bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract TP18: Iron Overload Induces Cognitive Deficits And Autophagy Impairment In The Brains Of Aged Mice In A Sex-Dependent Manner
Authors: Noh Brian , Rahman Syed M , McCullough Louise D , Moruno-Manchon Jose F
Published: February 2023

Iron deposits have been observed in postmortem brain samples from Alzheimer’s disease patients. Different iron overload models have been used to study neurodegeneration in young mice. However, the effects of excess iron with aging, which is a critical factor for Alzheimer’s disease and iron deposition in the brain, remain unexplored. In addition, iron metabolism is regulated by autophagy, a cellular process that degrades intracellular components. Brain autophagy is downregulated with aging and in females, and autophagy impairment occurs in many neurodegenerative disorders. Thus, we hypothesized that sex differences exist in autophagy in the brains of aged mice treated with iron overload. We treated aged (18-20 months) WT mice of both sexes with iron overload (5% ferric citrate, FC, 3 days/week/6 weeks), or a vehicle, by oral gavage. We found that aged females showed impairment in conditioning memory (Fear conditioning), but not aged males. Aged females treated with FC also showed significant increase of iron deposition in their brains, compared with control females. However, we did not find significant differences between control and treated male mice. Furthermore, the brain lysates of aged females treated with iron overload showed increased levels of the early autophagy marker BECLIN1 (in both sexes), and enhanced levels of the autophagy cargo p62 (in both sexes) and the lysosomal marker LAMP1 (only in females), compared with control mice. This suggests that iron overload induces the initial stage of autophagy in both sexes, but impairs late autophagy, mainly in female mice, compared with control mice. Furthermore, in primary culture of cerebral endothelial cells (CEC) isolated from aged mice, we found that CEC treated with ferric citrate exhibit enhanced DNA damage and reduced cell proliferation in a dose-dependent manner. Importantly, CEC derived from female mice showed reduced autophagy flux, compared with male CEC. This suggests that iron overload may be more detrimental in the cerebrovasculature of aged female mice than in males, and this could explain iron overload-induced memory deficiency in aged female mice, but not in males.
Abstract TP180: Clinical Features And Echocardiographic Changes Of Cryptogenic Perforator Infarction Patients With Patent Foramen Ovale
Authors: Tang Haiyan , Zhang Hui , Wu Fei , Yu Chun , Dong Qiang , Cao Wenjie
Published: February 2023

Background and Purpose:
We sought to investigate the clinical characteristics and echocardiographic changes of cryptogenic perforator infarction (PI) patient with patent foramen ovale (PFO).

Methods:
From Jan 2015 to Sep 2018, patients diagnosed with cryptogenic stroke were retrospectively included in our study. All patients received transthoracic echocardiography (TTE) and transcranial Doppler (TCD). PFO was diagnosed by the bubble test of TCD. Comparisons of clinical features and TTE findings were performed firstly in PI patients, between PFO and non-PFO, and then in patients with PFO, between PI and cortical infarction (CI).

Results:
A total of 228 patients were retrospectively analyzed. PFO was detected in 80 of the patients, of those, 23.7% (19/80) had pure PI (PFO-PI) and 76.3% (61/80) had CI (PFO-CI). In patients without PFO, 30.4% (40/148) had pure PI (non-PFO-PI). Compared with non-PFO-PI, PFO-PI associated with a higher proportion of vertebrobasilar circulation infarctions (VCI) (47.5% vs. 17.9%, p= 0.014), lower median left ventricle end-diastolic diameters (LVEDd) (46mm vs. 50mm, p= 0.016) and lower peak E-wave velocity (57cm/s vs. 72cm/s, p= 0.022). In PFO patients, PFO-PI was detected with a lower median aorta root diameters (32mm, p=0.004) and lower median LVEDd (46mm, p=0.024) than PFO-CI (34mm and 48mm, respectively).

Conclusions:
Higher proportion of VCI was found in PFO-PI. Lower LVEDd, lower Em and without aortic root dilatation were three echocardiographic features associated with PFO-PI in cryogenic PI patients.
Abstract TP186: Multidisciplinary Approach To Patent Foramen Ovale (pfo) Closure For Cryptogenic Stroke: Brain-heart Board Experience
Authors: Khan Muhib , McCarthy Philip , Haggerty Diana , Min Jiangyong , Miller Malgorzata , Tsai Jenny P , Merhi William , Berkompas Duane , Wees Nabil , Khan Nadeem I , Ahrar Asad , Evans Elizabeth , Dahu Musa , Gauri Andre , MOELKER TARAH A , chalfoun nagib
Published: February 2023

Background:
The prevalence of PFO is up to 40% in patients with cryptogenic stroke and TIA suggesting paradoxical embolism. PFO closure in carefully selected patients is an effective secondary preventive strategy in these patients. We report predictors of closure recommendation by multidisciplinary board for PFO closure and its impact on outcomes.

Methods:
Brain-Heart Board is comprised of vascular neurology, cardiology subspecialities (structural, electrophysiology and cardiac imaging). Adult patients referred to the board for consideration of PFO closure between October 2017 to March 2021 were included in this analysis. Demographics, comorbid conditions, infarct location, Risk of Paradoxical Embolism (RoPE) Score, event frequencies (recurrent TIA or stroke, intracranial hemorrhage (ICH), post-PFO closure cardiac arrythmias) and modified Rankin Scale (mRS at 1 year) were compared between the groups (PFO closure vs. medical management). Multivariable logistic regression was used to identify predictors of closure and chi-square tests to test differences in outcomes for patients according to management.

