This study was supported by grants from the American Medical Society for Sports Medicine. The authors would like to acknowledge and show our appreciation for our undergraduate research students including David Lawton, Drew Gardner, Mark Sundman, Kelsey Evans, Graham Cochrane, Jordan Cottle and Jack Halligan for their assistance in data collection over the past four years.

All the procedures described below have been approved by the Elon&#8217;s Institutional Review Board.
1. Computerized Neurocognitive Testing
<ol>
	<li>Ask the participants to be seated in front of the computer. Log participants on to system and instruct them to complete the computerized neuropsychological test which consists a demographic and background information section, self-reported symptom checklist, and 6 modules (word discrimination, design memory, X&#8217;s and O&#8217;s, symbol matching, color match, and three letters).</li>
	<li>Download summary report and enter four composite scores for verbal memory, visual memory, reaction time and motor processing speed.</li>
</ol>
2. Event Related Potentials
<ol>
	<li>Measure head circumference using a measuring tape to determine size of EEG net to be used. Determine placement of the EEG net by measuring anatomical landmarks.</li>
	<li>Soak the EEG net in solution of sodium chloride and baby shampoo for 5 min.</li>
	<li>Place the EEG net on the head of the participant. The system that we use includes 32 channels.</li>
	<li>Check impedance levels of the sites on the computer. For our system, an impedance below 100 kohm is deemed acceptable.</li>
	<li>Explain the cognitive tasks and let the participant practice each of the tasks.
	<ol>
		<li>Auditory Oddball Task: Instruct the participant to put on headphones and sit comfortably at a table. Inform them that they will hear a series of low and high tones and to respond as quickly and accurately as possible by clicking a button to a high frequency auditory tone.</li>
		<li>Flanker Task: Instruct participant to sit in front of a computer screen where they will be given a series of arrows projected onto a screen. Instruct participants to respond to the direction of the middle arrow by clicking the left mouse button if it was pointing left or clicking the right mouse button if it was pointing right as quickly and accurately as possible.&#160;</li>
	</ol>
	</li>
	<li>Instruct participant to complete two trials of both the auditory oddball task and the flanker task.</li>
	<li>Remove the EEG net from the participant and clean the EEG net by soaking in germicide disinfectant for 10 min.</li>
</ol>
3. Somatosensory Perceptual Responses
<ol>
	<li>Seat the participant comfortably with their left hand in a prone position resting on the sensory stimulus device and their fingers positioned along the contour of the device with the padded tips of digits 2 and 3 placed in contact with the stimulus probes.23</li>
	<li>Ask the participant to view task related instructions and cues on a computer monitor and enter responses using a two button computer mouse. Project 5 different tests: 2 simple single site reaction time tasks, a dual site amplitude discrimination,24 a dual site amplitude task with a single site adapting stimulus,24 and a temporal order judgment task.25&#160;&#160;</li>
	<li>Prior to the start of each test run, the cues on the computer will instruct the participant to complete practice trials to familiarize themselves with the task. The computer will give the participants performance feedback after each test (e.g. correct; incorrect).</li>
</ol>
4. Balance Protocol
<ol>
	<li>Instruct participants to put on slip-resistant socks and then stand on the balance system to become acquainted with instrument.</li>
	<li>Instruct participants to stand in a comfortable position, matching the center of pressure dot with the center dot on the screen. Researchers record this comfortable starting position such that all tests take place with the same foot position while standing on the device.</li>
	<li>Ask the participant to stand for 30 sec for each of the four conditions (eyes open/firm surface, eyes closed/firm surface, eyes open/foam surface, eyes closed/foam surface). Provide the participants a 10 sec rest between each condition and a 3 sec countdown before beginning of each recording.</li>
	<li>Repeat each of the conditions while completing a secondary task. Instruct them to count backwards by 7 starting from a random 3 digit number given to them (e.g., 843).</li>
	<li>Record a sway index score, a measure of the standard deviation of the amount of sway for each condition and center of pressure data to use later for further analysis.</li>
</ol>
5. Gait Assessment
<ol>
	<li>Assess participants&#8217; gait using a portable 15&#8217; long carpet instrumented gait analysis system with pressure sensors present throughout the length of the carpet to detect the participant&#8217;s footfalls.</li>
	<li>
	Ask the participants to walk across mat barefoot at a comfortable speed five times starting from a distance of 3&#8217; before the start of the mat and 3&#8217; after leaving the mat.
	</li>
	<li>
	Instruct the participants to complete five additional walking trials while counting backwards by seven from a random 3 digit number as a concurrent cognitive dual task.
	</li>
	<li>The dependent variables obtained as output from gait analysis include absolute and variability measures of several spatiotemporal parameters like velocity, cadence and step length.</li>
</ol>
6. Genetics
<ol>
	<li>Ask the participant to carefully remove the swab stick from its sterile container (being careful to only touch the stick end of the swab stick), move the swab into their mouth, and vigorously rub the swab inside of both cheeks for a total of 20 sec.</li>
	<li>Hand the swab stick to the technician who is wearing latex or nitrile gloves. Only hold the stick end and then place the swab tip into a sterile 1.7 ml tube (labeled with an identification number only), which is immediately placed on ice.</li>
	<li>Within 24 hr transfer the samples are to a -20 &#176;C freezer.</li>
	<li>Extract DNA using a standard DNA Purification Kit according to the manufacturer&#8217;s protocol. To obtain a more concentrated sample, after extraction, perform an isopropanol precipitation step and rehydrate DNA in 20 &#181;l of EDTA buffer (pH 8.0).</li>
	<li>Store the extracted DNA at -80 &#176;C until genotyping analysis using standard polymerase chain reaction (PCR) assays with fluorescent tags.</li>
	<li>&#8220;Call&#8221; the genotypes by PCR software and then verify manually by viewing PCR amplification plots.</li>
</ol>

