Blinded Scorer Research Articles

The effect of an antioxidant gel compared to chlorhexidine during the soft tissue healing process: An animal study.

Prolonged inflammation and oxidative stress can impede healing. To enhance healing efficiency, many solutions have been employed. This is an in vivo study comparing chlorhexidine (CHX) to a commercial antioxidant gel (AO). Envelope flaps were created in the lower incisor gingival region of 60 Sprague-Dawley rats, and acellular dermal matrix (ADM) was inserted. Animals were randomly assigned to postsurgical treatment application of AO gel or 0.12% CHX twice daily. A control group received no postsurgical treatment. Data collected (before surgery, 24 h, and 72 h) included surgical images, tissue samples, and weights. Blinded scorers assessed images using a wound healing scale. Real-time polymerase chain reaction (RT-PCR) was used for gene expression of tumor necrosis factor-alpha (TNFα), interleukin-1 (IL-1), myeloperoxidase (MPO), and superoxide dismutase (SOD). The AO group scored higher than the CHX and control groups in clinical evaluation (p<0.05). At 24h, TNFα expression was upregulated in the AO group compared to CHX (p=0.027) and controls (p=0.018). The AO group had significantly higher expression of antioxidant enzyme (SOD) at 24h compared to CHX (p=0.021). All animals lost weight in the first 24h. Animals treated with AO or CHX regained more weight at 72h than control animals (p=0.034 and 0.003, respectively). Animals treated with AO healed faster. AO led to earlier upregulation of TNFα and antioxidant enzyme SOD. We hypothesized that AO promoted an earlier inflammatory process while counteracting oxidative stress by increasing antioxidant responses via SOD.

RAPAMYCIN HAS A LIMITED EFFECT ON TENDON HEALING IN A RODENT MODEL

Tendon injuries occur frequently in athletes and the general population, with inferior healing leading to deposition of fibrotic scar tissue. New treatments are essential to limit fibrosis and enable tendon regeneration post-injury. In this study, we tested the hypothesis that rapamycin improves tendon repair and limits fibrosis by inhibiting the mTOR pathway.The left hindlimb of female adult Wistar rats was injured by needle puncture and animals were either given daily injections of rapamycin (2mg/kg) or vehicle. Animals were euthanized 1 week or 3 weeks post-injury (n=6/group). Left and right Achilles tendons were harvested, with the right limbs acting as controls. Tendon sections were stained with haematoxylin &amp; eosin, and scored by 2 blinded scorers, assessing alterations in cellularity, cell morphology, vascularity, extracellular matrix (ECM) organization and peritendinous fibrosis. Immunohistochemistry was performed for the tendon pan-vascular marker CD146 and the autophagy marker LC3.Injury resulted in significantly altered ECM organization, cell morphology and cellularity in both rapamycin and vehicle-treated groups, but no alterations in vascularity compared to uninjured tendons. Rapamycin had a limited effect on tendon repair, with a significant reduction in peritendinous fibrosis 3 weeks after injury (p=0.028) but no change in cell morphology, cellularity or ECM organization compared to vehicle treated tendons at either 1 week or 3 weeks post injury. CD146 labelling was increased at the site of injury, but there was no apparent difference in CD146 or LC3 labelling in rapamycin and vehicle treated tendons.The decrease in peritendinous fibrosis post-injury observed in rapamycin treated tendons indicates rapamycin as a potential therapy for tendon adhesions. However, the lack of improvement of other morphological parameters in response to rapamycin treatment indicates that rapamycin is not an effective therapy for injuries to the tendon core.Acknowledgements: This study was funded by Versus Arthritis (22607)

Poster 101: Evaluation of Novel Treatment Agents on Rotator Cuff Tendon and Muscle Pathology in a Murine Subacromial Impingement Model

Poster 101: Evaluation of Novel Treatment Agents on Rotator Cuff Tendon and Muscle Pathology in a Murine Subacromial Impingement Model

Polysomnographic versus video scoring of large muscle group movements during sleep in children with restless sleep

