bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract W MP118: In Situ B-cell Clonal Expansion and Recombinant Antibody Construction in Human Cerebral Cavernous Malformation
Authors: Shi Changbin , Shenkar Robert , Kinloch Andrew , Zheng Nai-Ying , Henderson Scott G , Shaaya Mark , Chong Anita S , Clark Marcus R , Wilson Patrick C , Awad Issam A
Published: March 2022

Background:
We had demonstrated immunocompetent cell infiltration and oligoclonal immune response in human cerebral cavernous malformation (CCM) lesions. However, T- and B- cell crosstalk, relationship between antibody and complement, and IgG repertoire in CCMs remain unknown. We hypothesize the synergetic role of T and B cells, antibody- and complement- mediated cytotoxicity, antigen-driven B cell clonal expansion, contributing to the CCM pathogenesis.

Methods:
This study enrolled a total of 10 patients with CCMs. B- and T- cells were immunostained in two excised human CCM lesion specimens. IgG and complement protein membrane attack complex (MAC) was co-localized in six other CCM lesions by immunofluorescence. Plasma cells in two CCM lesions were laser-microdissected, in which IgG heavy (H) and light (L) chain variable (V) region were amplified via nested PCR, cloned, sequenced, and aligned to a database IMGT®. The over-expressed IgGHV and IgGLV were used to construct the recombinant antibodies (r-abs).

Results:
In CCM lesions, T- and B- cells were clustered and co-localized, and plasma cells were clustered. IgG and MAC was co-localized in CCMs. IgG repertoire showed in situ B cell clonal expansion and ongoing somatic mutation. Four r-abs were constructed from clonally expanded plasma cells in CCM lesions.

Conclusions:
we showed the role of immunity in CCM pathogenesis via T- and B- cell crosstalk, antibody- and complement- mediated cytotoxicity, and clonal expansion. We gave the first demonstration that oligoclonal response is antigen-driven. The first synthesized r-abs will be used to identify disease relevant antigens. This would clarify the relationship of immune response to CCM lesion development, and may open novel therapeutic venues in CCMs.
Abstract W MP16: Lower Apparent Diffusion Coefficient Ratio Predicts Increased Swelling After Ischemic Stroke
Authors: Battey Thomas W , Karki Mahima , Wu Ona , Singhal Aneesh B , Sheth Kevin N , Kimberly W T
Published: March 2022

Introduction:
Swelling is common after acute ischemic stroke but little is known about predictors for its development. We explored several putative markers of swelling using brain magnetic resonance imaging (MRI).

Methods:
We retrospectively analyzed a cohort of acute stroke subjects with serial research brain MRI at baseline, 6 hours later, and then at 24 and 48 hours after the baseline MRI. We defined swelling as the expansion in stroke volume from baseline to the follow-up scan up about 2 days after stroke onset. Change in volume was also examined between each MRI interval. Apparent diffusion coefficient (ADC) and fluid-attenuated inversion recovery (FLAIR) signal intensity ratios were developed by normalizing the average signal intensity within the stroke lesion to that of the normal contralateral hemisphere. The association between ADC ratio and FLAIR ratio with expansion was assessed.

Results:
Twenty-one subjects had 71 scans available at four time points with a median baseline stroke volume of 98 mL [interquartile range (IQR) 66-135 mL]. The median increase in volume was 55 mL (IQR 35-84 mL) from baseline MRI. The changes from baseline to 24 hours (r=0.89, p<0.0001) and from 24 to 48 hrs (r=0.85, p<0.0001) were associated total expansion. FLAIR ratio exhibited a sharp elevation from baseline to 24 hours, and plateaued thereafter, but was not associated with expansion. ADC ratio decreased to a nadir between 6 and 24 hours, and then normalized at 48 hours. The nadir ADC ratio at 6 hours (r=-0.64, p=0.007) and 24 hours (r =-0.61, p=0.007) was associated with swelling.

Conclusions:
Acute stroke subjects exhibit differing trajectories of expansion, and those with greater total swelling show larger increases early on. The nadir ADC ratio, a marker of stroke severity and cytotoxic injury, correlated with total swelling. FLAIR ratio, a potential marker of vasogenic edema, was not correlated to lesion expansion in this cohort. Future studies are warranted to better understand the relationship between these imaging markers and clinical outcome.
Abstract W MP18: Baseline-24h CT ASPECTS Mismatch. A Predictor Variable of Successful Trhrombectomy
Authors: Cardona Pedro , Quesada Helena , Cano Luis , Lara Blanca , Mora Paloma , Aixut Sonia , Aja Lucia , Escrig Ana , De Miquel Maria Angeles , Rubio Francisco
Published: March 2022

BACKGROUND:
In acute thromboembolic stroke, mechanical recanalization with retrieval stents may result in immediate flow restoration. A selection criteria to treatment is a small volume of infarct in baseline CT scan, usually it is identified as an infarct <1/3 MCA or mild early ischemic changes (ASPECTS>7). We assess whether evaluation of basal and 24h CT using Alberta Stroke Program Early Computed Tomography Score (ASPECTS) have predictive value for functional independence after stroke on noncontrast CT (NCCT).

METHODS:
We performed an retrospective analysis of 257 consecutive patients treated with thrombectomy (Solitaire, Trevo) , from April of 2010 to June of 2013, that met inclusion criteria for intervention. Vessel patency pre and post procedural were measured by TIMI and TICI score.Successful recanalization (TICI 2b-3), functional and clinical good outcome (mRankin 0-2) and mortality at day 90 and symptomatic hemorrhage were prospectively assessed. We evaluated baseline NCCT, mean final infarct extension measuring by 24h-ASPECTS and change in ASPECTS from baseline to 24h CT.

