Kimbal Cooper

Prevalence of podiatric conditions seen in Special Olympics athletes: Structural, biomechanical and dermatological findings

BACKGROUND Podiatric conditions are frequently seen in persons with intellectual disabilities. Unfortunately, many are not detected or addressed. This study attempts to determine the prevalence of a variety of selected podiatric conditions in Special Olympics athletes and compare the findings to studies of a similarly matched general population. METHODS Screenings from 1580 Special Olympics athletes participating in a sampling of United States competitions in 2004-2006 were used for the analysis. The athletes voluntarily underwent a foot screening which followed the standardized Special Olympics Fit Feet protocol. RESULTS The most commonly identified conditions included overpronated gait, abducted gait, restriction in both the ankle and the first metatarsal phalangeal joint, pes planus, metatarsus adductus, brachymetatarsia, hallux abducto valgus, onychomycosis, onychocryptosis and tinea pedis. DISCUSSION/CONCLUSION The prevalence of all the structural conditions was notably higher than a similarly matched general population and both conditions and biomechanical/gait findings associated with ligamentous laxity like pes planus and hallux abducto valgus had especially high prevalence rates. Restricted motion in the ankle and first metatarsal phalangeal joint was surprising and may indicate some athletes have intellectual disabilities with contractures. The higher rates of dermatophyte related conditions may reflect increased exposure from the athletic competition environment, immunocompromise, issues with hygiene or both. Higher rates of onychocryptosis in the athletes may be due to ill fitting shoes and/or hygiene issues.

Zebrafish keratocyte explant cultures as a wound healing model system: differential gene expression & morphological changes support epithelial-mesenchymal transition.

The control of collective cell migration of zebrafish keratocyte sheets in explant culture is of interest for cell migration and epithelial wound healing and depends on the gene expression profile. In a zebrafish genome array, ∼17.5% of the probe sets were differentially expressed greater than two-fold (p≤0.003) between 1 and 7 days of explant culture. Among the differentially expressed genes were a variety of wound healing-related genes and many of the biomarkers for epithelial-mesenchymal transition (EMT), including a switch from keratin and E-cadherin to vimentin and N-cadherin expression and several EMT-related transcription factors were found to be differentially expressed. Supporting evidence for EMT is seen in both morphological change and rearrangement of the actin cytoskeleton and in expression of cadherins during explant culture with a visible disassembly of the cell sheet. TGFβ1 and TNFα expression were analyzed by qPCR at various time points and peak differential expression of both cytokines occurred at 3 days, indicating that the EMT process is ongoing under conditions routinely used in the study of fish keratocyte motility. These data establish that an EMT process is occurring during zebrafish keratocyte explant culture and support the use of this system as a wound healing model.

Mild aerobic exercise blocks elastin fiber fragmentation and aortic dilatation in a mouse model of Marfan syndrome associated aortic aneurysm

Regular low-impact physical activity is generally allowed in patients with Marfan syndrome, a connective tissue disorder caused by heterozygous mutations in the fibrillin-1 gene. However, being above average in height encourages young adults with this syndrome to engage in high-intensity contact sports, which unfortunately increases the risk for aortic aneurysm and rupture, the leading cause of death in Marfan syndrome. In this study, we investigated the effects of voluntary (cage-wheel) or forced (treadmill) aerobic exercise at different intensities on aortic function and structure in a mouse model of Marfan syndrome. Four-week-old Marfan and wild-type mice were subjected to voluntary and forced exercise regimens or sedentary lifestyle for 5 mo. Thoracic aortic tissue was isolated and subjected to structural and functional studies. Our data showed that exercise improved aortic wall structure and function in Marfan mice and that the beneficial effect was biphasic, with an optimum at low intensity exercise (55-65% V̇o2max) and tapering off at a higher intensity of exercise (85% V̇o2max). The mechanism underlying the reduced elastin fragmentation in Marfan mice involved reduction of the expression of matrix metalloproteinases 2 and 9 within the aortic wall. These findings present the first evidence of potential beneficial effects of mild exercise on the structural integrity of the aortic wall in Marfan syndrome associated aneurysm. Our finding that moderate, but not strenuous, exercise protects aortic structure and function in a mouse model of Marfan syndrome could have important implications for the medical care of young Marfan patients.NEW & NOTEWORTHY The present study provides conclusive scientific evidence that daily exercise can improve aortic health in a mouse model of Marfan syndrome associated aortic aneurysm, and it establishes the threshold for the exercise intensity beyond which exercise may not be as protective. These findings establish a platform for a new focus on promoting regular exercise in Marfan patients at an optimum intensity and create a paradigm shift in clinical care of Marfan patients suffering from aortic aneurysm complications.

