The effect of age on baseline neurocognitive performance.

OBJECTIVE: Evaluate the independent effect of age on baseline neurocognitive performance. STUDY DESIGN: Baseline ImPACT scores from tests taken by 7454 athletes aged 12-22 from 2009 to 2019 were split into three age cohorts: 12-14 years (3244), 15-17 years (3732), and 18-22 years (477). Linear regression analyses were used to evaluate the effect of age on ImPACT composite scores while controlling for demographic differences, medication-use, and symptom burden. Significance values have been set at p < 0.05. RESULTS: Linear regression analyses demonstrated that increased age does not significantly affect symptom score (ß = 0.06, p = 0.54) but does improve impulse control (ß = -0.45, p < 0.0001), verbal memory (ß = 0.23, p = 0.03), visualmotor (ß = 0.77, p < 0.0001), and reaction time (ß = -0.008, p < 0.0001) scores.  However, age did not have an effect on visual memory scores (ß = -0.25, p = 0.07). CONCLUSIONS: Age was shown to be an independent modifier of impulse control, verbal memory, visual motor, and reaction time scores but not visual memory or symptom scores.  This underscores the previous literature showing developmental differences as age increases among the adolescent athlete population.  This data also indicates the need for repeat neurocognitive baseline testing every other year as baseline scoring is likely to change as athletes become older.

Are Asian Americans BIPOC (Black, Indigenous, and People of Color)? Internalization of the model minority stereotype and COVID-19 racial bias on interracial solidarity toward Black Americans.

OBJECTIVES: Collective minoritized identities such as Black, Indigenous, and People of Color (BIPOC) may promote cross-racial solidarity. However, Asian Americans occupy a racially triangulated position as the "buffer minority" stereotyped as both the model minority and perpetual foreigner, complicating their inclusion in the BIPOC identity. The present study examined how the model minority and perpetual foreigner stereotypes relate to Asian Americans' perceived belongingness and identification with the BIPOC identity and, in turn, their interracial solidarity toward Black Americans. METHOD: A path analysis was conducted using cross-sectional data from a sample of 312 Asian Americans (Mage = 41.19, 69.87% college graduates, 46.79% foreign-born) to examine direct and indirect effects on Black community activism orientation (BCAO), with internalized model minority stereotype (MMS) and experiences of COVID-19 racial bias (CVRB) as exogenous predictors and perceived BIPOC inclusion and BIPOC self-identification as mediators. RESULTS: The BIPOC variables demonstrated a direct and positive association with BCAO. Furthermore, internalized MMS negatively related to BCAO through the sum effect of decreased perceived BIPOC inclusion and BIPOC self-identification, while experiences of CVRB positively related to BCAO through the sum effect of increased perceived BIPOC inclusion and BIPOC self-identification. CONCLUSIONS: Asian Americans' perceived BIPOC inclusion and BIPOC self-identification are shaped by their racially triangulated position, characterized by both the inhibiting effect of internalized MMS and the promoting effect of experiences of CVRB. This heightened or diminished BIPOC self-concept subsequently influenced their willingness to engage in interracial solidarity for Black Americans. (PsycInfo Database Record (c) 2024 APA, all rights reserved).

Navigating puberty, identity, and race among transnationally, transracially adopted Korean American adolescents.

This exploratory study examined the relation between pubertal timing and dimensions of ethnic-racial identity among adopted Korean Americans raised transracially in White families. The study also examined whether internalized racism moderated the association between pubertal timing and ethnic-racial identity. Adopted Korean American adolescents (N = 202; 108 females; ages 13-19 years) completed measures of pubertal development, ethnic-racial identity, and internalized racism in 2007. There was no significant main effect of pubertal timing for either male or female adolescents. Internalized racism moderated the relation between pubertal timing and ethnic-racial identity clarity (B = -.16, p = .015) among male adolescents. Specifically, earlier pubertal timing was significantly associated with lower ethnic-racial identity clarity for male adolescents with higher levels of internalized racism.

Bridging Neuroscience and Robotics: Spiking Neural Networks in Action.