RESULTS:
A total of 168 patients with ischemic stroke of the MCA territory (M1 segment or terminal internal carotid occlusion) were included in this study. We calculated ASPECTS mismatch as 24H NCCT ASPECTS/Baseline NCCT ASPECTS*100. Low percentage represents a reduced infarct and an higher efficacy of the neurointerventionism (TICI 2b-3, p:0.001) . Basal and 24h ASPECTS were dichotomized at >=8 vs <8 according with ROC analysis. Higher scores >=8 in NCCT ASPECTS implied a higher probability of an independent functional outcome at 90 days (mRankin 0-2) when compared to <8 ASPECTS group (OR 1.47, 95% CI 1.21-1.83) and likewise in 24h NCCT (OR 1.65, 95% CI 1.35-2.1).

The mismatch between basal ASPECTS in NCCT and 24h ASPECTS demonstrated important predictive value (OR 0.54, 95% CI 0.42-0.68) of clinical prognosis (cut-off value <20%, Sensivity 81%, Specificity 72%; AUC 0,74; p:0001), mainly in the group of 24h ASPECTS <8.

CONCLUSIONS:
The baseline and 24h NCCT ASPECTS were a strong predictor of functional outcome. Moreover lower mismatch between basal and 24h ASPECTS was also reliable as predictive value in good functional outcome.
Abstract W MP2: Daily Goals Poster Supports Effective Patient Education
Authors: Motz Deb S , Sharp Johanna A , Garcia Thelma , Moore Tracy , Huey Dicky , Austin Tommye
Published: March 2022

Background:
Patient education is a central component of any stroke program. Patients are educated on their stroke etiology, warning signs of stroke, activation of EMS, individualized stroke risk factors, medications prescribed at discharge and follow-up care. Patients are frightened and overwhelmed with the sudden change in their life leading to information overload. Education is further challenged due to the average LOS of 4 days during which patients are busy with tests, therapy and frequent nursing assessments.

Purpose:
The purpose of this process improvement is to develop a tool to support and facilitate effective stroke education.

Methods:
Stroke team members met to develop an interactive daily goals poster to address patient education and to serve as a communication tool between staff members, patients and families. This tool focused on the first four days, included all of the core measures and reflects documentation in the electronic medical record. The tool allows the staff to customize educational components to address patient-specific risk factors, is used as a teaching chart between the nurse and patient to track individual progress and remains with the patient from admission thru discharge. After discharge, the tool continues to serve as a reference for personal risk factors and prevention education.

Results:
This poster served as a communication tool to educate the patient and families on daily expectations and prompted questions of their individual treatment plan. After the tool was implemented, the stroke education core measure compliance improved from 75% in June 2011 to 86% in June 2013. Additionally, there was a marked improvement in patient satisfaction rating of 74% (n=322) in June 2011 to 88% (n=976) in June 2013. Indirectly, the tool served as a guide for new staff and fostered continuity of care between units.

Conclusion:
This tool proved to be an effective means to support and facilitate stroke patient and family education. Based on the new self- management standards implemented by The Joint Commission, this tool can easily be revised to successfully include an area to establish mutually agreed upon goals between patient and practitioners.
Abstract W MP23: About 30% of Wake-up Stroke Patients May be Candidate for the tPA Therapy Using Negative-FLAIR as a Tissue Clock
Authors: Nagai Koichiro , Kimura Kazumi , Aoki Junya , Shibazaki Kensaku , Ebata Takuya , Sato Takahiro , Saji Naoki , Uemura Junichi
Published: March 2022

Background and Purpose:
Most of the wake-up stroke (WUS) patients cannot be treated using intravenous thrombolysis (tPA) because of unknown onset time. Recent magnetic resonance imaging (MRI) studies reported that WUS patients may be the candidates for tPA therapy when showing no ischemia on the fluid-attenuated inversion recovery (negative-FLAIR), because negative FLAIR indicates the stroke onset to be within 4.5 h. The aim of the present study is 1) to investigate the frequency of WUS and the clinical and MRI findings of the WUS patients, and 2) calculate how many WUS patients with negative-FLAIR met the tPA criteria except the time parameter.

Methods:
Consecutive acute stroke patients with anterior-circulation within 12 of onset were enrolled. All patients were examined using diffusion-weighted imaging (DWI), and FLAIR on admission. WUS was defined as stroke on awaking. Neurological severity was assessed using the National Institutes of Health Stroke Scale (NIHSS) score. As a large infarction that is excluded from tPA therapy, the DWI-the Alberta Stroke Program Early Computed Tomography Score (DWI-ASPECTS) of ≤3 was defined.

Result:
A total of 619 consecutive patients were included in the study. The number of the WUS patients (WUS group) was 117 (19%), and the other (non-WUS group) was 502 (81%). Median NIHSS score on admission was 9 (interquartile range, 3-17) in the WUS group and 8 (3-16) in the non-WUS group (p=0.339). DWI-ASPECTS was similar between the WUS and the non-WUS groups (9 [7-10] vs. 9 [6-10], p=0.994). The number of patients treated using tPA was significantly smaller in WUS group compared to the non-WUS group (3 [3%] vs. 140 [28%], p<0.001). MRI study revealed that negative-FLAIR was seen in 50 (43%) of the 117 patients in the WUS group. Of the 50 patients with negative-FLAIR, 16 patients were excluded from the tPA therapy, the large infarction (6), platelet count <100,000 /mm (2), INR >1.7 (1), recent operation (1), and high blood pressure greater than 185/110 mmHg (6). Therefore, 34 (29%) of the 117 wake-up stroke patients met the tPA criteria.