TGFβ (transforming growth factor β) and keratocyte motility in 24 h zebrafish explant cultures

Fish keratocytes are used as a model system for the study of the mechanics of cell motility because of their characteristic rapid, smooth gliding motion, but little work has been done on the regulation of fish keratocyte movement. As TGFβ (transforming growth factor β) plays multiple roles in primary human keratinocyte cell migration, we investigated the possible involvement of TGFβ in fish keratocyte migration. Studying the involvement of TGFβ1 in 24 h keratocyte explant allows the examination of the cells before alterations in cellular physiology occur due to extended culture times. During this initial period, TGFβ levels increase 6.2‐fold in SFM (serum‐free medium) and 2.4‐fold in SFM+2% FBS (fetal bovine serum), while TGFβ1 and TGFβRII (TGFβ receptor II) mRNA levels increase ∼3‐ and ∼5‐fold respectively in each culture condition. Two measures of motility, cell sheet area and migration distance, vary with the amount of exogenous TGFβ1 and culture media. The addition of 100 ng/ml exogenous TGFβ1 in SFM increases both measures [3.3‐fold (P=4.5 × 10−5) and 26% (P=2.1 × 10−2) respectively]. In contrast, 100 ng/ml of exogenous TGFβ1 in medium containing 2% FBS decreases migration distance by 2.1‐fold (P=1.7 × 10−7), but does not affect sheet area. TGFβ1 (10 ng/ml) has little effect on cell sheet area in SFM cultures, but leads to a 1.8‐fold increase (P=1.5 × 10−2) with 2% FBS. The variable response to TGFβ1 may be, at least in part, explained by the effect of 2% FBS on cell morphology, mode of motility and expression of endogenous TGFβ1 and TGFβRII. Together, these results suggest that expression of TGFβ and its receptor are up‐regulated during zebrafish keratocyte explant culture and that TGFβ promotes fish keratocyte migration.

Prevalence of podiatric conditions seen in Special Olympics athletes: a comparison of USA data to an international population.

BACKGROUND Persons with intellectual disabilities frequently have podiatric conditions. Limited information exists on their prevalence in international cohorts of Special Olympics (SO) athletes. Findings from multiple United States (US) venues are compared to those from athletes screened at the 2011 Special Olympics World Summer Games in Athens, Greece (ATHENS). METHODS Data from Fit Feet screenings from 2096 ATHENS participants was compared to 7192 US participants. RESULTS Frequently noted in the ATHENS population were motion restriction in both the ankle and the first metatarsal phalangeal joint (1st MTPJ), pes planus, metatarsus adductus, brachymetatarsia, hallux abducto valgus (HAV), onychomycosis, onychocryptosis, and tinea pedis. ATHENS differed from the US cohort as HAV and restricted ankle joint and 1st MTPJ joint motion was less frequent. Significantly more tinea pedis, xerosis, and hyperhidrosis were present in the ATHENS population. DISCUSSION/CONCLUSION SO athletes have a higher prevalence of podiatric structural conditions compared to the general population, and some vary between ATHENS versus US. Less prevalent in ATHENS was HAV, and restricted motion in both the ankle and 1st MTPJ. This may reflect differences due to varied clinical observers. The higher rates of several dermatological conditions in ATHENS may reflect venue seasonal climate, or social factors.

Collective cell migration of primary zebrafish keratocytes

Fish keratocytes are an established model in single cell motility but little is known about their collective migration. Initially, sheets migrate from the scale at ~145 μm/h but over the course of 24h the rate of leading edge advance decreases to ~23 μm/h. During this period, leader cells retain their ability to migrate rapidly when released from the sheet and follower cell area increases. After the addition of RGD peptide, leader cell lamellae are lost, altering migratory forces within the sheet, resulting in rapid retraction. Leader and follower cell states interconvert within minutes with changes in cell-cell adhesions. Leader cells migrate as single cells when they detach from the leading edge and single cells appear to become leader cells if they rejoin the sheet. Follower cells rapidly establish leader cell morphology during closing of holes formed during sheet expansion and revert to follower cell morphology after hole-closure. Inhibition of Rho associated kinase releases leader cells and halts advancement of the leading edge suggesting an important role for the intercellular actomyosin cable at the leading edge. In addition, the presence of the stationary scale orients direction of sheet migration which is characterized by a more uniform advance of the leading edge than in some cell line systems. These data establish fish keratocyte explant cultures as a collective cell migration system and suggest that cell-cell interactions determine the role of keratocytes within the migrating sheet.