Results:
The board discussed 270 patients (229 stroke; 41 TIA). 119 (44%) patients were recommended for PFO closure of which 117 (98%) had infarct on neuroimaging. Age and RoPE score were similar in closure and medical management cohorts (age; 50±12 vs. 52±13, p>0.05 RoPE 4±3 vs. 6±2 p>0.05). In multivariable analysis, absence of infarct on neuroimaging was an independent predictor of medical management recommendation by the board (OR 0.05 95% CI 0.01-0.19 p<0.05). Event frequency was low in both cohorts (5.9% vs. 4.8% p>0.05) and were comprised primarily of cardiac arrhythmias (6 atrial fibrillation and 1 ICH in Closure group; 1 TIA and 1 recurrent stroke in medical management group). Excellent functional outcome (mRS 0-1) was similar in both cohorts (66% vs. 71% p>0.05) at 1 year.

Conclusion:
Multidisciplinary approach for selecting patients for PFO closure results in low frequency of complications (recurrent ischemic stroke, ICH and post-PFO closure cardiac arrythmias) and good outcomes. Presence of infarct on neuroimaging predicts closure recommendation by the multidisciplinary brain-heart board.
Abstract TP188: Ischemic Stroke While Bathing: Stroke Subtype And Association With Cerebral Small Vessel Disease
Authors: Ishikawa Takahiro , Sato Takeo , Okumura Motohiro , Kokubu Tatsushi , Takahashi Junichiro , Kitagawa Tomomichi , MAKI TANABE , ONDA ASAKO , Komatsu Teppei , Sakuta Kenichi , Sakai Kenichiro , Umehara Tadashi , Mitsumura Hidetaka , Iguchi Yasuyuki
Published: February 2023

Background and Purpose:
Ischemic stroke developed while bathing (ISB) occasionally causes fatal accident. Factors associated with ISB remain unclear. We aimed to estimate the clinical characteristics of ISB, especially the subtype of stroke and the association between cerebral small vessel disease (CSVD) markers.

Methods:
Consecutive patients with ischemic stroke ≤48 h from onset who underwent MRI during the hospitalization were included. We divided patients into ISB and non-ISB group to analyze differences in clinical characteristics, especially the stroke subtype based on TOAST classification and the association between CSVD markers. The CSVD markers were evaluated by MRI and defined as old lacunes, white matter hyperintensities scale (separately scored by periventricular hyperintensity [PVH] and deep and subcortical white matter hyperintensity), the number of cerebral microbleeds (CMBs) categorized as lobar, deep, and infratentorial lesions, and the scale of enlarged perivascular space in the basal ganglia and centrum semiovale.

Results:
We screened 2,112 consecutive patients with ischemic stroke including 1,517 patients (1,070 [71%] male, median age 69 years). Of these patients, 52 (3%) were included in ISB group. In univariable analysis, ISB group showed higher frequency of large-artery atherosclerosis (
p
= 0.004), lower frequency of cardioembolism (
p
= 0.008), smaller number of cerebral microbleeds in deep lesions (
p
= 0.025) and lower grade of PVH (
p
= 0.023). ISB group showed a lower NIHSS score on admission (
p
= 0.002). However, there was no difference in mRS score at 3 months from the onset. In the multivariable analysis (Figure), large-artery atherosclerosis (OR 2.46, 95% CI 1.21-5.01,
p
= 0.013) and CMBs in deep lesions (OR 0.75, 95% CI 0.58-0.98,
p
= 0.038) showed independently associated with ISB.

Conclusions:
ISB showed higher frequency of large-artery atherosclerosis, however, had a less association with the presence of CMBs in deep lesions.
Abstract TP193: The Protective Effects Of Exercise On Cerebral Ischemia Injury In Diabetes
Authors: Sawant Harshal , Sigdel Smara , Chen Shuzhen , Wang Jinju , Bihl Ji C
Published: February 2023

Ischemic stroke (IS) causes neurological dysfunction due to a loss of cerebral blood flow with exaggerated cerebral damage in diabetes. Exercise has shown beneficial effects on the vascular system and diabetes by mediating inter-organ communications. However, whether and how exercise contributes preventatively to the brain after IS in type 2 diabetes (T2D) is undefined. Here, we aimed to determine the effects of exercise on the metabolism, cerebral injury, neurological function, and protein expression in the brain after IS. T2D diabetic mice (db/db, 7-8 wks), and age/sex-matched controls (db/c) were subjected to exercise (10 m/min, 5 days/wk for 8-wks) or sedentary. Body weight and blood glucose were recorded once a week. One day after exercise, middle cerebral artery occlusion surgery was performed to induce IS. Sensorimotor deficits were assessed by the adhesive removal and corner tests two days after surgery. Afterward, the brain samples were collected for measuring the infarct size by cresyl violet staining. The proteins from the ipsilateral and contralateral brain were used for Western Blot to measure the levels of endothelial nitric oxide synthase (eNOS), neuronal NOS, NADPH oxidase (Nox2), Nox4, nuclear factor-kappa B (NF-κB), NF-κ light polypeptide gene enhancer in B-cells inhibitor, alpha (IκBα). We found: 1) exercise could stabilize the blood glucose in male db/db mice and have effects on preventing blood glucose increase at the early age of female db/db mice; 2) exercise could improve sensorimotor deficits by reducing tape contact and removal time and balancing the corner turning times (p< 0.05); 3) the infarct size is decreased in exercised group in both db/c (18.5 ± 2.2% and 22.2 ± 2.5%), and db/db mice (25.6 ± 3.1% and 35.6 ± 3.8%, exercise
vs.
sedentary, p< 0.05); 4) proteins related to oxidative stress and inflammation were downregulated in the brain of exercised diabetic IS mice. In conclusion, exercise could provide protective effects on stabilizing metabolism, improving neurological function, and reducing brain injury by downregulating the protein expression related to inflammation and oxidative stress in the brain.
Abstract TP194: The FLAIR Signal Intensity Ratio Predicts Early Neurologic Deterioration For Small Subcortical Infarcts
Authors: Fayad Fayez H , Shah Asghar , Fayad Noora , Chang Kelvin , Snow Ethan , Shu Liqi , Yaghi Shadi , Goldstein Eric D
Published: February 2023