Robots are becoming increasingly sophisticated in the execution of complex tasks. However, an area that requires development is the ability to act in dynamically changing environments. To advance this, developments have turned towards understanding the human brain and applying this to improve robotics. The present study used electroencephalogram (EEG) data recorded from 54 human participants whilst they performed a two-choice task. A build-up of motor activity starting around 400 ms before response onset, also known as the lateralized readiness potential (LRP), was observed. This indicates that actions are not simply binary processes but rather, response-preparation is gradual and occurs in a temporal window that can interact with the environment. In parallel, a robot arm executing a pick-and-place task was developed. The understanding from the EEG data and the robot arm were integrated into the final system, which included cell assemblies (CAs)-a simulated spiking neural network-to inform the robot to place the object left or right. Results showed that the neural data from the robot simulation were largely consistent with the human data. This neurorobotics study provides an example of how to integrate human brain recordings with simulated neural networks in order to drive a robot.

High altitude modulates concussion incidence, severity, and recovery in young athletes.

BACKGROUND: High altitude may affect concussion, but prior studies are limited . We tested whether high altitude affects sport-related concussion (SRC) incidence, severity, and recovery. METHODS: Twenty-five thousand eight hundred fifteen baseline and post-injury Immediate Post-Concussion Assessment and Cognitive Testing results were compiled from Florida and Colorado, low (27 m or 62 m) and high (1,640 m or 1,991 m) altitude locations, respectively. Incidence, severity, and recovery of injury were compared between altitudes. RESULTS: High altitude was associated with increased suspected concussion incidence (adjusted OR, 2.04 [95% CI, 1.86 to 2.24];P < .0001). However, high altitude was associated with lower concussion severity measured by Severity Index (SI) (adjusted OR, 0.42 [95% CI, 0.37 to 0.49];P < .0001). High altitude was associated with decreased recovery from post-concussive symptoms in the migraine (ß, -2.72 [95% CI, -3.31 to -2.13]; P < .0001), cognitive (ß, -1.88 [95% CI, -2.40 to -1.36]; P < .0001), and sleep symptom clusters (ß, -0.30 [95% CI, -0.52 to -0.08]; P = .007). Athletes with initial SI≥8 showed prolonged neurocognitive dysfunction at high altitude (HR, 1.38 [95% CI, 1.06 to 1.81]; P = .02). CONCLUSIONS: High altitude was associated with increased suspected concussions and prolonged recovery but less severe initial injury.

A Multivariable Analysis to Evaluate the Presence or Absence of Gender Differences in Baseline ImPACT Composite Scores and Symptom Severity Ratings in Student-Athletes Ages 12-18 Years.

INTRODUCTION: Gender differences in neurocognitive function have been reported over the past few decades. However, multiple studies that report gender differences in Immediate Post-Concussion Assessment and Cognitive Tests composite scores ignore potential confounders which may lead to inaccurate results. METHODS: A total of 4829 male and 2477 female baseline Immediate Post-Concussion Assessment and Cognitive Tests from 2009 to 2019 of subjects ages 12-18 years were used to evaluate gender differences in baseline neurocognitive scores and symptom severity ratings. Regression analyses were used to assess the effects of gender on neurocognitive performance at baseline while controlling for a number of potential confounders including symptom burden at the time of testing. RESULTS: Differences in 3 of 5 composite scores as well as severity rating scores were maintained in multivariate analysis. Females had increased Post-Concussion Symptom Scale (ß = 3.54, 95% confidence interval, 2.91 to 4.16, P < .0001) along with higher verbal memory (ß = 1.82, 95% confidence interval, 1.15 to 2.50, P < .0001) and visual motor (ß = 1.29, 95% confidence interval, 0.85-1.72, P < .0001) scores. CONCLUSIONS: Statistically significant gender differences were found in baseline neurocognitive function. This study clarifies for the first time that gender differences in these neurocognitive domains are not simply an artifact of differences in symptom burden. However, the small effect sizes call into question the clinical relevance of these differences.

