Thank you to the "Thim van der Laan" Foundation, Landquart, Switzerland, for the financial support. We also thank Thomas Konzett, University of Applied Sciences and Arts of Southern Switzerland, Landquart, Switzerland, and Ursula M. K&#252;ng, University College Physiotherapy, Landquart, Switzerland, for their assistance during the experiment, as well as Alexander Masselink, for his assistance with English.

Ethics Statement: The Swiss Cantonal Ethical Committee of Zurich approved this protocol (KEK-ZH No. 2015-0113)
1. Recruitment of Participants
<ol>
	<li>Recruit participants through social media and flyers.</li>
	<li>Screen and include participants who are university students between 18 and 30 years of age. Ensure that they can perform endurance sports a minimum of two times (minimum total time: 2 h) and a maximum of three times (maximum total time: 3 h) per week, that they are free of current pain symptoms, and that they have had no injuries in the torso or lower extremities in the previous 12 months.</li>
	<li>Screen and exclude participants if they are allergic to cold (including Raynaud&#39;s disease), suffer from cardiovascular diseases or any other disease, must take any medication, are pregnant, or have skeletal deviations.</li>
</ol>
2. Familiarization with the Testing Protocol (Day 1)
<ol>
	<li>One week prior to the experiment, instruct the participants to perform a maximum of 5 CMJs on a jump mat (see the Table of Materials), with a 1-min resting period between each jump12,27. Instruct them to start from an upright position and to place their hands on their hips to eliminate arm swing. Permit a self-determined range of motion and contraction time during the CMJs.</li>
	<li>Instruct the participants to return to the laboratory one week after the familiarization session in a pain-free state and without having performed exhaustive exercise 48 h prior to the experiment. 
	NOTE: The pain-free and exhaustion-free state will be assessed as the baseline measurements.</li>
</ol>
3. Baseline Measurements (Day 2)
<ol>
	<li>Measure the standing body-height, body-weight, and estimated lower-body fat percentage of the participants three times in a row and calculate the mean values28.</li>
	<li>Have the participants assess their individual DOMS of both frontal thighs on a visual analog scale (VAS; 0-10 cm)5 during a 3-s lasting squat position29 (90&#176; knee angle). Ask the participants: &#34;How sore are your frontal thighs?&#34; Record the number from the VAS in mm. 
	NOTE: The far-left endpoint of the scale (0 cm) represents &#34;no soreness,&#34; and the far-right endpoint of the scale (10 cm) represents &#34;severe soreness.&#34;</li>
	<li>Have the participants rate their individual perceived exertion in an upright standing position using a 6-20 BORG scale30. Ask the participants: &#34;What is your actual level of perceived exertion?&#34; Record the verbally communicated number. 
	NOTE: The participants rate this verbally by telling the researcher one number between 6 (no perceived exertion) and 20 (maximum perceived exertion).</li>
	<li>Have the participants perform a maximum of 3 CMJs12,27 (as practiced during the familiarization session), with a 30-s pause between the three jumps, on a jump mat. 
	NOTE: The jump heights from the CMJs are indirectly recorded as the time in flight by the jump mat system.</li>
	<li>Use the highest jump and calculate the peak power output (PPO) according to the formula used in the study of Sayers et al.31: 
	PPO = (60.7 x jump height [cm]) + (45.3 x body weight [kg]) &#8211; 2,055 
	NOTE: The subjective ratings and the objective assessments will be repeated directly after the recovery application (0 h) and 24, 48, and 72 h after the exhaustive jump protocol (see below).</li>
	<li>Randomly assign the participants either to the cold group or to the thermoneutral group by drawing lots.</li>
</ol>
4. Exhaustive Jump Protocol
<ol>
	<li>Directly after the randomization, instruct the participants to get ready to perform the exhaustive jump protocol, without any warm-up exercises.</li>
	<li>Have two researchers visually inspect the quality of the jumps (maximum jump performance and touching the ground with the fingers after each jump), verbally correct, and encourage the participants during the jump protocol.</li>
	<li>Have the participants perform a maximum of 3 x 30 CMJs or until exhaustion at their own speeds, with a 30 s pause between the sets. 
	NOTE: Allow the participants to sit down and drink water during the pause.</li>
</ol>
5. Recovery Application
<ol>
	<li>After the exhaustive jump protocol, have a blinded researcher apply either a cold cuff or a thermoneutral cuff (see the Table of Materials) to a participant&#39;s thighs.
	<ol>
		<li>Place the participant in supine position and apply both cuffs directly onto the skin of each thigh to ensure full contact, but using minimum pressure to avoid any compression effects. 
		NOTE: Optionally, refill the tank of a continuous programmable cooling and heating device with a ready-to-use, non-toxic lubrication mixture consisting of propylene glycol and demineralized water to the maximum level.</li>
	</ol>
	</li>
	<li>Start the device and apply either the cold modality (+ 8 &#176;C) or the thermoneutral modality (+ 32 &#176;C) for a duration of 20 min. 
	NOTE: Instruct the participants not to report information about their temperature sensation.</li>
	<li>Take off the thigh cuffs after the application and shut down the device.</li>
</ol>
6. Non-invasive Recovery Assessment After 0 h
NOTE: The researcher performing all recovery assessments should not be informed if the participants received a cold- or a thermoneutral-cuff application.
<ol>
	<li>Have the participants rate their individual DOMS and RPE, as described in steps 3.2 and 3.3.</li>
	<li>Have the participants perform a maximum of 3 CMJs and calculate the PPO, as described in steps 3.4 and 3.5.</li>
	<li>Instruct the participants to maintain their daily habits but to refrain from alcohol, sport, and recreational training during the 72 h recovery period. Instruct the participants to return to the laboratory exactly 24, 48, and 72 h after the exhaustive jump protocol.</li>
</ol>
7. Non-invasive Recovery Assessment After 24 h (Day 3), 48 h (Day 4), and 72 h (Day 5)
<ol>
	<li>Have the participants rate their individual DOMS and RPE, as described in steps 3.2 and 3.3.</li>
	<li>Have the participants perform a maximum of 3 CMJs and calculate the PPO, as described in steps 3.4 and 3.5. 
	NOTE: The protocol ends after the 72 h follow-up period (Day 5).</li>
</ol>