Introduction:
Early neurologic deterioration (END) occurs in nearly 40% of small subcortical infarct (SSI), with the majority occurring as a byproduct of completion of the initial infarct leading to significant morbidity. Presently there is not a robust predictive radiographic biomarker for END from SSI. The fluid-attenuated inversion recovery signal intensity ratio (FLAIR-SIR) ≥ 1.15 was shown to aid in differentiation of completed infarcts from those with penumbra allowing in identifying those that may benefit from alteplase. We aimed to use the FLAIR-SIR as a biomarker for predicting END from SSI. We hypothesized that those with a higher FLAIR-SIR had less risk of developing END due to completion of infarct.

Methods:
We retrospectively enrolled individuals who were seen at the Rhode Island Hospital between 2019 and 2022. Those included had a radiographic diagnosis of a SSI per STRIVE criteria. The primary outcome was END diagnosed as ≥ 2-point increase in NIH Stroke Scale score within 24 hours of admission. The primary exposure was FLAIR-SIR, defined as the ratio of ipsilateral FLAIR intensity within the region of infarct divided by the FLAIR of the contralateral identical anatomical location. Standard descriptive and unadjusted regression analyses were used.

Results:
A total of 195 individuals were included with a mean age 68 (± 13) years, and 59.5% (116/195) being male. END was identified in 3.6% (7/195). The mean overall FLAIR-SIR was 1.26 (± 0.30) with a mean of 1.26 (± 0.34) for those without END and 1.07 (± 0.25) for those with END (
p
= 0.66). Using Youden’s J index for FLAIR-SIR, the optimal cutoff was determined to be ≤ 1.15. For a FLAIR-SIR ≤ 1.15, unadjusted Cox regression finds a hazard ratio of 4.1 (95%CI 0.79 - 21.1,
p
= 0.092) for END within 24 hours.

Conclusions:
FLAIR-SIR measurement above 1.15 may serve as a radiographic predictor of less risk of END amongst SSI. Our findings corroborate prior studies identifying 1.15 as a threshold for at-risk and completed SSI and should receive thrombolytics to minimize risk of END. Future studies are needed to increase power and confirm our findings.
Abstract TP196: Real Life Utility Of PCSK9 Inhibitors Following Ischemic Stroke
Authors: Arons Danielle , Grimes Kathryn , Mehndiratta Prachi , Chaturvedi Seemant
Published: February 2023

Introduction:
Hyperlipidemia is a modifiable risk factor for stroke, with the magnitude of LDL-C reduction correlating to risk of recurrent ischemic stroke. A high-intensity statin is considered standard of care in addition to dietary modification. Proprotein convertase subtilisin/kexin type 9 (PCSK9) inhibitors are used if the LDL-C target is not achieved despite maximally tolerated statins with ezetimibe. We investigated the role for PCSK9 inhibitors in our ischemic stroke population.

Methods:
An IRB approved retrospective review of patients admitted for acute ischemic stroke between 2019 – 2020 was performed. We estimated the probability of benefit from PCSK9 treatment with the following criteria: LDL > 190 regardless of treatment, LDL 161-190 on any dose statin therapy, and LDL 131-160 on high intensity statin therapy.

Results:
774 patients with ischemic stroke were admitted during this time frame. Mean age 62.8 +/- 13.2 with 57% male (52% Black, 42% White). 584 (75%) of these patients had LDL above goal. One patient was on PCSK9 therapy on admission. Although the majority of our patients had uncontrolled cholesterol, only one additional patient would meet the strict prescription criteria for PCSK9 therapy of being adherent to maximal tolerated statin dose and ezetimibe with LDL above goal. Other potential patients with estimated benefit include LDL >190 (20 patients), LDL 161-190 (16 patients), LDL 131-160 (13 patients). In total this amounts to 6.7% of stroke patients who we would expect to need PCSK9 inhibitors in the future.

Conclusion:
Using conservative projections, we estimate that about 1 in 14 patients with ischemic stroke could benefit from PCSK9 therapy. Achieving LDL targets more consistently is a promising approach to decreasing the recurrent stroke rate. Further prospective data evaluating outcomes after initiating PCSK9 inhibitor treatment are needed to validate these findings.
Abstract TP199: Association Of Blood Pressure Parameters In Unsecured Aneurysmal Subarachnoid Hemorrhage And Outcomes
Authors: Haripottawekul Ariyaporn , Paracha Saba , Haque Deena , Persad-Paisley Elijah M , Shamshad Alizeh , Subramaniam Thanujaa , Reznik Michael , Furie Karen L , Yaghi Shadi , Mahta Ali
Published: February 2023

Background:
Elevated systolic blood pressure (SBP) has been linked to pre-procedural rebleeding risk and poor outcome in patients with aneurysmal subarachnoid hemorrhage (aSAH). However, the relationship between blood pressure parameters including mean arterial pressure (MAP) with rebleeding both prior to and during aneurysm securement remains unclear. This study seeks to determine the association between BP parameters and rebleeding events and outcomes in patients with aSAH.