<ol>
	<li>Kristman, V., 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=Does+the+Apolipoprotein+E4+allele+predispose+varsity+athletes+to+concussion?+A+prospective+cohort+study.">Does the Apolipoprotein E4 allele predispose varsity athletes to concussion? A prospective cohort study.</a> Clin J Sports Med. 18, 322-328 (2008).</li><li>McCrory, P., 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=Consensus+statement+on+concussion+in+sports+the+4th+International+Conference+on+Concussion+held+in+Zurich,+November+2012.">Consensus statement on concussion in sports the 4th International Conference on Concussion held in Zurich, November 2012.</a> Br J Sports Med. 47 (5), 250-258 (2012).</li><li>Giza, C. 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=Summary+of+evidence-based+guideline+update:+Evaluation+and+management+of+concussion+in+sports:+Report+of+the+Guideline+Development+Subcommittee+of+the+American+Academy+of+Neurology.">Summary of evidence-based guideline update: Evaluation and management of concussion in sports: Report of the Guideline Development Subcommittee of the American Academy of Neurology.</a> Neurology. 80 (24), 2250-2257 (2013).</li><li>Langlois, J. A., Rutland-Brown, W., Waid, 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=The+epidemiology+and+impact+of+traumatic+brain+injury:+a+brief+overview.">The epidemiology and impact of traumatic brain injury: a brief overview.</a> J Head Trauma Rehabil. 21, 375-378 (2006).</li><li>Faul, M., Xu, L., Wald, M. M., Coronado, V. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Traumatic+brain+injury+in+the+United+States:+emergency+department+visits,+hospitalizations,+and+deaths.">Traumatic brain injury in the United States: emergency department visits, hospitalizations, and deaths.</a> Centers for Disease Control and Prevention, National Center for Injury Prevention and Control. , (2010).</li><li>Broglio, S. P., Puetz, T. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+effect+of+sport+concussion+on+neurocognitive+function,+self-report+symptoms+and+postural+control:+a+meta-analysis.">The effect of sport concussion on neurocognitive function, self-report symptoms and postural control: a meta-analysis.</a> J Sports Med. 38, 53-67 (2008).</li><li>Cancelliere, 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=Protocol+for+a+systematic+review+of+prognosis+after+mild+traumatic+brain+injury:+an+update+of+the+WHO+Collaborating+Centre+Task+Force+findings.">Protocol for a systematic review of prognosis after mild traumatic brain injury: an update of the WHO Collaborating Centre Task Force findings.</a> Systematic Reviews. 1, 17 .</li><li>Guskiewicz, 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=Cumulative+effects+associated+with+recurrent+concussion+in+collegiate+football+players.">Cumulative effects associated with recurrent concussion in collegiate football players.</a> JAMA. 290, 2549-2555 (2003).</li><li>Makdissi, M., Darby, D., Maruff, P., Ugoni, A., Brukner, P., McCrory, P. 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W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Neuropsychological+assessment+of+sport-related+concussion.">Neuropsychological assessment of sport-related concussion.</a> Clin Sports Med. 30 (1), 78-88 (2011).</li><li>Broglio, S. P., Pontifex, M. B., O'Connor, P., Hillman, C. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+persistent+effects+of+concussion+on+neuroelectric+indices+of+attention.">The persistent effects of concussion on neuroelectric indices of attention.</a> J Neurotrauma. 26 (9), 1463-1470 (2009).</li><li>Holden, J. K., Nguyen, R. H., Francisco, E. M., Zhang, Z., Dennis, R. 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The authors declare that they have no competing financial interests.

The goal of this multidimensional approach to baseline concussion testing is twofold: 1) to better understand the impact of a concussion (acute and long term) on the neuromuscular system; 2) to help the sports medicine staff make return to play decisions (they primarily use neurocognitive testing as has been suggested by McCrory).26 This multifaceted approach to concussion evaluation provides valuable information related to the etiology, functional changes, and recovery from concussion. Little is understood ab...