None of the authors have competing or conflicting interests.

This protocol describes the execution of a standardized exhaustive jump protocol, a cryotherapy recovery modality, and a non-invasive assessment of recovery characteristics. The protocol of this randomized controlled trial provides several advantages compared to traditionally performed methods in the field of post-exercise recovery studies. Previous studies showed that jump protocols consisting of 100 jumps are a valid method to induce local muscle damage36,37. Ferreira-Junior et al. used a jump protocol consisting of five sets of 20 drop-jumps from a 0.6 m box, with 2 min rest intervals between the sets, to induce muscle damage of the frontal thighs36. Whereas other traditional exhaustion protocols require expensive or mechanical devices38, the current protocol induced muscular exhaustion without the use of any mechanical device. Researchers do not need to buy or rent expensive devices to induce muscular exhaustion or damage to the frontal thighs.
The recovery interventions were applied using a continuous programmable cooling and heating device that can deliver cold or heat to a specific area of the body. Two thigh cuffs were applied around both thighs in supine position for 20 min. Although CWI is one of the most promising methods of enhancing recovery39, the transportation and the necessary amount of water are two challenging factors for the execution of this intervention. Furthermore, to guarantee the ideal water temperature, an additional person is needed to monitor and add ice.
An advantage of the present protocol is the use of an automated and portable cooling and heating device that ensures a constant temperature during the cuff application. Another advantage of the described protocol is the possibility of assessing subjective and objective recovery characteristics without taking blood samples from the participants. The subjective ratings, DOMS (VAS; 0-10 cm) and RPE (BORG; 6-20), used after an exhaustive protocol have been described in numerous published articles24,40,41,42. The assessment of CMJs, as presented in the present protocol, shows a high test-retest reliability (intra-class correlation coefficient (ICC) between 0.48 and 0.88) and validity25,26.
There are some critical steps within the protocol that may present potential sources of error. Two researchers must observe, verbally encourage, and correct the participants during the exhaustive CMJs (3 x 30). The objective monitoring of maximum CMJs may be ensured using accelerometers or linear transducers. Another critical step is the application of the two thigh cuffs. To avoid any compression effects43,44, the cuffs must be wrapped around each thigh with a minimum level of pressure. The application of the cuffs with a minimum of pressure might take a couple of practice runs to master.
The present protocol has some limitations. The 30 s rest period between the sets of the exhaustive jump protocol is very short, and the cardiovascular impact is very high. Another limitation is the ready-to-use lubrication mixture for the recovery modality. It is important to take into account that the heat capacity of this lubrication mixture (i.e., propylene glycol and demineralized water) is slightly lower than that of normal water. The familiarization session of 5 jumps might be too small when the study population is not as physically active as described in the present protocol.
Finally, the assessment of CMJs presents the opportunity to assess objective recovery characteristics. Bishop et al. demonstrated a cheap and practicable method to assess jump height with the mobile-based app &#34;My Jump,&#34; which has been shown to be a reliable method for measuring this variable45,46. However, Rowsell et al. indicated that no apparent reductions in CMJ height during a 5-day follow-up period could be observed after exhaustive soccer tournament matches24. Rupp et al. observed similar results after an exhaustive endurance test34. These results are in line with the results of the presents study, showing that the assessment of CMJ height might not be sensitive enough to measure the amount of muscular exhaustion.
In this protocol, the cold application temperature was set at 8 &#176;C, whereas the temperature of the thermoneutral application was set at 32 &#176;C. It has been demonstrated that cold water temperatures are normally &#8804;20 &#176;C and that thermoneutral water temperatures have a range from 24 &#176;C to &#60;36 &#176;C47. It is important to consider that the amount of adipose tissue significantly affects the tissue cooling rate, with thicker skinfolds requiring longer application times48. Researchers should modify the cooling temperature and application duration according to their research populations.
Future studies should consider that the assessment of the maximum voluntary contractions of the knee extensors might be a more sensitive assessment of objective recovery characteristics compared to CMJs49. For this protocol to be effective, it is crucial that the participants perform a familiarization session for the CMJs. Future studies using a different study population than that described here should increase the number of jumps to guarantee a familiarization effect. Additionally, future studies might increase the resting time between the exhaustive CMJs to guarantee maximum jump performance, which would then be unaffected by the high cardiovascular demand.
In conclusion, the current exhaustive jump protocol is an easy and practical way to induce muscular exhaustion of the frontal thighs without the use of any mechanical devices. By combining subjective (i.e., DOMS and RPE) and objective (i.e., CMJ and PPO) parameters, recovery can be investigated without taking any blood samples during a 72 h recovery period. The local post-exercise cryotherapy application can be carried out nearly anywhere and guarantees constant cooling temperatures.