Epigenetically maintained SW13+ and SW13- subtypes have different oncogenic potential and convert with HDAC1 inhibition

BackgroundThe BRM and BRG1 tumor suppressor genes are mutually exclusive ATPase subunits of the SWI/SNF chromatin remodeling complex. The human adrenal carcinoma SW13 cell line can switch between a subtype which expresses these subunits, SW13+, and one that expresses neither subunit, SW13-. Loss of BRM expression occurs post-transcriptionally and can be restored via histone deacetylase (HDAC) inhibition. However, most previously used HDAC inhibitors are toxic and broad-spectrum, providing little insight into the mechanism of the switch between subtypes. In this work, we explore the mechanisms of HDAC inhibition in promoting subtype switching and further characterize the oncogenic potential of the two epigenetically distinct SW13 subtypes.MethodsSW13 subtype morphology, chemotaxis, growth rates, and gene expression were assessed by standard immunofluorescence, transwell, growth, and qPCR assays. Metastatic potential was measured by anchorage-independent growth and MMP activity. The efficacy of HDAC inhibitors in inducing subtype switching was determined by immunofluorescence and qPCR. Histone modifications were assessed by western blot.ResultsTreatment of SW13- cells with HDAC1 inhibitors most effectively promotes re-expression of BRM and VIM, characteristic of the SW13+ phenotype. During treatment, hyperacetylation of histone residues and hypertrimethylation of H3K4 is pronounced. Furthermore, histone modification enzymes, including HDACs and KDM5C, are differentially expressed during treatment but several features of this differential expression pattern differs from that seen in the SW13- and SW13+ subtypes. As the SW13- subtype is more proliferative while the SW13+ subtype is more metastatic, treatment with HDACi increases the metastatic potential of SW13 cells while restoring expression of the BRM tumor suppressor.ConclusionsWhen compared to the SW13- subtype, SW13+ cells have restored BRM expression, increased metastatic capacity, and significantly different expression of a variety of chromatin remodeling factors including those involved with histone acetylation and methylation. These data are consistent with a multistep mechanism of SW13- to SW13+ conversion and subtype stabilization: histone hypermodification results in the altered expression of chromatin remodeling factors and chromatin epigenetic enzymes and the re-expression of BRM which results in restoration of SWI/SNF complex function and leads to changes in chromatin structure and gene expression that stabilize the SW13+ phenotype.

Zebrafish keratocyte explants to study collective cell migration and reepithelialization in cutaneous wound healing.

Due to their unique motile properties, fish keratocytes dissociated from explant cultures have long been used to study the mechanisms of single cell migration. However, when explants are established, these cells also move collectively, maintaining many of the features which make individual keratocytes an attractive model to study migration: rapid rates of motility, extensive actin-rich lamellae with a perpendicular actin cable, and relatively constant speed and direction of migration. In early explants, the rapid interconversion of cells migrating individually with those migrating collectively allows the study of the role of cell-cell adhesions in determining the mode of migration, and emphasizes the molecular links between the two modes of migration. Cells in later explants lose their ability to migrate rapidly and collectively as an epithelial to mesenchymal transition occurs and genes associated with wound healing and inflammation are differentially expressed. Thus, keratocyte explants can serve as an in vitro model for the reepithelialization that occurs during cutaneous wound healing and can represent a unique system to study mechanisms of collective cell migration in the context of a defined program of gene expression changes. A variety of mutant and transgenic zebrafish lines are available, which allows explants to be established from fish with different genetic backgrounds. This allows the role of different proteins within these processes to be uniquely addressed. The protocols outlined here describe an easy and effective method for establishing these explant cultures for use in a variety of assays related to collective cell migration.

Reducing the Pain of Local 1% Lidocaine Infiltration with a Preceding Bacteriostatic Saline Injection A Double-blind Prospective Trial

BACKGROUND Lidocaine injection for local anesthesia is a common podiatric medical procedure. We tested the hypothesis that injection of bacteriostatic saline solution containing 0.9% benzyl alcohol before the lidocaine infiltration can reduce the burning caused by lidocaine injection. METHODS This double-blind prospective trial involved 45 participants who each received four injections in two areas of the dorsum of the foot and rated the perceived pain on a visual analog scale. The order of the injections was designed to disguise the control and intervention arms of the study. RESULTS The sensation of the lidocaine injection after the injection of saline was reduced significantly (P = .028). The percentage of lidocaine injections with visual analog scale scores of 0 increased by 36% after preinjection with bacteriostatic saline solution containing 0.9% benzyl alcohol. CONCLUSIONS The fact that 40% of the intervention visual analog scale pain scores for lidocaine injections were 0 suggests that a near painless lidocaine injection technique is an achievable goal and that the present technique is a simple and inexpensive method of reducing the pain of lidocaine injections.

Osteopathic Graduate Medical Education: New Research Standards Needed

The American Osteopathic Association (AOA) Council on Research, in their 2013-22 Research Stategic Plan for the Osteopathic Medical Profession1 and their subsequent article in The Journal of the American Osteopathic Association (JAOA),2 has called for setting research standards, vigorously encouraging research, and enforcing research standards at every college of osteopathic medicine (COM) and osteopathic postdoctoral training institution (OPTI). In light of this challenge, our group embarked on a review of the current state of research requirements in both osteopathic and allopathic graduate medical education (GME). for postdoctoral training regarding research