We would like to acknowledge all the colleagues in the NICU for their work.

This protocol was approved by the Institutional Review Board of Renji Hospital, Shanghai Jiaotong University School of Medicine.
1. Preparation of the Patient
NOTE: ICP sensor is placed in the patient by a surgical operation (Figure 1). The sensor is placed in the epidural space, subdural space, parenchyma, or ventricular system.
<ol>
	<li>Connect the ICP monitoring machine with the bedside monitor via a specific communication cable.</li>
	<li>Adjust the reference of patient&#39;s bedside monitor, so that the data from the bedside monitor are in accordance with the ICP monitoring machine.</li>
	<li>Perform an arterial line placement on patient&#39;s left or right radial artery20.</li>
	<li>Connect the arterial line with the baroreceptor.</li>
	<li>Connect the pressure transducer with patient&#39;s bedside monitor via a communication cable.</li>
	<li>Zero adjust the bedside monitor so that the measured blood pressure coincides with the actual value.</li>
</ol>
2. Recording of Physiological Parameters
<ol>
	<li>Connect the patient&#39;s bedside monitor with the data collecting and storing device via a network cable.</li>
	<li>Press the power button to turn on the data collecting device.</li>
	<li>Wait for a few seconds until the multimodality monitoring software runs automatically.</li>
	<li>Enter the information of the patient, including the name, ID, diagnosis, and initial Glasgow Coma Scale (GCS).</li>
	<li>Click the &#34;Save and start monitoring&#34; button to start data collecting and storing.</li>
	<li>Click the &#34;Turn off&#34; button on the screen at the end of the data acquisition (Figure 2).</li>
	<li>Enter the final GCS.</li>
</ol>
3. Parameter Analysis
<ol>
	<li>Dump the stored data to a U-disk through universal serial bus (USB) interfaces on the host.</li>
	<li>Enter a user name and password to log in to the analysis server.</li>
	<li>Click the &#34;Create new record&#34; button and select the data file in the U-disk to start uploading.</li>
	<li>Wait for around 10 to 20 min before the data are analyzed (depending on the data size).</li>
	<li>Click the &#34;Detail&#34; button of each record to see the result.</li>
	<li>Click the &#34;Chart viewing&#34; to see the final diagram.</li>
	<li>Drag the timeline to see more detail.</li>
	<li>Click the &#34;scattergram&#34; button to see the distribution of the parameters.</li>
</ol>

