This work was supported by Christopher and Dana Reeve Foundation (Grant CDRF OA2-0802-2), Kentucky Spinal Cord and Head Injury Research Trust (Grant 9-10A - KSCHIRT), Craig H. Neilsen Foundation (Grant 1000056824 - HN000PCG) and National Institutes of Health: National Heart Lung and Blood Institute (Grant 1R01HL103750-01A1).

1. Settings
<ol>
	<li>Surface electrode heads were placed over the muscle bellies of left (L) and right (R) respiratory muscles: sternocleidomastoid (SC), scalene (S), upper trapezius on midclavicular line (UT), clavicular portion of pectoralis on midclavicular line (P), diaphragm on parasternal line (D), intercostal at 6th intercostal space on anterior axillary line (IC), rectus abdominus at umbilical level (RA), obliquus abdominis on midaxillary line (O), lower trapezius paraspinally at midscapular level (LT), and paraspinal paraspinally on iliac intercrestal line (PS)6. The ground electrodes were placed over the acromion processes. A Motion Lab System Back Pack Unit, with attached electrodes, was connected to a Motion Lab EMG Desk Top Unit and Powerlab System (Figure 1).</li>
	<li>T-piece Monitoring Circuit to record the airway pressure was assembled as shown in Figure 2 and connected to the Low Pressure Transducer (MP45) using air tube.</li>
	<li>MP45 was connected to CD15 and Powerlab System (Figure 1 and Table 1).</li>
</ol>
2. RMCA Protocol
<ol>
	<li>The respiratory motor tasks consisted of Maximum Inspiratory Pressure Task (MIPT) and Maximum Expiratory Pressure Task (MEPT). To perform MIPT or MEPT, subjects were asked to produce maximum inspiratory effort from residual volume or expiratory efforts from total lung capacity for 5 sec using a T-piece Monitoring Circuit (Figures 1 and 2). Each maneuver was cued by an audible 5-sec long tone and repeated 3x. At least 1 min of rest was allowed between each effort.</li>
	<li>EMG input was amplified with a gain of 2,000; filtered at 30-1,000 Hz and sampled at 2,000 Hz. Airway pressure input was calibrated at 100 cm of water and sampled at 2,000 Hz. The EMG and airway pressure inputs were converted by the Powerlab acquisition system using 16-bit full scale ADC resolution. Airway pressure, sEMG and marker signals were recorded simultaneously9.</li>
</ol>
3. Data Analysis
<ol>
	<li>Multi-muscle activity distribution analysis windows of 5 sec each for MIPT or MEPT were determined from the event marker and airway pressure recorded with the cuing tone that signaled the subject when to begin and end the task (Figure 3). The sEMG activity for each muscle was calculated using a root mean square (RMS) algorithm6,12 (Figure 4). Three repeated trials for each task were averaged13 for each muscle (channel).</li>
	<li>The multi-muscle activation patterns were evaluated based on a vector analysis method known as the Voluntary Response Index (VRI)8 (Figures 4-6) using custom-made Matlab software (MathWorks). For each maneuver, the VRI calculation produces two values, a Magnitude and a Similarity Index (SI) (Figures 5-6). The Magnitude parameter, the amount of combined sEMG activity for all muscles within the specific time window, was calculated as a length of the Response Vector (RV) for specific task (Figure 7). The Similarity Index (SI) provides a value that expresses how similar the RV of SCI subject is to the Prototype Response Vector (PRV) obtained from healthy subjects during the same task. The SI value was computed for each task as a cosine of the angle between the SCI subject RV and PRV. The SI value ranges between 0 and 1.0 where value of 1.0 represents the best match for compared vectors9 (Figure 8) .</li>
</ol>