Concussions are occurring at alarming rates in the United States and have garnered quite a lot of attention as a public health concern.1-3 US Centers for Disease Control and Prevention (CDC) estimates that 1.6 to 3.8 million concussions occur in sports and recreational activities annually.4,5 Concussion as defined by the 2013 Concussion Consensus Statement2 &#8220;may be caused either by a direct blow to the head, face, neck or elsewhere on the body with an &#8216;impulsive&#8217; force transmitted to the head.&#8221; Concussion may result in neuropathological and/or substructural changes that may result in functional disturbances.2 These deficits may persist for several weeks. It is not uncommon for athletes to experience increased self-reported symptoms, decrements in postural control, and decreased neurocognitive function even 14 days following the initial injury.6 The prolonged nature of symptoms, the inconsistent identification of concussions, and the variability in preinjury abilities often lead to complex and non standardized return-to-play decisions by physicians, uncertain recovery times, and possibly long-term sequelae.7-9
Following a concussion, an individual may experience both short-term and long-term effects. The short-term effects of sport related concussions may include changes in playing ability, confusion, memory disturbance, the loss of consciousness, slowing of reaction time, loss of coordination, headaches, dizziness, vomiting, changes in sleep patterns and mood changes. These symptoms typically resolve in a matter of days.2,10 However, while some individuals recover from a single concussion rather quickly, many experience lingering effects that can last for weeks or months following the injury.10,11, 12 These symptomatic disturbances to daily function can be quantified using cognitive and performance related tests. While no one single test should determine diagnosis of a concussion, a battery of tests and known relationships between tests can aid the medical staff in making diagnoses, return to classroom, and return to play decisions.2
There is great variability in an individual&#8217;s risk for concussion and their corresponding recovery.11 The factors related to concussion susceptibility and recovery time course are not well known or understood. Several factors have been suggested that may impact an individual&#8217;s concussion susceptibility and recovery. These factors include the individual&#8217;s concussion history, the severity of the initial injury, history of migraines, history of learning disabilities, history of psychiatric comorbidities, and possibly genetic factors.7, 9, 13, 14
Many studies have individually investigated specific factors for both the short-term and long-term effects of concussions, recovery time course, and genetics as a factor of concussions.4,8,15-17 What has not been clearly established is an effective multifaceted approach to concussion evaluation that would yield valuable information related to the etiology, functional changes, and recovery from concussion. Due to the variety of symptoms and the uncertain time course of recovery, a multifaceted approach to concussion evaluation is warranted and this should include baseline testing of all athletes prior to participation in practice and competition as well as timely evaluation post injury. A recent review suggests that neurocognitive assessments may be more sensitive to recovery from a concussion than monitoring symptoms alone.18 It may be that there are other objective measures that may be better indicators of recovery from concussion.
For this protocol, we use several tasks to assess various components of the system to see how they are impacted by a concussion. A computerized neurocognitive test can assess memory, processing speed, problem solving skills, cognitive efficiency and impulse control.6 EEG with auditory and visual processing tasks can be used to assess neuroefficiency through the examination of event related potentials.19 A somatosensory discrimination task can be used to assess peripheral and central sensory processing capabilities.20 Balance and gait measures can be used to assess functional performance capabilities.6,21 In addition, we assess various genotypes that may have relations to concussion history, concussion recovery and cognitive function.22 We baseline test our varsity student-athletes on this battery of tests and repeat tests if they incur a concussion when asymptomatic.
The purpose of this project is to assess potential short-term and long-term decrements in performance as a result of concussions using genetic, neurocognitive, electrophysiological, behavioral, somatosensory, balance and gait measures. Understanding the potential mechanisms that may be related to various symptoms and impairments that occur with a concussion are important in furthering our knowledge about concussion. Greater knowledgle about these changes may in the future aid in concussion diagnosis as well as concussion management as it relates to return to play and return to academics.
All measures described below are taken at baseline (before student-athlete participation in sport). Our current protocol is to complete the computerized neurocognitive testing at 48 hr along with the balance protocol because we believe that these provide useful information regarding recovery and possible return-to-play and return-to-academics. When the student-athlete reports asymptomatic they again return to the laboratory where all baseline measures are again conducted, except for genetic testing. The full protocol, baseline and asymptomatic, takes approximately 90 min to complete in one testing period.

