps of stromal vascular fraction (SVF)-enriched fat grafting as a ...

Taste Exam: A Brief and Validated Test

The emerging importance of taste in medicine and biomedical research, and new knowledge about its genetic underpinnings, has motivated us to supplement classic taste-testing methods in two ways. First, we explain how to do a brief assessment of the mouth, including the tongue, to ensure that taste papillae are present and to note evidence of relevant disease. Second, we draw on genetics to validate taste test data by comparing reports of perceived bitterness intensity and inborn receptor genotypes. Discordance between objective measures of genotype and subjective reports of taste experience can identify data collection errors, distracted subjects or those who have not understood or followed instructions. Our expectation is that fast and valid taste tests may persuade researchers and clinicians to assess taste regularly, making taste testing as common as testing for hearing and vision. Finally, because many tissues of the body express taste receptors, taste responses may provide a proxy for tissue sensitivity elsewhere in the body and, thereby, serve as a rapid, point-of-care test to guide diagnosis and a research tool to evaluate taste receptor protein function.

This protocol measures human taste responses and includes a brief anatomical assessment, a short taste test, and a validation method using the subject&#39;s reported sensation and taste receptor genotype.

The emerging importance of taste in medicine and biomedical research, and new knowledge about its genetic underpinnings, has motivated us to supplement ...

Traditional Trail Making Test Modified into Brand-new Assessment Tools: Digital and Walking Trail Making Test

The Trail Making Test (TMT) is a well-accepted tool for evaluating executive function. The standard TMT was invented more than 60 years ago and has been modified into many versions. With the development of digital technologies, TMT is now modified to a digitized version. The present study demonstrated digital TMT (dTMT) performed on a computer, and Walking TMT (WTMT) on the floor. Both revealed more information compared with the traditional version of TMT.

Here, we present a protocol to show how to perform two types of cognitive assessment tools derived from the paper-pencil version of the Trail Making Test.

The Trail Making Test (TMT) is a well-accepted tool for evaluating executive function. The standard TMT was invented more than 60 years ago and has been ...

Intratracheal Administration of Dry Powder Formulation in Mice

In the development of inhalable dry powder formulations, it is essential to evaluate their biological activities in preclinical animal models. This paper introduces a noninvasive method of intratracheal delivery of dry powder formulation in mice. A dry powder loading device that consists of a 200 &#181;L gel loading pipette tip connected to an 1 mL syringe via a three-way stopcock is presented. A small amount of dry powder (1-2 mg) is loaded into the pipette tip and dispersed by 0.6 mL of air in the syringe. Because pipette tips are disposable and inexpensive, different dry powder formulations can be loaded into different tips in advance. Various formulations can be evaluated in the same animal experiment without device cleaning and dose refilling, thereby saving time and eliminating the risk of cross-contamination from residual powder. The extent of powder dispersion can be inspected by the amount of powder remaining in the pipette tip. A protocol of intubation in mouse with a custom-made light source and a guiding cannula is included. Proper intubation is one of the key factors that influences the intratracheal delivery of dry powder formulation to the deep lung region of the mouse.

Dry powder formulations for inhalation have great potential in treating respiratory diseases. Before entering human studies, it is necessary to evaluate the efficacy of the dry powder formulation in preclinical studies. A simple and noninvasive method of the administration of dry powder in mice through the intratracheal route is presented.

In the development of inhalable dry powder formulations, it is essential to evaluate their biological activities in preclinical animal models. This paper ...