<ol>
	<li>Hawthorne, C., Piper, I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Monitoring+of+intracranial+pressure+in+patients+with+traumatic+brain+injury.">Monitoring of intracranial pressure in patients with traumatic brain injury.</a> Front Neurol. 5, 121 (2014).</li><li>Cooper, D. J., 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=Decompressive+craniectomy+in+diffuse+traumatic+brain+injury.">Decompressive craniectomy in diffuse traumatic brain injury.</a> N Engl J Med. 364 (16), 1493-1502 (2011).</li><li>Jiang, J. Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Head+trauma+in+China.">Head trauma in China.</a> Injury. 44 (11), 1453-1457 (2013).</li><li>Hu, X., Xu, P., Asgari, S., Vespa, P., Bergsneider, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Forecasting+ICP+elevation+based+on+prescient+changes+of+intracranial+pressure+waveform+morphology.">Forecasting ICP elevation based on prescient changes of intracranial pressure waveform morphology.</a> IEEE Trans Biomed Eng. 57 (5), 1070-1078 (2010).</li><li>Di Ieva, A., Schmitz, E. M., Cusimano, M. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Analysis+of+intracranial+pressure:+past,+present,+and+future.">Analysis of intracranial pressure: past, present, and future.</a> Neuroscientist. 19 (6), 592-603 (2013).</li><li>Czosnyka, M., 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=Intracranial+pressure:+more+than+a+number.">Intracranial pressure: more than a number.</a> Neurosurg Focus. 22 (5), 10 (2007).</li><li>Lu, C. W., 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=Complexity+of+intracranial+pressure+correlates+with+outcome+after+traumatic+brain+injury.">Complexity of intracranial pressure correlates with outcome after traumatic brain injury.</a> Brain. 135 (8), 2399-2408 (2012).</li><li>Nucci, C. G., 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=Intracranial+pressure+wave+morphological+classification:+automated+analysis+and+clinical+validation.">Intracranial pressure wave morphological classification: automated analysis and clinical validation.</a> Acta Neurochir (Wien). 158 (3), 581-588 (2016).</li><li>Eide, P. K., Sorteberg, W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Association+among+intracranial+compliance,+intracranial+pulse+pressure+amplitude+and+intracranial+pressure+in+patients+with+intracranial+bleeds.">Association among intracranial compliance, intracranial pulse pressure amplitude and intracranial pressure in patients with intracranial bleeds.</a> Neurol Res. 29 (8), 798-802 (2007).</li><li>Czosnyka, M., Czosnyka, Z. H., Whitfield, P. C., Donovan, T., Pickard, J. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Age+dependence+of+cerebrospinal+pressure-volume+compensation+in+patients+with+hydrocephalus.">Age dependence of cerebrospinal pressure-volume compensation in patients with hydrocephalus.</a> J Neurosurg. 94 (3), 482-486 (2001).</li><li>Howells, T., Lewen, A., Skold, M. K., Ronne-Engstrom, E., Enblad, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=An+evaluation+of+three+measures+of+intracranial+compliance+in+traumatic+brain+injury+patients.">An evaluation of three measures of intracranial compliance in traumatic brain injury patients.</a> Intensive Care Med. 38 (6), 1061-1068 (2012).</li><li>Schuhmann, M. U., 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=Value+of+overnight+monitoring+of+intracranial+pressure+in+hydrocephalic+children.">Value of overnight monitoring of intracranial pressure in hydrocephalic children.</a> Pediatr Neurosurg. 44 (4), 269-279 (2008).</li><li>Czosnyka, M., Pickard, J. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Monitoring+and+interpretation+of+intracranial+pressure.">Monitoring and interpretation of intracranial pressure.</a> J Neurol Neurosurg Psychiatry. 75 (6), 813-821 (2004).</li><li>Sorrentino, E., 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=Critical+thresholds+for+cerebrovascular+reactivity+after+traumatic+brain+injury.">Critical thresholds for cerebrovascular reactivity after traumatic brain injury.</a> Neurocrit Care. 16 (2), 258-266 (2012).</li><li>Carrera, E., 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=What+shapes+pulse+amplitude+of+intracranial+pressure.">What shapes pulse amplitude of intracranial pressure.</a> J Neurotrauma. 27 (2), 317-324 (2010).</li><li>Eide, P. 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=Pressure-derived+versus+pressure+wave+amplitude-derived+indices+of+cerebrovascular+pressure+reactivity+in+relation+to+early+clinical+state+and+12-month+outcome+following+aneurysmal+subarachnoid+hemorrhage.">Pressure-derived versus pressure wave amplitude-derived indices of cerebrovascular pressure reactivity in relation to early clinical state and 12-month outcome following aneurysmal subarachnoid hemorrhage.</a> J Neurosurg. 116 (5), 961-971 (2012).</li><li>Lazaridis, C., 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=Patient-specific+thresholds+of+intracranial+pressure+in+severe+traumatic+brain+injury.">Patient-specific thresholds of intracranial pressure in severe traumatic brain injury.</a> J Neurosurg. 120 (4), 893-900 (2014).</li><li>Czosnyka, M., 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=Monitoring+and+interpretation+of+intracranial+pressure+after+head+injury.">Monitoring and interpretation of intracranial pressure after head injury.</a> Acta Neurochir Suppl. 96, 114-118 (2006).</li><li>Steiner, L. A., 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=Continuous+monitoring+of+cerebrovascular+pressure+reactivity+allows+determination+of+optimal+cerebral+perfusion+pressure+in+patients+with+traumatic+brain+injury.">Continuous monitoring of cerebrovascular pressure reactivity allows determination of optimal cerebral perfusion pressure in patients with traumatic brain injury.</a> Crit Care Med. 30 (4), 733-738 (2002).</li><li>Tegtmeyer, K., Brady, G., Lai, S., Hodo, R., Braner, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Videos+in+Clinical+Medicine.+Placement+of+an+arterial+line.">Videos in Clinical Medicine. Placement of an arterial line.</a> N Engl J Med. 354 (15), 13 (2006).</li><li>Chesnut, R. M., 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+trial+of+intracranial-pressure+monitoring+in+traumatic+brain+injury.">A trial of intracranial-pressure monitoring in traumatic brain injury.</a> N Engl J Med. 367 (26), 2471-2481 (2012).</li></ol>

No financial support was received.