<table border="0" cellpadding="0" cellspacing="0" width="858">
	<tbody>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Name of Material/ Equipment</td>
			<td style="width:356px;">Company</td>
			<td style="width:119px;">Catalog Number</td>
			<td style="width:167px;">Comments/Description</td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:216px;">ImPACT</td>
			<td style="width:356px;">ImPACT, Pittsburgh, PA</td>
			<td style="width:119px;"></td>
			<td>Neurocognitive concussion testing</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">EEG</td>
			<td style="width:356px;">EGI, Eugene, OR</td>
			<td style="width:119px;"></td>
			<td>EEG 32-channel system</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Stim2</td>
			<td style="width:356px;">Compumedics Neuroscan, Charlotte, NC</td>
			<td style="width:119px;"></td>
			<td>Software for task presentation for flanker task and auditory oddball</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">NetStation</td>
			<td style="width:356px;">EGI, Eugene, OR</td>
			<td style="width:119px;"></td>
			<td>Software for data collection and analysis of EEG</td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;width:216px;">Sensory Device</td>
			<td style="width:356px;">Cortical Metrics</td>
			<td style="width:119px;"></td>
			<td>Sensory testing</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Balance System SD</td>
			<td>Biodex Medical Systems, Inc., Shirley, NY</td>
			<td style="width:119px;"></td>
			<td>balance testing</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Force Plate</td>
			<td>AMTI Corp., Watertown, MA, USA</td>
			<td style="width:119px;"></td>
			<td>Gait Initiation</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Movement Analysis</td>
			<td>Qualisys AB, Gothenburg, Sweden</td>
			<td style="width:119px;"></td>
			<td>Gait Analysis</td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;">GAITRite&#160;</td>
			<td style="width:356px;">CIR systems, Inc., Sparta, NJ, USA</td>
			<td style="width:119px;"></td>
			<td>Gait analysis</td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;width:216px;">PCR</td>
			<td style="width:356px;">Applied Biosystems, Foster City, CA</td>
			<td style="width:119px;"></td>
			<td>Genetic Analysis</td>
		</tr>
		<tr height="24">
			<td height="24" style="height:24px;">Matlab</td>
			<td style="width:356px;">Mathworks, Natick, MA, USA</td>
			<td style="width:119px;"></td>
			<td>Gait and balance analysis</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:216px;">Visual 3D</td>
			<td style="width:356px;">C Motion, Inc, Germantown, MD, USA</td>
			<td style="width:119px;"></td>
			<td>3D analysis&#160;</td>
		</tr>
	</tbody>
</table>

a neuroscientific approach to the examination of concussions in student-athletes

Concussions are occurring at alarming rates in the United States and have become a serious public health concern. The CDC estimates that 1.6 to 3.8 million concussions occur in sports and recreational activities annually. Concussion as defined by the 2013 Concussion Consensus Statement &#8220;may be caused either by a direct blow to the head, face, neck or elsewhere on the body with an &#8216;impulsive&#8217; force transmitted to the head.&#8221; Concussions leave the individual with both short- and long-term effects. The short-term effects of sport related concussions may include changes in playing ability, confusion, memory disturbance, the loss of consciousness, slowing of reaction time, loss of coordination, headaches, dizziness, vomiting, changes in sleep patterns and mood changes. These symptoms typically resolve in a matter of days. However, while some individuals recover from a single concussion rather quickly, many experience lingering effects that can last for weeks or months. The factors related to concussion susceptibility and the subsequent recovery times are not well known or understood at this time. Several factors have been suggested and they include the individual&#8217;s concussion history, the severity of the initial injury, history of migraines, history of learning disabilities, history of psychiatric comorbidities, and possibly, genetic factors. Many studies have individually investigated certain factors both the short-term and long-term effects of concussions, recovery time course, susceptibility and recovery. What has not been clearly established is an effective multifaceted approach to concussion evaluation that would yield valuable information related to the etiology, functional changes, and recovery. The purpose of this manuscript is to show one such multifaceted approached which examines concussions using computerized neurocognitive testing, event related potentials, somatosensory perceptual responses, balance assessment, gait assessment and genetic testing.

Computerized Neurocognitive Testing
An example of results for the computerized neurocognitive test can be seen in Figure 1. The computer program elicits composite scores on Verbal Memory, Visual Memory, Visual Motor Speed and Reaction Time which are often used to make return-to-play and return-to-learn concussion management protocols. The verbal and visual memory composites evaluate attentional processes, learning and memory. Visual motor speed measures visual...

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A Neuroscientific Approach to the Examination of Concussions in Student-Athletes

There is great variability in an individual&#8217;s risk for concussion and their corresponding recovery. A multifaceted approach to concussion evaluation is warranted; including baseline testing of athletes before participation in sport and timely evaluation post injury. The goal of this protocol is to provide an appropriate multifaceted approach to examine concussions.

Concussions are occurring at alarming rates in the United States and have become a serious public health concern. The CDC estimates that 1.6 to 3.8 ...

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12.6K Views.  Elon University. There is great variability in an individual’s risk for concussion and their corresponding recovery. A multifaceted approach to concussion evaluation is warranted; including baseline testing of athletes before participation in sport and timely evaluation post injury. The goal of this protocol is to provide an appropriate multifaceted approach to examine concussions.