<ol>
	<li>Schilero, G. J., Spungen, A. M., Bauman, W. A., Radulovic, M., Lesser, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Pulmonary+function+and+spinal+cord+injury.">Pulmonary function and spinal cord injury.</a> Respir. Physiol. Neurobiol. 166, 129-141 (2009).</li><li>Winslow, C., Rozovsky, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effect+of+spinal+cord+injury+on+the+respiratory+system.">Effect of spinal cord injury on the respiratory system.</a> Am. J. Phys. Med. Rehabil. 82, 803-814 (2003).</li><li>Garshick, 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=A+prospective+assessment+of+mortality+in+chronic+spinal+cord+injury.">A prospective assessment of mortality in chronic spinal cord injury.</a> Spinal Cord. 43, 408-416 (2005).</li><li>Jain, N. B., Brown, R., Tun, C. G., Gagnon, D., Garshick, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Determinants+of+forced+expiratory+volume+in+1+second+(FEV1),+forced+vital+capacity+(FVC),+and+FEV1/FVC+in+chronic+spinal+cord+injury.">Determinants of forced expiratory volume in 1 second (FEV1), forced vital capacity (FVC), and FEV1/FVC in chronic spinal cord injury.</a> Arch. Phys. Med. Rehabil. 87, 1327-1333 (2006).</li><li>Stolzmann, K. L., Gagnon, D. R., Brown, R., Tun, C. G., Garshick, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Longitudinal+change+in+FEV1+and+FVC+in+chronic+spinal+cord+injury.">Longitudinal change in FEV1 and FVC in chronic spinal cord injury.</a> Am. J. Respir. Crit. Care Med. 177, 781-786 (2008).</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=American+Thoracic+Society/European+Respiratory+Society.+ATS/ERS+Statement+on+respiratory+muscle+testing.">American Thoracic Society/European Respiratory Society. ATS/ERS Statement on respiratory muscle testing.</a> Am. J. Respir. Crit. Care Med. 166, 518-624 (2002).</li><li>Sherwood, A. M., McKay, W. B., Dimitrijevic, M. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Motor+control+after+spinal+cord+injury:+assessment+using+surface+EMG.">Motor control after spinal cord injury: assessment using surface EMG.</a> Muscle Nerve. 19, 966-979 (1996).</li><li>Lee, D. 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=Toward+an+objective+interpretation+of+surface+EMG+patterns:+a+voluntary+response+index+(VRI).">Toward an objective interpretation of surface EMG patterns: a voluntary response index (VRI).</a> J. Electromyogr. Kinesiol. 14, 379-388 (2004).</li><li>Ovechkin, A., Vitaz, T., de Paleville, D. T., Aslan, S., McKay, W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Evaluation+of+respiratory+muscle+activation+in+individuals+with+chronic+spinal+cord+injury.">Evaluation of respiratory muscle activation in individuals with chronic spinal cord injury.</a> Respir. Physiol. Neurobiol. 173, 171-178 (2010).</li><li>Lim, H. K., Sherwood, A. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Reliability+of+surface+electromyographic+measurements+from+subjects+with+spinal+cord+injury+during+voluntary+motor+tasks.">Reliability of surface electromyographic measurements from subjects with spinal cord injury during voluntary motor tasks.</a> J. Rehabil. Res. Dev. 42, 413-422 (2005).</li><li>Lim, H. 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=Neurophysiological+assessment+of+lower-limb+voluntary+control+in+incomplete+spinal+cord+injury.">Neurophysiological assessment of lower-limb voluntary control in incomplete spinal cord injury.</a> Spinal Cord. 43, 283-290 (2005).</li><li>Sherwood, A. M., Graves, D. E., Priebe, M. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Altered+motor+control+and+spasticity+after+spinal+cord+injury:+subjective+and+objective.">Altered motor control and spasticity after spinal cord injury: subjective and objective.</a> 37, 41-52 (2000).</li><li>McKay, W. B., Lim, H. K., Priebe, M. M., Stokic, D. S., Sherwood, A. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Clinical+neurophysiological+assessment+of+residual+motor+control+in+post-spinal+cord+injury+paralysis.">Clinical neurophysiological assessment of residual motor control in post-spinal cord injury paralysis.</a> Neurorehabil. Neural Repair. 18, 144-153 (2004).</li><li>Marino, R. 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=International+standards+for+neurological+classification+of+spinal+cord+injury.">International standards for neurological classification of spinal cord injury.</a> J. Spinal. Cord. Med. 26, S50-S56 (2003).</li><li>American Spinal Injury Association and International Spinal Cord Society. <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 Standards for Neurological Classification of Spinal Cord Injury. , (2006).</li></ol>

No conflict of interest to declare.

