bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract P675: Cardiovascular and Cerebrovascular Events in Status Migrainosus and Migraine With Aura- A National Estimate
Authors: Patel Priyadarshee , Patel Urvish K , Grewal Prabhsharn , Reddy Gowry , Sosale Vasudha , Yar Khan Nibras , Mijares Aaron , Martin Mehwish , Urhoghide Eseosa , Patel Jahanvi , Bhushan Tanushri , Shah Shamik
Published: March 2021

Objective:
Primary aim of this study is to compare the prevalence of vascular events [VEs - acute ischemic stroke (AIS), transient ischemic attack (TIA), acute myocardial infarction (AMI) and angina] among patients with status migrainosus and intractable migraine (SM), migraine with aura (MWA), migraine without aura (MWOA) and non-migraineurs. Secondary aim of this study is to evaluate the linkage of VEs and associated disability with different migraine subtypes.

Methods:
A cross-sectional study was performed using National Inpatient Sample [2016-2017] for adult hospitalizations. We extracted a cohort of secondary diagnoses of specific migraine subtypes amongst which primary diagnoses of VEs were identified using ICD 10 code. Weighted analysis using chi-square and multivariable survey logistic regression was performed to identify prevalence of VEs and evaluate relationship and disability (using APRDRG Risk Severity Score) between VEs and migraine subtypes.

Results:
Out of total 58,259,589 US hospitalizations, 0.1%, 0.08% and 0.03% had SM, MWA and MWOA, respectively. Prevalence of AIS (5.5 vs 0.96 vs 1.96 vs 1.71%) and TIA (3.99 vs 1.07 vs 0.49 vs 0.38%) was higher amongst MWA in comparison with SM, MWOA and non-migraineurs, respectively. (p<.0001) Prevalence of AMI was lower amongst migraineurs (SM:0.40 vs MWA:0.86 vs MWOA:0.86 vs non-migraineurs:2.15%) but prevalence of angina was higher amongst MWOA (0.01 vs 0.07 vs 0.21 vs 0.06%). (p<.0001) MWA and MWOA were associated with higher odds of AIS [MWA - aOR 3.7; 95%CI 3.4-4.1; MWOA - 1.7 (1.3-2.1)] and TIA [MWA - 11.8 (10.5-13.1); MWOA - 4.0 (2.9-5.6)] compared to non-migraineurs. Angina was associated with MWOA [3.0 (1.4-6.8)]. AIS hospitalization with SM had 52% [1.52 (1.2-1.9)] higher odds of extreme/severe disability compared to MWA. TIA patients with SM had higher odds of disability compared to MWA [3.0 (2.1-4.3)] and non-migraineurs [1.9 (1.4-2.6)]. Angina with MWOA had a significant disability [1.5 (1.04-2.2)] compared to non-migraineurs.

Conclusion:
SM and MWA had a significant role not only to predict AIS, TIA and angina but also increase disability. Early identification and management of these migraine subtypes may prevent VEs and mitigate disability associated with them.
Abstract P687: Acute Kidney Failure in Patients With Malignant Middle Cerebral Artery Infarction Undergoing Osmotherapy With Mannitol
Authors: Moustafa Haidar , Schoene Daniela , Pallesen Lars-Peder , Prakapenia Alexandra , Siepmann Timo , Puetz Volker , Barlinn Kristian , Barlinn Jessica
Published: March 2021

Introduction:
To explore kidney safety profile of osmotic diuretic mannitol in patients with malignant middle cerebral artery (MCA) infarction.

Methods:
We analyzed data from consecutive patients with malignant MCA infarction (01/2008-12/2017). Malignant MCA infarction was defined according to clinical and radiographic DESTINY criteria. Clinical and laboratory variables were collected for all patients. We compared clinical endpoints including acute kidney failure (AKF; according to Kidney Disease: Improving Global Outcomes [KDIGO] definition) and hemodialysis between patients who received mannitol and those who did not. Multivariable model was built to explore predictor variables of AKF, in-hospital death and functional outcome at discharge.

Results:
Overall, 228 patients with malignant MCA infarction were analyzed: median age 67 years (IQR, 56-76), 58% men, median NIHSS 23 (17-32) points. Decompressive craniectomy was performed in 103/228 (45.2%) patients. Mannitol was administered in 100/228 (43.9%) patients over an average of 85 (30.3-139.3) hours. Average dosage was 700 (250-1050) g. Patients treated with mannitol more frequently suffered from AKF (40% vs. 7.9%; p<0.0001) and needed hemodialysis (7.4% vs. 0.9%; p=0.024) than patients without mannitol. At discharge, kidney function completely recovered in 38.5% and 20%, respectively (p=0.459). In multivariable model adjusted for age, history of chronic kidney disease, nephrotoxic premedication, admission systolic blood pressure, concurrent urinary tract infection, contrast agent exposure and peak osmolality, mannitol therapy emerged as single predictor of AKF (OR 4.14, 95%CI 1.2-14.2; p=0.024). Neither AKF nor mannitol therapy was associated with in-hospital death or short-term functional outcome (p>0.05).

Conclusions:
Acute kidney failure appears to be a frequent complication of osmotic diuretic mannitol in patients with malignant MCA infarction. Given the lack of evidence supporting effectiveness of mannitol in these patients, its use should be carefully considered.
Abstract P700: Stroke in Lupus Spectrum Disorder: A Single-Center Case Series in the Kentucky Appalachian Region
Authors: Grewal Parneet , Pettigrew Luther C
Published: March 2021

Background:
Lupus spectrum disorders (LSDs) encompass systemic lupus erythematosus (SLE), lupus nephritis, antiphospholipid syndrome (APS), and mixed connective tissue disease. Neuro-vascular disorders associated with LSDs include headache, transient ischemic attack, vascular cognitive impairment, immune-mediated cerebral vasculitis, thrombo-embolic stroke, cortical venous sinus thrombosis (CVST), and intracranial hemorrhage. We conducted a retrospective survey of neuro-vascular complications of LSDs in a university hospital-based stroke service within Kentucky Appalachia.