BackgroundRestless sleep is a common complaint in children with sleep and other disorders, when it occurs as an isolated symptom it can depict a separate condition called restless sleep disorder (RSD). A key diagnostic feature of RSD is the polysomnographic (PSG) scoring of five or more large muscle movements (LMM) per hour of sleep. Initial publications on RSD used video-PSG (vPSG) analysis to count LMM. However, recently, scoring criteria for LMM on PSG signals alone have been published, but their correlation with vPSG scoring has not been done. In this study we compare vPSG to PSG scoring of LMM in a group of children with restless sleep. MethodsVideo-PSG studies from 20 children (14 boys and six girls, mean age 11.3 years, SD 4.17) were reviewed by a blinded scorer. First, scorer used a video analysis (video-LMM), then on a different day the scorer used PSG criteria for LMM without video (PSG-LMM). ResultsPSG-LMM were found to be significantly more numerous than video-LMM (mean LMM index 8.6 ± 2.95 SD vs. 6.9 ± 2.40 SD, respectively) while their duration was not statistically different (mean LMM duration 11.6 ± 2.33 s vs. 11.6 ± 2.99 s, respectively). 95.4% of video-LMM were also detected by PSG. Both LMM indexes and durations obtained by the two methods correlated significantly between them. A cut off of 6.24 LMM/hour by PSG-LMM was calculated to be equivalent to 5 LMM by video-LMM. ConclusionsPSG-LMM scoring criteria detects >95% of LLM scored by video-PSG. PSG-LMM lead to the detection of a higher number of movements, indicating their higher sensitivity in picking up LMM in PSG recording. This further supports that these criteria need adequate consideration for their eventual inclusion into the current sleep scoring manual. A higher index may be needed to diagnose RSD.

Towards a new approach to detect sleepiness: Validation of the objective sleepiness scale under simulated driving conditions

The Objective Sleepiness Scale (OSS) was developed to detect and quantify sleepiness on the basis of two direct and reliable sleepiness indicators: EEG and EOG. The present study aims to test whether the OSS can be used to detect sleepiness episodes that impair performance on driving and vigilance tasks accurately and with a good time synchronization. Forty-three healthy volunteers performed monotonous driving sessions on a simulator and the psychomotor vigilance task (PVT) in a normal sleep condition and after partial sleep deprivation. OSS reliability and time synchronization for sleepiness detection were tested on driving (standard deviation of vehicle lateral position and off-road duration) and PVT (reaction time and lapses). Inter-rater reliability of the scale was evaluated by two blinded scorers. Results show that the OSS score indicates higher sleepiness in sleep deprivation conditions (p < 0.001) and with time-on-task. Differences of performance between OSS score calculated with multiple pairwise comparisons, indicate OSS score increase when driving performance (SDLP and off-road duration) decreases (p < 0.001 for comparisons between OSS stages 0 vs 2, 0 vs 3, 1 vs 2 and p < 0.05 for 1 vs 3). Reaction time during PVT is also related to the OSS score (p < 0.05 for OSS values from 0 to 2, 0 to 3, 1 to 2 and 1 to 3). There is no proportional relation between OSS score and performance impairment, but a threshold effect between levels 1 and 2 of the scale is observed. Positive outcomes are also obtained for time synchronization of the OSS assessed on driving performance (p < 0.001 for both SDLP and off-road duration). Finally, inter-rater agreement is found to be considerable. The results allow us to consider using the Objective Sleepiness Scale as a tool for research on sleepiness.

Extracellular biomolecular free radical formation during injury

ObjectiveDetermine if oxidative damage increases in articular cartilage as a result of injury and matrix failure and whether modulation of the local redox environment influences this damage.Osteoarthritis is an age associated disease with no current disease modifying approaches available. Mechanisms of cartilage damage in vitro suggest tissue free radical production could be critical to early degeneration, but these mechanisms have not been described in intact tissue. To assess free radical production as a result of traumatic injury, we measured biomolecular free radical generation via immuno-spin trapping (IST) of protein/proteoglycan/lipid free radicals after a 2 J/cm2 impact to swine articular cartilage explants. This technique allows visualization of free radical formation upon a wide variety of molecules using formalin-fixed, paraffin-embedded approaches. Scoring of extracellular staining by trained, blinded scorers demonstrated significant increases with impact injury, particularly at sites of cartilage cracking. Increases remain in the absence of live chondrocytes but are diminished; thus, they appear to be a cell-dependent and -independent feature of injury. We then modulated the extracellular environment with a pulse of heparin to demonstrate the responsiveness of the IST signal to changes in cartilage biology. Addition of heparin caused a distinct change in the distribution of protein/lipid free radicals at sites of failure alongside a variety of pertinent redox changes related to osteoarthritis. This study directly confirms the production of biomolecular free radicals from articular trauma, providing a rigorous characterization of their formation by injury.