Conclusion:
About 30% of the WUS patients may be candidate for the tPA therapy using the negative-FLAIR findings.
Abstract W MP25: Stroke Outcomes in Patients Receiving Extracorporeal Membranous Oxygenation
Authors: Nasr Deena , Rabinstein Alejandro
Published: March 2022

Objective:
Using the Nationwide Inpatient Sample (NIS), we determined morbidity, mortality and trends in ischemic and hemorrhagic stroke complications in patients receiving Extracorporeal Membranous Oxygenation (ECMO).

Materials and Methods:
Using the NIS from 2001-2011, patients receiving ECMO were identified. We analyzed their ischemic and hemorrhagic stroke complication rates and compared discharge status between these patients and those without cerebrovascular complications. Morbidity was defined as discharge to a long-term facility. Ischemic and hemorrhagic stroke rates were compared between two time periods (2001-2006 and 2007-2011). Continuous variables were compared with a student’s t-test and categorical variables were compared with the chi-squared test.

Results:
A total of 23,950 ECMO patients were included in this study of which 22108 patients (92.3%) had no cerebrovascular complications, 980 patients (4.1%) had ischemic stroke and 862 patients (3.6%) had hemorrhagic stroke. Of those without cerebrovascular complications there was a 49.9% mortality rate and a 6.8% morbidity rate. Patients with ischemic stroke had a mortality rate of 44.3% (P=0.0006) and a morbidity rate of 14.6% (P<0.0001). Patients with hemorrhagic stroke had a mortality rate of 59.7% (P<0.0001) and a morbidity rate of 9.5% (P=0.002). The proportion of patients with ischemic stroke increased from 2.8% in 2001-2006 to 5.1% in 2007-2011 (P<0.0001) and patients with hemorrhagic stroke increased from 2.4% in 2001-2006 to 4.5% in 2007-2011(P<0.0001).

Conclusion:
ECMO patients complicated with ischemic and hemorrhagic stroke had higher morbidity than those with no cerebrovascular complications. Patients with hemorrhagic stroke had a 10% worsened mortality rate than those with no cerebrovascular complications. The proportion of ECMO patients with cerebrovascular complications has nearly doubled between 2001-2011.
Abstract W MP3: Modifiable Risk Factors of Stroke and Psychosocial Health Factors Among Community-Residing Adults With Prehypertension
Authors: Song Hee-Young
Published: March 2022

Introduction:
Prehypertension that is systolic blood pressure (BP) over 120 mmHg or diastolic BP over 80 mmHg is known to be a predictor for the future development of hypertension and to be associated with increased risk for stroke. Although the impact of traditional risk factors such as hypercholesterolemia, overweight/obesity, lack of physical activity and high-fat diets on the development of hypertension and stroke has been delineated in people with prehypertension, recent research also suggests a need to examine further the relationship between psychosocial health factors such as stress, depression, and social engagement and BP.

Objectives:
This study was undertaken to assess modifiable risk factors of stroke,social support and participation, and depression, and to explore the relationship between these factors and BP among community-residing adults with prehypertension.

Methods:
A total of 178 participants aged 46.8 years old were recruited by a convenient sampling in two urban areas in South Korea. Trained interviewers interviewed participants with a structured questionnaire including demographic characteristics, lifestyle risk factors, stress, social support and participation, and depression. BP and anthropometric measurement and fasting blood sampling were performed to measure physiological risk factors. Data were analyzed using SPSS statistics 20.0.

Results:
Mean systolic BP of participants was 131.04 mmHg and mean diastolic BP, 80.70 mmHg. Participants were over-weighted and only 28% of them did regular physical activity, showing mean Beck depression inventory scores of 29.61 which fell under the category of severe depression. After controlling for age and sex, stepwise multiple regression showed that BMI and depression were predictors of higher systolic BP and depression, a predictor of higher diastolic BP.

Conclusions:
These results suggest the need to address psychosocial health factors, along with well- established risk factors such as BMI and physical activity, for BP management in community-residing prehypertensive adults. Further studies are warranted to fully understand and establish psychosocial factors and the mechanisms which contribute to BP elevation, and to develop interventions that decrease depression and BP.
Abstract W MP33: Brain and Aortic Aneurysm Screening - Cost-Effectiveness of Screening for Intracranial Aneurysms
Authors: Jiang Boxiang , Ball Benjamin Z , Gaughen John R , Brown Robert D , Torner James C , Stukenborg George J , Southerland Andrew M , Worrall Bradford B
Published: March 2022

Introduction:
Intracranial aneurysm (IA) and abdominal aortic aneurysm (AAA) rupture carry high mortality (40% and 75% respectively) and substantial morbidity in survivors. Recent research suggests that IA and AAA share genetic risk in addition to environmental factors. Retrospective data from our institution identified a population harboring both IA and AAA and showed a high rupture rate for IA (44.2%). We previously demonstrated that screening for AAA in patients with IA is cost-effective based on estimated and published data. We hypothesize that screening for IA in patients with AAA is also cost-effective.

Method:
We developed a decision-making model using TreeAge Pro 2013 software assuming a hypothetical population with known AAA and truncated screening versus no screening for IA using magnetic resonance angiography. Based on IA size and location, patients were either treated endovascularly or placed on surveillance. Proportions, risks, utilities, and costs for each decision arm were derived from literature.

Results:
At a given rupture rate, incremental effectiveness varied linearly with coprevalence of AAA and IA. For an assumed rupture rate of 5%/year, screening was effective for coprevalence >6.8% (top graph) and was cost-effective for coprevalence >7.7%. The incremental cost-effectiveness ratio (ICER) favored screening (ICER=$54,193/QALY) at a 7.8% coprevalence and dropped to $27,841.50/QALY at 8.8% coprevalence. Our model was sensitive to IA rupture rate as well as coprevalence (bottom graph).