Methods:
Consecutive hospital admissions and consultations were retrospectively surveyed from January 2012 to January 2017. Inpatient medical records were reviewed to accumulate laboratory data and neuro-imaging studies. Diagnoses of SLE and other LSDs conformed to Systemic Lupus International Collaborating Clinics criteria.

Results:
During this 5-year survey, 10,953 cases of ischemic or hemorrhagic stroke were admitted or consulted and 75 patients (0.7%) met criteria for LSD. Female:male ratio favored women over men (4:1) and the median age was 48 years (range 22 - 87). The most common LSD was SLE and/or APS (n = 61; 81.3% total). The most frequent neuro-vascular complication was acute thrombo-embolic stroke (n = 30; 40%). Among 42 (56%) ischemic stroke patients, 17 had multifocal infarcts, 11 had isolated cortical strokes, and 7 had lacunes in the subcortical structures or brainstem. The majority of ischemic stroke patients (n = 48; 64%) did well and were ultimately discharged to home; 19 (25.3%) were re-admitted with recurrent stroke. Four patients had vasculitis (5.3%) and three had moyamoya syndrome (4%). Intracranial hemorrhage affected four patients (5.3%); hemorrhagic cerebral micro-angiopathy occurred in three (4%). Two patients had CVST (2.7%).

Conclusion:
Neuro-vascular complications of LSDs are heterogeneous and affect youthful patients with female preponderance. The large proportion of thrombo-embolic stroke in our population may be attributed to immune-mediated arterial coagulopathy or cardiac structural disease. Most patients with LSD complicated by ischemic stroke had good outcome, but there was high risk of readmission due to recurrence.
Abstract P706: Age and Elevated Coronary Artery Calcium Score as a Predictor of Cerebral Ischemic Events
Authors: Singh Aditya , Stys Tom , Thorp Andrew , Bares Valerie , Wilson Jeffrey , Stys Adam
Published: March 2021

Introduction:
Coronary artery calcium (CAC) score is a good predictor of coronary plaque burden and cardiovascular events, however its role in ischemic stroke and transient ischemic attacks (TIA) has been sparsely studied.

Methods:
Participants age ≥18 years with heart screen done from Nov 2008- Feb 2019 were selected and were assessed for documented Ischemic events (Ischemic stroke and TIA) after their heart screen. Only the most recent heart screen per person was considered. Patients were further divide into two groups based on their age. Comparisons between the two age groups was done using chi-square for categorical variables. Logistic regression was used to assess age and CAC as predictors of ischemic events and ROC curves were formed.

Results:
A total of 330896 individuals were included in the study over a span of 10 years, of which 53.9% were females. 894 (0.2%) patients were observed to have a documented diagnosis of ischemic stroke or TIA. Older patients (Age ≥50) were found to have significantly higher percentage of ischemic events as compared to young patients (3.5% vs 0.9%, p<0.001), which was also observed among various CAC groups. There was also a significant difference in distribution of CAC where older patients had higher CAC even when splitting the cohort into distinct CAC categories (P <0.001). On regression analysis elevated CAC and age were found to be a good predictors of ischemic events. Predictability of cerebrovascular ischemic events was significantly higher when elevated CAC score and patient age were considered together as compared age and CAC alone (P<0.001).

Conclusion:
Elevated coronary artery calcium score and age are a good predictor of ischemic stroke and TIA.
Abstract P717: Central Midbrain Strokes Are More Liable for Respiratory Failure Then Other Brain Stem Strokes
Authors: Hussein Omar M , Caceres Eder , saba kasser , Kamdar Hera , Sawalha khalid , Hamed Mohammad , Abd Elazim Ahmed , Alsarah Ali A , Davis Herbert
Published: March 2021

Background:
Respiratory centers are known to be present in the central medulla oblongata and pons. There are multiple complex respiratory networks involving these centers. The midbrain periaqueductal grey is believed to act as a regulator of the respiratory function. The effect of brain-stem strokes on respiration remains understudied. There is a lack of clear understanding of the anatomical influence of such strokes on respiration. We attempted to identify brain-stem locations with the highest liability for respiratory failure in case of stroke.

Methods:
We included all ischemic and hemorrhagic brain-stem strokes from our stroke-registry between 2016 and 2018 then performed univariate/multivariate regression-analyses on variables that might predict respiratory failure and the need for intubation. The brain stem was divided into nine locations (right lateral, central, left lateral in each of the midbrain, pons, and medulla oblongata).

Results:
Out of 128 brain-stem strokes of different sizes and etiologies, central midbrain strokes were the only significant and independent affected location associated with respiratory failure and endotracheal intubation (coefficient= 0.1256, 95%-CI= 0.0175, 0.2338, p= 0.023). R-squared was equal to 15% when only central midbrain strokes stayed in the model.

Conclusions:
While one might assume that central medullary and pontine strokes have the most impact on respiration; our results show that central midbrain is the most impactful, accounting for about 15% of respiratory instability associated with brain-stem strokes. This can be explained by the adaptive nature of respiratory circuits within the medulla and pons. Central periaqueductal grey within the midbrain controls the rate and depth of respiration and might not have the same flexibility present elsewhere.
Abstract P719: Diet Counseling in Post-Stroke Patients in the United States
Authors: Patel Vishal , Gandrakota Nikhila , Moore Miranda , Polu Ashok , Shah Megha
Published: March 2021

Introduction:
Research demonstrates dietary modifications in post-stroke patients facilitated by nutrition consultations improve post stroke recovery and reduce risk of recurrent stroke. Racial/ethnic differences in post-stroke nutrition practices demonstrate that focused dietary counseling of minority groups may be needed. It is unknown to what extent clinicians are providing dietary counseling in ambulatory office visits.