The behavioral and physiological effects of dog appeasing pheromone on canine behavior during separation from the owner

Behavioral problems in the domestic dog (Canis familiaris) increase the likelihood of the dog being rehomed or relinquished to a rescue shelter. Problem behaviors that result in relinquishment include unwanted elimination, destructive behavior, and excessive vocalization when owners are absent. Dog appeasing pheromone (DAP) is currently marketed via veterinarians as a stress relief product and purported to help dogs cope in stressful situations and as a potential solution to reduce anxiety. This study investigated whether a DAP diffuser affected behavioral and physiological stress parameters in 10 dogs in a laboratory environment. A repeated measures design with and without the use of DAP and in the presence and absence of the owner was used. Behavioral responses, such as barking, passive behavior, scratching, whining, orienting behavior, exploration, and locomotion, were recorded in real time and video recorded using a focal instantaneous sampling technique. To control for potential bias, 10% of the videos were scored using a second blinded scorer to assess interrater reliability. Heart rate (HR), heart rate variability using standard deviation of normal to normal beats, and eye temperature and ear temperature (°C) were also collected to assess dogs' physiological state. When dogs were separated from their owner, there was a significant increase in orienting behavior during both the DAP and without DAP application trial phase, suggesting arousal due to owner absence rather than any discernible effect of DAP. A significant increase was recorded in core eye temperature when the owner was absent and the DAP diffuser was not switched on; however, eye temperature also increased when the owners were present after the DAP condition, suggesting that it may be the owner's presence and the dog's arousal levels that affect core eye temperature rather than any effect of DAP. There was no significant effect of DAP on HR or ear temperature. Overall, our results suggest that the application of a DAP diffuser did not markedly influence the behavior, HR, and eye or ear temperature of dogs.

Use of a Transformed ECG Signal to Detect Respiratory Effort During Apnea.

To evaluate the ability of a transformed electrocardiography (ECG) signal recorded using standard electrode placement to detect inspiratory bursts from underlying surface chest wall electromyography (EMG) activity and the utility of the transformed signal for apnea classification compared to uncalibrated respiratory inductance plethysmography (RIP). Part 1: 250 consecutive adult studies without regard to respiratory events were retrospectively reviewed. The ECG signal was transformed with high pass filtering and viewed with increased sensitivity and channel clipping to determine the fraction of studies with inspiratory burst visualization as compared to chest wall EMG (right thorax). Part 2: 445 consecutive studies were reviewed to select 40 with ≥ 10 obstructive and ≥ 10 mixed or central apneas (clinical scoring). Five obstructive and 5 central or mixed apneas were randomly selected from each study. A blinded scorer classified the apneas using either RIP or a transformed ECG signal using high pass filtering and QRS blanking. The agreement between the two classifications was determined by kappa analysis. Part 1: Inspiratory burst visualization was noted in the transformed ECG signals and chest wall EMG signals in 83% and 71% of the studies (P < .001). Part 2: The percentage agreement between RIP and transformed ECG signal classification was 88.5%, the kappa statistic was 0.81 (95% CI 0.76 to 0.86) and interclass correlation was 0.84, showing good agreement. A transformed ECG signal can exhibit inspiratory bursts in a high proportion of patients and is potentially useful for detecting respiratory effort and apnea classification.

Crossed Cerebellar Atrophy in Perinatal Stroke.