Effects of a history of headache and migraine treatment on baseline neurocognitive function in young athletes.

OBJECTIVE/BACKGROUND: Despite the prevalence of concussions in young athletes, the impact of headaches on neurocognitive function at baseline is poorly understood. We analyze the effects of a history of headache treatment on baseline ImPACT composite scores in young athletes. METHODS: A total of 11,563 baseline ImPACT tests taken by 7,453 student-athletes ages 12-22 between 2009 and 2019 were reviewed. The first baseline test was included. There were 960 subjects who reported a history of treatment for headache and/or migraine (HA) and 5,715 controls (CT). The HA cohort included all subjects who self-reported a history of treatment for migraine or other type of headache on the standardized questionnaire. Chi-squared tests were used to compare demographic differences. Univariate and multivariate regression analyses were used to assess differences in baseline composite scores between cohorts while controlling for demographic differences and symptom burden. RESULTS: Unadjusted analyses demonstrated that HA was associated with increased symptoms (ß=2.30, 95% CI: 2.18-2.41, p<.0001), decreased visual memory (ß=-1.35, 95% CI: -2.62 to -0.43, p=.004), and increased visual motor speed (ß=0.71, 95% CI: 0.23-1.19, p=.004) composite scores. Baseline scores for verbal memory, reaction time, and impulse control were not significantly different between cohorts. Adjusted analyses demonstrated similar results with HA patients having greater symptom burden (ß=1.40, 95% CI: 1.10-1.70, p<.0001), lower visual memory (ß=-1.25, 95% CI: -2.22 to -0.27, p=.01), and enhanced visual motor speed (ß=0.60, 95% CI: 0.11-1.10, p=.02) scores. CONCLUSION: HA affected symptom, visual motor speed, and visual memory ImPACT composite scores. Visual memory scores and symptom burden were significantly worse in the HA group while visual motor speed scores were better, which may have been due to higher stimulant use in the HA group. The effects of HA on visual motor speed and visual memory scores were independent of the effects of the increased symptom burden.

Effects of Recurrent Mild Traumatic Brain Injuries on Incidence, Severity, and Recovery of Concussion in Young Student-Athletes.

OBJECTIVE: To examine the effects of recurrent concussions on the incidence, severity, and recovery of significant neurocognitive dysfunction (SND) in young athletes. SETTING: Various US youth sports organizations that utilize Immediate Post-Concussion Assessment and Cognitive Testing (ImPACT) for baseline and postinjury concussion testing. PARTICIPANTS: Data from 11 563 ImPACT baseline evaluations of US student-athletes aged 12 to 22 years were separated into 2 cohorts: subjects reporting 2 or more previous concussions (PC; n = 976 baseline evaluations) at baseline and a control group reporting zero previous concussions (CT; n = 7743 baseline evaluations). Subjects reporting 1 prior concussion were excluded. DESIGN: Retrospective cohort. MAIN MEASURES: Differences in SND incidence, severity, and recovery between the 2 cohorts were assessed using chi-squared tests, t tests, survival analyses, and multivariate regressions. RESULTS: The PC cohort had a higher incidence of head injury leading to ImPACT (436.7 per 1000 person-years vs 194.4 per 1000 person-years, P < .0001) and a higher incidence of SND (140.4 vs 71.8, P < .0001) than controls. However, the Severity Index (SI) demonstrated that SND severity was lower in the PC group (7.55 vs 8.59, P = .04). Adjusted analyses similarly demonstrated that the PC cohort had increased SND incidence (odds ratio = 1.93; 95% CI, 1.61 to 2.31; P < .0001), decreased SI (ß = -1.37; 95% CI, -2.40 to -0.34; P = .009), and equivalent recovery (hazard ratio = 0.98; 95% CI, 0.76 to 1.72; P = .90). CONCLUSION: Participants with a history of concussion have a higher incidence of SND but present with lower severity SND, which may be a result of increased concussion education or symptom awareness. Recurrent concussion has no significant impact on acute neurocognitive recovery. Together, these results provide evidence against the supposition that a history of concussion increases the severity of future SND.