This aim of this article is to introduce the new multimodality monitoring and analyzing system for neurosurgical critical patients, which can be used to monitor physiological parameters, analyze the current clinical status and, hopefully, predict the prognosis of the neurosurgical critical patients. Nowadays, the focus of ICP monitoring is mainly on the mean ICP value while ignoring other parameters, which may carry risks of inaccuracy or delay4,5,21. On the other hand, the ICP derived parameters such as ICP waveform, AMP, RAP, PRx, and IAAC are reported to have great significance in predicting prognosis and guiding proper treatment5,16,17,18,19. However, all these parameters require multimodality monitoring and big data acquisition and analysis, which can take up a lot of time and effort. Besides, there are few specific devices that are suitable for clinicians in their daily clinical work currently.
In order to solve this problem, we recently developed a multimodality monitoring system. It uses an automatic data collecting and storing device to continuously acquire patients&#39; physiological information before sending the data to the online server for analysis work to investigate the meaning of these ICP derived parameters. As is mentioned previously, the multimodality monitoring system has the following advantages. First of all, it can collect real-time data at a high frequency. The frequency of data collection can reach up to 120 Hz. Secondly, it can record multiple parameters, i.e., ICP waveform, PRx, RAP, and IAAC. All of them are critical in the analysis of current clinical status and in the prediction of prognosis. Moreover, it can achieve long term continuous monitoring. It can record all the details throughout the treatment of patients. Last but not least, it is portable and easy to learn. Clinicians can easily upload the data to a specific analysis server, which will perform the calculation and graphic construction automatically. There are only two critical steps in the protocol. One is the placement of ICP sensor. The other is the arterial line placement and arterial blood pressure monitoring. Once correctly connected and operated, the multimodality monitoring system can collect and store real-time data before analyzing and graphing them automatically, which is suitable for clinicians.
However, it also has some limitations. This system only includes ICP and invasive blood pressure and lacks other parameters such as EKG, EEG, or TCD. Another limitation is that the automatic analysis mode is not abundant, and can only do certain calculations and graphic constructions.
We have been continuously modifying this system and have fixed several problems that occurred during the long-term use. We are confident that all the limitations will be addressed in the future and it will be more applicable.
We hope that the multimodality monitoring system will be accepted as a key measure to monitor physiological parameters, to analyze the current clinical status, and to predict the prognosis of neurosurgical critical patients in the future.

ICP monitoring is widely used to evaluate intracranial status in neurosurgery department, especially in neurosurgical critical patients1,2,3. Besides mean ICP value, the ICP derived parameters such as ICP waveform, AMP, RAP, PRx, IAAC, and so on, can reflect the status of intracerebral circulation, cerebrospinal compensatory reserve, and brain compliance4,5,6,7,8,9,10,11,12,13. They can be indicative of impending neurological deterioration and even outcome of patients14,15,16,17,18. They can also be used as guidance of proper treatment19. However, most of the clinicians focus only on the mean ICP value while ignoring these parameters. This is partly because there are few specific devices that are suitable for clinicians in their daily clinical work.
To address these drawbacks, we have recently developed a multimodality monitoring system. We use an automatic data collecting and storing device to continuously acquire patients&#39; physiological information such as blood pressure, ICP, and oxygen saturation, and analyze these physiological parameters in order to reveal the current clinical status and, hopefully, predict the prognosis of the neurosurgical critical patients. This multimodality monitoring system has several advantages: (1) it can collect real-time data at high frequency, (2) it can record multiple parameters, i.e., ICP waveform, PRx, RAP, and IAAC, (3) it can achieve long term continuous monitoring, and (4) it is portable and easy to learn.
The objective of this article, therefore, is to show a detailed method of how to use the multimodality monitoring system to record various physiological parameters in neurosurgical critical patients.

<table style="border-collapse:collapse;width:639pt" width="852">
	<colgroup>
		<col style="width:136pt" width="181" />
		<col style="width:171pt" width="228" />
		<col style="width:134pt" width="179" />
		<col style="width:198pt" width="264" />
	</colgroup>
	<tbody>
		<tr height="21" style="height:15.75pt">
			<td height="21" style="width:136pt;height:15.75pt" width="181">Bedside monitor</td>
			<td style="width:171pt" width="228">Philips</td>
			<td style="width:134pt" width="179">IntelliVue MP40 M8003A</td>
			<td style="width:198pt" width="264">With interfacing module</td>
		</tr>
		<tr height="21" style="height:15.75pt">
			<td height="21" style="height:15.75pt">ICP monitoring machine</td>
			<td>&#160;Johnson &#38; Johnson or Sophysa</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="21" style="height:15.75pt">
			<td height="21" style="height:15.75pt">Arterial cannula</td>
			<td>BD</td>
			<td>REF682245</td>
			<td></td>
		</tr>
		<tr height="21" style="height:15.75pt">
			<td height="21" style="height:15.75pt">Pressure transducer</td>
			<td>Haisheng Medical</td>
			<td>DBPT-0103</td>
			<td></td>
		</tr>
		<tr height="21" style="height:15.75pt">
			<td height="21" style="height:15.75pt">Data collection device</td>
			<td>Shanghai Haoju</td>
			<td>Neumatic</td>
			<td></td>
		</tr>
		<tr height="21" style="height:15.75pt">
			<td height="21" style="height:15.75pt">Computer</td>
			<td></td>
			<td></td>
			<td>Requires Windows operating system</td>
		</tr>
	</tbody>
</table>

a detailed protocol for physiological parameters acquisition and analysis in neurosurgical critical patients