Methods:
We used data from the 2011 to 2016 National Ambulatory Medical Care Surveys to assess trends in post-stroke diet consultations during routine office visits across all races and ethnicities. We use multivariate logistic regression models to assess variation in the provision of dietary counseling by patient and provider characteristics.

Results:
There were 4,960 patients with a history of stroke among a total of 249,282 in the study period. The proportion of patients with stroke over the age of 60 who received diet counseling increased from 6% in 2012 (95% Confidence Interval (CI)=4%, 9%) to 14% in 2016 (95% CI= 7%, 21%). Adjusted models showed that other, non-Hispanic patients had a higher likelihood of receiving diet counseling, compared with whites (Odds Ratio (OR)=1.80, 95% CI=0.85, 3.82). Patients in the South were more likely to receive diet counseling than patients from the West (OR=1.47, 95% CI=0.81, 2.67). Patients in metropolitan areas had (OR=1.84, 95% CI=1.09, 3.09) a higher likelihood of counseling compared to patients in rural areas (OR=1.84, 95% CI=1.09, 3.09). Neurologists were less likely to provide counseling when compared to family medicine physicians (OR=0.14, 95% CI=0.07, 0.28).

Conclusion:
Major gaps in post-stroke dietary counseling exist; though the proportion of Americans over 60 receiving dietary counseling in ambulatory visits has been increasing in the past half-decade. Future interventions should focus on addressing gaps in counseling.
Abstract P721: Parkinson’s Disease Patients With Ischemic Stroke Have Shorter Hospital Stay Compared to Those Without Parkinson’s Disease: A National Inpatient Sample Study 1998-2017
Authors: Moreno Gomez Veronica , Shoaib Maria , Khaliq Muhammad Farhan , Xu Chao , Copeland Timothy , Ortiz Garcia Jorge G
Published: March 2021

Introduction:
A paucity of studies exists about the influence of Parkinson’s Disease (PD) in the length of stay (LOS) among patients admitted with acute ischemic stroke (AIS). We seek to identify if a prior history of PD impacts the LOS of AIS patients, and to establish the factors associated it.

Methodology:
We analyzed data of the NIS from the Healthcare Cost and Utilization Project (HCUP) from 1998 to 2017, using a negative binomial regression to assess sociodemographic factors and hospital characteristics associated with LOS in AIS patients with PD.

Results:
A total of 835,380,672 hospitalizations were identified with discharge diagnosis of AIS. The cohort was divided into 2 groups based on presence of history of PD during the same admission. 12,799,437 of AIS patients had discharge records with a secondary diagnosis of PD. Non-Parkinson’s disease cohort had longer hospital stay (Incidence Rate Ratio IRR 1.02 (95% CI 1.01-1.03). An increased LOS was observed in males compared to females (IRR 1.03 (1.00-1.03)); Black (IRR 1.17 (95% CI 1.16-1.18)), Hispanics (IRR 1.13 (95% CI 1.11-1.14)) and Asians (IRR 1.22 (95% CI 1.19-1.26)) when compared to whites, Medicaid when compared to Medicare (IRR 1.37 (95% CI 1.34-1.41)). Higher Elixhauser index corresponded to longer LOS (IRR 1.14 (CI 1-14-1.14)). In terms of hospital factors, Urban teaching hospital was associated with longer LOS when compared to rural hospital (IRR 1.18 (95% CI 1.16-1.19)), Northeast compared to West region hospital areas (IRR 1.3 (95% CI 1.27-1.33)). Decreasing LOS was seen over the course of last 20 years.

Conclusion:
Admission for ischemic stroke tended to result in longer length of stay among non-Parkinson’s patients. Patient characteristics are a greater source of variability in length of stay than Parkinson’s disease status.
Abstract P738: Matrix Metalloproteinase-3 (mmp3) Activation Impairs Endothelial Integrity and Increases Vascular Injury After Ischemic Stroke in Female Diabetic Rats
Authors: Abdul Yasir , JAMIL SARAH , Li Weiguo , Ergul Adviye
Published: March 2021

Introduction:
Females patients with diabetes suffer from poor outcomes of ischemic stroke but underlying reasons are not fully understood. We have shown that 1) adult female diabetic rats develop greater infarct and hemorrhagic transformation (HT) leading to poorer outcomes after ischemic stroke, and 2) MMP3 activity is increased to a greater extent in female endothelial cells as well as the microvasculature of diabetic female rats than in males. This led us to hypothesize that MMP3 mediates disruption of endothelial integrity amplifying vascular injury in female diabetic rats.

Methods:
Diabetic female Wistar rats, subjected to 60 min MCAO, received a single dose of MMP3 inhibitor (UK356618; 15mg/kg; iv) or vehicle after reperfusion. On Day 3, adhesive removal time (ART), behavioral composite score, brain infarct size, edema and macroscopic HT were recorded. Primary brain microvascular endothelial cells (BMVECs) isolated from female rats were cultured in 25mM glucose plus 100μM sodium palmitate for 48 hours followed by hypoxia insult for 24 hours in the presence/absence of UK356618 (20nM). Endothelial phenotype and integrity were assessed.

Results:
Brain edema and HT scores were lower and outcomes were improved with MMP3 inhibition but infarct size was not different between the groups (Table). Hypoxia decreased tight junction protein occludin-1 while increasing transforming growth factor receptor-1 (TGF-R1), a key modulator of endothelial phenotype. Treatment with inhibitor UK356618 reversed these responses.