Cryotherapy is a frequently used treatment modality to enhance post-exercise recovery1,2. The vasoconstrictive response of the body after exposure to cold is one of the main mechanisms of reducing the inflammatory processes3,4. Frequently used cryotherapy procedures include cold packs5 or cuffs6, cold air7,8, cold water immersion (CWI)9, whole-body cryotherapy10,11, cooling vests12, and internal cooling methods13,14. However, internal cooling procedures have demonstrated contradictory results15,16.
Controversial results on recovery have also been reported after external local cooling applications. One study reported that post-exercise cryotherapy did not enhance vertical jump performance (VJP) but decreased subjective ratings of perceived exertion (RPE) compared to active recovery procedures17. In contrast, another study demonstrated that cryotherapy had no influence on post-exercise subjective RPE5. A meta-analysis by Hohenauer et al. indicated that post-exercise cryotherapy might have a positive effect on subjective recovery characteristics, with no effects on inflammatory markers1.
The majority of studies that determine the effects of cryotherapy on recovery involve expensive devices to induce muscular exhaustion7,18,19 and invasive procedures to assess blood plasma markers or cytokines19,20,21. The aims of the present protocol are to induce similar muscular exhaustion without the use of any device and to provide a non-invasive method to determine the effects of local cryotherapy cuff application on subjective and objective recovery characteristics. The rationale behind this method is that it provides a tool for other researchers or coaches to induce muscular exhaustion, without using any expensive devices; to implement local cooling strategies based on this protocol; and to measure both subjective and objective recovery, without using invasive methods. This might help to assess subjective and objective recovery characteristics following an exhaustive jump protocol in a sport-realistic, non-invasive setting.
Published studies show that validated, non-invasive methods for evaluating subjective recovery characteristics (i.e., delayed-onset muscle soreness (DOMS)22 and RPE)23 have been used with success. Countermovement jump (CMJ) performance17,24, with a high test-retest reliability25,26, can also be used as a non-invasive method to assess objective recovery characteristics.

<table border="0" cellpadding="0" cellspacing="0" width="601">
	<tbody>
		<tr height="147">
			<td height="147" style="height:147px;width:216px;">&#160;Anthropometer 100</td>
			<td style="width:99px;">GPM Anthropometric Instruments (URL:http://www.seritex.com/)</td>
			<td style="width:119px;">not applicable&#160;</td>
			<td style="width:167px;">Standing body-height can be measured with other accurate systems</td>
		</tr>
		<tr height="105">
			<td height="105" style="height:105px;width:216px;">TANITA TBF 611</td>
			<td style="width:99px;">TANITA corporation (URL:http://www.tanita.com/en/)</td>
			<td style="width:119px;">500314M</td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="168">
			<td height="168" style="height:168px;width:216px;">Just Jump System&#160;</td>
			<td style="width:99px;">Probotics Inc. (URL:http://www.probotics.org/JustJump/JustJump.htm)&#160;</td>
			<td style="width:119px;">23056311</td>
			<td style="width:167px;">This system includes the jump mat and the jump height recorder. Other accurate systems, measuring vertical jump height may be used alternatively</td>
		</tr>
		<tr height="126">
			<td height="126" style="height:126px;width:216px;">Zamar Therapy ZT Clinic&#160;</td>
			<td style="width:99px;">Zamar Medical D.O.O (URL:http://www.zamar.care/)</td>
			<td style="width:119px;">MG675AA00F&#160;</td>
			<td style="width:167px;">This is a continous programmable cooling and heating device&#160;</td>
		</tr>
		<tr height="84">
			<td height="84" style="height:84px;width:216px;">Zamar Large Thigh Thermal Wraps</td>
			<td style="width:99px;">Zamar Care (URL:http://www.zamar.care/sport.html)</td>
			<td style="width:119px;">not applicable&#160;</td>
			<td style="width:167px;">2 Thigh Thermal Wraps are needed</td>
		</tr>
		<tr height="63">
			<td height="63" style="height:63px;width:216px;">Zamar Equi Insulated 4.7 m &#34;V&#34;t Pipe &#38; Safety Connector</td>
			<td style="width:99px;">http://www.zamar.care/clinic.html</td>
			<td style="width:119px;">ZAM-1ACS410</td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="105">
			<td height="105" style="height:105px;width:216px;">Non Tox Freeze 4&#160;</td>
			<td style="width:99px;">Pakelo Motor Oil S.r.l. (URL:http://www.pakelo.com/)</td>
			<td style="width:119px;">0131.34.47</td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="147">
			<td height="147" style="height:147px;width:216px;">Schmerzskala (VAS 0 - 10 cm)</td>
			<td style="width:99px;">Mundipharma Medical Company (URL:http://www.mundipharma.ch/index.php?id=73)</td>
			<td style="width:119px;">not applicable&#160;</td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="168">
			<td height="168" style="height:168px;width:216px;">BORG scale (6 - 20)</td>
			<td style="width:99px;">URL:http://www.mesics.de/fileadmin/user/literature/Allgemein/Borg-Skala_Loellgen.pdf</td>
			<td style="width:119px;">not applicable&#160;</td>
			<td style="width:167px;"></td>
		</tr>
	</tbody>
</table>