Intracranial pressure (ICP) monitoring is now widely used in neurosurgical critical patients. Besides mean ICP value, the ICP derived parameters such as ICP waveform, amplitude of pulse (AMP), the correlation of ICP amplitude and ICP mean (RAP), pressure reactivity index (PRx), ICP and arterial blood pressure (ABP) wave amplitude correlation (IAAC), and so on, can reflect intracranial status, predict prognosis, and can also be used as guidance of proper treatment. However, most of the clinicians focus only on the mean ICP value while ignoring these parameters because of the limitations of the current devices. We have recently developed a multimodality monitoring system to address these drawbacks. This portable, user-friendly system will use a data collecting and storing device to continuously acquire patients&#39; physiological parameters first, i.e., ABP, ICP, and oxygen saturation, and then analyze these physiological parameters. We hope that the multimodality monitoring system will be accepted as a key measure to monitor physiological parameters, to analyze the current clinical status, and to predict the prognosis of the neurosurgical critical patients.

This new multimodality monitoring system was applied on 22 neurosurgical critical patients (15 males). 12 of them (54.55%) had suffered from traumatic brain injury (TBI), 9 of them (40.91%) had intracranial hemorrhage and 1 of them (4.55%) had severe cerebral infarction. The total monitoring time is more than 1,900 h (about 88 h per patient). After successful surgery, we continuously monitored and analyzed their ICP, BP, CPP, PRx, RAP, and IAAC. Figure 3 shows representative data from this monitoring system. We can adjust the timeline to see more detailed information (Figure 4). Besides, we are able to analyze the relationship between any two parameters, which is demonstrated in Figure 5.
<img alt="Figure 1" class="xfigimg" src="/files/ftp_upload/56388/56388fig1.jpg" /> 
Figure 1: ICP sensor placement. The patient undergoes an operation for ICP sensor placement before monitoring. The sensor can be placed in the epidural space, subdural space, parenchyma, or ventricular system. This figure shows ventricular puncture and sensor placement. The ICP sensor is located at the tip of the catheter. <a href="https://cloudflare-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/56388/56388fig1large.jpg" target="_blank">Please click here to view a larger version of this figure.</a>
<img alt="Figure 2" class="xfigimg" src="/files/ftp_upload/56388/56388fig2.jpg" /> 
Figure 2: Screen of monitoring system. The upper left area shows current ICP waveform and blood pressure. The lower left area shows current RAP, PRx, IAAC, CPP, and AMP. The right part of the screen shows the history. Shown at the bottom are some of the functional buttons such as output, screen shot, and adding markers. <a href="https://cloudflare-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/56388/56388fig2large.jpg" target="_blank">Please click here to view a larger version of this figure.</a>
<img alt="Figure 3" class="xfigimg" src="/files/ftp_upload/56388/56388fig3.jpg" /> 
Figure 3: Representative data of a cerebral hemorrhage patient. The parameters such as ICP, BP, CPP, PRx, RAP, and IAAC are recorded and plotted. The X axis is the timeline and the Y axes are the separate parameters. <a href="https://cloudflare-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/56388/56388fig3large.jpg" target="_blank">Please click here to view a larger version of this figure.</a>
<img alt="Figure 4" class="xfigimg" src="/files/ftp_upload/56388/56388fig4.jpg" /> 
Figure 4: Detailed information of representative data. As we adjust the timeline, we can see more detail. The X axis is the timeline and the Y axes are the separate parameters. <a href="https://cloudflare-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/56388/56388fig4large.jpg" target="_blank">Please click here to view a larger version of this figure.</a>
<img alt="Figure 5" class="xfigimg" src="/files/ftp_upload/56388/56388fig5.jpg" /> 
Figure 5: Relationship between ICP and PRx. We can change the parameters on the X axis and Y axis to analyze the relationship between any two parameters. In this figure, the X axis is the ICP and the Y axis is the PRx. Each red dot represents a real-time record. We can see that, in this patient, his PRx has a variation from -0.7 to 1 and the lowest PRx is reached when his ICP is 10 mmHg.&#160;<a href="https://cloudflare-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/56388/56388fig5large.jpg" style="font-size: 14px;" target="_blank">Please click here to view a larger version of this figure.</a>