Standard clinical tests to evaluate respiratory motor function after SCI and other disorders include the pulmonary function tests and the American Spinal Injury Association Impairment Scale (AIS) evaluation14,15. However, these tools are not designed for quantitative evaluation of the trunk and respiratory motor control. In our previously published work9, we have shown that the RMCA is a valid method to quantitatively evaluate the respiratory motor function affected by SCI. We have demonstrated that...

<table border="1">
 <tbody>
 <tr>
 <td>Name of the Device</td>
 <td>Vendor</td>
 <td>Product #</td>
 <td>Comments</td>
 </tr>
 <tr>
 <td>PowerLab System 16/35</td>
 <td>ADInstruments</td>
 <td>PL3516</td>
 <td>Number of units depends on number of channels recorded</td>
 </tr>
 <tr>
 <td>EMG System MA 300</td>
 <td>Motion Lab Systems</td>
 <td>MA300-XVI</td>
 <td>Number of units depends on number of channels recorded</td>
 </tr>
 <tr>
 <td>Low Pressure Transducer MP45</td>
 <td>Validyne</td>
 <td>MP45-40-871</td>
 <td></td>
 </tr>
 <tr>
 <td>Basic Carrier Demodulator CD15</td>
 <td>Validyne</td>
 <td>CD15-A-2-A-1</td>
 <td></td>
 </tr>
 <tr>
 <td>Air Pressure Manometer</td>
 <td>Boehringer</td>
 <td>4103</td>
 <td>Needed for MP45 calibration</td>
 </tr>
 <tr>
 <td>Event Marker</td>
 <td></td>
 <td></td>
 <td>Hand held switch that when pressed gives a DC voltage and sound output (including 5-sec long mark)</td>
 </tr>
 <tr>
 <td>Alcohol Wipes</td>
 <td>Henry Schein</td>
 <td>1173771</td>
 <td>Needed for electrodes placement</td>
 </tr>
 <tr>
 <td>Electrode Gel</td>
 <td>Lectron II</td>
 <td>36-3000-25</td>
 <td>Needed for electrodes placement</td>
 </tr>
 <tr>
 <td>Tagaderm</td>
 <td>Henry Schein</td>
 <td>7779152</td>
 <td>Needed for electrodes placement</td>
 </tr>
 <tr>
 <td>Noseclip </td>
 <td>Henry Schein</td>
 <td>1089460</td>
 <td></td>
 </tr>
 <tr>
 <td>T-piece Ventilator Monitoring Circuit with One-way Valves </td>
 <td>Alleglance (Airlife)</td>
 <td>1504</td>
 <td></td>
 </tr>
 <tr>
 <td>Air Tube </td>
 <td>UnoMedical</td>
 <td>400E</td>
 <td></td>
 </tr>
 <tr>
 <td></td>
 <td></td>
 <td></td>
 <td>Table 1. List of specific equipment and supplies used for the Respiratory Motor Control Assessment.</td>
 </tr>
 </tbody>
 </table>

evaluation of respiratory muscle activation using respiratory motor control assessment (rmca) in individuals with chronic spinal cord injury

During breathing, activation of respiratory muscles is coordinated by integrated input from the brain, brainstem, and spinal cord. When this coordination is disrupted by spinal cord injury (SCI), control of respiratory muscles innervated below the injury level is compromised1,2 leading to respiratory muscle dysfunction and pulmonary complications. These conditions are among the leading causes of death in patients with SCI3. Standard pulmonary function tests that assess respiratory motor function include spirometrical and maximum airway pressure outcomes: Forced Vital Capacity (FVC), Forced Expiratory Volume in one second (FEV1), Maximal Inspiratory Pressure (PImax) and Maximal Expiratory Pressure (PEmax)4,5. These values provide indirect measurements of respiratory muscle performance6. In clinical practice and research, a surface electromyography (sEMG) recorded from respiratory muscles can be used to assess respiratory motor function and help to diagnose neuromuscular pathology. However, variability in the sEMG amplitude inhibits efforts to develop objective and direct measures of respiratory motor function6. Based on a multi-muscle sEMG approach to characterize motor control of limb muscles7, known as the voluntary response index (VRI)8, we developed an analytical tool to characterize respiratory motor control directly from sEMG data recorded from multiple respiratory muscles during the voluntary respiratory tasks. We have termed this the Respiratory Motor Control Assessment (RMCA)9. This vector analysis method quantifies the amount and distribution of activity across muscles and presents it in the form of an index that relates the degree to which sEMG output within a test-subject resembles that from a group of healthy (non-injured) controls. The resulting index value has been shown to have high face validity, sensitivity and specificity9-11. We showed previously9 that the RMCA outcomes significantly correlate with levels of SCI and pulmonary function measures. We are presenting here the method to quantitatively compare post-spinal cord injury respiratory multi-muscle activation patterns to those of healthy individuals.