Conclusion:
Our model suggests the cost-effectiveness of screening for IA in patients with AAA, below the accepted societal threshold of an ICER $60,000/QALY at a coprevalence ≥7.8% and IA rupture rate ≥5%. These are plausible assumptions for this genetically high-risk population. Further prospective study is warranted to establish coprevalence of AAA and IA, rupture rates, and determine the benefit of dedicated screening.
Abstract W MP36: The Phenomenon of Delayed Pipeline Embolization Device Migration: Technical Considerations, Complication Avoidance, and Management.
Authors: Chalouhi Nohra , Tjoumakaris Stavropoula , Gonzalez L Fernando , Hasan David , Pema Peter , Gould Grahame , Rosenwasser Robert , Jabbour Pascal
Published: March 2022

Introduction:
The Pipeline Embolization Device (PED) is a dedicated flow diverter designed to treat intracranial aneurysms. New safety concerns are emerging as neurointerventionalists accumulate experience with the device. In this study, we describe the phenomenon of spontaneous delayed migration of the PED and discuss important technical considerations for prevention and management of this complication.

Methods:
A total of 155 patients were treated with the PED at our institution from November 2010 to January 2013. Patients who had spontaneous delayed migration of their devices during the study period were prospectively identified.

Results:
Five patients (3.2%) were found to have spontaneous delayed migration of their PEDs. All 5 cases occurred late during the study period (second half of the cohort). The PED migrated proximally in 4 patients and distally in 1 patient. The migration distance was 11 mm on average (range, 5-20 mm). Mismatch in arterial diameter between inflow and outflow vessels was a constant finding. One patient suffered a subarachnoid hemorrhage and expired as a result of PED migration while one patient presented with complete middle cerebral artery occlusion and was left severely disabled. In the 3 remaining cases, PED migration led to renewed aneurysm filling. PED migration was managed conservatively in 1 patient, with parent vessel occlusion in 1, and with additional placement of PEDs in 3.

Conclusion:
Spontaneous delayed migration of the PED is a serious and potentially fatal complication. The device may migrate proximally or distally and result in aneurysm rupture or thrombotic events. Obtaining complete expansion of the PED, using a longer PED, increasing vessel coverage, adjunctive aneurysm coiling, and avoiding dragging and stretching of the PED are important preventive measures. Neurointerventionalists should be aware of this serious complication and take all necessary preventive measures.
Abstract W MP37: A Novel Homocysteine Inhibitor Enhances Neurological Recovery after Cerebral Ischemia in Mice and Non-Human Primates
Authors: Suzuki Hiroko , Fukudome Daisuke , Yuki Satoshi
Published: March 2022

Background:
Hyperhomocyteinemia is associated with cardiovascular and cerebrovascular diseases. Many studies have investigated the deleterious effects of intercellular homocysteine (Hcy) accumulation on vascular and brain tissue. AEB-577 is a potent Hcy synthesis inhibitor based on a non-adenosine analog. We hypothesized that AEB-577 could normalize Hcy content in brain tissue, reduce cytotoxicity of Hcy and ameliorate neurological dysfunction after stroke.

Method:
Male C57BL/6N mice were subjected to photochemically induced thrombosis, and treated orally with AEB-577 (1, 3, 10 mg/kg) or vehicle 10 minutes after the infarction. Infarct volume was assessed by tetrazolium staining and motor incoordination was evaluated using rota-rod test at 24 hours after infarction. In the second experiment, Hcy content in peri-infarct area was measured at 3 days after infarction by HPLC. In a non-human primate experiment, male cynomolgus monkeys were subjected to permanent middle artery occlusion (pMCAO), and divided into 3 groups based on infarction volume assessed by MRI images 18 hours after the onset. And then, vehicle or AEB-577 (4, 12 mg/body) was orally administered once a day for 84 days. The neurological deficits including paralysis, sensory dysfunction, and deficiencies of consciousness and motility were assessed at 3, 5, 7, 11, 14, 28, 56, 84 days after the onset of MCA occlusion. Plasma Hcy level and brain Hcy content were assessed at 84 days.

Result:
AEB-577 reduced infarct volume and improved motor deficit compared with vehicle treated mice. Hcy content in peri-infarct area significantly increased. AEB-577 decreased elevated Hcy content to normal level. In the monkey study, AEB-577 treatment starting 24 hours after stroke onset significantly promoted neurological recovery. AEB-577 decreased Hcy level in plasma and brain tissue compared with the placebo group at 84 days after stroke onset.

Conclusion:
Our results indicate that the novel Hcy synthesis inhibitor, AEB-577, can improve functional recovery after cerebral infarction modulating not only plasma Hcy level also Hcy content in brain tissue.
Abstract W MP39: Basic Fibroblast Growth Factor after Ischemic Stroke in Diabetic Patients - Research for Biomarkers in Ischemic Stroke (REBIOS)
Authors: Nakamura Kuniyuki , Ago Tetsuro , Arimura Koichi , Makihara Noriko , Nishimura Ataru , Matsuo Ryu , Wakisaka Yoshinobu , Kuroda Junya , Isomura Tetsu , Awano Hideto , Suzuki Kazuo , Okada Yasushi , Kamouchi Masahiro , Ooboshi Hiroaki , Kitazono Takanari
Published: March 2022

Background:
In patients with diabetes mellitus (DM), functional outcome after ischemic stroke is poor. However, the precise mechanisms remain unclear. We hypothesized that basic fibroblast growth factor (bFGF), a neurotrophic/growth factor, does not function after ischemic stroke in DM patients compared with in non-DM ones.