Conclusions:
The lack of a difference in infarct size with the treatment suggests that MMP3 inhibition improves short term outcomes in female diabetic rats via preservation of endothelial integrity. The decrease in TGF-R1 needs to be further investigated for long term effects of MMP3 inhibition on vascular restoration and recovery after stroke in diabetes.
Abstract P749: Functional and Molecular Characterization of Endothelial and Vascular Smooth Muscle Cells Derived From Moyamoya Disease-Induced Pluripotent Stem Cells
Authors: Rao Shailaja , Uchino Haruto , Pendharkar Arjun V , Zhang Qian , Cheng Michelle Y , Steinberg Gary
Published: March 2021

Background:
Moyamoya disease (MMD) is a rare, progressive steno-occlusive cerebrovascular disorder of the internal carotid artery, leading to stroke. Affected arteries exhibited thickened intima with depleted elastic lamina and media, indicating a dysfunction of the vascular smooth muscle cells (VSMCs) and endothelial cells (ECs). However, the pathogenesis of the disease is still unclear. We aim to address this gap in knowledge by using patient derived induced pluripotent stem cells (iPSCs), to generate VSMCs and ECs.

Methods:
Peripheral blood mononuclear cells (PBMCs) from controls and MMD patients (n=3 per group) were used for generating iPSCs. Functional properties of differentiated ECs and VSMCs in normoxia/hypoxia model (1%O
2
) were assessed for cell proliferation by BrDU incorporation, migration by scratch assays and apoptosis by exposure to hydrogen peroxide (H2O2).
In vitro
angiogenic tube formation was assessed with ECs alone, as well as ECs and VSMCs as a co-culture. Hypoxia inducible factor 1α (HIF1α) and Intercellular adhesion molecule-1 (ICAM-1) activation was determined using qPCR and western blot in VSMCs.

Results:
Functional proliferative assays showed that MMD ECs proliferated faster than control ECs. Migration assays showed that MMD ECs migrate slower in response to VEGF after hypoxia. MMD ECs were found to be more sensitive to insults such as H2O2 treatment and exhibited more apoptosis. In contrast, MMD VSMCs proliferate and migrate similar to controls, but exhibited elevated levels of HIF1α, ICAM-1 mRNA and protein expression. MMD and control ECs showed similar levels of tube formation in single cultures, however, when co-culturing with VSMCs, MMD VSMCs failed to support EC tubes beyond 24 h, resulting in tube destabilization.

Conclusions:
Our preliminary results indicate that both MMD VSMCs and ECs are dysfunctional and may be related to the elevated expression of HIF1α and ICAM-1, possibly contributing to MMD pathology. Current ongoing studies include investigating the interactions between MMD VSMCs and ECs using co-cultures, as well as transcriptome analysis of these differentiated cells, which will provide important insights into the cellular and molecular mechanisms underlying MMD.
Abstract P750: 3K3A-APC Restores Oligodendrocyte Pools in Models of White Matter Stroke via PAR1 Signaling
Authors: Nikolakopoulou Angeliki M , Wang Yaoming , Dai Zhonghua , Huuskonen Mikko , Montagne Axel , Lazic Divna , Sagare Abhay , Zhao Zhen , Griffin John H , Zlokovic Berislav V
Published: March 2021

A recombinant variant of activated protein C, 3K3A-APC, was recently tested in a successful phase II clinical trial for ischemic stroke (RHAPSODY), which demonstrated that the treatment may reduce hemorrhages and is safe in patients that were treated with tPA and/or thrombectomy. Animal models of large ischemic stroke (middle cerebral artery occlusion) and neurodegeneration have shown that the effects of 3K3A-APC are mediated by preserved APC cell signaling activities (anti-apoptotic neuronal protection, protection of blood-brain barrier (BBB) integrity and anti-inflammatory activities). Because recent studies have suggested a role of protease activated receptor 1 (PAR1) in oligodendrocyte recovery after white matter damage, we wanted to test whether biased PAR1 signaling mediated by 3K3A-APC would protect oligodendrocytes in
in vivo
and
ex vivo
models of ischemic white matter stroke. First, we used a mouse model of white matter stroke induced by injecting a vasoconstrictor
N
5
-(1-Iminoethyl)-L-ornithine (L-NIO) into the corpus callosum. Our results show that treatment with recombinant murine 3K3A-APC (0.2 mg/kg, starting 4h after stroke) protected oligodendrocytes (OLS, Olig2+CNPase+) and oligodendrocyte precursor cells (OPCs, Olig2+PDGFRα+) from cell death at 1 and 7 days after stroke.
In vitro
, 3K3A-APC protected primary mouse oligodendrocytes from apoptosis when exposed to OGD conditions at nanomolar concentrations (EC50 concentration was 5.2 nM). Improved oligodendrocyte viability
in vivo
was associated by about ~67% reduction in overall cell death (cresyl violet staining, T2w and diffusion tensor MRI) and reduced BBB leakage (fibrin staining and DCE-MRI) as well as improved functional recovery (adhesive removal and grid walking tests). Protective effects of 3K3A-APC were abolished by silencing of PAR1 with small interfering RNA (siRNA)
in vivo
and
in vitro
. In conclusion, PAR1 mediated signaling initiated by 3K3A-APC protects oligodendrocyte pools in models of ischemic white matter damage with preservation of neurovascular unit integrity.
Abstract P753: Comparison of Calibration vs Non-Calibration Techniques in the Automated Capture of Eye Movement Data: Initial Validation of the Roadie Device for Detecting Posterior Circulation Stroke
Authors: Hassan Mohamed , Aldridge Chad , Zhuang Yan , McMurry Timothy L , Rohde Gustavo , Southerland Andrew M
Published: March 2021

Background:
Posterior circulation stroke (PCS) accounts for ~20% of ischemic strokes. Existing EMS screening tools lack accuracy in the diagnosis of PCS. We aim to develop an automated screening tool to detect abnormal eye movements in patients presenting with PCS.

Methods:
As an initial step, we built a portable platform called RoADIE (Rolling Apparatus to Detect Impairment of the Eyes), equipped with eye-tracking software to acquire gaze data from patients presenting with PCS, acute vestibular syndrome, and normal controls. We first performed a validation study in 19 healthy controls comparing calibration (C) vs non-calibration (NC) techniques. Eye movements were captured using a standard H-Test exam. The NC condition was performed first for each volunteer to avoid a learning effect. Correlation between C and NC tests was determined using the Spearman coefficient (r).