Background and Purpose- Perinatal stroke causes most hemiparetic cerebral palsy and lifelong disability. Crossed cerebellar atrophy (CCA) is chronic cerebellar volume loss following contralateral motor pathway injury. We hypothesized that CCA is quantifiable in perinatal stroke and associated with poor motor outcome. Methods- Term-born children with perinatal stroke, magnetic resonance imaging beyond 6 months of age, and no additional neurological disorders were recruited. Blinded scorers measured cerebellar volumes expressed as ratios (contralesional/ipsilesional), with values <1 suggesting CCA. Motor outcomes including perinatal stroke outcome measure (PSOM) motor and cognitive scores (good/poor), Assisting Hand Assessment, and Melbourne Assessment were compared with cerebellar volume measures. Results- Seventy-three children met criteria (53% male). Mean cerebellar ratios were <1.0 (0.975±0.04; range, 0.885-1.079; P<0.001) suggesting occurrence of CCA. Cerebellar ratios did not differ between stroke types or across PSOM motor outcomes. Larger ipsilesional cerebellar volume was associated with poor PSOM cognitive outcome (P=0.042), possibly with poor PSOM motor outcome (P=0.063), and overall PSOM score (P=0.034). Conclusions- CCA occurs in perinatal stroke but is not strongly associated with motor outcome. However, ipsilesional cerebellar volume is associated with poor cognitive and overall outcomes.

The role of excessive body weight and meniscal instability in the progression of osteoarthritis in a rat model

BackgroundThe main objective of the study was to determine if excessive body weight and meniscal destabilization cause knee articular cartilage damage and changes in meniscal histology indicative of knee osteoarthritis (OA). MethodsForty female Lewis rats were randomly assigned to regular diet sham surgery, regular diet medial meniscus destabilization (DMM) surgery, high-fat diet sham surgery, or high-fat diet DMM surgery. Surgical intervention occurred four weeks after diet initiation. Rats were sacrificed four weeks post-surgery. Progression of OA was evaluated in femoral condyles and tibial plateaus using the Osteoarthritis Research Society International (OARSI) scoring system by two blinded scorers. Ossification of menisci was evaluated with digitized histological slides using a grid. ResultsDMM surgery had greater OARSI scores than sham surgery for all knee surfaces in both diet groups (all P < 0.004). The high-fat diet sham surgery group had greater OARSI scores than the regular diet sham surgery group on all knee surfaces (all P < 0.02). Histological evaluation showed the lateral meniscus had more bone than the medial meniscus, but there was no relationship between diet or surgery groups and amount of bone in the meniscus. ConclusionsResults suggested excessive body weight may contribute to spontaneous onset of OA and emphasized the importance of preventative care in those at risk of developing OA.

Use of Chest Wall EMG to Classify Hypopneas as Obstructive or Central.

To compare classification of hypopneas as obstructive or central based on an effort signal derived from surface chest wall electromyography (CW-EMG-EF) coupled with airflow amplitude versus classification using The AASM Manual for the Scoring of Sleep and Associated Events: Rules, Terminology and Technical Specifications (AASM Scoring Manual) criteria; and to characterize hypopneas classified as obstructive versus central using a resistance surrogate. CW-EMG was recorded in the eighth intercostal space at the right midaxillary line. Five hypopneas were randomly selected from 65 consecutive adult clinical positive airway pressure titration studies meeting study criteria. A blinded scorer classified the hypopneas based on two groups of signals: Group 1: positive airway pressure flow (PAP flow), chest and abdominal effort, and snoring; or Group 2: smoothed PAP flow (for blinding amplitude but not flattening visible) and effort (CW-EMG-EF). A resistance surrogate (CW-EMG-EF / PAP flow) normalized to a pre-event breath was compared between obstructive and central hypopneas classified by AASM Scoring Manual criteria. The percentage agreement (Group 1 versus Group 2) was 92% and the kappa was 0.75 (95% confidence interval 0.65 to 0.85). The resistance surrogate was significantly higher in obstructive hypopneas versus central hypopneas during the first and second half of hypopneas. The resistance surrogate (mean ± standard deviation) for the second half of hypopnea was obstructive: 7.59 ± 7.24 versus central: 1.27 ± 0.56, P < .001). The resistance surrogate increased from the first to second half of hypopnea only for obstructive hypopneas. CW-EMG provides a useful complementary signal for hypopnea classification and a resistance surrogate based on CW-EMG is much higher in hypopneas classified as obstructive by AASM Scoring Manual criteria.