non-invasive assessments of subjective and objective recovery characteristics following an exhaustive jump protocol

Fast recovery after strenuous exercise is important in sports and is often studied via cryotherapy applications. Cryotherapy has a significant vasoconstrictive effect, which seems to be the leading factor in its effectiveness. The resulting enhanced recovery can be measured by using both objective and subjective parameters. Two commonly measured subjective characteristics of recovery are delayed-onset muscle soreness (DOMS) and ratings of perceived exertion (RPE). Two important objective recovery characteristics are countermovement jump (CMJ) performance and peak power output (PPO). Here, we provide a detailed protocol to induce muscular exhaustion of the frontal thighs with a self-paced, 3 x 30 countermovement jump protocol (30-s rest between each set). This randomized controlled trial protocol explains how to perform local cryotherapy cuff application (+ 8 &#176;C for 20 min) and thermoneutral cuff application (+ 32 &#176;C for 20 min) on both thighs as two possible post-exercise recovery modalities. Finally, we provide a non-invasive protocol to measure the effects of these two recovery modalities on subjective (i.e., DOMS of both frontal thighs and RPE) and objective recovery (i.e., CMJ and PPO) characteristics 24, 48, and 72 h post-application. The advantage of this method is that it provides a tool for researchers or coaches to induce muscular exhaustion, without using any expensive devices; to implement local cooling strategies; and to measure both subjective and objective recovery, without using invasive methods. Limitations of this protocol are that the 30 s rest period between sets is very short, and the cardiovascular demand is very high. Future studies may find the assessment of maximum voluntary contractions to be a more sensitive assessment of muscular exhaustion compared to CMJs.

The schematic representation of the test protocol is presented in Figure 1. This section illustrates that this protocol was successful at inducing muscular exhaustion and at monitoring subjective and objective recovery characteristics during a 72 h follow-up period, without using invasive techniques (Figure 2 and Figure 3).
Goodall et al. studied DOMS in a 90&#176; squat position after a muscle damage protocol29. Likewise, the present study also assessed DOMS in this position. As described in previously published articles, the DOMS elevated directly after the recovery application (0 h) and after 48 h in both groups (Figure 2A)29,32. However, in both groups, these values did not return to baseline values after a 72 h recovery period32.
This protocol also allows for the observation of changes in overall perceived exertion (Figure 2B). In the present study, a 6-20 BORG scale was used as in the study of Rowsell et al.24. In accordance to the study of Minett et al., the RPE was elevated in both groups directly after the recovery application12. However, these values returned to baseline after 24 h in the thermoneutral group and remained at this level for up to 72 h. In the cold group, the RPE values increased again after 48 h and remained at this level for up to 72 h.
Figure 3A and Figure 3B show that the exhaustive jump protocol induced a decrease in jump height (CMJ) and PPO in both conditions directly after the recovery application (0 h)12,33. Both the jump height (CMJ) and PPO increased after 24 h and again decreased after 48 and 72 h in the cold and thermoneutral groups. These results are in line with previously published articles24,33,34.

<img alt="Figure 1" src="/files/ftp_upload/55612/55612fig1.jpg" /> 
Figure 1: Schematic representation of the test protocol. Downward-pointing arrows denote the time points when recovery characteristics were measured. DOMS = delayed-onset muscle soreness, RPE = rated perceived exertion, CMJ = countermovement jump, PPO = peak power output. This figure has been modified from Hohenauer et al.35. <a href="https://cloudflare-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/55612/55612fig1large.jpg" target="_blank">Please click here to view a larger version of this figure.</a>