Results:
Conjugate gaze (i.e. ability of eyes to track in unison) demonstrated strong correlation along the horizontal [r = 0.976 (C), 0.922 (NC)] and vertical axes [r= 0.866 (C), 0.881(NC)]. Smooth pursuit (i.e. ability of each eye to track moving target) also showed strong correlation in the horizontal plane: right eye [r = 0.945 (C), 0.946 (NC), left eye [r=0.945 (C), 0.943 (NC)]. Vertical tracking showed moderate correlation: right eye [r=0.652 (C), 0.575 (NC), left eye [r=0.678 (C), 0.550 (NC)].

Conclusion:
In this initial validation test of a portable eye tracking platform, we demonstrated strong correlation for both conjugate gaze and smooth pursuit in the horizontal axis, and moderate correlation for eye tracking in the vertical axis. Given similar performance in non-calibrated tests, a calibration procedure may not be necessary for future data capture. Next steps will include validation of the eye tracking device in prospective patients presenting with PCS and acute vestibular syndrome.
Acknowledgement:
Funding support through AHA Innovative Project Award 19IPLOI34760692
Abstract P755: Perlecan Lg3 is Neuroprotective and Functionally Restorative in Experimental Ischemic Stroke
Authors: Biose Ifechukwude Joachim , Rutkai Ibolya , Adkisson Davis , Clossen Bryan , Gage Gary , Bix Gregory J
Published: March 2021

Despite promising advancements, ischemic stroke remains a leading cause of death and disability. We previously determined that human recombinant ~85-kDa C-terminal protein fragment of the vascular basement membrane proteoglycan perlecan termed domain V (DV) is neuroprotective, pro-angiogenic, neurogenic and functionally restorative when administered after experimental stroke (transient and permanent middle cerebral artery occlusion -MCAO) in mice and rats. As previous studies suggest, the 23-kDa subdomain of DV called laminin globular domain 3 (LG3) may confer most of DV’s biological activity and is more amenable to production, we tested whether human recombinant perlecan DV LG3 (also termed “DV
LG3
”) might be neuroprotective and functionally restorative in the transient MCAO (intraluminal filament model).

Methods:
Fifty male mice (C57BL/6J, 10-12 weeks old, 24–30 g) underwent MCAO for 60 min and were survived for seven days. At reperfusion mice were randomly allocated to one of three treatment groups and immediately received either human recombinant DV treatment (positive control, n=16 2 mg/kg, i.p.), LG3 (n=19 6 mg/kg, i.p.), or equivalent volume of PBS vehicle (n=15) ; and again on post-MCAO days 2 and 4. The DV:LG3 dose ratio was selected based on a preliminary
in vivo
dose evaluation study of therapeutic equivalence. Functional outcome measures (grip strength and grid-hang tests) were determined at baseline and on post-MCAO days 1, 3 and 7. Body weight was measured daily as per welfare. To determine infarct size on post-MCAO day 7, whole brain samples were harvested, sectioned and stained with 2,3,5- Triphenyltetrazolium chloride. The induction of MCAO, DV, DV
LG3
or PBS treatments, functional tests and analyses were performed, blindly, by different individuals. While DV treatment significantly reduced mean infarct volume as expected (p<0.022, compared to PBS control group), DV
LG3
further reduced infarct volumes (p<0.0001), weight loss (p<0.0002) and improved functional outcome measures (p<0.01) compared to PBS group. Our findings show that human recombinant DV
LG3
is neuroprotective and functionally restorative, may be even more effective than DV, and is worthy of more study as a readily producible therapy for ischemic stroke.
Abstract P766: Ischemic Injury in a Dmcao Model of Stroke Demonstrates Long-Term Immune Cell Diapedesis Into the Brain Parenchyma
Authors: Ujas Thomas A , Malone Mary K , Torres Vanessa O , Messmer Sarah , Turchan-Cholewo Jadwiga , Stowe Ann M
Published: March 2021

Background:
Stroke injury following a middle cerebral artery occlusion (MCAo) induces a rapid migration of leukocytes into the injured brain that lasts for weeks. The current study focuses on whether a focal cortical stroke using the distal MCAo (dMCAo) model induces similar long-term immune cell diapedesis into the brain parenchyma as seen following transient (t)MCAo stroke.

Methods:
Cells were isolated from spleens and brain hemispheres of adult 1-year old male C57BL/6J (B6; Jackson Labs) mice 30 days after a dMCAo (n=10). Sham animals (n=5) received a craniotomy without the distal MCA ligation. Peripheral migration was assessed in the spleen and brain using flow cytometry (FACSymphony) to identify viable (Ghost dye 780) CD3, CD4, CD8b, CD11b, Ly6C/Ly6G, CD19, CD45, and NK1.1 leukocytes. Populations were analyzed with FlowJo and assessed via repeated measures two-way ANOVA, Sidak post-hoc test (Graphpad Prism). Significance was p<0.05.

Results:
CD45
+
leukocytes were elevated in the ipsilesional (ipsi) hemisphere compared to the contralesional hemisphere (p=0.01) after dMCAo, though a group hemispheric effect (F(1,13)=5.4; p=0.04) suggests long-term inflammation in sham-treated mice. Hemispheric effects also occurred for CD8
+
T cells (p=0.046), B cells (p=0.03), monocytes (p=0.01), and macrophages (p=0.06), with elevations in both dMCAo and sham-treated mice in the ipsi vs. contralesional hemispheres. Only monocyte populations were significantly elevated (p=0.03) in dMCAo vs. sham mice.