Watch this Scientific Journal Video about A Detailed Protocol for Physiological Parameters Acquisition and Analysis in Neurosurgical Critical Patients at JoVE.com

A Detailed Protocol for Physiological Parameters Acquisition and Analysis in Neurosurgical Critical Patients

Recently, we have developed a portable multimodality monitoring system for the monitoring of various physiological parameters in neurosurgical critical patients. Detailed protocols on how to use this multimodality monitoring system are presented here.

Intracranial pressure (ICP) monitoring is now widely used in neurosurgical critical patients. Besides mean ICP value, the ICP derived parameters such as ...

a-detailed-protocol-for-physiological-parameters-acquisition-analysis

Nanyang Technological University

Research

JoVE Journal

Medicine

6.8K Views.  Shanghai Jiaotong University School of Medicine. Recently, we have developed a portable multimodality monitoring system for the monitoring of various physiological parameters in neurosurgical critical patients. Detailed protocols on how to use this multimodality monitoring system are presented here.

Video: A Detailed Protocol for Physiological Parameters Acquisition and Analysis in Neurosurgical Critical Patients

Intra-Operative Behavioral Tasks in Awake Humans Undergoing Deep Brain Stimulation Surgery

Deep brain stimulation (DBS) is a surgical procedure that directs chronic, high frequency electrical stimulation to specific targets in the brain through implanted electrodes. Deep brain stimulation was first implemented as a therapeutic modality by Benabid et al. in the late 1980s, when he used this technique to stimulate the ventral intermediate nucleus of the thalamus for the treatment of tremor 1. Currently, the procedure is used to treat patients who fail to respond adequately to medical management for diseases such as Parkinson's, dystonia, and essential tremor. The efficacy of this procedure for the treatment of Parkinson's disease has been demonstrated in well-powered, randomized controlled trials 2. Presently, the U.S. Food and Drug Administration has approved DBS as a treatment for patients with medically refractory essential tremor, Parkinson's disease, and dystonia. Additionally, DBS is currently being evaluated for the treatment of other psychiatric and neurological disorders, such as obsessive compulsive disorder, major depressive disorder, and epilepsy. 
DBS has not only been shown to help people by improving their quality of life, it also provides researchers with the unique opportunity to study and understand the human brain. Microelectrode recordings are routinely performed during DBS surgery in order to enhance the precision of anatomical targeting. Firing patterns of individual neurons can therefore be recorded while the subject performs a behavioral task. Early studies using these data focused on descriptive aspects, including firing and burst rates, and frequency modulation 3. More recent studies have focused on cognitive aspects of behavior in relation to neuronal activity 4,5. This article will provide a description of the intra-operative methods used to perform behavioral tasks and record neuronal data with awake patients during DBS cases. Our exposition of the process of acquiring electrophysiological data will illuminate the current scope and limitations of intra-operative human experiments.

Deep brain stimulation surgery offers a unique opportunity to examine information encoding in the awake human brain. This article will describe intra-operative methods used to perform cognitive and behavioral tasks while simultaneously acquiring physiological data such as EMG, single-unit neuronal activity and/or local field potentials.

Deep brain stimulation (DBS) is a surgical procedure that directs chronic, high frequency electrical stimulation to specific targets in the brain through ...

Manual Muscle Testing:  A Method of Measuring Extremity Muscle Strength Applied to Critically Ill Patients

Survivors of acute respiratory distress syndrome (ARDS) and other causes of critical illness often have generalized weakness, reduced exercise tolerance, and persistent nerve and muscle impairments after hospital discharge.1-6 Using an explicit protocol with a structured approach to training and quality assurance of research staff, manual muscle testing (MMT) is a highly reliable method for assessing strength, using a standardized clinical examination, for patients following ARDS, and can be completed with mechanically ventilated patients who can tolerate sitting upright in bed and are able to follow two-step commands. 7, 8 
This video demonstrates a protocol for MMT, which has been taught to &ge;43 research staff who have performed &gt;800 assessments on &gt;280 ARDS survivors. Modifications for the bedridden patient are included. Each muscle is tested with specific techniques for positioning, stabilization, resistance, and palpation for each score of the 6-point ordinal Medical Research Council scale.7,9-11 Three upper and three lower extremity muscles are graded in this protocol: shoulder abduction, elbow flexion, wrist extension, hip flexion, knee extension, and ankle dorsiflexion. These muscles were chosen based on the standard approach for evaluating patients for ICU-acquired weakness used in prior publications. 1,2.