Use of Contralateral Knee as Control in the Destabilization of Medial Meniscus Osteoarthritis Rat Model

Destabilization of the medial meniscus (DMM) is a common surgical procedure used to induce osteoarthritis (OA) in a rodent model. However, there is a lack of evidence on using the contralateral limb as a control. Contralateral rat knees have been used as the control in noninvasive OA pain models, but not DMM models; and the contralateral limb of guinea pigs has been used to study the development of OA after DMM surgery. In a previous study, we examined the effects of DMM and obesity on the development and progression of OA in a rat model. In that study, 42 female Lewis rats were randomly assigned to 4 groups: Group 1 received a sham surgery (control) and was on a regular diet, Group 2 received a DMM surgery and was on a regular diet, Group 3 received a sham surgery and was on a high‐fat diet, and Group 4 received a DMM surgery and was on a high‐fat diet. The rats were on their respective diets for 4 weeks, underwent surgery, and were sacrificed 4 weeks after surgery. The knee joints of surgical and contralateral limbs were harvested. The Osteoarthritis Research Society International (OARSI) scoring system was used to score the damage to the articular cartilage of the knee. In the surgical limbs, DMM resulted in significantly greater OARSI scores than the sham surgery in the lateral and medial compartments of the knee in both diet groups. After that study, the contralateral knee tissues were stored in 10% formalin. For the current study, we used those stored tissues to determine whether the contralateral limbs could serve as a control in future studies that involve DMM in a rat model. Knees were decalcified, embedded in paraffin, and sectioned coronally at 10 μm. Slides were stained using Safranin O and fast green for counterstain. Four articular surfaces of the knee (medial and lateral femoral condyles and medial and lateral tibial plateaus) were histologically evaluated for degenerative changes by a blinded scorer. A generalized linear mixed model was used to analyze differences between the rat knee groups. Comparison of sham and DMM rat knees across both diet groups showed no significant difference in OARSI scores (P=.61). When comparing within each of the diet groups, there was no significant difference between the DMM and sham scores in the high‐fat diet groups (P&gt;.99) or regular diet groups (P=.49). After comparing the knee surface scores for all groups, the medial tibial plateau had significantly lower scores than the other 3 surfaces (P&lt;.001). Based on results of the current study, we believe the contralateral limb can be used as a control in short‐term (4 weeks or less) OA studies using DMM in a rat model. This will save time and resources by achieving an adequate sample size with fewer animals. Our results also showed that the medial tibial plateau was significantly less susceptible to the development of spontaneous OA compared to the other 3 articular surfaces studied.Support or Funding InformationSupport for this project was received from Dr. Peter Kondrashov, Department of Anatomy, A.T. Still University, Kirksville College of Osteopathic Medicine.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

0608 ASSESSMENT OF OBSTRUCTIVE SLEEP APNEA IN PATIENTS WITHCHRONIC PAIN SYNDROME: HOW DOES PORTABLE RESPIRATORY RECORDING COMPARE TOPOLYSOMNOGRAPHY?

Obstructive Sleep Apnea (OSA) is a common condition seen in veterans with Chronic Pain Syndrome (CPS). Portable respiratory recording (PRT) is cheaper and more readily available than polysomnography (PSG). The validity of PRT in the diagnosis of OSA in patients with CPS is not well characterized. Veterans with CPS admitted to a referral pain rehabilitation center underwent PSG along with supervised PRT in the sleep laboratory on presentation. Studies were scored independently by a blinded scorer using AASM scoring criteria, hypopneas were scored based on oxygen desaturation and arousal criteria in PSG while they were scored based on oxygen desaturation on PRT. Generated reports were compared for Apnea Hypopnea Index (AHI), Oxygen Saturation Nadir (OSN), Oxygen Desaturation Index (ODI), Time Supine (TS), and Mean Heart Rate (MHR) using mean, standard deviation and Parson Correlation Coefficient (PCC). Twenty-five veterans with CPS were enrolled (22 male, 3 females with an average BMI of 30.15). Average AHI was 23 +/-15 via PSG and 18.2 +/-16.5 via PRT, with PCC 0.69. Twenty three patients were found to have OSA based on PSG while nineteen patients were found to have OSA based on PRT. Ten patients had severe OSA based on PSG while four patients had severe OSA based on PRT. OSN was 86.1 +/- 22.8% via PSG and 85.8 +/-22.7% via PRT, PCC 0.54. ODI was 13.6 +/- 13.6 via PSG and 14.3 +/- 11.1 via PRT, PCC 0.82. TS was 192.7 +/- 109 min via PSG vs 223 +/- 124.4 min via PRT, PCC 0.81. MHR was 67.6 +/- 19.8 bpm via PSG vs 63.8 +/- 18.6 bpm via PRT, PCC 0.97. The results were not statistically significant. The sensitivity and severity characterization of PRT is suboptimal when compared to the PSG gold standard with chronic pain. Oximetry and positional measurements are similar between PRT and PSG. Further analysis of PRT sensitivity based on patient characteristics may help identify more suitable candidates for PRT testing. None.