Video: A Neuroscientific Approach to the Examination of Concussions in Student-Athletes

Bronchial Thermoplasty:  A Novel Therapeutic Approach to Severe Asthma

Bronchial thermoplasty is a non-drug procedure for severe persistent asthma that delivers thermal energy to the airway wall in a precisely controlled manner to reduce excessive airway smooth muscle. Reducing airway smooth muscle decreases the ability of the airways to constrict, thereby reducing the frequency of asthma attacks. Bronchial thermoplasty is delivered by the Alair System and is performed in three outpatient procedure visits, each scheduled approximately three weeks apart. The first procedure treats the airways of the right lower lobe, the second treats the airways of the left lower lobe and the third and final procedure treats the airways in both upper lobes. After all three procedures are performed the bronchial thermoplasty treatment is complete.
Bronchial thermoplasty is performed during bronchoscopy with the patient under moderate sedation. All accessible airways distal to the mainstem bronchi between 3 and 10 mm in diameter, with the exception of the right middle lobe, are treated under bronchoscopic visualization. Contiguous and non-overlapping activations of the device are used, moving from distal to proximal along the length of the airway, and systematically from airway to airway as described previously. Although conceptually straightforward, the actual execution of bronchial thermoplasty is quite intricate and procedural duration for the treatment of a single lobe is often substantially longer than encountered during routine bronchoscopy. As such, bronchial thermoplasty should be considered a complex interventional bronchoscopy and is intended for the experienced bronchoscopist. Optimal patient management is critical in any such complex and longer duration bronchoscopic procedure. This article discusses the importance of careful patient selection, patient preparation, patient management, procedure duration, postoperative care and follow-up to ensure that bronchial thermoplasty is performed safely.
Bronchial thermoplasty is expected to complement asthma maintenance medications by providing long-lasting asthma control and improving asthma-related quality of life of patients with severe asthma. In addition, bronchial thermoplasty has been demonstrated to reduce severe exacerbations (asthma attacks) emergency rooms visits for respiratory symptoms, and time lost from work, school and other daily activities due to asthma.

Bronchial thermoplasty is a non-drug procedure for severe persistent asthma that delivers thermal energy to the airway wall in a precisely controlled manner to reduce excessive airway smooth muscle. Reducing airway smooth muscle decreases the ability of the airways to constrict, thereby reducing the frequency of asthma attacks.

Bronchial thermoplasty is a non-drug procedure for severe persistent asthma that delivers thermal energy to the airway wall in a precisely controlled ...

A Strategy to Identify de Novo Mutations in Common Disorders such as Autism and Schizophrenia

There are several lines of evidence supporting the role of de novo mutations as a mechanism for common disorders, such as autism and schizophrenia. First, the de novo mutation rate in humans is relatively high, so new mutations are generated at a high frequency in the population. However, de novo mutations have not been reported in most common diseases. Mutations in genes leading to severe diseases where there is a strong negative selection against the phenotype, such as lethality in embryonic stages or reduced reproductive fitness, will not be transmitted to multiple family members, and therefore will not be detected by linkage gene mapping or association studies. The observation of very high concordance in monozygotic twins and very low concordance in dizygotic twins also strongly supports the hypothesis that a significant fraction of cases may result from new mutations. Such is the case for diseases such as autism and schizophrenia. Second, despite reduced reproductive fitness1 and extremely variable environmental factors, the incidence of some diseases is maintained worldwide at a relatively high and constant rate. This is the case for autism and schizophrenia, with an incidence of approximately 1% worldwide. Mutational load can be thought of as a balance between selection for or against a deleterious mutation and its production by de novo mutation. Lower rates of reproduction constitute a negative selection factor that should reduce the number of mutant alleles in the population, ultimately leading to decreased disease prevalence. These selective pressures tend to be of different intensity in different environments. Nonetheless, these severe mental disorders have been maintained at a constant relatively high prevalence in the worldwide population across a wide range of cultures and countries despite a strong negative selection against them2. This is not what one would predict in diseases with reduced reproductive fitness, unless there was a high new mutation rate. Finally, the effects of paternal age: there is a significantly increased risk of the disease with increasing paternal age, which could result from the age related increase in paternal de novo mutations. This is the case for autism and schizophrenia3. The male-to-female ratio of mutation rate is estimated at about 4&#x2013;6:1, presumably due to a higher number of germ-cell divisions with age in males. Therefore, one would predict that de novo mutations would more frequently come from males, particularly older males4. A high rate of new mutations may in part explain why genetic studies have so far failed to identify many genes predisposing to complexes diseases genes, such as autism and schizophrenia, and why diseases have been identified for a mere 3% of genes in the human genome. Identification for de novo mutations as a cause of a disease requires a targeted molecular approach, which includes studying parents and affected subjects. The process for determining if the genetic basis of a disease may result in part from de novo mutations and the molecular approach to establish this link will be illustrated, using autism and schizophrenia as examples.

Molecular genetic strategy for finding de novo mutations causing common disorders such as autism and schizophrenia.

There are several lines of evidence supporting the role of de novo mutations as a mechanism for common disorders, such as autism and schizophrenia. First, ...