Conclusions:
Our study shows that immune cells remain elevated in the injured hemisphere at 30 days after a focal stroke confined to the neocortex, but inflammation occurred in both sham and dMCAo-treated animals. Only monocytes were differentially affected by dMCAo, and infiltrating cell numbers are not as robust as after tMCAo. This demonstrates a long-term injury response from the craniotomy in the dMCAo model that should be considered for long-term studies using this model.
Abstract P780: Myosin Light Chain Dephosphorylation by Ppp1r12c Promotes Atrial Hypocontractility in Atrial Fibrillation
Authors: Gonzalez-Gonzalez Francisco J , Srikanth Perike , Capote Andrielle E , Katherina M Alsina , Levin Benjamin , Sridhar Arvind , Jacob Grand , Martin Jody , Akar Joseph , Warren Chad M , Solaro John , Darbar Dawood , Goergen Craig J , Wolska Beata M , Wehrens Xander H , McCauley Mark
Published: March 2021

Atrial fibrillation (AF) is the most common sustained arrhythmia, with an estimated prevalence in the U.S.of 6.1 million. AF increases the risk of a thromboembolic stroke in five-fold. Although atrial hypocontractility contributes to stroke risk in AF, the molecular mechanisms reducing myofilament contractile function in AF remains unknown. We have recently identified protein phosphatase 1 subunit 12c (PPP1R12C) as a key molecule targeting myosin light-chain phosphorylation in AF.

Objective:
We hypothesize that the overexpression of PPP1R12C causes hypophosphorylation of atrial myosin light-chain 2 (MLC2a), thereby decreasing atrial contractility in AF.

Methods and Results:
Left and right atrial appendage tissues were isolated from AF patients versus sinus rhythm (SR). To evaluate the role of the PP1c-PPP1R12C interaction in MLC2a de-phosphorylation, we utilized Western blots, co-immunoprecipitation, and phosphorylation assays. In patients with AF, PPP1R12C expression was increased 3.5-fold versus SR controls with an 88% reduction in MLC2a phosphorylation. PPP1R12C-PP1c binding and PPP1R12C-MLC2a binding were significantly increased in AF. In vitro studies of either pharmacologic (BDP5290) or genetic (T560A), PPP1R12C activation demonstrated increased PPP1R12C binding with both PP1c and MLC2a, and dephosphorylation of MLC2a. Additionally, to evaluate the role of PPP1R12C expression in cardiac function, mice with lentiviral cardiac-specific overexpression of PPP1R12C (Lenti-12C) were evaluated for atrial contractility using echocardiography, versus wild-type and Lenti-controls. Lenti-12C mice demonstrated a 150% increase in left atrium size versus controls, with reduced atrial strain and atrial ejection fraction. Also, programmed electrical stimulation was performed to evaluate AF inducibility in vivo. Pacing-induced AF in Lenti-12C mice was significantly higher than controls.

Conclusion:
The overexpression of PPP1R12C increases PP1c targeting to MLC2a and provokes dephosphorylation, associated with a reduction in atrial contractility and an increase in AF inducibility. All these discoveries suggest that PP1 regulation of sarcomere function at MLC2a is a main regulator of atrial contractility in AF.
Abstract P784: A Novel Rat Model of Embolic Cerebral Ischemia Using a Cell-Implantable Radiopaque Hydrogel Micro Fiber
Authors: Komatsu Teppei , Ohta Hiroki , Takakura Naoki , Kitagawa Tomomichi , Hata Junichi , Terawaki Koshiro , Koizumi Makoto , Muta Kanako , Kurashina Yuta , Onoe Hiroaki , Okano Hirotaka J , Iguchi Yasuyuki
Published: March 2021

Introduction:
The failure of neuroprotective treatment-related clinical trials may be partially caused by unestablished animal models. We aimed to develop a novel focal stroke model using a cell-implantable radiopaque hydrogel micro fiber. The micro fiber is made of barium alginate hydrogel containing zirconia, and fabricated in a dual coaxial laminar flow microfluidic device.

Methods:
Using male Sprague Dawley rats (n=10), a catheter (ID 0.42mm, OD 0.55mm) was navigated from the caudal ventral artery to the left internal carotid artery using digital subtraction angiography. A radiopaque hydrogel micro fiber (0.4 mm in diameter, 1 mm in length) was advanced in the catheter by slow injection of heparinized physiological saline to establish local occlusion. 9.4-T MRI and 2% 2,3,5-triphenyl tetrazolium chloride staining were performed 24 hours after the creation of the stroke model.

Results:
The anterior cerebral artery-middle cerebral artery bifurcation was selectively embolized at all rats. The median operating time was 8.5 min (interquartile range; 6.25-12 min). The median infarct volume (mm
3
) was 262 (interquartile range; 260-274) 24 hours after occlusion.

Conclusions:
We present a novel rat model for focal infarct only in the middle cerebral artery territory using a radiopaque hydrogel micro fiber. Furthermore, this model can be used for regeneration research of cerebral infarction because the hydrogel micro fiber can encapsulate stem cell.Figure Legends A: Puncture of the caudal ventral artery in a rat. B: Cerebral angiography of a rat. C: Occlusion of the anterior cerebral artery-middle cerebral artery bifurcation by a radiopaque hydrogel fiber (arrowhead). D: MR angiography detected selective occlusion of left middle cerebral artery. E: T2-weighted image showed infarct only in the middle cerebral artery after 24 hours occlusion. A radiopaque fiber did not affect the 9.4-T MRI. F: Radiopaque hydrogel fiber encapsulated HeLa cells.
Abstract P792: Therapeutic Catalytic Nanoantioxidants Derived From Activated Charcoal
Authors: Derry Paul J , McHugh Emily A , Liopo Anton V , Vo Anh T , Gnanasekaran Aswini , Tour James M , Kent Thomas A
Published: March 2021

Introduction:
Our laboratories’ focus is synthesis of potential carbon nanoparticle (CNP) therapeutics for acute injury. To enhance the likelihood of regulatory approval, we tested whether medicinal grade activated charcoal (AC) could serve as the initial source. Taken up avidly and localizing to organelles, e.g. mitochondria, CNP’s broad redox potential could facilitate electron transfer. Here, we test whether CNPs from AC are cytoprotective in a brain endothelial cell line and their restoration of cerebral autoregulation in a traumatic brain injury model.