Survivors of acute respiratory distress syndrome (ARDS) and critical illness frequently develop long-lasting muscle weakness. Manual muscle testing (MMT) is a standardized clinical examination commonly used to measure strength of peripheral skeletal muscle groups. This video demonstrates MMT using the 6-point Medical Research Council scale.

Survivors of acute respiratory distress syndrome (ARDS) and other causes of critical illness often have generalized weakness, reduced exercise tolerance, ...

Parabiosis in Mice: A Detailed Protocol

Parabiosis is a surgical union of two organisms allowing sharing of the blood circulation. Attaching the skin of two animals promotes formation of microvasculature at the site of inflammation. Parabiotic partners share their circulating antigens and thus are free of adverse immune reaction. First described by Paul Bert in 18641, the parabiosis surgery was refined by Bunster and Meyer in 1933 to improve animal survival2. In the current protocol, two mice are surgically joined following a modification of the Bunster and Meyer technique. Animals are connected through the elbow and knee joints followed by attachment of the skin allowing firm support that prevents strain on the sutured skin. Herein, we describe in detail the parabiotic joining of a ubiquitous GFP expressing mouse to a wild type (WT) mouse. Two weeks after the procedure, the pair is separated and GFP positive cells can be detected by flow cytometric analysis in the blood circulation of the WT mouse. The blood chimerism allows one to examine the contribution of the circulating cells from one animal in the other.

Parabiotic joining of two organisms leads to the development of a shared circulatory system. In this protocol, we describe the surgical steps to form a parabiotic connection between a wild-type mouse and a constitutive GFP-expressing mouse.

Parabiosis is a surgical union of two organisms allowing sharing of the blood circulation. Attaching the skin of two animals promotes formation of ...

Clinical Assessment of Spatiotemporal Gait Parameters in Patients and Older Adults

Spatial and temporal characteristics of human walking are frequently evaluated to identify possible gait impairments, mainly in orthopedic and neurological patients1-4, but also in healthy older adults5,6. The quantitative gait analysis described in this protocol is performed with a recently-introduced photoelectric system (see&#160;Materials table) which has the potential to be used in the clinic because it is portable, easy to set up (no subject preparation is required before a test), and does not require maintenance and sensor calibration. The photoelectric system consists of series of high-density floor-based photoelectric cells with light-emitting and light-receiving diodes that are placed parallel to each other to create a corridor, and are oriented perpendicular to the line of progression7. The system simply detects interruptions in light signal, for instance due to the presence of feet within the recording area. Temporal gait parameters and 1D spatial coordinates of consecutive steps are subsequently calculated to provide common gait parameters such as step length, single limb support and walking velocity8, whose validity against a criterion instrument has recently been demonstrated7,9. The measurement procedures are very straightforward; a single patient can be tested in less than 5 min&#160;and a comprehensive report can be generated in less than 1&#160;min.

This protocol is used to evaluate spatial and temporal gait variables of neurological/orthopedic patients and older persons by means of a recently-introduced floor-based photocell system.

Spatial and temporal characteristics of human walking are frequently evaluated to identify possible gait impairments, mainly in orthopedic and ...

A Simple Critical-sized Femoral Defect Model in Mice

While bone has a remarkable capacity for regeneration, serious bone trauma often results in damage that does not properly heal. In fact, one tenth of all limb bone fractures fail to heal completely due to the extent of the trauma, disease, or age of the patient. Our ability to improve bone regenerative strategies is critically dependent on the ability to mimic serious bone trauma in test animals, but the generation and stabilization of large bone lesions is technically challenging. In most cases, serious long bone trauma is mimicked experimentally by establishing a defect that will not naturally heal. This is achieved by complete removal of a bone segment that is larger than 1.5 times the diameter of the bone cross-section. The bone is then stabilized with a metal implant to maintain proper orientation of the fracture edges and allow for mobility. 
Due to their small size and the fragility of their long bones, establishment of such lesions in mice are beyond the capabilities of most research groups. As such, long bone defect models are confined to rats and larger animals. Nevertheless, mice afford significant research advantages in that they can be genetically modified and bred as immune-compromised strains that do not reject human cells and tissue.
Herein, we demonstrate a technique that facilitates the generation of a segmental defect in mouse femora using standard laboratory and veterinary equipment. With practice, fabrication of the fixation device and surgical implantation is feasible for the majority of trained veterinarians and animal research personnel. Using example data, we also provide methodologies for the quantitative analysis of bone healing for the model.