The Measurement and Treatment of Suppression in Amblyopia

Amblyopia, a developmental disorder of the visual cortex, is one of the leading causes of visual dysfunction in the working age population. Current estimates put the prevalence of amblyopia at approximately 1-3%1-3, the majority of cases being monocular2. Amblyopia is most frequently caused by ocular misalignment (strabismus), blur induced by unequal refractive error (anisometropia), and in some cases by form deprivation.
Although amblyopia is initially caused by abnormal visual input in infancy, once established, the visual deficit often remains when normal visual input has been restored using surgery and/or refractive correction. This is because amblyopia is the result of abnormal visual cortex development rather than a problem with the amblyopic eye itself4,5 . Amblyopia is characterized by both monocular and binocular deficits6,7 which include impaired visual acuity and poor or absent stereopsis respectively. The visual dysfunction in amblyopia is often associated with a strong suppression of the inputs from the amblyopic eye under binocular viewing conditions8. Recent work has indicated that suppression may play a central role in both the monocular and binocular deficits associated with amblyopia9,10 .
Current clinical tests for suppression tend to verify the presence or absence of suppression rather than giving a quantitative measurement of the degree of suppression. Here we describe a technique for measuring amblyopic suppression with a compact, portable device11,12 . The device consists of a laptop computer connected to a pair of virtual reality goggles. The novelty of the technique lies in the way we present visual stimuli to measure suppression. Stimuli are shown to the amblyopic eye at high contrast while the contrast of the stimuli shown to the non-amblyopic eye are varied. Patients perform a simple signal/noise task that allows for a precise measurement of the strength of excitatory binocular interactions. The contrast offset at which neither eye has a performance advantage is a measure of the "balance point" and is a direct measure of suppression. This technique has been validated psychophysically both in control13,14 and patient6,9,11 populations.
In addition to measuring suppression this technique also forms the basis of a novel form of treatment to decrease suppression over time and improve binocular and often monocular function in adult patients with amblyopia12,15,16 . This new treatment approach can be deployed either on the goggle system described above or on a specially modified iPod touch device15.

Amblyopia is a developmental disorder of the visual cortex that is often accompanied by strong suppression of one eye. We present a new technique for measuring and treating interocular suppression in patients with amblyopia that can be deployed using virtual reality goggles or a portable iPod Touch device.

Amblyopia, a developmental disorder of the visual cortex, is one of the leading causes of visual dysfunction in the working age population. Current ...

Clinical Examination Protocol to Detect Atypical and Classical Scrapie in Sheep

The diagnosis of scrapie, a transmissible spongiform encephalopathy (TSEs) of sheep and goats, is currently based on the detection of disease-associated prion protein by post mortem tests. Unless a random sample of the sheep or goat population is actively monitored for scrapie, identification of scrapie cases relies on the reporting of clinical suspects, which is dependent on the individual&#39;s familiarization with the disease and ability to recognize clinical signs associated with scrapie. Scrapie may not be considered in the differential diagnosis of neurological diseases in small ruminants, particularly in countries with low scrapie prevalence, or not recognized&#160;if it presents as nonpruritic form like atypical scrapie. To aid in the identification of clinical suspects, a short examination protocol is presented to assess the display of specific clinical signs associated with pruritic and nonpruritic forms of TSEs in sheep, which could also be applied to goats. This includes assessment of behavior, vision (by testing of the menace response), pruritus (by testing the response to scratching), and movement (with and without blindfolding). This may lead to a more detailed neurologic examination&#160;of reporting animals as scrapie suspects. It&#160;could also be used in experimental TSE studies of sheep or goats to evaluate disease progression or to identify clinical end-point.

A short clinical examination protocol is presented to aid in the detection of classical and atypical scrapie in sheep and goats.

The diagnosis of scrapie, a transmissible spongiform encephalopathy (TSEs) of sheep and goats, is currently based on the detection of disease-associated ...

Diagnosis of Musculus Gastrocnemius Tightness - Key Factors for the Clinical Examination

Common foot and ankle pathologies have been linked to isolated Musculus gastrocnemius tightness (MGT). Various examination techniques have been described to assess MGT. Still, a standardized examination procedure is missing. Literature argues for weightbearing examination but the degree of knee flexion needed to eliminate the restraining effect of the M. gastrocnemius on ankle dorsiflexion (ADF) is unknown. This manuscript investigates the effect of knee flexion on ankle dorsiflexion and provides a detailed description of a standardized examination protocol. Examination on 20 healthy individuals revealed, that 20&#176; of knee flexion is sufficient to fully eliminate the influence of the M. gastrocnemius on ADF. This builds the prerequisite for a standardized examination for MGT. Non-weightbearing and weightbearing examination of ADF has to be conducted with the knee fully extended and at least 20&#176; flexed. Two investigators should conduct non-weightbearing testing with the subject in supine position. In order to obtain reliable results, the axis of the fibula should be marked. One examiner can conduct weightbearing examination with the subject in lunge stance. Isolated MGT is present if ADF is impaired with the knee fully extended and knee flexion results in a significant ADF increase. The herein presented standardized examination is the prerequisite for future studies aiming at establishing norm values.