Figure 3 represents the electromyogram and airway pressure (on top) simultaneously recorded during MEPT from a non-injured (left) and SCI (right) individuals. Note decreased airway pressure and absence of sEMG activity in expiratory muscles in an SCI subject when compared to a non-injured individual (marked with gray ellipses). Note also that start of the task, as marked on the bottom, is associated with increased sEMG activity and raising airway pressure.
Figure 4</st...

Watch this Scientific Journal Video about Evaluation of Respiratory Muscle Activation Using Respiratory Motor Control Assessment RMCA in Individuals with Chronic Spinal Cord Injury at JoVE.com

Evaluation of Respiratory Muscle Activation Using Respiratory Motor Control Assessment RMCA in Individuals with Chronic Spinal Cord Injury

The purpose of this publication is to present our original work on a multi-muscle surface electromyographic approach to quantitatively characterize respiratory muscle activation patterns in individuals with chronic spinal cord injury using vector-based analysis.

Evaluation of Respiratory Muscle Activation Using Respiratory Motor Control Assessment (RMCA) in Individuals with Chronic Spinal Cord Injury

During breathing, activation of respiratory muscles is coordinated by integrated input from the brain, brainstem, and spinal cord. When this coordination ...

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10.2K visualizações.  University of Louisville. O objetivo desta publicação é apresentar o nosso trabalho original sobre uma abordagem eletromiografia de superfície multi-muscular para caracterizar quantitativamente os padrões de ativação dos músculos respiratórios em indivíduos com lesão medular crônica usando análise baseada em vetor.

Vídeo: Evaluation of Respiratory Muscle Activation Using Respiratory Motor Control Assessment RMCA in Individuals with Chronic Spinal Cord Injury

Longitudinal Evaluation of Mouse Hind Limb Bone Loss After Spinal Cord Injury using Novel, in vivo, Methodology

Spinal cord injury (SCI) is often accompanied by osteoporosis in the sublesional regions of the pelvis and lower extremities, leading to a higher frequency of fractures 1. As these fractures often occur in regions that have lost normal sensory function, the patient is at a greater risk of fracture-dependent pathologies, including death. SCI-dependent loss in both bone mineral density (BMD, grams/cm2) and bone mineral content (BMC, grams) has been attributed to mechanical disuse 2, aberrant neuronal signaling 3 and hormonal changes 4. The use of rodent models of SCI-induced osteoporosis can provide invaluable information regarding the mechanisms underlying the development of osteoporosis following SCI as well as a test environment for the generation of new therapies 5-7 (and reviewed in 8). Mouse models of SCI are of great interest as they permit a reductionist approach to mechanism-based assessment through the use of null and transgenic mice. While such models have provided important data, there is still a need for minimally-invasive, reliable, reproducible, and quantifiable methods in determining the extent of bone loss following SCI, particularly over time and within the same cohort of experimental animals, to improve diagnosis, treatment methods, and/or prevention of SCI-induced osteoporosis.
An ideal method for measuring bone density in rodents would allow multiple, sequential (over time) exposures to low-levels of X-ray radiation. This study describes the use of a new whole-animal scanner, the IVIS Lumina XR (Caliper Instruments) that can be used to provide low-energy (1-3 milligray (mGy)) high-resolution, high-magnification X-ray images of mouse hind limb bones over time following SCI. Significant bone density loss was seen in the tibiae of mice by 10 days post-spinal transection when compared to uninjured, age-matched control (na&iuml;ve) mice (13% decrease, p&lt;0.0005). Loss of bone density in the distal femur was also detectable by day 10 post-SCI, while a loss of density in the proximal femur was not detectable until 40 days post injury (7% decrease, p&lt;0.05). SCI-dependent loss of mouse femur density was confirmed post-mortem through the use of Dual-energy X-ray Absorptiometry (DXA), the current "gold standard" for bone density measurements. We detect a 12% loss of BMC in the femurs of mice at 40 days post-SCI using the IVIS Lumina XR. This compares favorably with a previously reported BMC loss of 13.5% by Picard and colleagues who used DXA analysis on mouse femurs post-mortem 30 days post-SCI 9. Our results suggest that the IVIS Lumina XR provides a novel, high-resolution/high-magnification method for performing long-term, longitudinal measurements of hind limb bone density in the mouse following SCI.