Ym155 Induces Oxidative Stress-Mediated DNA Damage and Cell Cycle Arrest, and Causes Programmed Cell Death in Anaplastic Thyroid Cancer Cells.

Anaplastic thyroid cancer (ATC) is one of the most lethal malignancies with a median survival time of about 4 months. Currently, there is no effective treatment, and the development of new therapies is an important and urgent issue for ATC patients. YM155 is a small molecule that was identified as the top candidate in a high-throughput screen of small molecule inhibitors performed against a panel of ATC cell lines by the National Cancer Institute. However, there were no follow-up studies investigating YM155 in ATC. Here, we determined the effects of YM155 on ATC and human primary benign thyroid cell (PBTC) survival with alamarBlue assay. Our data show that YM155 inhibited proliferation of ATC cell lines while sparing normal thyroid cells, suggesting a high therapeutic window. YM155-induced DNA damage was detected by measuring phosphorylation of γ-H2AX as a marker for DNA double-strand breaks. The formamidopyrimidine-DNA glycosylase (FPG)-modified alkaline comet assay in conjunction with reactive oxygen species (ROS) assay and glutathione (GSH)/glutathione (GSSG) assay suggests that YM155-mediated oxidative stress contributes to DNA damage. In addition, we provide evidence that YM155 causes cell cycle arrest in S phase and in the G2/M transition and causes apoptosis, as seen with flow cytometry. In this study, we show for the first time the multiple effects of YM155 in ATC cells, furthering a potential therapeutic approach for ATC.

Standing by: The spatial organization of coercive institutions in China.

How do authoritarian states organize their coercive institutions over space? We argue that autocrats maximize the utility of their limited coercive resources by clustering them with perceived threats in society, i.e., segments of the population that are ideologically distant and have mobilizational potential. We test this proposition using a dataset that covers the universe of police stations (N = 147,428) and religious sites (N = 115,394) in China. We find that police stations are more likely to be located within walking distance of foreign religious sites (churches) than other sites (temples), even after controlling for the estimated population within 1 km of each site and a set of key site attributes. This finding is robust to using alternative model specifications, different variable measurements, and multiple data sources. Moving beyond the clustering pattern, we also address the temporal order issue and show that the Chinese state has allocated more new coercive resources around existing foreign religious sites than native sites, i.e., after these sites are already in place. This study enriches our understanding of how autocrats rule and further opens up an emerging new methodological avenue for research on authoritarian politics.

What drives consumer activism during trade disputes? Experimental evidence from Canada.

What drives consumer activism during trade disputes? We investigate this important and timely question using a survey experiment in the context of the recent Canada-US trade dispute. We find that Canadians are more likely to express willingness to take punitive actions in the form of boycotting during a trade conflict when they learn that Americans are taking such actions (retaliation), when many fellow citizens are taking such actions (peer pressure), and when they are rallied by their government (elite cue). Among the three conditions, peer pressure has the largest effect. These findings contribute to our understanding of the microfoundations of consumer activism during international trade disputes. They also have important policy implications in a world where both protectionism and populism are rising.

Psychological correlates of interest in genetic testing among Korean American adoptees and their parents.

Adopted persons increasingly have turned to genetic testing to obtain health information or to search for birth family. The present study investigated psychological factors that may contribute to interest among adoptees and their parents in genetic testing for the adoptee, including adoptees' ethnic identity development, their thoughts or curiosity about birth family (birth family thoughts), and the interaction of these two factors. Data were drawn from the second wave of a longitudinal study, conducted in 2014, on transracially, transnationally adopted Korean American adolescents and their adoptive parents. In a sample of 106 adolescent-parent dyads, 2 adolescents (1.89%) had undergone genetic testing. Among the dyads in which adolescents had not sought genetic testing, 47.12% of adolescents and 43.27% of parents indicated interest in genetic testing for the adolescent adoptee. Adolescents' interest in genetic testing was independent from parents' interest. Neither adolescent psychological adjustment nor physical health was related to interest in genetic testing in either adolescents or parents. Adolescents' birth family thoughts were related to adolescents' interest in genetic testing, but not to parents' interest in genetic testing for their child. This study showed ethnic identity exploration and resolution moderated the relationship between birth family thoughts and adolescents' interest in genetic testing. Results point to the relevance of birth family thoughts and identity development to genetic testing in transnational and transracial adolescent adoptees.