Methods and Results:
We recruited 171 patients with ischemic stroke from the Fukuoka Stroke Registry, a prospective multi-centered study for acute stroke in Japan, and age- and sex- matched healthy subjects. Blood samples were obtained at five time points after the onset, day 0 (within 24 hours), 3, 7, 14 and 90. Plasma bFGF values were measured by Human MAP® v1.6, a multiplexed immunoassay (Myriad RBM, Inc.). Plasma bFGF was significantly higher in stroke patients at day 0 than in the controls (162.1 ± 21.0 vs. 91.0 ± 8.1 pg/ml,
p
= 0.001). The concentration of bFGF remained high during 90 days after onset. Unexpectedly, bFGF was significantly higher in patients with DM than in those without DM (180.8 ± 19.5 vs. 127.5 ± 13.1 pg/ml,
p
= 0.033), while the presence of DM did not associate with bFGF levels in healthy controls. Plasma bFGF was correlated positively with HbA1c (
p
= 0.037), causal plasma glucose (
p
= 0.048), and ICAM-1 (
p
= 0.032) / VCAM-1 (
p
= 0.086), representative adhesion molecules expressed in endothelial cells. In a mouse middle cerebral artery occlusion (MCAO) stroke model, bFGF expression in streptpzotocin-induced diabetic mice was significantly greater in ischemic hemisphere than in contralateral hemisphere. Immunohistochemical examination demonstrated that bFGF was expressed highly in vWF
+
endothelial cells, desmin
+
pericytes, and MAP2
+
neurons, in peri-infarct areas after MCAO.

Conclusion:
Plasma bFGF was increased after ischemic stroke, which was prominent in DM patients. The presence of hyperglycemia in acute ischemic stroke may affect the expression of bFGF in various cell types in peri-infarct areas, which in turn may be associated with endothelial function, including the expression of adhesion molecules.
Abstract W MP46: Beyond the Brain: Stroke Induces Vascular Breakdown and Unique Profiles of Peripheral Inflammatory Response in the Eye and Spinal Cord
Authors: Pan Sarah , Ritzel Rodney , Patel Anita , Lieberman Richard , McCullough Louise
Published: March 2022

Ischemic stroke, a leading cause of death and long-term disability, has far-reaching effects across the CNS beyond the region of infarct. As extensions of the brain, neural networks of the eye and spinal cord are affected by cerebral ischemia through the transmission of inflammatory and degenerative changes. Although neuropathological effects of MCAO have been reported in both regions, the contribution of the peripheral inflammatory response to the evolution of injury in these CNS regions is unknown. We evaluated local glial responses, vascular integrity, leukocyte infiltration, and cytokine expression after stroke in the eye and spinal cord.

Adult male C57Bl/6 WT or GFP+ bone marrow chimeric mice were subjected to 90-min MCAO or sham surgery (n=3-6). Imaging of the retinal vasculature was performed by ophthalmoscopy. At 8h, 24 and/or 36h, 72h and 7d post-stroke mice were perfused with PBS and sacrificed. Organs were harvested for immunohistochemistry, Evans blue extravasation, flow cytometry, and RT-qPCR.

Early (8h) after MCAO, over four-fold increases in TNFα and IL-1β gene expression were observed in the ipsilateral eye relative to sham (4.64±1.21, 1±0.17; 11.03±7.75, 1±0.21). Microglial activation indicated by increased CD11b, CD45, and cell granularity at 36h was accompanied by an influx of peripheral leukocytes predominantly comprised of neutrophils (81%). In contrast to BBB breakdown which was highest at 24h, blood-retinal barrier breakdown peaked in the ipsilateral eye at 36h, shown by a twelve-fold increase in Evans blue concentration (sham, 24h, 36h: 0.16, 0.41, 1.95 ug/mg tissue wet weight). Similarly, major blood-spinal cord barrier breakdown occurred at 36h (0.32 vs 0.052 ug/mg). This was followed by a largely monocytic response with corresponding increases in TNFα, IL-1β, and IFNγ gene expression.

Together, our data show robust unilateral peripheral inflammatory responses after stroke in CNS regions outside of the brain. In addition, blood-parenchymal barrier permeability peaks later in the eye and spinal cord than in the brain. Given the distinct time course and inflammatory components associated with each region, these findings may have wide implications for treatments attenuating CNS-wide inflammatory damage after stroke.
Abstract W MP48: Post-stroke Administration of Exogenous Inter-alpha Inhibitor Proteins is Neuroprotective.
Authors: Venna Venugopal R , Mancini Nickolas , Scranton Victoria , Patrizz Anthony , Lim Yow-Pin , McCullough Louise D
Published: March 2022

Background and Purpose:
Current stroke therapies approved for human use are limited by time constraints and carry significant hemorrhagic risk. Down regulation of pro-inflammatory cytokines, modulation of peripheral leukocyte infiltration and reduced microglial activation benefit the ischemic brain. A novel immunomodulator protein family extracted from human plasma (Inter-alpha Inhibitor Proteins - IAIP) has been developed that has desirable anti-inflammatory properties in addition to an advantageous long half-life (8-12 hours).

Methods:
C57BL/6 mice were subjected to transient focal ischemia by middle cerebral artery occlusion (MCAO) for 90 minutes. One cohort was given IAIP (30mg/kg) immediately at reperfusion (90 minutes after stroke). A second “delayed treatment” cohort was given IAIP 6 hours after stroke onset. Both cohorts were given a second dose 24 hours post-stroke. Motor and sensory deficits were assessed using the neurological deficit score (NDS). Infarct was assessed at either 48 hours or 7 day time points. Infarct data presented as mean ±SEM, NDS data are presented as median±IQR.