Diagnosis of Musculus Gastrocnemius Tightness - Key Factors for the Clinical Examination

Isolated Musculus gastrocnemius tightness is a common cause for foot and ankle pathologies. Currently no standardized examination procedure exists. This manuscript demonstrates that 20 degree of knee flexion eliminates the restraining effect of the M. gastrocnemius on ankle dorsiflexion and presents a video description of a standardized examination protocol.

Common foot and ankle pathologies have been linked to isolated Musculus gastrocnemius tightness (MGT). Various examination techniques have been described ...

Echocardiographic and Histological Examination of Cardiac Morphology in the Mouse

An increasing number of genetically modified mouse models has become available in recent years. Moreover, the number of pharmacological studies performed in mice is high. Phenotypic characterization of these mouse models also requires the examination of cardiac function and morphology. Echocardiography and magnetic resonance imaging (MRI) are commonly used approaches to characterize cardiac function and morphology in mice. Echocardiographic and MRI equipment specialized for use in small rodents is&#160;expensive and requires a dedicated space. This protocol describes cardiac measurements in mice using a clinical echocardiographic system with a 15 MHz human vascular probe. Measurements are performed on anesthetized adult mice. At least three image sequences are recorded and analyzed for each animal in M-mode in the parasternal short-axis view. Afterwards, cardiac histological examination is performed, and cardiomyocyte diameters are determined on hematoxylin-eosin- or wheat germ agglutinin (WGA)-stained paraffin sections. Vessel density is determined morphometrically after Pecam-1 immunostaining. The protocol has been applied successfully to pharmacological studies and different genetic animal models under baseline conditions, as well as after experimental myocardial infarction by the permanent ligation of the left anterior descending coronary artery (LAD). In our experience, echocardiographic investigation is limited to anesthetized animals and is feasible in adult mice weighing at least 25 g.

Echocardiographic examination is frequently used in mice. Expensive high-resolution ultrasound devices have been developed for this purpose. This protocol describes an affordable echocardiographic procedure combined with histological morphometric analyses to determine cardiac morphology.

An increasing number of genetically modified mouse models has become available in recent years. Moreover, the number of pharmacological studies performed ...

The 4-vessel Sampling Approach to Integrative Studies of Human Placental Physiology In Vivo

The human placenta is highly inaccessible for research while still in utero. The current understanding of human placental physiology in vivo is therefore largely based on animal studies, despite the high diversity among species in placental anatomy, hemodynamics and duration of the pregnancy. The vast majority of human placenta studies are ex vivo perfusion studies or in vitro trophoblast studies. Although in vitro studies and animal models are essential, extrapolation of the results from such studies to the human placenta in vivo is uncertain. We aimed to study human placenta physiology in vivo at term, and present a detailed protocol of the method. Exploiting the intraabdominal access to the uterine vein just before the uterine incision during planned cesarean section, we collect blood samples from the incoming and outgoing vessels on the maternal and fetal sides of the placenta. When combining concentration measurements from blood samples with volume blood flow measurements, we are able to quantify placental and fetal uptake and release of any compound. Furthermore, placental tissue samples from the same mother-fetus pairs can provide measurements of transporter density and activity and other aspects of placental functions in vivo. Through this integrative use of the 4-vessel sampling method we are able to test some of the current concepts of placental nutrient transfer and metabolism in vivo, both in normal and pathological pregnancies. Furthermore, this method enables the identification of substances secreted by the placenta to the maternal circulation, which could be an important contribution to the search for biomarkers of placenta dysfunction.

The 4-vessel Sampling Approach to Integrative Studies of Human Placental Physiology In Vivo

We present a detailed method to study human placental physiology in vivo at term. The method combines blood sampling from the incoming and outgoing vessels on the maternal and fetal sides of the placenta with ultrasound measurements of volume blood flow and placental tissue sampling.

The human placenta is highly inaccessible for research while still in utero. The current understanding of human placental physiology in vivo is therefore ...

An In Vitro Approach to Photodynamic Therapy

Photodynamic therapy (PDT) is a medical procedure that involves incubation of an exogenously applied photosensitizer (PS) followed by visible light photoactivation to induce cell apoptosis. The Federal Drug Administration has approved PDT for the treatment of actinic keratosis, and clinical guidelines recommend PDT as a treatment for certain non-melanoma skin cancers and acne vulgaris. PDT is an advantageous therapeutic modality as it is low cost, non-invasive, and associated with minimal adverse events and scaring. In the first step of PDT, a PS is applied and allowed to accumulate intracellularly. Subsequent light irradiation induces reactive oxygen species formation, which may ultimately lead to cell apoptosis, membrane disruption, mitochondrial damage, immune modulation, keratinocyte proliferation, and collagen turnover. Herein, we present an in vitro method to study PDT in an adherent cell line. This treatment protocol is designed to simulate PDT and may be adjusted to studying the use of PDT with various cell lines, photosensitizers, incubation temperatures, or photoactivation wavelengths. Squamous cell carcinoma cells were incubated with 0, 0.5, 1.0, and 2 mM 5-aminolevulinic acid (5-ALA) for 30 min and photoactivated with 417 nm blue light for 1,000 s. The primary outcome measure was apoptosis and necrosis, as measured by annexin-V and 7-aminoactinomycin D flow cytometry. There was a dose-dependent increase in cell apoptosis following thirty-minute incubation of 5-ALA. To achieve high inter-test validity, it is important to maintain consistent incubation and light parameters when performing in vitro PDT experiments. PDT is a useful clinical procedure and in vitro research may allow for the development of novel PSs, optimization of protocols, and new indications for PDT.