<img alt="Figure 2" src="/files/ftp_upload/55612/55612fig2.jpg" /> 
Figure 2: Change in DOMS and RPE over time. (A) Delayed-onset muscle soreness (DOMS; (median &#177; interquartile ranges). A repeated-measures ANOVA revealed a time effect (P = 0.003) and a group effect (P = 0.03), but no group x time interaction (P = 0.89). Post-hoc differences between the groups were P &#62;0.05 for all time points. (B) Rated perceived exertion (RPE; median &#177; interquartile ranges). A repeated-measures ANOVA revealed a time effect (P &#60;0.001) and a group effect (P = 0.09), but no group x time interaction (P = 0.29). Post-hoc differences between the groups were P &#62;0.05 for all time points. * indicates a significant time effect (p &#60;0.05). # indicates significant differences from baseline values within groups (p &#60;0.05). This figure has been modified from Hohenauer et al.35. <a href="https://cloudflare-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/55612/55612fig2large.jpg" target="_blank">Please click here to view a larger version of this figure.</a>
<img alt="Figure 3" src="/files/ftp_upload/55612/55612fig3.jpg" /> 
Figure 3: Change in CMJ and PPO over time. (A) Jump height during countermovement jump (CMJ; mean &#177; SD). A repeated-measures ANOVA revealed a time effect (P &#60;0.001), but no group effect (P = 0.35) and time x group interaction (P = 0.35). Post-hoc differences between the groups were P &#62;0.05 for all time points. (B) Peak power output (PPO; mean &#177; SD). A repeated-measures ANOVA revealed a time effect (P &#60;0.001), but no group effect (P = 0.96) and time x group interaction (P = 0.35). Post-hoc differences between groups were P &#62;0.05 for all time points. * indicates a significant time effect (P &#60;0.05). This figure has been modified from Hohenauer et al.35. <a href="https://cloudflare-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/55612/55612fig3large.jpg" target="_blank">Please click here to view a larger version of this figure.</a>

Watch this Scientific Journal Video about Non-invasive Assessments of Subjective and Objective Recovery Characteristics Following an Exhaustive Jump Protocol at JoVE.com

Non-invasive Assessments of Subjective and Objective Recovery Characteristics Following an Exhaustive Jump Protocol

This protocol describes the procedure for non-invasive recovery assessments during a 72 h recovery period. This protocol induces muscular exhaustion of the frontal thighs using countermovement jumps and uses either local cold-cuff or thermoneutral-cuff application as a recovery modality.

Fast recovery after strenuous exercise is important in sports and is often studied via cryotherapy applications. Cryotherapy has a significant ...

non-invasive-assessments-subjective-objective-recovery

Nanyang Technological University

Research

JoVE Journal

Medicine

7.6K Views.  University of Applied Sciences and Arts of Southern Switzerland. This protocol describes the procedure for non-invasive recovery assessments during a 72 h recovery period. This protocol induces muscular exhaustion of the frontal thighs using countermovement jumps and uses either local cold-cuff or thermoneutral-cuff application as a recovery modality.

Video: Non-invasive Assessments of Subjective and Objective Recovery Characteristics Following an Exhaustive Jump Protocol

An in vivo Rodent Model of Contraction-induced Injury and Non-invasive Monitoring of Recovery

Muscle strains are one of the most common complaints treated by physicians. A muscle injury is typically diagnosed from the patient history and physical exam alone, however the clinical presentation can vary greatly depending on the extent of injury, the patient's pain tolerance, etc. In patients with muscle injury or muscle disease, assessment of muscle damage is typically limited to clinical signs, such as tenderness, strength, range of motion, and more recently, imaging studies. Biological markers, such as serum creatine kinase levels, are typically elevated with muscle injury, but their levels do not always correlate with the loss of force production. This is even true of histological findings from animals, which provide a "direct measure" of damage, but do not account for all the loss of function. Some have argued that the most comprehensive measure of the overall health of the muscle in contractile force. Because muscle injury is a random event that occurs under a variety of biomechanical conditions, it is difficult to study. Here, we describe an in vivo animal model to measure torque and to produce a reliable muscle injury. We also describe our model for measurement of force from an isolated muscle in situ. Furthermore, we describe our small animal MRI procedure.

An in vivo Rodent Model of Contraction-induced Injury and Non-invasive Monitoring of Recovery

An in vivo animal model of injury is described. The method takes advantage of the subcutaneous position of the fibular nerve. Velocity, timing of muscle activation, and arc of motion are all pre-determined and synchronized using commercial software. Post injury changes are monitored in vivo using MR imaging/spectroscopy.

Muscle strains are one of the most  common complaints treated by physicians. A muscle injury is typically diagnosed from the patient history and  physical ...

Non-invasive Optical Measurement of Cerebral Metabolism and Hemodynamics in Infants