Design/Methods:
We performed a retrospective analysis of a prospectively collected cohort of consecutive patients with aSAH admitted to an academic center between July 2016 and March 2021. All BP parameters, which were recorded on an hourly basis from admission, were reviewed. Per our institutional protocol, the SBP target is <140 mmHg for all unsecured aSAH patients. Rebleeding was defined as radiographic worsening of hemorrhage prior to or immediately following aneurysm securement, as seen on imaging and determined by a radiologist. Binary regression analysis was used to determine association between maximum recorded BP parameters and outcomes including rebleeding and poor functional outcome defined as modified Rankin Scale 4-6 at 3 months post-discharge.

Results:
The cohort included 295 patients (mean age 57 years [SD 13.4], 61% female, 87% received endovascular treatments, 13% surgical clipping). Two or more consecutive SBP values >140 were seen in 41% and >160 in 15% of patients. Rebleeding prior or during securing aneurysms including intra-procedural bleeding occurred in 53 patients (18%). There was no association between either maximum recorded SBP (169.7 mmHg [36.2] vs. 166.9 [32.7], p=0.57) or MAP (121 mmHg [22.9] vs. 117.8 [22.9], p=0.42) and rebleeding. However, maximum recorded MAP was associated with poor outcome (OR 1.01 for 1 mmHg increase in MAP, 95% CI: 1.006-1.02, p=0.039).

Conclusions:
Elevated MAP peri-securement of a ruptured cerebral aneurysm can be associated with poor outcome. Multicenter prospective studies should further examine the association between MAP cut offs and outcomes to be considered in guidelines.
Abstract TP2: Iv-thrombolysis Improves Perfusion Profiles In Patients With Anterior Circulation Occlusion Regardless Of Recanalization
Authors: Olive-Gadea Marta , Requena Manuel , Garcia-Tornel Alvaro , Rizzo Federica , Rubiera Marta , Muchada Marian , Juega Jesus , PAGOLA JORGE , Rodriguez-Luna David , Rodriguez Villatoro Noelia , tomasello alejandro , Molina Carlos A , Ribo Marc
Published: February 2023

Introduction:
Recent publications suggest a positive added effect of iv-thrombolysis (IVT) in patients that undergo endovascular treatment (EVT). It is hypothesized that thrombolytics might have a beneficial impact on microcirculation beyond recanalization. We aim to analyze the potential impact of IVT in patients who underwent a repeated CTP after interhospital transfer.

Methods:
We retrospectively screened 116 patients transferred to our comprehensive stroke center from a primary stroke center capable to perform CTP from June 2021 to August 2022. We collected clinical and radiological data of patients that underwent CTP at both centers, which were analyzed with Rapid software. A neurointerventionalist assessed the occlusion location in CTA.

Results:
Twenty-eight patients with anterior circulation occlusion underwent two multimodal studies, median time between CTPs was 168[142-190]min. Fifteen (53.6%) patients received IVT in the primary stroke center and 13(46.4%) received EVT. Reperfusion occurred in 3(10.7%) cases, migration to distal segments in 5(17.9%). Among the 20 patients without changes in the occlusion location, CTP volumes remained stable (Tmax>6s 61[41-141]ml vs 62[24-178], p=0.72; Tmax>10s 19[8-96] vs 13[0-105], p=0.55; CBF30 0[0-66] vs 0[0-65], p=0.99). CTP volumes tended to decrease with IVT, but a modest increase was observed in patients that did not receive IVT (Tmax>6s -17[-29,+8]ml vs +22[-8,+42], p=0.08; Tmax>10s -8[-16,+6] vs +6[0,+37], p=0.10; CBF<30% 0[-6,0] vs 0[0,+4], p=0.14) (figure). In a linear regression to predict CTP volumes in the second center, interaction between baseline values and IVT was significant for Tmax>10s (p=0.06) and CBF<30% (p<0.05).

Conclusion:
In acute stroke patients with a LVO, no substantial changes are expected in CTP volumes in the absence of recanalization. However, patients that receive IVT tend to present reduced hypoperfusion volumes as compared with patients that do not receive IVT.
Abstract TP20: Validation Of Bedside Applicability Of Manual Versus Automated Measurements Of Brain Arterial Diameters
Authors: Garzon Mancera Nicolas D , Khasiyev Farid , Spagnolo Allende Antonio J , Rundek Tatjana , Wright Clinton B , Elkind Mitchell , Sacco Ralph L , Del Brutto Victor J , DEL BRUTTO OSCAR , Gutierrez Jose
Published: February 2023

Introduction:
Dolichoectasia of brain arteries has significant clinical implications. It can be measured with automated imaging software such as LKEB Automated Vessel Analysis (LAVA). However, this is not currently used as part of standard clinical practice. Non-automated alternatives such as a Picture Archiving Communication System like RadiAnt are more widely accessible in clinical settings Method: In this validation study, we included 600 participants of the Northern Manhattan Study (NOMAS) and 260 participants of the Atahualpa project. Both studies obtained 1.5T, 3-dimensional time-of-flight brain MRAs. Using RadiAnt (version 2022.1), three independent readers (general practitioner (GP), neurology resident (NR), and vascular neurologist (VN)) measured the diameter of the internal carotid artery (ICA); the most proximal 5 mm of the middle cerebral artery (MCA) and anterior cerebral artery (ACA); and the most proximal portion of the basilar artery (BA). The same vessels were also measured by LAVA. We calculated the intraclass correlation of each rater’s diameters versus those obtained with LAVA (version 5.10.0). GP and NR measured the NOMAS cohort. VN measured the Atahualpa project cohort.