Longitudinal Evaluation of Mouse Hind Limb Bone Loss After Spinal Cord Injury using Novel, in vivo, Methodology

A longitudinal examination of bone loss in the femurs and tibiae of adult mice was performed following spinal cord injury using sequential low-dose X-ray scans. Tibia bone loss was detected throughout the study, while bone loss in the femur was not detected until 40 days post injury.

Spinal cord injury (SCI) is often accompanied by osteoporosis in the sublesional regions of the pelvis and lower extremities, leading to a higher ...

Acute and Chronic Tactile Sensory Testing after Spinal Cord Injury in Rats

Spinal cord injury (SCI) impairs sensory systems causing allodynia1-8. To identify cellular and molecular causes of allodynia, sensitive and valid sensory testing in rat SCI models is needed. However, until recently, no single testing approach had been validated for SCI so that standardized methods have not been implemented across labs. Additionally, available testing methods could not be implemented acutely or when severe motor impairments existed, preventing studies of the development of SCI-induced allodynia3. Here we present two validated sensory testing methods using von Frey Hair (VFH) monofilaments which quantify changes in tactile sensory thresholds after SCI4-5. One test is the well-established Up-Down test which demonstrates high sensitivity and specificity across different SCI severities when tested chronically5. The other test is a newly-developed dorsal VFH test that can be applied acutely after SCI when allodynia develops, prior to motor recovery4-5. Each VFH monofilament applies a calibrated force when touched to the skin of the hind paw until it bends. In the up-down method, alternating VFHs of higher or lower forces are used on the plantar L5 dermatome to delineate flexor withdrawal thresholds. Successively higher forces are applied until withdrawal occurs then lower force VFHs are used until withdrawal ceases. The tactile threshold reflects the force required to elicit withdrawal in 50% of the stimuli. For the new test, each VFH is applied to the dorsal L5 dermatome of the paw while the rat is supported by the examiner. The VFH stimulation occurs in ascending order of force until at least 2 of 3 applications at a given force produces paw withdrawal. Tactile sensory threshold is the lowest force to elicit withdrawal 66% of the time. Acclimation, testing and scoring procedures are described. Aberrant trials that require a retest and typical trials are defined. Animal use was approved by Ohio State University Animal Care and Use Committee.

We describe two tactile sensory testing methods for acute or chronic periods of spinal cord injury in rats. These validated procedures can detect the development and maintenance of allodynia-like sensations.

Spinal cord injury (SCI) impairs sensory systems causing allodynia1-8. To identify cellular and molecular causes of allodynia, sensitive and valid sensory ...

A Murine Model of Cervical Spinal Cord Injury to Study Post-lesional Respiratory Neuroplasticity

A cervical spinal cord injury induces permanent paralysis, and often leads to respiratory distress. To date, no efficient therapeutics have been developed to improve/ameliorate the respiratory failure following high cervical spinal cord injury (SCI). Here we propose a murine pre-clinical model of high SCI at the cervical 2 (C2) metameric level to study diverse post-lesional respiratory neuroplasticity. The technique consists of a surgical partial injury at the C2 level, which will induce a hemiparalysis of the diaphragm due to a deafferentation of the phrenic motoneurons from the respiratory centers located in the brainstem. The contralateral side of the injury remains intact and allows the animal recovery. Unlike other SCIs which affect the locomotor function (at the thoracic and lumbar level), the respiratory function does not require animal motivation and the quantification of the deficit/recovery can be easily performed (diaphragm and phrenic nerve recordings, whole body ventilation). This pre-clinical C2 SCI model is a powerful, useful, and reliable pre-clinical model to study various respiratory and non-respiratory neuroplasticity events at different levels (molecular to physiology) and to test diverse putative therapeutic strategies which might improve the respiration in SCI patients.