Perinatal brain injury remains a significant cause of infant mortality and morbidity, but there is not yet an effective bedside tool that can accurately screen for brain injury, monitor injury evolution, or assess response to therapy. The energy used by neurons is derived largely from tissue oxidative metabolism, and neural hyperactivity and cell death are reflected by corresponding changes in cerebral oxygen metabolism (CMRO2). Thus, measures of CMRO2 are reflective of neuronal viability and provide critical diagnostic information, making CMRO2 an ideal target for bedside measurement of brain health.
Brain-imaging techniques such as positron emission tomography (PET) and single-photon emission computed tomography (SPECT) yield measures of cerebral glucose and oxygen metabolism, but these techniques require the administration of radionucleotides, so they are used in only the most acute cases.
Continuous-wave near-infrared spectroscopy (CWNIRS) provides non-invasive and non-ionizing radiation measures of hemoglobin oxygen saturation (SO2) as a surrogate for cerebral oxygen consumption. However, SO2 is less than ideal as a surrogate for cerebral oxygen metabolism as it is influenced by both oxygen delivery and consumption. Furthermore, measurements of SO2 are not sensitive enough to detect brain injury hours after the insult 1,2, because oxygen consumption and delivery reach equilibrium after acute transients 3. We investigated the possibility of using more sophisticated NIRS optical methods to quantify cerebral oxygen metabolism at the bedside in healthy and brain-injured newborns. More specifically, we combined the frequency-domain NIRS (FDNIRS) measure of SO2 with the diffuse correlation spectroscopy (DCS) measure of blood flow index (CBFi) to yield an index of CMRO2 (CMRO2i) 4,5.
With the combined FDNIRS/DCS system we are able to quantify cerebral metabolism and hemodynamics. This represents an improvement over CWNIRS for detecting brain health, brain development, and response to therapy in neonates. Moreover, this method adheres to all neonatal intensive care unit (NICU) policies on infection control and institutional policies on laser safety. Future work will seek to integrate the two instruments to reduce acquisition time at the bedside and to implement real-time feedback on data quality to reduce the rate of data rejection.

We combined frequency-domain near-infrared spectroscopy measures of cerebral hemoglobin oxygenation with diffuse correlation spectroscopy measures of cerebral blood flow index to estimate an index of oxygen metabolism. We tested the utility of this measure as a bedside screening tool to evaluate the health and development of the newborn brain.

Perinatal brain injury remains a significant cause of infant mortality  and morbidity, but there is not yet an effective bedside tool that can  accurately ...

Non-invasive Assessment of Microvascular and Endothelial Function

The authors have utilized capillaroscopy and forearm blood flow techniques to investigate the role of microvascular dysfunction in pathogenesis of cardiovascular disease. Capillaroscopy is a non-invasive, relatively inexpensive methodology for directly visualizing the microcirculation. Percent capillary recruitment is assessed by dividing the increase in capillary density induced by postocclusive reactive hyperemia (postocclusive reactive hyperemia capillary density minus baseline capillary density), by the maximal capillary density (observed during passive venous occlusion). Percent perfused capillaries represents the proportion of all capillaries present that are perfused (functionally active), and is calculated by dividing postocclusive reactive hyperemia capillary density by the maximal capillary density. Both percent capillary recruitment and percent perfused capillaries reflect the number of functional capillaries. The forearm blood flow (FBF) technique provides accepted non-invasive measures of endothelial function: The ratio FBFmax/FBFbase is computed as an estimate of vasodilation, by dividing the mean of the four FBFmax values by the mean of the four FBFbase values. Forearm vascular resistance at maximal vasodilation (FVRmax) is calculated as the mean arterial pressure (MAP) divided by FBFmax. Both the capillaroscopy and forearm techniques are readily acceptable to patients and can be learned quickly.
The microvascular and endothelial function measures obtained using the methodologies described in this paper may have future utility in clinical patient cardiovascular risk-reduction strategies. As we have published reports demonstrating that microvascular and endothelial dysfunction are found in initial stages of hypertension including prehypertension, microvascular and endothelial function measures may eventually aid in early identification, risk-stratification and prevention of end-stage vascular pathology, with its potentially fatal consequences.

Capillaroscopy is a non-invasive, relatively inexpensive methodology for directly visualizing the microcirculation. The forearm blood flow technique provides accepted non-invasive measures of endothelial function.

The authors have utilized capillaroscopy and forearm blood flow techniques to investigate the role of microvascular dysfunction in pathogenesis of ...

Lesion Explorer: A Video-guided, Standardized Protocol for Accurate and Reliable MRI-derived Volumetrics in Alzheimer's Disease and Normal Elderly

Obtaining in vivo human brain tissue volumetrics from MRI is often complicated by various technical and biological issues. These challenges are exacerbated when significant brain atrophy and age-related white matter changes (e.g. Leukoaraiosis) are present. Lesion Explorer (LE) is an accurate and reliable neuroimaging pipeline specifically developed to address such issues commonly observed on MRI of Alzheimer's disease and normal elderly. The pipeline is a complex set of semi-automatic procedures which has been previously validated in a series of internal and external reliability tests1,2. However, LE's accuracy and reliability is highly dependent on properly trained manual operators to execute commands, identify distinct anatomical landmarks, and manually edit/verify various computer-generated segmentation outputs.
LE can be divided into 3 main components, each requiring a set of commands and manual operations: 1) Brain-Sizer, 2) SABRE, and 3) Lesion-Seg. Brain-Sizer's manual operations involve editing of the automatic skull-stripped total intracranial vault (TIV) extraction mask, designation of ventricular cerebrospinal fluid (vCSF), and removal of subtentorial structures. The SABRE component requires checking of image alignment along the anterior and posterior commissure (ACPC) plane, and identification of several anatomical landmarks required for regional parcellation. Finally, the Lesion-Seg component involves manual checking of the automatic lesion segmentation of subcortical hyperintensities (SH) for false positive errors.
While on-site training of the LE pipeline is preferable, readily available visual teaching tools with interactive training images are a viable alternative. Developed to ensure a high degree of accuracy and reliability, the following is a step-by-step, video-guided, standardized protocol for LE's manual procedures.