Results:
The intraclass correlations between diameters obtained by GP and LAVA were stronger for the BA, left ICA, and right ICA (table 1). For VN and LAVA, the same correlations were also significant. For NR and LAVA, correlations were more significant for BA and left MCA. Rater-LAVA correlations for the remaining vessels failed to reach 60%. There is a strong correlation between diameters obtained by GP and NR for the BA and right ACA.

Conclusion:
Our results suggest that manual measurements of the diameter of the ICAs and BA regardless of training status are reliable and could be used to identify dilated brain arteries using diameter cutoffs for possible interventions. The associations with the remaining vessels were too variable to support the same conclusion.
Abstract TP209: Role Of Endoglin In The Regulation Of Organic Anion Transporting Polypeptide 1a4 At The Blood-brain Barrier: Relevance To The Treatment Of Ischemic Stroke
Authors: Betterton Robert , Williams Erica I , Stanton Joshua , Lochhead Jeffrey , Davis Tom , Ronaldson Patrick
Published: February 2023

Objectives:
We have shown that transforming growth factor-β (TGF-β)/activin-like kinase 1 (ALK1) signaling at the blood-brain barrier (BBB) can regulate organic anion transporting polypeptide 1a4 (Oatp1a4), a transporter involved in brain uptake of drugs relevant to stroke pharmacotherapy (i.e., 3-hydroxy-3-methylglutaryl coenzyme A (HMG CoA) reductase inhibitors (statins)). To date, the involvement of endoglin (ENG), a co-receptor that can modulate the TGF-β/ALK1 pathway, in regulation of Oatp1a4 expression/activity has not been elucidated. The objective of this study is to evaluate whether ENG plays a role in the control of BBB transport mechanisms for stroke therapeutics.

Methods:
Our studies were conducted using the immortalized mouse brain endothelial cell line bEnd.3. Endoglin knockdown mutants were generated through lentiviral transfection of ENG-shRNA and validated using confocal microscopy and western blot analysis. Cells were then treated with bone morphogenetic protein-9 (BMP-9; 1 nM), a known TGF-β/ALK1 agonist. Oatp1a4 protein expression and transport activity were measured using western blot analysis and
in vitro
uptake assays using the known Oatp substrate drug [
3
H]atorvastatin. Additionally, ENG expression in bEnd.3 cells was assessed under normoxic and oxygen/glucose deprivation (OGD) conditions (i.e., 1% O
2
in glucose-free medium).

Results:
BMP-9 treatment in non-transfected bEnd.3 cells resulted in increased Oatp1a4 protein expression and increased cellular uptake of [
3
H]atorvastatin. These enhancements in Oatp1a4 functional expression were attenuated in ENG-shRNA knockdown bEnd.3 cells treated with BMP-9. Confocal microscopy and western blot analysis confirmed decreased ENG protein expression following shRNA transfection. Under OGD conditions, expression of ENG in bEnd.3 cells was significantly increased.

Conclusions:
Our data provide the first evidence for involvement of ENG in regulation of Oatp1a4 by TGF-β/ALK1 signaling in brain microvessel endothelial cells. These observations have clear implications for CNS delivery of drugs relevant to treatment of stroke. Studies are ongoing in our laboratory to fully evaluate ENG regulation of BBB transport mechanisms in the setting of ischemic stroke.
Abstract TP213: Aging Promotes The Rupture Of Intracranial Aneurysm In Mice
Authors: Uchikawa Hiroki , Sato Hiroki , Ai Jinglu , Lawton Michael , Eguchi Satoru , Hashimoto Tomoki
Published: February 2023

Introduction:
Aging is a known risk factor for intracranial aneurysm rupture in human clinical studies. However, clinical studies only showed the association, but not the causality of aging and aneurysm rupture. Using our well-established mouse model, we directly tested the hypothesis that aging increases the rupture rate of aneurysms.

Methods:
We used both male and female C57BL/6J mice at 10 weeks and 18 months of age. We induced intracranial aneurysms by a combination of elastase injection and DOCA-salt hypertension. We compared the formation and rupture rates of intracranial aneurysms and the survival curve between young mice and aged mice of each sex.

Results:
Aged female mice had a higher rupture rate (86.7% vs 38.9%; P<0.05, Fisher’s exact test, Fig. A and B), and a higher formation rate (88.2% vs 62.1%; P<0.05, Fisher’s exact test) than young mice. Aged female mice had a lower survival rate than young mice (Fig. C). Aged male mice had both a higher rupture rate (94.1% vs 62.5%; P<0.05, Fig. D and E) and a higher formation rate (94.4% vs 64.0%; P<0.05) than young mice. There is a trend of a worse survival rate in aged male mice (Fig. F).

Conclusions:
Aging promotes both the formation and rupture of intracranial aneurysms in both male and female mice. These data agree with clinical observations and will serve as a basis for further investigation of the roles of the aging-related biological changes in the development of aneurysm rupture.
Abstract TP221: Protective Effects Of Galectin-3 On Endothelial Progenitor Cells Under Ischemic Conditions
Authors: Wesley Umadevi V , Dempsey Robert J
Published: February 2023