A Novel Assessment Tool for Quantitative Evaluation of Science Literature Search Performance: Application to First-Year and Senior Undergraduate Biology Majors

Expertise in searching and evaluating scientific literature is a requisite skill of trained scientists and science students, yet information literacy instruction varies greatly among institutions and programs. To ensure that science students acquire information literacy skills, robust methods of assessment are needed. Here, we describe a novel tool for longitudinal, crossover assessment of literature-searching skills in science students and apply it to a cross-sectional assessment of literature-searching performance in 145 first-year and 43 senior biology majors. Subjects were given an open-ended prompt requiring them to find multiple sources of information addressing a particular scientific topic. A blinded scorer used a rubric to score the resources identified by the subjects and generate numerical scores for source quality, source relevance, and citation quality. Two versions of the assessment prompt were given to facilitate eventual longitudinal study of individual students in a crossover design. Seniors were significantly more likely to find relevant, peer-reviewed journal articles, provide appropriate citations, and provide correct answers to other questions about scientific literature. This assessment tool accommodates large numbers of students and can be modified easily for use in other disciplines or at other levels of education.

A Novel Assessment Tool for Quantitative Evaluation of Science Literature Search Performance: Application to First-Year and Senior Undergraduate Biology Majors

Expertise in searching and evaluating scientific literature is a requisite skill of trained scientists and science students, yet information literacy instruction varies greatly among institutions and programs. To ensure that science students acquire information literacy skills, robust methods of assessment are needed. Here, we describe a novel tool for longitudinal, crossover assessment of literature-searching skills in science students and apply it to a cross-sectional assessment of literature-searching performance in 145 first-year and 43 senior biology majors. Subjects were given an open-ended prompt requiring them to find multiple sources of information addressing a particular scientific topic. A blinded scorer used a rubric to score the resources identified by the subjects and generate numerical scores for source quality, source relevance, and citation quality. Two versions of the assessment prompt were given to facilitate eventual longitudinal study of individual students in a crossover design. Seniors were significantly more likely to find relevant, peer-reviewed journal articles, provide appropriate citations, and provide correct answers to other questions about scientific literature. This assessment tool accommodates large numbers of students and can be modified easily for use in other disciplines or at other levels of education.

Point‐of‐care cTnI Tests Accurately Predict Heat Stroke Severity; Proof of Concept for a Heat Stroke Field Test

Heat stroke (HS) remains a significant health threat for both civilians and the U.S. Armed Forces. Despite the substantial public health concern posed by HS, there is still no field or clinical test of HS severity. We suggested previously that cardiac troponin (cTnI) could serve as a robust biomarker of HS severity. In the current study, we hypothesized that cTnI band intensity on a small, portable, Lifesign MI© point‐of‐care strip (POC strip) could be used to predict overall HS severity and HS induced organ damage. In brief, conscious male rats (N=24) continuously monitored for HR, BP, and core temperature (Tc) (radiotelemetry) were taken to a TcMax of 41.9 ± 0.1°C and then allowed to recover for 24h at Ta of 20°C. Blood samples (~120µL) were taken at Tc,Max and 24hrs‐post heat via submandibular bleed and 4 drops of blood were placed on POC strips. POC cTnI band intensity was ranked using a simple 4 point scale via 2 blinded scorers, and then compared to cTnI levels measured by a clinical grade blood analyzer (iSTAT®). Blood was also analyzed for biomarkers of systemic organ damage (VetScan® Liver Profile). HS severity, as previously defined using in‐heat heart rate &amp; blood pressure as well as 24‐h recovery Tc profile, correlated strongly with cTnI (r= 0.82) &amp; markers of systemic organ damage (albumin r= 0.90, alanine aminotransferase r= 0.87, blood urea nitrogen r= 0.71). POC cTnI band intensity ranking accurately predicted cTnI levels (r= 0.80) and overall HS severity (r = 0.91). This suggests that qualitative cTnI POC tests can accurately determine HS severity and could serve as simple, portable, cost effective HS tests in the field. Author views not official US Army or DoD policy.