Lesion Explorer (LE) is a semi-automatic, image-processing pipeline developed to obtain regional brain tissue and subcortical hyperintensity lesion volumetrics from structural MRI of Alzheimer's disease and normal elderly. To ensure a high level of accuracy and reliability, the following is a video-guided, standardized protocol for LE's manual procedures.

Obtaining in vivo human brain tissue volumetrics from MRI is often complicated by various technical and biological issues. These challenges are ...

Non-invasive Imaging and Analysis of Cerebral Ischemia in Living Rats Using Positron Emission Tomography with 18F-FDG

Stroke is the third leading cause of death among Americans 65 years of age or older1. The quality of life for patients who suffer from a stroke fails to return to normal in a large majority of patients2, which is mainly due to current lack of clinical treatment for acute stroke. This necessitates understanding the physiological effects of cerebral ischemia on brain tissue over time and is a major area of active research. Towards this end, experimental progress has been made using rats as a preclinical model for stroke, particularly, using non-invasive methods such as 18F-fluorodeoxyglucose (FDG) coupled with Positron Emission Tomography (PET) imaging3,10,17. Here we present a strategy for inducing cerebral ischemia in rats by middle cerebral artery occlusion (MCAO) that mimics focal cerebral ischemia in humans, and imaging its effects over 24 hr using FDG-PET coupled with X-ray computed tomography (CT) with an Albira PET-CT instrument. A VOI template atlas was subsequently fused to the cerebral rat data to enable a unbiased analysis of the brain and its sub-regions4. In addition, a method for 3D visualization of the FDG-PET-CT time course is presented. In summary, we present a detailed protocol for initiating, quantifying, and visualizing an induced ischemic stroke event in a living Sprague-Dawley rat in three dimensions using FDG-PET.

Non-invasive Imaging and Analysis of Cerebral Ischemia in Living Rats Using Positron Emission Tomography with 18F-FDG

Brain damage resulting from cerebral ischemia may be non-invasively imaged and studied in rats using pre-clinical positron emission tomography coupled with the injectable radioactive probe, 18F-fluorodeoxyglucose. Further, the use of modern software tools that include volume of interest (VOI) brain templates dramatically increase the quantitative information gleaned from these studies.

Stroke is the third leading cause of death among Americans 65 years of age or older1. The quality of life for patients who suffer from a stroke fails to ...

Murine Model for Non-invasive Imaging to Detect and Monitor Ovarian Cancer Recurrence

Epithelial ovarian cancer is the most lethal gynecologic malignancy in the United States. Although patients initially respond to the current standard of care consisting of surgical debulking and combination chemotherapy consisting of platinum and taxane compounds, almost 90% of patients recur within a few years. In these patients the development of chemoresistant disease limits the efficacy of currently available chemotherapy agents and therefore contributes to the high mortality. To discover novel therapy options that can target recurrent disease, appropriate animal models that closely mimic the clinical profile of patients with recurrent ovarian cancer are required. The challenge in monitoring intra-peritoneal (i.p.) disease limits the use of i.p. models and thus most xenografts are established subcutaneously. We have developed a sensitive optical imaging platform that allows the detection and anatomical location of i.p. tumor mass. The platform includes the use of optical reporters that extend from the visible light range to near infrared, which in combination with 2-dimensional X-ray co-registration can provide anatomical location of molecular signals. Detection is significantly improved by the use of a rotation system that drives the animal to multiple angular positions for 360 degree imaging, allowing the identification of tumors that are not visible in single orientation. This platform provides a unique model to non-invasively monitor tumor growth and evaluate the efficacy of new therapies for the prevention or treatment of recurrent ovarian cancer.

We describe a non-invasive animal imaging platform that allows the detection, quantification, and monitoring of ovarian cancer growth and recurrence. This intra-peritoneal xenograft model mimics the clinical profile of patients with ovarian cancer.

Epithelial ovarian cancer is the most lethal gynecologic malignancy in the United States. Although patients initially respond to the current standard of ...

A Multi-Modal Approach to Assessing Recovery in Youth Athletes Following Concussion

Concussion is one of the most commonly reported injuries amongst children and youth involved in sport participation. Following a concussion, youth can experience a range of short and long term neurobehavioral symptoms (somatic, cognitive and emotional/behavioral) that can have a significant impact on one&#8217;s participation in daily activities and pursuits of interest (e.g., school, sports, work, family/social life, etc.). Despite this, there remains a paucity in clinically driven research aimed specifically at exploring concussion within the youth sport population, and more specifically, multi-modal approaches to measuring recovery. This article provides an overview of a novel and multi-modal approach to measuring recovery amongst youth athletes following concussion. The presented approach involves the use of both pre-injury/baseline testing and post-injury/follow-up testing to assess performance across a wide variety of domains (post-concussion symptoms, cognition, balance, strength, agility/motor skills and resting state heart rate variability). The goal of this research is to gain a more objective and accurate understanding of recovery following concussion in youth athletes (ages 10-18 years). Findings from this research can help to inform the development and use of improved approaches to concussion management and rehabilitation specific to the youth sport community.