Stroke remains the major cause of long term functional deficits and death with limited treatment options. Ischemic stroke leads to endothelial dysfunction, blood brain barrier disruption, poor angiogenesis, inflammation, and neuronal cell death. Stem/Progenitor cells including mesenchymal, neural, and endothelial progenitor cells (EPC) can enhance tissue regeneration and repair. However, the beneficial effects of progenitor cells have been modest presumably due to their poor recruitment and survival in an ischemic microenvironment. Galectin-3 (Gal3), a β-galactoside-binding protein controls cell proliferation, survival, angiogenesis, and migration/recruitment. We have shown that the expression of Gal3 is significantly up-regulated in ischemic rat brain, and that blocking of Gal3 decreases the cerebral blood vessel density. We recently demonstrated that Gal3 reduces cerebral infarction and enhances recovery of motor/sensory functions in an animal stroke model. In this study we examined the effects of Gal3 on bone marrow derived EPC. EPCs were characterized by the uptake of acetylated LDL and expression of CD34 and VEGFR2/KDR. We further demonstrate that Gal3 enhances the proliferation, angiogenesis, migration, and survival of bone marrow derived EPCs in an in vitro ischemic conditions following oxygen glucose deprivation. Gal3 effects were associated with up-regulation of chemoattractant SDF1 and VEGF, further emphasizing the role of Gal3 in stem/progenitor function. Overall, these results indicate a key role for Gal3 in neuro-vascular protection and functional recovery following ischemic stroke through modulation of angiogenic and apoptotic pathways in progenitor cells.
Abstract TP227: Identification Of Somatostatin (sst) Interneuron-specific Micrornas And Transcriptome Associated With Cerebral Ischemia
Authors: Liu Xianshuang , Chopp Michael , Pan Wanlong , Wang Xinli , Golembieski William , Zhang Zheng Gang
Published: February 2023

Background:
Somatostatin (SST) lineage neurons, the second largest family of GABAergic interneurons in the neocortex, play a critical role in cortical activity and neuroplasticity after stroke. microRNAs (miRNAs) play an important role in mediating ischemic brain repair. Elucidating changes of miRNAs in SST neurons after stroke may provide molecular insight into SST neuron function.

Methods:
To systematically determine miRNA expression in SST neurons, we employed a miRNA tagging and affinity-purification (miRAP) method to select specific SST neurons via the Cre-loxp binary system in male SST-tAgo2 (tAgo2: a GFP-MYC-Ago2 fusion protein) mice subjected to 7 day middle cerebral artery occlusion (MCAO) (N=3/group).

Results:
Using miRNA sequencing, we found a total of 523 known and novel miRNAs with distinct profiles that were expressed in SST neurons isolated from ischemic hemispheres compared to miRNAs from sham operated mice. Among them, 70 and 7 miRNAs showed significantly increased and decreased expression, respectively, with a fold change above 2.0 and over 100 copy numbers. Top 3 upregulated miRNAs included miR-381-3p, miR-543-3p and miR-433-3p, while the top 3 downregulated ones were miR-9-5p, miR-423-5p and miR-486b-5p. Furthermore, using miRAP method in combination with RNA-sequencing, we uncovered over 1000 differential gene expression with at least 1.5-fold enrichment in ischemic SST neurons. Integrative analysis of miR-mRNA network by IPA algorithm revealed that 23 miRNAs were predicted to target deregulated mRNAs. Enriched pathways targeted by miRNAs were involved in NTRK3 (neurotrophic receptor tyrosine kinase 3), insulin, PI3K, EGFR1, mTOR signaling, etc, which are associated with neurodevelopment, neuronal growth and survival. These data highlight the importance of these gene regulatory mechanisms to SST neuron function after stroke.

Summary/Conclusion:
Taken together, our study provides comprehensive miRNA and transcriptional signatures for SST neurons after stroke. These data will facilitate a deeper understanding of the neuroplasticity after brain injury, potentially leading to new therapeutic targets against ischemic stroke.

Funding:
NIDDK Diacomp 21AU4165 and AHA/ASA 18IPA34170331 (X.S.L).
Abstract TP230: A Novel Mouse Model Of IVH Reveals Local, But Not Global Inflammation, And No Cognitive Impairment Over Multiple Time Points
Authors: Demel Stacie L , Walsh Kyle B , Woo Daniel , Luo Agnes
Published: February 2023

Background:
Intracerebral hemorrhage is the most severe stroke subtype, and extension of blood into the ventricles (IVH) is associated with increased morbidity and mortality. The mechanisms that contribute to poor clinical outcomes are unclear. We set out to establish a novel IVH mouse model for the purpose of measuring motor and cognitive function in acute and chronic stages.

Methods:
Mice underwent either an autologous blood (40ul; IVH) or normal saline (40 ul; SHAM) injection into a lateral ventricle. Motor function was tested in an open field automated locomotion test at 3- and 7-days post-surgery, and weekly thereafter until 90 days after injection. Anxiety-like behavior was measured using Elevated Plus Maze at 14, 30- and 120-days post injection, and cognitive function was tested using Barnes Maze 30 days post injection. Neuroinflammation was assessed utilizing Iba1+ for activated microglia and GFAP+ immunoreactivity for reactive astrocytes. Adult neurogenesis was assessed at the subventricular and subgranular zones using tamoxifen treated nestin-creER-Ai9-tdTomato mice that were harvested at 30 days post injection.

Results:
MRI revealed blood in the ventricles up to 7 days post injection; confirmed via histology. Hydrocephalus occurred in the IVH, but not SHAM group (p = 0.04). There was no difference in cognitive function or anxiety-like behavior at any time point post injection. At 30 days, mild astrocyte reactivity was noted near the lateral ventricular wall in the IVH, but not SHAM group. There was no difference in reactive astrocytes between the IVH and SHAM groups in the hippocampus or cortex or newly born immature neurons.