Animal models are frequently employed to mimic serious bone injury in biomedical research. Due to their small size, establishment of stabilized bone lesions in mice are beyond the capabilities of most research groups. Herein, we describe a simple method for establishing and analyzing experimental femoral defects in mice.

While bone has a remarkable capacity for regeneration, serious bone trauma often results in damage that does not properly heal. In fact, one tenth of all ...

A Detailed Protocol for Perspiration Monitoring Using a Novel, Small, Wireless Device

Perspiration monitoring can be utilized for the detection of certain diseases, such as thermoregulation and mental disorders, particularly when the patients are unaware of such disorders or are having difficulty expressing their symptoms. Until now, several devices for perspiration monitoring have been developed; however, such devices tend to have a relatively large exterior, considerable power consumption, and/or less sensitivity.
Recently, we developed a small, wireless device for perspiration monitoring. The device consists of a temperature/relative humidity (T/RH) sensor, battery-driven small data logger, and silica gel as a desiccant in a small cylindrical exterior. The T/RH sensor is placed between the detection windows (through which the water vapor from the skin enters) and the silica gel. The underlying principle of the perspiration monitoring device is based on Fick&#39;s law of diffusion, which means that water vapor flux from the skin to the silica gel (i.e.&#160;transepidermal water loss and perspiration) can be captured by change in humidity at the T/RH sensor. In addition, a baseline subtraction method was adopted to distinguish perspiration and transepidermal water loss.
As shown in the previous report, the developed device can monitor the perspiration at any sites of the body in an easy, wireless manner. However, detailed methods of how to use the device have not been disclosed yet. In this article, therefore, we would like to show the point-by-point tutorials of how to use the device for perspiration monitoring, by showing the sympathetic activity test with the sympathetic skin response monitoring as an example.

Recently, we developed a small wireless device for perspiration monitoring. In this article, we present detailed protocols on how to use the device for perspiration monitoring with an example of the sympathetic activity test.

Perspiration monitoring can be utilized for the detection of certain diseases, such as thermoregulation and mental disorders, particularly when the ...

Objective Nociceptive Assessment in Ventilated ICU Patients: A Feasibility Study Using Pupillometry and the Nociceptive Flexion Reflex

The concept of objective nociceptive assessment and optimal pain management have gained increasing attention. Despite the known negative short- and long-term consequences of unresolved pain or excessive analgosedation, adequate nociceptive monitoring remains challenging in non-communicative, critically ill adults. In the intensive care unit (ICU), routine nociceptive evaluation is carried out by the attending nurse using the Behavior Pain Scale (BPS) in mechanically ventilated patients. This assessment is limited by medication use (e.g., neuromuscular blocking agents) and the inherent subjective character of nociceptive evaluation by third parties.
Here, we describe the use of two nociceptive reflex testing devices as tools for objective pain evaluation: the pupillary dilation reflex (PDR) and nociception flexion reflex (NFR). These measurement tools are non-invasive and well tolerated, providing clinicians and researchers with objective information regarding two different nociceptive processing pathways: (1)&#160;the pain-related autonomic reactivity&#160;and (2) the ascending component of the somatosensory system. The use of PDR and NFR measurements are currently limited to specialized pain clinics and research institutions because of impressions that these are technically demanding or time-consuming procedures, or even because of a lack of knowledge regarding their existence.
By focusing on the two abovementioned nociceptive reflex assessments, this study evaluated their feasibility as a physiological pain measurement method in daily practice. Pursuing novel technologies for evaluating the analgesia level in unconscious patients may further improve individual pharmacological treatment and patient related outcome measures. Therefore, future research must include large well-designed clinical trials in a real-life environment.

Pain assessment in anesthetized patients who cannot communicate with the outside world in any way remains challenging despite the development of innovative objective pain evaluation tools. In this project, the pupillary dilation reflex and the nociception flexion reflex are assessed in critically ill, mechanically ventilated adult patients.

The concept of objective nociceptive assessment and optimal pain management have gained increasing attention. Despite the known negative short- and ...