Intradiscal injection of simvastatin results in radiologic, histologic, and genetic evidence of disc regeneration in a rat model of degenerative disc disease

Background contextA large percentage of back pain can be attributed to degeneration of the intervertebral disc (IVD). Bone morphogenetic protein 2 (BMP-2) is known to play an important role in chondrogenesis of the IVD. Simvastatin is known to upregulate expression of BMP-2. Thus, we hypothesized that intradiscal injection of simvastatin in a rat model of degenerative disc disease (DDD) would result in retardation of DDD. PurposeThe purpose of the present study was to develop a novel conservative treatment for DDD and related discogenic back pain. Study design/settingThe setting of this study is the laboratory investigation. MethodsDisc injury was induced in 272 rats via 21-ga needle puncture. After 6 weeks, injured discs were treated with simvastatin in a saline or hydrogel carrier. Rats were sacrificed at predetermined time points. Outcome measures assessed were radiologic, histologic, and genetic. Radiologically, the magnetic resonance imaging (MRI) index (number of pixels multiplied by the corresponding image densities) was determined. Histologically, disc spaces were read by three blinded scorers using a previously described histologic grading scale. Genetically, nuclei pulposi were harvested, and polymerase chain reaction was run to determine relative levels of aggrecan, collagen type II, and BMP-2 gene expression. ResultsRadiologically, discs treated with 5 mg/mL of simvastatin in hydrogel or saline demonstrated MRI indices that were normal through 8 weeks after treatment, although this was more sustained when delivered in hydrogel. Histologically, discs treated with 5 mg/mL of simvastatin in hydrogel demonstrated improved grades compared with discs treated at higher doses. Genetically, discs treated with 5 mg/mL of simvastatin in hydrogel demonstrated higher gene expression of aggrecan and collagen type II than control. ConclusionsDegenerate discs treated with 5 mg/mL of simvastatin in a hydrogel carrier demonstrated radiographic and histologic features resembling normal noninjured IVDs. In addition, the gene expression of aggrecan and collagen type II (important constituents of the IVD extracellular matrix) was upregulated in treated discs. Injection of simvastatin into degenerate IVDs may result in retardation of disc degeneration and represents a promising investigational therapy for conservative treatment of DDD.

200: Pharmacodynamics of immunotherapy for opsoclonus–myoclonus syndrome: Impact on cytokines/chemokines

Pediatric opsoclonus–myoclonus syndrome (OMS) is a neuroblastoma-associated neuroinflammatory disorder. The purpose of this study was to evaluate the capacity of immunotherapies used in treating OMS to normalize several putative biomarkers of disease activity. Soluble BAFF and APRIL and a variety of chemokine ligands for CXCR3, CXCR5, CCR4, CCR7 were measured in cerebrospinal fluid (CSF) and serum by ELISA in 433 children (296 OMS, 109 controls, 28 other inflammatory neurological disorders/OIND). A blinded scorer rated motor severity on the validated OMS evaluation scale. In untreated OMS, significant elevations of CSF BAFF (+57%), CXCL10 (+2.7-fold), CXCL13 (+16.5-fold) and serum CCL22 (+55%) and CCL17 (+121%) concentrations were found ( P P = 0.0003). In conclusion, striking distinctions in immunotherapy effects on BAFF/APRIL, CXCL13/CXCR5, CCL22/CCR4, and CCL21/CCR7 signaling were found. The contrasting pharmacodynamic patterns may offer utility as treatment biomarkers.