Results:
At 48 hours mice given IAIP at reperfusion had significantly smaller cortical (59.26%±1.80% versus 31.58%±8.53%, n=8, p<.05) and hemisphere (46.05%±1.55% vs. 22.53%±6.55%, n=8, p<.05) infarct volumes. Delayed IAIP treated mice also had significantly smaller cortical (52.44%±1.73% vs. 22.99%±4.59%, n=6, p<.05) and hemispheric infarcts (43.86%±2.11% vs. 19.45%±3.59%, n=6, p<.05) at 48 hours. Analysis at 7 days showed significantly smaller cortical (23.30%±2.53% vs. 3.46%±0.46%, n=6, p<.05) and total hemisphere (22.33%±2.87% vs 5.90%±0.86%, n=6, p<.05) infarcts in mice treated with IAIP 6 hrs after stroke suggesting durability of neuroprotective effects. Significant improvement in NDS (1±1, n=19 versus 2±1, n=18) was also seen in mice treated with IAIP vs. controls. Similar beneficial effects of IAIP were seen in aged mice.

Conclusions:
Delayed administration of IAIP at 6 and 24 hours post-stroke results in significantly smaller infarcts and improved functional outcomes, indicating IAIP could potentially be translated into an important novel treatment strategy for adults with brain ischemia.
Abstract W MP49: Carriage of the PlA2 Allele of Glycoprotein IIIa is Associated With an Increased Risk of Ischaemic Stroke: A Meta-Analysis
Authors: Ellis Ben H , Floyd Christopher N , Ferro Albert
Published: March 2022

Introduction:
The PlA1/A2 polymorphism of glycoprotein IIIa has been associated with an increased risk of stroke, but with poor agreement between studies. Here we present a meta-analysis to assess the hypothesis that carriage of the PlA2 allele increases stroke risk.

Methods:
Electronic databases (MEDLINE and EMBASE) were searched for all articles evaluating carriage of the PlA2 allele and the incidence of stroke. Pooled odds ratios (ORs) were calculated using fixed-effect and random-effect models.

Results:
A total of 34 articles were eligible for statistical analysis (5,475 cases and 7,247 controls). For carriage of the PlA2 allele, OR 1.15 (n=12,115; 95% CI=1.05-1.26; p=0.002) was observed for total adult stroke, with sub-group analyses identifying an association with ischaemic (n=9,460; OR 1.22, 95% CI=1.10-1.35; p=0.0001) but not haemorrhagic stroke (n=2,470; OR 0.90, 95% CI=0.73-1.10; p=0.300). Significance was lost for ischaemic stroke in both young adults (18-60yr) (n=1,746; OR 1.26, 95% CI=0.97-1.65; p=0.09) and children (n=334; OR 1.24, 95% CI=0.74-2.08; p=0.41). Significant heterogeneity was observed for total (I2=60.1; p=0.0001) and ischaemic (I2=60.6; p=0.0003) stroke. Significance was lost for total stroke when analysed using a random-effect model (OR 1.15, 95% CI=0.99-1.34; p=0.06), but persisted in the ischaemic stroke sub-group (OR 1.23, 95% CI=1.03-1.46; p=0.02). Egger’s regression test suggested a low probability of publication bias (p>0.1 for all groups).

Conclusions:
This meta-analysis supports the carriage of the PlA2 allele of glycoprotein IIIa as a risk factor for ischaemic stroke in adults. The totality of published data does not enable sub-group analyses based on sex, ethnicity or traditional cardiovascular risk factors, and so the need for further studies remains.
Abstract W MP59: Spanish Language Consent Process Increases Prehospital Clinical Trial Enrollment in the Hispanic Community
Authors: Cherin Samantha B , Sanossian Nerses , Ali Latisha , Restrepo Lucas , Valdes- Sueiras Miguel , Hamilton Scott , Conwit Robin , Eckstein Marc , Pratt Franklin , Stratton Samuel , Starkman Sidney , Saver Jeffrey L ,   For the FAST-MAG Investigators and Coordinators
Published: March 2022

Background:
At over 45.5 million and growing, Hispanics are the United States’ fastest emerging ethnic group. Hispanics have a different prevalence of risk factors for stroke when compared with non-Hispanic whites, including increased frequency of hypertension, diabetes and obesity. Adequate ethnic representation in clinical trials is important to assure generalizability of results. Methods to enhance the Hispanic participants include Spanish language consent processes. We describe Hispanic participants in a prehospital clinical trial using Spanish as well as English consent forms.

Methods:
The NIH Field Administration of Stroke Therapy - Magnesium (FAST-MAG) Phase 3 Trial is a randomized study of prehospital initiated neuroprotection versus placebo in patients evaluated by paramedics less than two hours from symptom onset. After paramedics screened potential subjects, they dialed English or Spanish language enrollment lines, using voice-over-internet simulring numbers to directly connect to the cellphones of 4 Spanish-speaking or 4 English- speaking physician-investigators. Spanish speaking patients were able to learn about the study and be consented into the trial in their primary language.

Results:
There were 399 subjects enrolled in FAST-MAG who identified as Hispanic, 23% of the total cohort of 1700. Approximately half (48%) were enrolled using the Spanish- language consent process. Hispanic participants in FAST-MAG differed from non-Hispanic participants in several baseline characteristics, including being younger (65 vs. 71, p<0.001), more likely to have intracranial hemorrhage stroke subtype (33% vs. 20%, p<0.001), more often diabetic (32% vs. 19%, P<0.001), more severe baseline deficits (NIHSS 13 vs. 11, p=0.001) and higher systolic blood pressure (163 vs. 159, p=0.008). The two groups did not vary in gender, history of hypertension, time to paramedic evaluation, or rates of thrombolysis in cerebral ischemia.