This article provides an overview of a multi-modal approach to assessing recovery following concussion in youth athletes. The described protocol uses pre- and post-concussion assessment of performance across a wide variety of domains and can inform the development of improved concussion rehabilitation protocols specific to the youth sport community.

Concussion is one of the most commonly reported injuries amongst children and youth involved in sport participation. Following a concussion, youth can ...

A Methodological Approach to Non-invasive Assessments of Vascular Function and Morphology

The endothelium is the innermost lining of the vasculature and is involved in the maintenance of vascular homeostasis. Damage to the endothelium may predispose the vessel to atherosclerosis and increase the risk for cardiovascular disease. Assessments of peripheral endothelial function are good indicators of early abnormalities in the vascular wall and correlate well with assessments of coronary endothelial function. The present manuscript details the important methodological steps necessary for the assessment of microvascular endothelial function using laser Doppler imaging with iontophoresis, large vessel endothelial function using flow-mediated dilatation, and carotid atherosclerosis using carotid artery ultrasound. A discussion on the methodological considerations for each of the techniques is also presented, and recommendations are made for future research.

The present article describes the methodological considerations for several non-invasive assessments of vascular function and morphology that are commonly used in medical research to assess different stages of atherosclerosis.

The endothelium is the innermost lining of the vasculature and is involved in the maintenance of vascular homeostasis.  Damage to the endothelium may ...

Spinal Cord Lateral Hemisection and Asymmetric Behavioral Assessments in Adult Rats

Incomplete spinal cord injury (SCI) often leads to impairments of sensorimotor functions and is clinically the most frequent type of SCI. Human Brown-S&#233;quard syndrome&#160;is a common type of incomplete SCI caused by a lesion to one half of the spinal cord which results in paralysis and loss of&#160;proprioception&#160;on the same (or ipsilesional) side as the injury, and loss of pain and temperature sensation on the opposite (or contralesional) side. Adequate methodologies for producing a spinal cord lateral hemisection (HX) and assessing neurological impairments are essential to establish a reliable animal model of Brown-S&#233;quard syndrome. Although lateral hemisection model plays a pivotal role in basic and translational research, standardized protocols for creating such a hemisection and assessing unilateralized function are lacking. The goal of this study is to describe step-by-step procedures to produce a rat spinal lateral HX at the 9th thoracic (T9) vertebral level. We, then, describe a combined behavior scale for HX (CBS-HX) that provides a simple and sensitive assessment of asymmetric neurological performance for unilateral SCI. The CBS-HX, ranging from 0 to 18, is composed of 4 individual assessments which include unilateral hindlimb stepping (UHS), coupling, contact placing, and grid walking. For CBS-HX, the ipsilateral and contralateral hindlimbs are assessed separately. We found that, after a T9 HX, the ipsilateral hindlimb showed impaired behavior function whereas the contralateral hindlimb showed substantial recovery. The CBS-HX effectively discriminated behavioral functions between ipsilateral and contralateral hindlimbs and detected temporal progression of recovery of the ipsilateral hindlimb. The CBS-HX components can be analyzed separately or in combination with other measures when needed. Although we only provided visual descriptions of the surgical procedures and behavioral assessments of a thoracic HX, the principle may be applied to other incomplete SCIs and at other levels of the injury.

Here we describe surgical procedures to produce a reliable spinal cord lateral hemisection (HX) at the 9th thoracic level in adult rats and neurobehavioral assessments designed for detecting asymmetric deficits after such a unilateral injury.

Incomplete spinal cord injury (SCI) often leads to impairments of sensorimotor functions and is clinically the most frequent type of SCI. Human ...

Non-invasive Assessment of Dorsiflexor Muscle Function in Mice

Assessment of skeletal muscle contractile function is an important measurement for both clinical and research purposes. Numerous conditions can negatively affect skeletal muscle. This can result in a loss of muscle mass (atrophy) and/or loss of muscle quality (reduced force per unit of muscle mass), both of which are prevalent in chronic disease, muscle-specific disease, immobilization, and aging (sarcopenia). Skeletal muscle function in animals can be evaluated by a range of different tests. All tests have limitations related to the physiological testing environment, and the selection of a specific test often depends on the nature of the experiments. Here, we describe an in vivo, non-invasive technique involving a helpful and easy assessment of force frequency-curve (FFC) in mice that can be performed on the same animal over time. This permits monitoring of disease progression and/or efficacy&#160;of a potential therapeutic treatment.

Measurement of rodent skeletal muscle contractile function is a useful tool that can be used to track disease progression as well as efficacy of therapeutic intervention. We describe here the non-invasive, in vivo assessment of the dorsiflexor muscles that can be repeated over time in the same mouse.

Assessment of skeletal muscle contractile function is an important measurement for both clinical and research purposes. Numerous conditions can negatively ...