Respiratory failure is the leading cause of death following a cervical spinal cord injury. Having a reproducible, quantifiable, and reliable pre-clinical animal model of respiratory failure induced by a partial cervical injury will help to understand the subsequent respiratory and non-respiratory neuroplasticity and allow testing putative repair strategies.

A cervical spinal cord injury induces permanent paralysis, and often leads to respiratory distress. To date, no efficient therapeutics have been developed ...

Controlled Cortical Impact Model for Traumatic Brain Injury

Every year over a million Americans suffer a traumatic brain injury (TBI). Combined with the incidence of TBIs worldwide, the physical, emotional, social, and economical effects are staggering. Therefore, further research into the effects of TBI and effective treatments is necessary. The controlled cortical impact (CCI) model induces traumatic brain injuries ranging from mild to severe. This method uses a rigid impactor to deliver mechanical energy to an intact dura exposed following a craniectomy. Impact is made under precise parameters at a set velocity to achieve a pre-determined deformation depth. Although other TBI models, such as weight drop and fluid percussion, exist, CCI is more accurate, easier to control, and most importantly, produces traumatic brain injuries similar to those seen in humans. However, no TBI model is currently able to reproduce pathological changes identical to those seen in human patients. The CCI model allows investigation into the short-term and long-term effects of TBI, such as neuronal death, memory deficits, and cerebral edema, as well as potential therapeutic treatments for TBI.

Traumatic brain injuries (TBIs) remain a serious health problem. Using the controlled cortical impact surgery model, research on the effects of TBI and possible treatment methods may be performed.

Every year over a million Americans suffer a traumatic brain injury (TBI).  Combined with the incidence of TBIs worldwide, the physical, emotional, ...

A Radio-telemetric System to Monitor Cardiovascular Function in Rats with Spinal Cord Transection and Embryonic Neural Stem Cell Grafts

High thoracic or cervical spinal cord injury (SCI) can lead to cardiovascular dysfunction. To monitor cardiovascular parameters, we implanted a catheter connected to a radio transmitter into the femoral artery of rats that underwent a T4 spinal cord transection with or without grafting of embryonic brainstem-derived neural stem cells expressing green fluorescent protein. Compared to other methods such as cannula insertion or tail-cuff, telemetry is advantageous to continuously monitor blood pressure and heart rate in freely moving animals. It is also capable of long term multiple data acquisitions. In spinal cord injured rats, basal cardiovascular data under unrestrained condition and autonomic dysreflexia in response to colorectal distension were successfully recorded. In addition, cardiovascular parameters before and after SCI can be compared in the same rat if a transmitter is implanted before a spinal cord transection. One limitation of the described telemetry procedure is that implantation in the femoral artery may influence the blood supply to the ipsilateral hindlimb.

We present a protocol for using a radio-telemetric system to record cardiovascular parameters in T4 spinal cord transected rats eight weeks after embryonic brainstem neural stem cell grafting into the lesion site. Telemetry is an advanced technique to accurately evaluate cardiovascular function in conscious freely moving spinal cord injured rats.

High thoracic or cervical spinal cord injury (SCI) can lead to cardiovascular dysfunction. To monitor cardiovascular parameters, we implanted a catheter ...

Calibrated Forceps Model of Spinal Cord Compression Injury

Compression injuries of the murine spinal cord are valuable animal models for the study of spinal cord injury (SCI) and spinal regenerative therapy. The calibrated forceps model of compression injury is a convenient, low cost, and very reproducible animal model for SCI. We used a pair of modified forceps in accordance with the method published by Plemel et al. (2008) to laterally compress the spinal cord to a distance of 0.35 mm. In this video, we will demonstrate a dorsal laminectomy to expose the spinal cord, followed by compression of the spinal cord with the modified forceps. In the video, we will also address issues related to the care of paraplegic laboratory animals. This injury model produces mice that exhibit impairment in sensation, as well as impaired hindlimb locomotor function. Furthermore, this method of injury produces consistent aberrations in the pathology of the SCI, as determined by immunohistochemical methods. After watching this video, viewers should be able to determine the necessary supplies and methods for producing SCI of various severities in the mouse for studies on SCI and/or treatments designed to mitigate impairment after injury.