Endotoxin Activity Assay for the Detection of Whole Blood Endotoxemia in Critically Ill Patients

Lipopolysaccharide, also known as endotoxin, is a fundamental component of gram-negative bacteria and plays a crucial role in the development of sepsis and septic shock. The early identification of an infectious process that is rapidly evolving to a critical illness might prompt a quicker and more intensive treatment, thereby potentially leading to better patient outcomes. The Endotoxin Activity (EA) assay can be used at the bedside as a reliable biomarker of systemic endotoxemia. The detection of elevated endotoxin activity levels has been repeatedly shown to be associated with an increased disease severity in patients with sepsis and septic shock. The assay is quick and easy to perform. Briefly, after sampling, an aliquot of whole blood is mixed with an anti-endotoxin antibody and with added LPS. Endotoxin activity is measured as the relative oxidative burst of primed neutrophils as detected by chemioluminescence. The assay&#39;s output is expressed on a scale from 0 (absent) to 1 (maximal) and categorized as &#8220;low&#8221; (&#60;0.4 units), &#8220;intermediate&#8221; (0.4&#8211;0.59 units), or &#8220;high&#8221; (&#8805;0.6 units). The detailed methodology and rationale for the implementation of the EA assay are reported in this manuscript.

We hereby present a protocol to measure at the bedside the endotoxin activity of human whole blood samples. The Endotoxin Activity assay is a simple test to perform and may be a useful biomarker in critically ill patients with sepsis.

Lipopolysaccharide, also known as endotoxin, is a fundamental component of gram-negative bacteria and plays a crucial role in the development of sepsis ...

A Real-World High-Intensity Interval Training Protocol for Cardiorespiratory Fitness Improvement

High-Intensity Interval Training (HIIT) has emerged as an interesting time-efficient approach to increase exercise adherence and improve health. However, few studies have tested the efficiency of HIIT protocols in a &#34;real world&#34; setting, e.g., HIIT protocols designed for outdoor spaces without specialized equipment. This study presents a &#34;real world&#34; training protocol, named &#34;beep training&#34;, and compares the efficiency of a HIIT regiment versus a traditional long-duration Moderate-Intensity Continuous Training (MICT) regiment using this beep training protocol on VO2 max of overweight untrained men. Twenty-two subjects performed outdoor running with MICT (n = 11) or HIIT (n = 11). Cardiorespiratory fitness was assessed before and after training protocols using a metabolic analyzer. Both training protocols were performed 3 days a week for 8 weeks using the Beep Test results. The MICT group performed the exercise program at 60%-75% of the maximum speed of the 20 m shuttle test (Vmax) and with a progression of the distance of 3,500-5,000 m. The HIIT group performed the interval exercise with 7-10 bouts of 200 m at 85%-100% of the maximum speed of the 20 m shuttle test (Vmax), interspersed with 1 min of passive recovery. Although the HIIT group presented a significantly lower training volume than the MICT group (p &#60; 0.05) after 8 weeks of beep training, HIIT was superior to MICT in improving VO2 max (MICT: ~4.1%; HIIT: ~7.3%; p &#60; 0.05). The &#34;real world&#34; HIIT regiment based on beep training protocol is a time-efficient, low-cost, and easy-to-implement protocol for overweight untrained men.

This study presents a low-cost and easy-to-implement "real world" high-intensity interval training (HIIT) protocol for scientific research and discusses its efficiency for cardiorespiratory fitness.

High-Intensity Interval Training (HIIT) has emerged as an interesting time-efficient approach to increase exercise adherence and improve health. However, ...

Protocol for Developing a Femur Osteotomy Model in Wistar Albino Rats

Fracture healing is a physiological process resulting in the regeneration of bone defects by the coordinated action of osteoblasts and osteoclasts. Osteoanabolic drugs have the potential to augment the repair of fractures but have constraints like high costs or undesirable side effects. The bone healing potential of a drug can initially be determined by in vitro studies, but in vivo studies are needed for the final proof of concept. Our objective was to develop a femur osteotomy rodent model that could help researchers understand the development of callus formation following fracture of the shaft of the femur and that could help establish whether a potential drug has bone healing properties. Adult male Wistar albino rats were used after Institutional Animal Ethics Committee clearance. The rodents were anesthetized, and under aseptic conditions, complete transverse fractures at the middle one-third of the shafts of the femurs were created using open osteotomy. The fractures were reduced and internally fixed using intramedullary K-wires, and secondary fracture healing was allowed to take place. After surgery, intraperitoneal analgesics and antibiotics were given for 5 days. Sequential weekly x-rays assessed callus formation. The rats were sacrificed based on radiologically pre-determined time points, and the development of the fracture callus was analyzed radiologically and using immunohistochemistry.

Here, we present a protocol to iatrogenically fracture the shaft of the femur of Wistar albino rats and follow up on the development of the callus. This femur osteotomy model can help researchers evaluate the process of fracture healing and to study how a drug could influence fracture healing.

Fracture healing is a physiological process resulting in the regeneration of bone defects by the coordinated action of osteoblasts and osteoclasts. ...