An In Vitro Approach to Photodynamic Therapy

Photodynamic therapy (PDT) is a medical procedure that involves incubation of an exogenously applied photosensitizer (PS) followed by visible light photoactivation to induce apoptosis. We present an in vitro PDT protocol designed to simulate PDT that may be used to study differences in PS incubation and light treatment parameters.

Photodynamic therapy (PDT) is a medical procedure that involves incubation of an exogenously applied photosensitizer (PS) followed by visible light ...

A Metadata Extraction Approach for Clinical Case Reports to Enable Advanced Understanding of Biomedical Concepts

Clinical case reports (CCRs) are a valuable means of sharing observations and insights in medicine. The form of these documents varies, and their content includes descriptions of numerous, novel disease presentations and treatments. Thus far, the text data within CCRs is largely unstructured, requiring significant human and computational effort to render these data useful for in-depth analysis. In this protocol, we describe methods for identifying metadata corresponding to specific biomedical concepts frequently observed within CCRs. We provide a metadata template as a guide for document annotation, recognizing that imposing structure on CCRs may be pursued by combinations of manual and automated effort. The approach presented here is appropriate for organization of concept-related text from a large literature corpus (e.g., thousands of CCRs) but may be easily adapted to facilitate more focused tasks or small sets of reports. The resulting structured text data includes sufficient semantic context to support a variety of subsequent text analysis workflows: meta-analyses to determine how to maximize CCR detail, epidemiological studies of rare diseases, and the development of models of medical language may all be made more realizable and manageable through the use of structured text data.

We present a protocol and associated metadata template for the extraction of text describing biomedical concepts in clinical case reports. The structured text values produced through this protocol can support deep analysis of thousands of clinical narratives.

Clinical case reports (CCRs) are a valuable means of sharing observations and insights in medicine. The form of these documents varies, and their content ...

A Novel Approach to Monitoring Graft Neovascularization in the Human Gingiva

Laser speckle contrast imaging (LSCI) is a novel method for measuring superficial blood perfusion over large areas. Since it is non-invasive and avoids direct contact with the measured area, it is suitable for monitoring blood flow changes during wound healing in human patients. Vestibuloplasty is periodontal surgery to the oral vestibule, aiming to restore vestibular depth with simultaneous enlargement of the keratinized gingiva. In this special clinical case, a split thickness flap was elevated at the first upper premolar and a xenogenic collagen matrix was adapted to the resulting recipient bed. LSCI was used to monitor the re- and neovascularization of the graft and the surrounding mucosa for one year. A protocol is introduced for the correct adjustment of microcirculation measurement in the oral mucosa, highlighting difficulties and possible failures.
The clinical case study presented demonstrated that &#8212; following the appropriate protocol &#8212; LSCI is a suitable and reliable method for following up microcirculation in a healing wound in the human oral mucosa and gives useful information on graft integration.

This study introduces a protocol for measuring microcirculation in human oral mucosa by laser speckle contrast imaging. The monitoring of wound healing after vestibuloplasty combined with a xenogenic collagen graft is presented on a clinical case.

Laser speckle contrast imaging (LSCI) is a novel method for measuring superficial blood perfusion over large areas. Since it is non-invasive and avoids ...

A Neuroscientific Approach to the Examination of Concussions in Student-Athletes

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Bronchial Thermoplasty: A Novel Therapeutic Approach to Severe Asthma

A Strategy to Identify de Novo Mutations in Common Disorders such as Autism and Schizophrenia

The Measurement and Treatment of Suppression in Amblyopia

Clinical Examination Protocol to Detect Atypical and Classical Scrapie in Sheep

Diagnosis of Musculus Gastrocnemius Tightness - Key Factors for the Clinical Examination

Echocardiographic and Histological Examination of Cardiac Morphology in the Mouse

The 4-vessel Sampling Approach to Integrative Studies of Human Placental Physiology In Vivo

An In Vitro Approach to Photodynamic Therapy

A Metadata Extraction Approach for Clinical Case Reports to Enable Advanced Understanding of Biomedical Concepts

A Novel Approach to Monitoring Graft Neovascularization in the Human Gingiva