Spinal cord injury models should be highly reproducible. We demonstrate that the calibrated forceps compression model of spinal cord injury is an easy to use surgical method for generating reproducible injuries to the murine spinal cord.

Compression injuries of the murine spinal cord are valuable animal models for the study of spinal cord injury (SCI) and spinal regenerative therapy. The ...

Contrast Enhanced Ultrasound Imaging for Assessment of Spinal Cord Blood Flow in Experimental Spinal Cord Injury

Reduced spinal cord blood flow (SCBF) (i.e., ischemia) plays a key role in traumatic spinal cord injury (SCI) pathophysiology and is accordingly an important target for neuroprotective therapies. Although several techniques have been described to assess SCBF, they all have significant limitations. To overcome the latter, we propose the use of real-time contrast enhanced ultrasound imaging (CEU). Here we describe the application of this technique in a rat contusion model of SCI. A jugular catheter is first implanted for the repeated injection of contrast agent, a sodium chloride solution of sulphur hexafluoride encapsulated microbubbles. The spine is then stabilized with a custom-made 3D-frame and the spinal cord dura mater is exposed by a laminectomy at ThIX-ThXII. The ultrasound probe is then positioned at the posterior aspect of the dura mater (coated with ultrasound gel). To assess baseline SCBF, a single intravenous injection (400 &#181;l) of contrast agent is applied to record its passage through the intact spinal cord microvasculature. A weight-drop device is subsequently used to generate a reproducible experimental contusion model of SCI. Contrast agent is re-injected 15 min following the injury to assess post-SCI SCBF changes. CEU allows for real time and in-vivo assessment of SCBF changes following SCI. In the uninjured animal, ultrasound imaging showed uneven blood flow along the intact spinal cord. Furthermore, 15 min post-SCI, there was critical ischemia at the level of the epicenter while SCBF remained preserved in the more remote intact areas. In the regions adjacent to the epicenter (both rostral and caudal), SCBF was significantly reduced. This corresponds to the previously described &#8220;ischemic penumbra zone&#8221;. This tool is of major interest for assessing the effects of therapies aimed at limiting ischemia and the resulting tissue necrosis subsequent to SCI.

Contrast Enhanced Ultrasound imaging is a reliable in-vivo tool for quantifying spinal cord blood flow in an experimental rat spinal cord injury model. This paper contains a comprehensive protocol for application of this technique in association with a contusion model of thoracic spinal cord injury.

Reduced spinal cord blood flow (SCBF) (i.e., ischemia) plays a key role in traumatic spinal cord injury (SCI) pathophysiology and is accordingly an ...

A Component-resolved Diagnostic Approach for a Study on Grass Pollen Allergens in Chinese Southerners with Allergic Rhinitis and/or Asthma

Sensitization to grass pollen imposes a global risk for allergic airway diseases. Although prevention relies on local investigation of the pollen allergens, data on this topic are limited in southern China. Any available data were obtained by self-report questionnaires, skin prick tests, and total or specific IgE tests using crude extracts. For many reasons, these methods are unreliable. Serum sIgE reactivity to Bermuda grass, Timothy grass, and Humulus scandens allergens in a cohort of patients from Greater Guangzhou (southern China&#39;s largest city and its outskirts) with allergic rhinitis and/or asthma were examined using a fully-automated immunoassay analyzer as a component-resolved diagnostics (CRD) tool.
For the first time, a considerably high prevalence of Bermuda grass sIgE positivity was demonstrated in Chinese southerners with allergic rhinitis and/or asthma. In these patients, a subtle prevalence of sensitization to Timothy grass and Humulus scandens was also noted, which may arise from cross-reactivity, as the latter two are not common in the region. This was also supported by the detection of allergen components.
Fully-automated immunoassay analyzers may offer satisfactory consistency between regions, laboratories, and institutions and over time. The automaticity of the instrument may enable a standardized detection that would not have been readily revealed before the advent of CRD.
This is a study that uses a CRD approach to investigate sensitization to grass pollen allergens in southern China. It adds to current evidence in the literature. Future studies are needed to validate these findings. However, although CRD is a useful tool, the findings made with the fully-automated immunoassay analyzer should not substitute for other laboratory investigations, clinical evaluations, and physician expertise.