Conclusion:
While global neuroinflammation was not detected in our novel mouse model of pure IVH, local inflammation continued out to 30 days post IVH. This long-term activation was not accompanied by cognitive deficits, suggesting that the model does not emulate clinical findings, but supports study of local inflammation after IVH.
Abstract TP232: The Change Of Long-term Motor Functions After Post-stroke In Neuronal Cell Specific Sirt1 Knock-in Mice
Authors: Jun Yonghyun , Lim Chiyeon , Lim Sehyun , Lee Seeun , Cho Suin , Lee Kyungjin , Kim Hyung-Hwan
Published: February 2023

Background and Rationale:
Ischemic stroke is one of the leading causes of morbidity and mortality. Sirt1 is an NAD+ dependent deacetylase known as an important regulator for stroke research. During ischemia, the expression of total Sirt1 is regulated down. To investigate the change of motor function in neuronal-specific Sirt1 overexpression, we generated the neuronal-specific Sirt1 knock-in (nSirt1 KI) mice and examined its behavior effect after post-stroke.

Methods:
To determine the role of Sirt1 in ischemic stroke, we tested the pharmacological effects of metformin and sirtinol, and to investigate the genetic role of neuronal Sirt1 in the brain, we generated the nSirt1 KI mice and induced the Ischemia/Reperfusion (I/R: 30 min. middle cerebral artery occlusion (MCAO)/1D-14D reperfusion after 30 min. MCAO) injury model. Infarction volume was measured by TTC staining and relative cerebral blood flow was measured by Laser Doppler. We also determined neurological deficit score (NDS), wire-hanging test, and cylinder test at 1D-14D after 30 min. MCAO. Protein levels from brain lysates in the cerebral cortex and hippocampus were studied by western blotting.

Results:
nSirt1 KI mice exhibited increased rCBF and reduced infarction volume compared to that of littermate control in I/R injury model. Based on western blotting analysis of brain tissue lysates in nSirt1 KI mice, the expression of Sirt1 was significantly increased by ~5 times. Overexpression of Sirt1 reduced infarction volume was reversed by sirtinol. The daily locomotor changes of neurological deficit score for 14 days were significantly changed compared to the littermate control. The Wire-hanging test shows that locomotor abnormality was improved in nSirt1 KI mice compared to littermate control. The Cylinder test shows that locomotor abnormality was improved in nSirt1 KI mice. We found that brain shrinkages at 14 days after post-stroke were not severe in nSirt1 KI mice compared to the littermate control.

Conclusion:
These findings indicate that the long-term enhancement of motor functions in neuronal Sirt1 overexpression may be an important therapeutic target for post-stroke and long survival.
Abstract TP233: Cerebroprotective Role Of M
6
A Demethylase Fat Mass And Obesity-associated Protein After Experimental Stroke
Authors: Vemuganti Raghu , Chokkalla Anil K , Jeong Soomin , Mehta Suresh L , Morris-Blanco Kahlilia , Bathula Saivenkateshkomal , Qureshi Simran
Published: February 2023

Fat mass and obesity-associated protein (FTO) demethylates
N
6
-methyladenosine (m
6
A), which is a critical epitranscriptomic regulator of neuronal function. We previously reported that ischemic stroke induces m
6
A hypermethylation with a simultaneous decrease in FTO expression in the neurons. Currently, we evaluated the functional significance of restoring FTO with an adeno-associated virus 9 (AAV9), and thus reducing m
6
A methylation in the post-stroke brain damage. Adult male and female C57BL/6J mice were injected with FTO AAV9 (intracerebral) at 21 days prior to inducing transient middle cerebral artery occlusion. Post-stroke brain damage (infarction, atrophy and white matter integrity) and neurobehavioral deficits (motor function, cognition, depression and anxiety-like behaviors) were evaluated between days 1 and 28 of reperfusion. FTO overexpression significantly decreased the post-stroke m
6
A hypermethylation. More importantly, exogenous FTO substantially decreased post-stroke grey and white matter damage and improved motor function recovery, cognition and depression-like behavior in both sexes. These results demonstrate that FTO-dependent m
6
A demethylation minimizes long-term sequelae of stroke independent of sex.
Abstract TP24: Importance Of Staff Engagement And Performance Improvement Model In A High Performing Emergency Department Stroke Center
Authors: Mojares Joseph C , Maria Shannon N
Published: February 2023

Background and Purpose:
As a high-performing Advanced Primary Stroke Center (HPAPSC) an opportunity to explore learnings from local existing strategies on improving door-to-needle times (DTN) will assist in sustaining performance. With the use of Teleneurology technology and an early adopter of Tenecteplase, the APSC has exceeded national benchmarks on DTN. The challenge for a low-volume, HPAPSC is how to sustain performance while increasing the reliability of evidence practices, enhancing staff engagement, and reducing patient harm.

Methods:
With the availability of evidence-based interventions, the HPAPSC facility adopted the engage, educate, execute, and evaluate improvement concepts. Every stroke alert (SA) event is evaluated. Using a rapid improvement process in real-time, the team debriefs to identify learning opportunities (LO) for each event. The nurse’s involvement in the team debriefing process increased staff proficiency and engagement in the SA process. Each LO is translated to new training, coaching, and workflow modification. The ED team regularly assesses workflows and identifies unintended consequences. Finally, The SA team is recognized for their excellent practice.

Results:
From January 2020 to June 2022, we continue to meet DTN for less than 30 minutes at 89%. Staff engagement led to process changes including a new targeted SA workflow, increase recognition of posterior strokes in triage, reduced SA response time on walk-in patients, and increase compliance in post-thrombolytic assessment. Since the start of the project in January 2022 the Tenecteplase treatment rate is at 2.9% of acute ischemic stroke patients based on the number of stroke alerts. The bleed rate is at 4.2%.