Cerebellar Atrophy in Childhood Arterial Ischemic Stroke

Crossed cerebellar atrophy is uncommon in childhood arterial ischemic stroke. Acute corticospinal tract diffusion-weighted imaging (CST-DWI) changes occur in stroke of all ages. Contralateral CST-DWI is unexplained but approximates corticopontocerebellar pathways. We hypothesized that cerebellar atrophy can be quantified on clinical neuroimaging in childhood arterial ischemic stroke and is predicted by contralesional CST-DWI. Consecutive children (>28 days-18 years) were included with the following features: (1) acute, unilateral, middle cerebral artery arterial ischemic stroke, (2) DWI <14 days from stroke onset, (3) anatomic T1 MRI >6 months, and (4) Pediatric Stroke Outcome Measure >12 months. Blinded scorers measured cerebellar volumes (left/right/hemisphere/vermis/total) using Osirix software. Cerebellar volumes ratios (nonstroke/stroke) were expressed as asymmetry indices (AI), with chronic/acute ratio <1.0 suggesting crossed atrophy. Acute brain stem and cerebellum (peduncle, hemisphere) DWI ratios were scored. Software measures were compared with visual inspection. Associations between AI, motor outcome (good/poor), and contralesional CST-DWI were sought. Rater reliabilities were assessed. Twenty-three children were studied (median age, 6.3±4.4 years; 62% male). Baseline cerebellar volumes were comparable (right=56.9 cm(3), left=57.1 cm(3)). Cerebellar atrophy was suggested across the sample with overall AI <1.0 (0.973±0.05; P=0.009). Visual atrophy detection was specific (≈100%) but insensitive (54%). Children with poor motor outcome did not have lower AI (0.983±0.027 versus 0.965±0.068; P=0.40); however, children with acute contralesional CST-DWI did (0.928±0.078 versus 986±0.040; P=0.03). Acute cerebellar DWI did not predict atrophy. Rater reliabilities were excellent (>0.92). Cerebellar atrophy can be demonstrated on MRI in childhood arterial ischemic stroke. Association with acute contralesional pontine DWI signal suggests early degeneration of corticopontocerebellar connections. The clinical significance of cerebellar atrophy in childhood stroke remains to be determined.

BAFF/APRIL system in pediatric OMS: relation to severity, neuroinflammation, and immunotherapy

BackgroundB-cell dysregulation has been implicated but not fully characterized in pediatric opsoclonus-myoclonus syndrome (OMS), a neuroblastoma-associated neuroinflammatory disorder.ObjectiveTo assess the role of B-cell activating factor (BAFF) and a proliferation-inducing ligand (APRIL), two critical B cell-modulating cytokines, as potential biomarkers of disease activity and treatment biomarkers in OMS.MethodsSoluble BAFF and APRIL were measured in cerebrospinal fluid (CSF) and serum by ELISA in 433 children (296 OMS, 109 controls, 28 other inflammatory neurological disorders (OIND)). BAFF-R receptors on circulating CD19+ B cells were measured by flow cytometry. A blinded scorer rated motor severity on the OMS Evaluation Scale. Immunotherapies were evaluated cross-sectionally and longitudinally.ResultsThe mean CSF BAFF concentration, which was elevated in untreated OMS and OIND, correlated with OMS severity category (P = 0.006), and reduction by adrenocorticotropic hormone or corticotropin (ACTH) (−61%) or corticosteroids (−38%) was seen at each level of severity. In contrast, CSF APRIL was normal in OMS and OIND and unaffected by immunotherapy. When the entire OMS dataset was dichotomized into ‘high’ versus ‘normal’ CSF BAFF concentration, the phenotype of the high group included greater motor severity and number of CSF oligoclonal bands, and a higher concentration of inflammatory chemokines CXCL13 and CXCL10 in CSF and CXCL9 and CCL21 in serum. Serum APRIL was 6.7-fold higher in the intravenous immunoglobulins (IVIg) group, whereas serum BAFF was 2.6-fold higher in the rituximab group. The frequency of B cell BAFF-R expression was similar in untreated and treated OMS. Longitudinal studies of CSF BAFF revealed a significant decline in ACTH-treated patients (with or without rituximab) (P < 0.0001). Longitudinal studies of serum APRIL showed a 2.9-fold increase after 1 to 2 g/kg IVIg monotherapy (P = 0.0003).ConclusionsStriking distinctions in BAFF/APRIL signaling were found. OMS displayed heterogeneity in CSF BAFF expression, which met many but not all criteria as a potential biomarker of disease activity. We speculate that CSF BAFF may have more utility in a biomarker panel than as a stand-alone biomarker, and that the selective upregulation of both serum APRIL by IVIg and BAFF by rituximab, as well as downregulation of CSF BAFF by ACTH/steroids, may have utility as treatment biomarkers.