Materials and Methods:
5-30 nM oxidized AC (OAC) were synthesized from medicinal AC, treated with oleum and 90% nitric acid, then covalently PEGylated. Cell number was assessed in bEnd.3 cells incubated 18 hrs with 50 μM H
2
O
2
w/wo 8 mg/L PEG-OACs. An electron transport assay was performed with NADH (e
-
donor) and resazurin (e
-
acceptor) using kinetic UV-vis spectroscopy. 2 mg/kg PEG-OACs were administered IV to rats upon resuscitation and at 200 minutes after mild fluid percussion injury model with superimposed 80 minutes of hypotensive shock, a model known to impair reperfusion. Cerebral perfusion was monitored by laser doppler flowmetry.

Results:
PEG-OAC improved viability (Fig. 1a; 47% vs. 80%; p=.005). Oxidation of NADH was 3.8x faster than w/o OAC (Fig. 1b). Cerebral perfusion was restored by PEG-OAC after direct cortical injury compared to PBS vehicle (n=10; Fig. 1c; p=.0008).

Conclusion:
PEG-OACs protect cultured cells from oxidative stress caused by H
2
O
2
, a byproduct of ischemia, exhibit electron shuttle-like properties and restore cerebral reperfusion in-vivo when given at a clinically relevant time point.
Abstract P802: Potential Sex Differences in Endothelial Cell Death Pathways: Relevance to Stroke Recovery
Authors: Abdul Yasir , JAMIL SARAH , Li Weiguo , Chandran Raghavendar , Ergul Adviye
Published: March 2021

Introduction:
We have shown that iron chelation with deferoxamine (DFX) prevents stroke-induced cerebral vasoregression and improves sensorimotor and cognitive deficits after ischemic stroke in diabetic male rats. Diabetic female rats, on the other hand, develop greater hemorrhagic transformation (HT) but no vasoregression and DFX improves outcomes. HT breakdown products like hemin contribute significantly in secondary inflammation and neuronal death. Given the importance of brain microvascular endothelial cells (BMVECs) on neurovascular restoration in stroke recovery and our previous findings of increased ferroptotic cell death in endothelial cells from male animals, the objectives of this study were to determine 1) the impact of hemin on ferroptotic death in BMVECs of female and male origin, and 2) effect of DFX on ferroptosis.

Methods:
Male (HBEC5i) and female (BMECd3) human endothelial cell lines were grown in normal conditions. In some experiments cells were treated with hemin (50μM) for 6 hours with and without ferroptosis inhibitor ferrostatin-1 (20μM) or DFX (100μM). Cell viability (MTT and CytoSelect assays), ferroptosis marker proteins and lipid peroxidation (LPO) were assessed. by immunoblotting and, respectively.

Results:
GPX4 and NRF2 content was decreased with hemin in both sexes while 4HNE immunofluroscence for LPO, a key feature of ferroptosis, was increased. DFX did not prevent hemin-induced changes in GPX4 and ferritin heavy chain in female cells while it was effective in male cells. DFX and ferrostatin-1 improved cell viability in both male and female BMVECs. There were differences in ferrostatin 1 and DFX effects (Table).

Conclusions:
These results suggest there may be sex differences in how ferroptosis occurs in response to hemin. Better understanding of endothelial ferroptotic cell death regulation and the effect of iron chelation on cell survival can provide novel insights with respect to sex differences in stroke recovery.
Abstract P808: The Main Peptide Ssubstrates of Neurolysin Enhance Brain Microvascular Permeability in a Human
in vitro
Model
Authors: Al-Ahmad Abraham , Karamyan Vardan T
Published: March 2021

Increased brain microvascular permeability and disruption of blood-brain barrier (BBB) function are among hallmarks of ischemic stroke. Recently, peptidase neurlysin (Nln) has been identified as a compensatory and cerebroprotective mechanism in the post-stroke brain that functions to process a diverse group of extracellular neuropeptides, including bradykinin (BK), neurotensin (NT) and substance P (SP). A number of studies suggest involvement of BK, NT and SP in BBB impairment and edema formation after stroke, however there is paucity of data in regards to the direct effects of these peptides on the brain microvascular endothelial cells (BMECs) and BBB. The purpose of this study was to evaluate the direct effects of BK, NT and SP on permeability of BBB in an
in vitro
model based on human, induced pluripotent stem cell (iPSC)-derived BMECs. Our data indicate that all three peptides increase BBB permeability in a concentration-dependent manner in an
in vitro
model formed from two different iPSC lines (CTR90F and CTR65M) and widely used hCMEC/D3 human BMECs. Combination of BK, NT and SP at a sub-effective concentration also resulted in increased BBB permeability in the iPSC-derived model. Furthermore, we observed abrogation of BK, NT and SP effects with pretreatment of pharmacological blockers targeting their specific receptors or in presence of recombinant neurolysin (Nln). This is the first experimental study to document increased permeability of BBB in response to direct action of NT in an
in vitro
model. In addition, our study confirms the expected but not well-documented, direct effect of SP on BBB permeability and adds to the well-recognized actions of BK on BBB. Lastly, we demonstrate that peptidase Nln can neutralize the effects of these peptides on BBB, suggesting potential therapeutic implications.
Abstract P829: Bi-Directional Quality Review in a Large Telestroke Network: Improving Stroke Care Through Mutual Learning
Authors: Majersik Jennifer J , Ekstrom Erin , Johnson Jaleen R , Wicks Heather , Dart Chona , Christensen Lacey , Reynolds Tawnie , Chung Lee S , Hannon Peter , Reddy Vivek , Harman Tyler D
Published: March 2021

Introduction:
Our hub and spoke telestroke (TS) network consists of 26 sites across 6 states, with >1000 consults/year. Without a shared EMR, patient outcomes are often unknown. We aimed to improve communication of patient outcomes and patient care by implementing multi-disciplinary quality review across multiple health systems.