Conclusions:
The ED SA team learned that transparency and staff involvement provide continued engagement, improved team dynamics, and a culture of continuous improvement. Thus, sustaining door-to-needle times, improving clinical outcomes and quality of care.
Abstract TP240: The Change Of Motor Functions After Post-stroke In 5xfad
Authors: Jun Yonghyun , Lim Chiyeon , Lim Sehyun , Lee Seeun , Kim Hyung-Hwan
Published: February 2023

Background and Rationale:
Recent Alzheimer’s disease (AD) studies with 5XFAD, a mouse model of Alzheimer’s disease, were mainly focused on cognitive functions. Here, to investigate the change of motor functions after post-stroke in the AD mouse model, we generated the 5XFAD mice and examined their long-term behavior changes.

Methods:
To determine the role of 5XFAD after post-stroke, we generated the 5XFAD mice, induced the Ischemia/Reperfusion (30-minute middle cerebral artery occlusion (MCAO)/14 days reperfusion after 30-minute MCAO induction) injury model in mice, and tested the several behavior effects of 5XFAD (2 months vs. 6 months). Relative cerebral blood flow was measured by Laser Doppler. We daily investigated neurological deficit score, wire-hanging test with both time latency and grip score, and cylinder test for 14 days.

Results:
We found that CBF changes and brain shrinkages at 14 days after post-stroke were not changed in 5XFAD mice compared to the littermate control. The daily locomotor changes of neurological deficit score for 14 days were not significantly worsened compared to the littermate control. The Cylinder test shows that locomotor abnormalities were not changed in 5XFAD mice. The Wire-hanging test shows that locomotor abnormalities both latency and grip scores were worsened in 5XFAD mice compared to littermate control.

Conclusion:
These findings indicate that the long-term abnormalities of locomotor in 5XFAD mice may be an important therapeutic target for post-stroke in AD.
Abstract TP27: Data Drives Outcomes: Optimizing Smoking Cessation Education And Documentation When Transitioning To A New Electronic Medical Record
Authors: Rexford Tracy , Saas Alissa M , Campbell Margie L , Lee Jessica D
Published: February 2023

Background and Purpose:
In June 2021, the University of Kentucky transitioned to the EPIC documentation platform. With the change in electronic medical record (EMR), the documentation location of smoking cessation changed and compliance decreased. "Smoking cessation" per Get with the Guidelines (GWTG) is the percent of patients with ischemic or hemorrhagic stroke or transient ischemic attack (TIA) with a history of smoking cigarettes, who are, or whose care givers are, given smoking cessation advice or counseling during hospital stay. There is a four-month delay in GWTG data abstraction & implications. As a result, the medical center was unaware of declining smoking cessation compliance rates. In November of 2021, it was reported that smoking cessation had dropped to 69.0% compliance over 5 months starting in June 2021 with a low of 60.0% in October 2021. The mean compliance rate for smoking cessation 4 months prior to June 2021 was 93.0% with 100.0% compliance for the month of April 2021. As an accredited Comprehensive Stroke Center, sustaining the GWTG Stroke Quality Award is of utmost importance.

Methods:
Two initiatives were introduced in response to decreased compliance in smoking cessation education documentation from June-October 2021. First, Stroke Transfer Coordinators responsibilities were modified to include day and night shift clinical coaching with bedside staff on importance of smoking cessation education and documentation. Second, the Medical Director of the Comprehensive Stroke Center communicated the importance of smoking cessation to provider team and partnered with informatics to optimize clinical progress notes documentation in EPIC.

Results:
After implementation of Stroke Transfer Coordinator support along with provider communication and EPIC smart phase addition smoking cessation compliance rose to 100% in November 2021 and sustained 100% through February 2022. Current smoking cessation compliance rate for the first two quarters of 2022 is 99.1%.

Conclusion:
It is imperative that data continues to drive clinical initiatives in stroke care and data is analyzed frequently to improve quality practice. An interdisciplinary team approach resulted in positive outcomes for smoking cessation education.
Abstract TP28: Support For The Stroke Program Coordinator
Authors: Okongo Casey , Brown Harold E , Elkins Kelley , Sumner Jessica , Zielke Shelly , Bellamy Lisa M
Published: February 2023

Introduction:
The role of stroke program coordinator (SPC) has historically been underestimated and not well supported. SPCs generally have overall responsibility and accountability for the program yet are rarely afforded the appropriate training and support. A network that is focused on improving stroke prevention and care in three midwestern states recognized the turn-over in this integral role and prioritized efforts to provide better support by offering a variety of services, resources, and education.

Methods:
Network staff provide new SPCs an initial personal orientation to the network and to their role. There is also a full-day training session that is offered twice a year to learn more in-depth information about stroke and program management. The network hosts a SPC teleconference quarterly to provide information about important programmatic and clinical updates, upcoming events, and to encourage sharing of best practices. One resource offered to affiliate hospitals of the network, the Stroke Program Management Guidebook was created with over 200 deliverables that are designed to help efficiently and effectively build/maintain a program, and to help with certification goals. New affiliate hospitals receive a comprehensive “Initial Program Curriculum”, including a review of these deliverables, strategic planning, advisory services, education about program development/management, community education, continuous quality improvement, and multi-disciplinary clinical care.

Conclusion:
An evaluation of how these support services affect the hospital and their stroke program coordinator has not been formally completed. However, in 2014 the network had 24 hospitals, 25% of those had achieved some level of stroke certification. Today there are 37 hospitals and 51% of these hospitals have achieved some level of stroke certification. Of those hospitals, 10 have reached acute stroke ready certification, seven primary stroke certification and two have obtained comprehensive stroke certification. From 2014 to today this network has trained 166 stroke program coordinators within affiliate hospitals. Further research is needed to determine the true impact of the additional SPC support provided by the network.