Platelet CD40 Mediates Leukocyte Recruitment and Neointima Formation after Arterial Denudation Injury in Atherosclerosis-Prone Mice.

The role of platelets in the development of thrombosis and abrupt closure after angioplasty is well recognized. However, the direct impact of platelets on neointima formation after arterial injury remains undetermined. Herein, we show that neointima formation after carotid artery wire injury reduces markedly in CD40-/- apolipoprotein E-deficient (apoE-/-) mice but only slightly in CD40 ligand-/-apoE-/- mice, compared with apoE-/- mice. Wild-type and CD40-deficient platelets were isolated from blood of apoE-/- and CD40-/-apoE-/- mice, respectively. The i.v. injection of thrombin-activated platelets into CD40-/-apoE-/- mice was performed every 5 days, starting at 2 days before wire injury. Injection of wild-type platelets promoted neointima formation, which was associated with increased inflammation by stimulating leukocyte recruitment via up-regulation of circulating platelet surface P-selectin expression and the formation of platelet-leukocyte aggregates. It was also associated with further promoting the luminal deposition of platelet-derived regulated on activation normal T cell expressed and secreted/chemokine (C-C motif) ligand 5 and expression of monocyte chemoattractant protein-1 and vascular cell adhesion molecule 1 in wire-injured carotid arteries. Remarkably, all these inflammatory actions by activated platelets were abrogated by lack of CD40 on injected platelets. Moreover, injection of wild-type platelets inhibited endothelial recovery in wire-injured carotid arteries, but this effect was also abrogated by lack of CD40 on injected platelets. Results suggest that platelet CD40 plays a pivotal role in neointima formation after arterial injury and might represent an attractive target to prevent restenosis after vascular interventions.

ATP1A3 mosaicism in families with alternating hemiplegia of childhood.

Alternating hemiplegia of childhood (AHC) is a rare and severe neurodevelopmental disorder characterized by recurrent hemiplegic episodes. Most AHC cases are sporadic and caused by de novo ATP1A3 pathogenic variants. In this study, the aim was to identify the origin of ATP1A3 pathogenic variants in a Chinese cohort. In 105 probands including 101 sporadic and 4 familial cases, 98 patients with ATP1A3 pathogenic variants were identified, and 96.8% were confirmed as de novo. Micro-droplet digital polymerase chain reaction was applied for detecting ATP1A3 mosaicism in 80 available families. In blood samples, four asymptomatic parents, including two paternal and two maternal, and one proband with a milder phenotype were identified as mosaicism. Six (7.5%) parental mosaicisms were identified in multiple tissues, including four previously identified in blood and two additional cases identified from paternal sperms. Mosaicism was identified in multiple tissues with varied mutant allele fractions (MAFs, 0.03%-33.03%). The results suggested that MAF of mosaicism may be related to phenotype severity. This is the first systematic report of ATP1A3 mosaicism in AHC and showed mosaicism as an unrecognized source of previously considered "de novo" AHC. Identifying ATP1A3 mosaicism provides more evidence for estimating recurrence risk and has implications in genetic counseling of AHC.

Inhibition of CD147 improves oligodendrogenesis and promotes white matter integrity and functional recovery in mice after ischemic stroke.