Methods:
To determine interest in and format of the review, we first conducted interviews with coordinators from 3 highly engaged sites of varying sizes and capabilities. We had strong consensus that a bi-directional quality review would be helpful and that group-based discussion was preferred over single site review. Coordinators chose review flags of long process metrics, missed treatments, thrombectomies, complications, and mortalities with sentinel events reviewed immediately, off-cycle. A case review sheet is uploaded to HIPAA-compliant Box drive and neuroimaging to PACS. Hub and spoke present cases over Zoom using standard SOAR format: Situation, Outcome, Assessment, Recommendation. The group grades each case as standard of care (SOC) met, exceeded, or not met, +/- opportunities for improvement (OFI). Discussion includes recommendations to the individual hospital or network.

Results:
From 4/2019-7/2020, we conducted 8 bi-directional case reviews of 21 TS cases with 9 spokes (mean 4.3 spokes/review, range 2-6). Of 47 spoke participants, 43% were stroke coordinators, 34% were ED managers, with rare hospitalists present. Hub participants were 8 vascular neurologists/fellows and 3 TS coordinators/managers. Case content was 33% recognition, 49% acute treatment, and 19% disposition. SOC was rated as met or exceeded in 86% of cases with OFI noted in 100%. The Table details discussion themes.

Conclusion:
A bi-directional quality review can share knowledge and best practices across a large TS network, improving inter-facility communication and site engagement. We hope next to increase physician attendance and engage more sites.
Abstract P83: Acute Stroke Presentations During the Course of the COVID-19 Pandemic
Authors: Vanguru Husitha , Kolikonda Murali , Brown Dena R , Collins Dana , Buletko Andrew B , Russman Andrew N , Hussain Muhammad S , Uchino Ken
Published: March 2021

Background:
Decline in presentations of acute stroke during the early period of COVID-19 pandemic have been reported. We aimed to investigate the stroke presentations during the subsequent months as the pandemic evolved into a second wave.

Methods:
Data was obtained from a health system with 19 emergency departments (EDs) in northeast Ohio in the United States. Baseline period from January 1 to February 29, 2020, was compared with the individual months during COVID-19 period from March through July. Variables included were numbers of daily stroke alerts across the EDs, thrombolysis, thrombectomy, time to presentation, stroke severity, time from door-to-needle in thrombolysis, and door-to puncture in thrombectomy. The time periods were compared using nonparametric statistics and Poisson regression with month, weekend, and daily COVID cases as independent variables.

Results:
A total of 2264 stroke alerts from EDs were analyzed between January 1 to July 31, 2020. Total daily stroke alerts decreased from a median of 10 (interquartile range [IQR]:10-13) in January and February to 9 (IQR:6-11, p=0.001) in March, 8(IQR:7-10, p=0.0001) in April, 10 (IQR:8-11, p=0.04) in May, and returned similar to baseline in June (12, IQR:10-13, p=0.5) and July (13, IQR:11-14,p=0.1). In Poisson regression, stroke alert numbers showed no significant association with daily COVID-19 counts, but significant association with months, with rate ratios of 0.74 (95%CI 0.64-0.85) for March, 0.71 (95%CI 0.61-0.82) for April, and 0.86 (95%CI 0.75-0.98) for May, but not with June and July. Time to presentation and stroke severity were unchanged throughout the study period. Thrombolysis volume decreased in March and May but thrombectomy volume was unchanged.

Conclusion:
We observed a decrease in stroke presentations across emergency departments by about 30% during the early period of COVID-19 pandemic, followed by return to baseline frequency despite a second wave of COVID-19 cases.
Abstract P848: A Better Way to NIHSS
Authors: GRAVES Alex M , Jones Jamie , Bragg Alyssa
Published: March 2021

Introduction:
“A Better Way to NIHSS” evaluated feasibility and effectiveness of in-person National Institutes of Health Stroke Scale (NIHSS) training compared to traditional training across a 17-hospital system. Stroke practitioners are encouraged and sometime required to be NIHSS certified. Despite the ubiquity of the scale, it presents challenges for care providers. Items with poor reliability, improper technique, and scoring errors may lead to a lack of recognition of neurologic decline. Standard computer-based training may not effectively prepare RNs. An updated in-person course was needed to improve practitioner satisfaction and scoring accuracy.

Hypothesis:
We hypothesized that we could improve practitioner satisfaction, scoring accuracy and system efficacy by developing and implementing an in-person NIHSS course.

Methods:
Pitfalls of traditional computer training were identified by a stroke APN. In collaboration with national and regional subject matter experts, evidence-based curriculum was developed. Emphasis was placed upon live demonstration of exam technique, and student return-demonstration for psychomotor competency. Tools were provided to meet visual and read/write learning styles. Over 1-year, a hospital system collaborated to standardize the curriculum, HR certification and tracking process, and instructor training. Instructors received 1-on-1 training and mentorship to effectively deliver content. Instructors in training often co-taught classes until they were comfortable, had speaker notes to assist them, and training sessions geared to their needs. Standardized evaluation tools, stakeholder input, data, and random chart audits were utilized to evaluate effectiveness.

Results:
Over 18 months, 14 instructors were trained and 896 RNs certified. Evaluations showed improved learner knowledge, skills, and confidence. Educator training led to improved instructor evaluations. Documentation showed improved accuracy of scoring during random chart audits. Items with greatest improvement were limb ataxia, facial droop, visual fields, and aphasia.

Conclusion:
Systems should consider implementing and offering in-person NIHSS course to reduce the incidence of improper exam technique and scoring errors.