This work describes a protocol that uses a component-resolved approach to study sensitization to grass pollen allergens in a cohort of patients from southern China with allergic rhinitis and/or asthma.

Sensitization to grass pollen imposes a global risk for allergic airway diseases. Although prevention relies on local investigation of the pollen ...

Generation of a Chronic Obstructive Pulmonary Disease Model in Mice by Repeated Ozone Exposure

Chronic obstructive pulmonary disease (COPD) is characterized by persistent airflow limitation and lung parenchymal destruction. It has a very high incidence in aging populations. The current conventional therapies for COPD focus mainly on symptom-modifying drugs; thus, the development of new therapies is urgently needed. Qualified animal models of COPD could help to characterize the underlying mechanisms and can be used for new drug screening. Current COPD models, such as lipopolysaccharide (LPS) or the porcine pancreatic elastase (PPE)-induced emphysema model, generate COPD-like lesions in the lungs and airways but do not otherwise resemble the pathogenesis of human COPD. A cigarette smoke (CS)-induced model remains one of the most popular because it not only simulates COPD-like lesions in the respiratory system, but it is also based on one of the main hazardous materials that causes COPD in humans. However, the time-consuming and labor-intensive aspects of the CS-induced model dramatically limit its application in new drug screening. In this study, we successfully generated a new COPD model by exposing mice to high levels of ozone. This model demonstrated the following: 1) decreased forced expiratory volume 25, 50, and 75/forced vital capacity (FEV25/FVC, FEV50/FVC, and FEV75/FVC), indicating the deterioration of lung function; 2) enlarged lung alveoli, with lung parenchymal destruction; 3) reduced fatigue time and distance; and 4) increased inflammation. Taken together, these data demonstrate that the ozone exposure (OE) model is a reliable animal model that is similar to humans because ozone overexposure is one of the etiological factors of COPD. Additionally, it only took 6 - 8 weeks, based on our previous work, to create an OE model, whereas it requires 3 - 12 months to induce the cigarette smoke model, indicating that the OE model might be a good choice for COPD research.

This study describes the successful generation of a new chronic obstructive pulmonary disease (COPD) animal model by repeatedly exposing mice to high concentrations of ozone.

Chronic obstructive pulmonary disease (COPD) is characterized by persistent airflow limitation and lung parenchymal destruction. It has a very high ...

Evaluation of Coronary Flow Reserve After Myocardial Ischemia Reperfusion in Rats

Coronary artery disease is the leading cause of death worldwide. After an acute myocardial infarction, early and successful myocardial intervention via recanalization of the coronary artery is the most effective strategy for reducing the size of ischemic myocardium. The coronary microvasculature cannot be visualized and imaged&#160;in vivo, but there are several invasive and noninvasive techniques that can be used to assess parameters which depend directly on coronary microvascular function. The endothelial function after ischemia reperfusion can be assessed also at the level of the coronary circulation via the coronary flow reserve (CFR).&#160;In this study, peak velocity of left anterior descending (LAD) coronary arteries was measured in rats in vivo via Transthoracic Doppler Echocardiography during resting and stress challenge (induced by Dobutamine). A normal heart can increase its coronary blood flow up to four times above the resting values during stress induction. Following ischemia reperfusion, we found a significantly diminished CFR, which can be used as a marker of coronary microvascular dysfunction. CFR has opened a window on the importance of microvascular dysfunction and has been shown to predict cardiovascular risk independent of whether the severe obstructive disease is present.

Coronary flow reserve (CFR), is defined as the ratio of maximal coronary blood flow to the resting coronary blood flow. We present a protocol for evaluating CFR in rats via ultrasound, which offers the opportunity to predict cardiovascular risk factors in the absence of obstructive coronary disease.

Coronary artery disease is the leading cause of death worldwide. After an acute myocardial infarction, early and successful myocardial intervention via ...