White matter damage is an important contributor to long-term neurological deficit after stroke. Our previous study has shown that inhibition of CD147 ameliorates acute ischemic stroke in mice. In this study, we aimed to investigate whether inhibition of CD147 promotes white matter repair and long-term functional recovery after ischemic stroke.Male adult C57BL/6 mice were subjected to transient (1-h) middle cerebral artery occlusion (tMCAO). Anti-CD147 function-blocking antibody (αCD147) was injected intravenously once daily for 3 days beginning 4 h after onset of ischemia. Sensorimotor and cognitive functions were evaluated up to 28 days after stroke. We found that αCD147 treatment not only prevented neuronal and oligodendrocyte cell death in the acute phase, but also profoundly protected white matter integrity and reduced brain atrophy and tissue loss in the late phase, leading to improved sensorimotor and cognitive functions for at least 28 days after stroke. Mechanistically, we found that αCD147 treatment increased the number of proliferating NG2(+)/PDGFRα(+) oligodendrocyte precursor cells (OPCs) and newly generated mature APC(+)/Sox10(+) oligodendrocytes after stroke, possibly through upregulation of SDF-1/CXCR4 axis in OPCs. In conclusion, inhibition of CD147 promotes long-term functional recovery after stroke, at least in part, by enhancing oligodendrogenesis and white matter repair.

Reconstruction of Symptomatic Congenital Anterior Cruciate Ligament Insufficiency.

BACKGROUND: Hypoplasia or congenital absence of the anterior cruciate ligament (ACL) is a rare disorder occurring in ∼1 in every 6000 births. Although some patients with hypoplasia or agenesis of the ACL may not complain of instability, others desire to participate in more demanding activities that require the stability of a competent ACL. There are limited reports of surgical treatment of this patient population. The purpose of this study was to report ACL reconstruction in a case series of patients with symptomatic congenital ACL deficiency. METHODS: A retrospective medical record review of the surgical treatment of 14 knees (13 patients) with congenital absence of the ACL at a tertiary care institution from 1995 to 2012 was performed. Patients with a minimum of 1 year of clinical follow-up were eligible for inclusion. RESULTS: The mean age at time of surgery was 12.6 (range, 3 to 22), including 6 patients <12 years of age. Mean follow-up was 2.9 years (range, 1 to 6.6). Nine of 13 patients (69%) had underlying congenital abnormalities/associated syndromes. Preoperative Lachman and pivot shift examination was International Knee Documentation Committee grade C or D in all but 1 knee. ACL reconstruction was performed with combined intra-articular/extra-articular physeal sparing reconstruction with iliotibial band (n=5), autograft hamstring (n=2) or bone-patellar tendon-bone (n=3), or allograft (n=4). Multiligament reconstruction of associated ligamentous deficiency was performed in 7 knees (50%). Postoperative Lachman and pivot shift testing was International Knee Documentation Committee (IKDC) grade A or B in all but 1 knee. One patient with congenital absence of multiple knee ligaments required revision ACL reconstruction surgery, with concurrent first-time posterior cruciate ligament reconstruction, due to persistent instability. None required revision surgery due to graft tear at a minimum of 1-year follow-up. CONCLUSIONS: Surgical stabilization of symptomatic congenital ACL insufficiency, with associated ligamentous reconstruction as required on a case-by-case basis, results in improved stability at early clinical follow-up, with low complication rates. LEVEL OF EVIDENCE: Level IV-retrospective case series.

Taurine Reduces tPA (Tissue-Type Plasminogen Activator)-Induced Hemorrhage and Microvascular Thrombosis After Embolic Stroke in Rat.