Conclusion:
Better representation of Hispanics in clinical research is aided by availability of Spanish language consent processes. Dual language phone lines for consent elicitation are an effective strategy to increase enrollment of Hispanic participants in prehospital clinical research.
Abstract W MP68: MRI Infarct Patterns Do Not Predict Long-Term Cardiac Rhythm Recorder Findings
Authors: Phoa Keith , Kate Mahesh , Shuaib Ashfaq , Yaseen Ibrahim , Butcher Kenneth , Buck Brian
Published: March 2022

Intro:
It has been hypothesized that stroke/TIA patients with occult paroxysmal atrial fibrillation (PAF) have characteristic embolic patterns of cerebral infarction. We tested this hypothesis in patients studied with extended cardiac monitoring and MRI.

Methods:
Acute ischemic stroke patients with a negative 24-hour Holter monitor were prospectively enrolled in a study of event-triggered external loop recordings for 21 days. Fifty-five of 88 patients had acute MRI scans. Lesion volumes were calculated planimetrically on diffusion-weighted MRI (DWI) and fluid-attenuated inversion recovery (FLAIR) sequences. Leukoaraiosis volumes (FLAIR), acute and chronic lesion number and distribution were also assessed.

Results:
PAF was detected in 34.5% (19/55) of patients. Duration of PAF was <30s in 52.6% (10/19) of patients. Median(IQR) DWI lesion volumes (ml) were similar between patients with (1.74 (4.57)) and without PAF (1.57 (13.39), p=0.70). Median(IQR) chronic ischemic lesion volume on FLAIR (ml) in patients with (0.34 (3.28)) and without PAF (0.28(3.01)) was similar (
P=
0.82). Leukoaraiosis volumes were also similar (8.46 (12.51) vs. 3.42 (5.25),
P
=0.06), as were the total number of lesions (4 (6) vs. 5 (5),
P
=0.48), and total number of arterial territories affected (2 (2) vs. (2 (2),
P
=0.09). Patients with PAF duration <30s and >30s had similar median DWI volumes (2.21 (4.08) vs. 0.13 (6.24),
P
=0.61), FLAIR volumes (0.34 (2.87) vs. 0.88 (12.24),
P
=0.89), leukoaraiosis volumes (7.69 (13.11) vs. 8.46 (26.79),
P
=1.00), total number of lesions (6.5 (6) vs. 3 (7),
P
=0.72), and total number of arterial territories affected (3 (2) vs. 2 (3),
P
=0.84).

Conclusion:
Ischemic lesion volume, number and distribution in patients with occult PAF are similar to those with other mechanisms of infarction. In addition, patients with even very brief runs of PAF (<30s) have similar ischemic lesion loads to those with longer duration episodes. Our data suggest all ischemic stroke patients should be investigated for PAF.
Abstract W MP71: Quantification of Cerebral Vascular Reserve Mapping from Dynamic Susceptibility Contrast Magnetic Resonance Imaging
Authors: Jeong Yong I , Qian Renee , Vakil Parmede , Hurley Michael C , Ansari Sameer A , Bendok Bernard R , Batjer Hunt , Carroll Timothy J
Published: March 2022

Accurate measurement of cerebral vascular reserve (CVR) can be used as an indicator of cerebral hemodynamic impairment. The NIH/NINDS Stroke Progress Review Group has identification of CVR imaging as a top research priority. The Bookend Technique is a dynamic susceptibility contrast perfusion imaging technique that can quantify absolute cerebral blood flow (qCBF, ml/100g/min) and cerebral blood volume (qCBV, ml/100g). We report on first results using bookend perfusion scans to quantify CVR. We hypothesize that quantitative images of cerebral perfusion can be used to quantitate cerebral vascular reserve over the whole brain. We conducted an IRB approved, retrospective study with seven patients (4 male, 3 female, ages 61 ± 13.1 years) selected for angiographically confirmed occlusive vessel disease. The water exchange response curve, which mitigates the compartmentalization effects of the Gadolinium, was found to introduce a bias in multi-injection exams. We developed a new water exchange response curve that accounts for residual T1-shortening from the pre-stress images. Quantitative perfusion images were acquired before and after injection of vasodilator acetazolamide (ACZ) and the computed qCBF maps were coregistered and subtracted to generate CVR maps. We successfully measured the reactivity of cerebral vasculature. The CVR increase was found to be +10.3% ± 14.2% in unimpaired regions, and a net decrease of -8.63% ± 17.1% was observed in affected territories. We found the expected linear relationship between CVR and Cerebral Perfusion Pressure (CVR = 58.2*(CPP)-14.5). In conclusion, our results indicate that CVR can be successfully quantified from MR perfusion images using ACZ induced vascular reactivity and the Bookend Technique. Therefore, the resulting CVR maps can describe and present stages of hemodynamic failure of patients.
Abstract W MP80: Stroke Alters Neutrophil and Monocyte Function
Authors: Ruhnau Johanna , Schulze Karsten , Gaida Bernadette , Langner Sönke , Kessler Christof , Bröker Barbara , Dressel Alexander , Vogelgesang Antje
Published: March 2022

Objective:
Stroke-induced immune alterations predispose patients to infections. While the relationship between stroke and the adaptive immune system has been investigated in detail, to date it is unknown whether the innate immune system, which forms the first line of antibacterial defense, is also impaired in stroke patients. We therefore investigated chemotaxis, phagocytosis, oxidative burst, degranulation of defensins, and NETosis in monocytes and in neutrophil granulocytes.