BACKGROUND AND PURPOSE: Taurine (2-aminoethansulfolic amino acid) exerts neuroprotective actions in experimental stroke. Here, we investigated the effect of taurine in combination with delayed tPA (tissue-type plasminogen activator) on embolic stroke. METHODS: Rats subjected to embolic middle cerebral artery occlusion were treated with taurine (50 mg/kg) at 4 hours in combination with tPA (10 mg/kg) at 6 hours. Control groups consisted of ischemic rats treated with either taurine (50 mg/kg) or saline at 4 hours or tPA (10 mg/kg) alone at 2 or 6 hours after middle cerebral artery occlusion. RESULTS: We found that combination treatment with taurine and tPA robustly reduced infarct volume and neurological deficits 3 days after stroke, whereas treatment with taurine alone had a less-significant protective effect. tPA alone at 6 hours had no effects on infarct volume but instead induced intracerebral hemorrhage. The combination treatment with taurine prevented tPA-associated hemorrhage and reduced intravascular deposition of fibrin/fibrinogen and platelets in downstream microvessels and hence improved microvascular patency. These protective effects are associated with profound inhibition of CD147 (cluster of differentiation 147)-dependent MMP-9 (matrix metalloproteinase-9) pathway in ischemic brain endothelium by taurine. Notably, targeted inhibition of CD147 by intracerebroventricular injection of the rat CD147 siRNA profoundly inhibited ischemia-induced and tPA-enhanced MMP-9 activity in ischemic brain endothelium and blocked tPA-induced cerebral hemorrhage. Finally, the combination treatment with taurine and tPA improved long-term outcome at least 45 days after stroke compared with saline-treated group. CONCLUSIONS: Our results suggest that taurine in combination with tPA may be a clinically feasible approach toward future attempts at combination stroke therapy.

Profiling the RNA editomes of wild-type C. elegans and ADAR mutants.

RNA editing increases transcriptome diversity through post-transcriptional modifications of RNA. Adenosine deaminases that act on RNA (ADARs) catalyze the adenosine-to-inosine (A-to-I) conversion, the most common type of RNA editing in higher eukaryotes. Caenorhabditis elegans has two ADARs, ADR-1 and ADR-2, but their functions remain unclear. Here, we profiled the RNA editomes of C. elegans at different developmental stages of wild-type and ADAR mutants. We developed a new computational pipeline with a "bisulfite-seq-mapping-like" step and achieved a threefold increase in identification sensitivity. A total of 99.5% of the 47,660 A-to-I editing sites were found in clusters. Of the 3080 editing clusters, 65.7% overlapped with DNA transposons in noncoding regions and 73.7% could form hairpin structures. The numbers of editing sites and clusters were highest at the L1 and embryonic stages. The editing frequency of a cluster positively correlated with the number of editing sites within it. Intriguingly, for 80% of the clusters with 10 or more editing sites, almost all expressed transcripts were edited. Deletion of adr-1 reduced the editing frequency but not the number of editing clusters, whereas deletion of adr-2 nearly abolished RNA editing, indicating a modulating role of ADR-1 and an essential role of ADR-2 in A-to-I editing. Quantitative proteomics analysis showed that adr-2 mutant worms altered the abundance of proteins involved in aging and lifespan regulation. Consistent with this finding, we observed that worms lacking RNA editing were short-lived. Taken together, our results reveal a sophisticated landscape of RNA editing and distinct modes of action of different ADARs.

Three-Dimensional Printing of Cell Exclusion Spacers (CES) for Use in Motility Assays.

PURPOSE: Cell migration/invasion assays are widely used in commercial drug discovery screening. 3D printing enables the creation of diverse geometric restrictive barrier designs for use in cell motility studies, permitting on-demand assays. Here, the utility of 3D printed cell exclusion spacers (CES) was validated as a cell motility assay. METHODS: A novel CES fit was fabricated using 3D printing and customized to the size and contour of 12 cell culture plates including 6 well plates of basal human brain vascular endothelial (D3) cell migration cells compared with 6 well plates with D3 cells challenged with 1uM cytochalasin D (Cyto-D), an F-actin anti-motility drug. Control and Cyto-D treated cells were monitored over 3 days under optical microscopy. RESULTS: Day 3 cell migration distance for untreated D3 cells was 1515.943µm ± 10.346µm compared to 356.909µm ± 38.562µm for the Cyt-D treated D3 cells (p < 0.0001). By day 3, untreated D3 cells reached confluency and completely filled the original voided spacer regions, while the Cyt-D treated D3 cells remained significantly less motile. CONCLUSIONS: Cell migration distances were significantly reduced by Cyto-D, supporting the use of 3D printing for cell exclusion assays. 3D printed CES have great potential for studying cell motility, migration/invasion, and complex multi-cell interactions.