Methods:
Patients suffering from acute ischemic stroke were recruited within 12 h of symptom onset; blood was sampled on admission and on days 1, 3, 5, and 7. Cell migration, phagocytosis, and oxidative bursting of phagocytes were determined by the MIGRATEST Kit®, the Phagotest Kit®, and the Phagoburst Kit®, respectively. Human neutrophil peptides 1-3 and serum metanephrine levels were measured by enzyme-linked immunosorbent assay, and NETosis was quantified by immunohistochemistry.

Results:
The key mechanisms required for bacterial killing, oxidative burst and NETosis, were significantly reduced in samples taken from stroke patients whereas migration, phagocytic function, and defensin production remained unimpaired in monocytes and granulocytes from these patients.

Interpretation:
Stroke-induced immune alterations include impairment of the first-line defense carried out by specialized phagocytes against bacteria. The hypothesis that these alterations enhance susceptibility to acquired infections is supported by our observation that oxidative bursting in monocytes was more impaired in stroke patients with stroke-associated infections.
Abstract W MP82: Role of OPG/RANKL/RANK System in Ischemic Brain in Mice
Authors: Shimamura Munehisa , Nakagami Hironori , Osako Mariana K , Kurinami Hitomi , Wakayama Kouji , Koriyama Hiroshi , Morishita Ryuichi
Published: March 2022

Background & Purpose:
Although recent clinical studies showed that the high serum osteoprotegerin (OPG, decoy receptor for RANKL) level was associated with unfavorable outcome in ischemic stroke, its mechanism is unknown. Since OPG/ RANKL (receptor activator of nuclear factor-β ligand)/ RANK (receptor for RANKL) system has critical roles in immune systems as well as osteoclast biology, we hypothesized the OPG/RANKL/RANK system might play roles in post-ischemic inflammation and immune response. To explore the possibility, we examined their expressions and functions in mice.

Methods:
The expression and the functional analysis of OPG/RANKL/RANK system were examined in 70 min-transient cerebral artery occlusion model in C57BL6/J (WT) mice or OPG
-/-
mice, whose serum RANKL level is higher than that in WT mice. The effects of RANKL/RANK signaling were also examined in mixed glial-neuron cultures stimulated by lipopolysaccharide (LPS).

Results:
RANK mRNA was increased from 4 hrs after ischemia, whereas OPG and RANKL mRNA were up-regulated from 5 hrs after ischemia. The RANKL and RANK were expressed in activated macrophage/microglia and OPG was partly expressed in activated macrophage/microglia. OPG
-/-
mice showed reduced infarct volume 72 hrs after ischemia, whereas the infarct volume was not reduced in OPG
-/-
mice treated with RANK/Fc chimera. Also, RANKL-treated wild type mice showed the reduction in infarct volume. The expression of IL-6, TNF-α, and IL-1β was less in OPG
-/-
mice and RANKL-treated mice. Additionally, the LPS-stimulated neuron-glia mixed culture showed less neuronal death in the group of RANKL-treatment cultures through inhibition of inflammatory cytokines production.

Conclusions:
These data indicate that RANKL/RANK signaling might work as anti-inflammation in the ischemic brain through inhibiting TLR signaling pathways. High serum OPG level in clinical situations might inhibit RANKL/RANK signaling and exacerbate the post-ischemic inflammation, resulting in poor outcomes. Further studies on the OPG/RANKL/RANK system might shed light on the clarification of molecular mechanisms in post-ischemic inflammation.
Abstract W MP83: Transcutaneous Vagus Nerve Stimulation for Treatment of Acute Cerebral Ischemia in Rats
Authors: Ay Ilknur , Ay Hakan
Published: March 2022

Introduction:
Direct electrical stimulation of the vagus nerve in the neck using surgically implanted electrodes reduces infarct volume in rats. But the invasive nature of the direct stimulation makes it impractical in acute human stroke. We tested the effect of various non-invasive or minimally invasive vagus nerve stimulation (VNS) techniques on infarct size, physiological parameters, and clinical outcome after transient middle cerebral artery (MCA) occlusion in rats.

Methods:
We induced cerebral ischemia by filament occlusion of the right MCA for 2h, followed by reperfusion in adult male spontaneously hypertensive rats (n=52). We performed: (1) auricular transcutaneous VNS (atVNS) via application of a microelectrode array to the auricular concha, (2) cervical transcutaneous VNS via application of HNS12 transcutaneous nerve stimulator (ctVNS-1) and Gammacore non-invasive cervical vagus nerve stimulator (ctVNS-2) put on to the skin overlying the vagus nerve in the neck. We applied tVNS for 1h starting 30 min after the induction of ischemia. We measured neurological deficit daily; at the end of 1 week we euthanized the animals to determine cerebral infarct volume. In an additional group of animals, we performed c-Fos immunohistochemistry to verify the activation of brainstem vagal centers after tVNS.

Results:
There was no difference in physiological parameters between control and tVNS-treated animals. c-Fos immunohistochemistry showed activation of nucleus tractus solitarius indicating that tVNS was able to activate the same brain stem nuclei activated by direct VNS. The effect of different tVNS techniques on infarct volume was comparable; infarct volume was 28% smaller with atVNS, 30% with ctVNS-1, and 33% with ctVNS-2 as compared with control animals (p<0.05 for all 3 techniques; n=6 per group). The effect of tVNS on tissue outcome was associated with better neurological scores and grip strength measurements (p<0.05 for all 3 techniques).

Conclusion:
Non-invasive stimulation of the vagus nerve for a brief period of time provides significant protection against ischemic injury in rats. Given its simplicity and feasibility at the setting of acute ischemic stroke, tVNS holds promise for treatment of human stroke.