bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract TP167: Segmentation of leukoaraiosis on noncontrast brain using CT-MRI paired data without human annotation
Authors: Kim Chi Kyung , Oh Kyungmi
Published: January 2025

Purpose:
White matter hyperintensities (WMH), also known as leukoaraiosis (LA), are common brain abnormalities in elderly individuals. Evaluating LA on CT is challenging due to the less distinguishable hypoattenuation against white matter. We aimed to develop and validate a segmenting algorithm of LA using CT-MRI paired data.

Methods:
We included 744 CT-MRI paired dataset of patients with ischemic stroke from 4 stroke centers. An external test set comprised 411 patients from non-overlapping hospitals. WMH on MRI was segmented using validated software and the segmentation mask was registered onto NCCT space. We compared predicted LA versus ground-truth registered LA and WMH on MRI using Dice similarity coefficient (DSC) and concordance correlation coefficient (ρ).

Results:
Mean age (SD) for training and external test datasets were 68.1 (SD 12.7) and 69.2 (SD 13.5) years and 33.2% and 47.9% were female, respectively. In the internal validation dataset, the algorithm achieved a DSC of 0.53, with a volumetric correlation (ρ) of 0.848 with registered LA volume on CT. External validation showed a DSC of 0.527, with ρ values of 0.919 and 0.760 for predicted LA volumes compared to registered LA and WMH volumes on MRI, respectively. Subgroup analysis demonstrated consistent performance across different CT vendors and infarct volumes.

Conclusion:
Our deep learning algorithm offers a significant advancement in LA segmentation on CT, bridging the gap between CT and MRI assessments. It improves patient care by providing a consistent, accessible, and accurate method for evaluating LA, supporting both clinical practice and large-scale research.
Abstract TP169: Presence of a Mismatch Perfusion Profile in Large Core Ischemic Stroke Over Time Significantly Varies by Mismatch Definition
Authors: McCullough-Hicks Margy , Seners Pierre , Mlynash Michael , Olivot Jean Marc , Heldner Mirjam , Mordasini Pasquale , Strambo Davide , Michel Patrik , Carrera Emmanuel , Heit Jeremy , Lansberg Maarten , Albers Gregory
Published: January 2025

Introduction:
The prevalence of Mismatch (MM) profiles on admission perfusion imaging in large core ischemic strokes, stratified by time since last known normal (LKN) and MM profile definition, is not well described.

Methods:
We used a retrospective multicenter study of patients with anterior circulation large vessel occlusion, baseline MR or CT perfusion (MRP, CTP), and definitive LKN within 24 hours. We studied four MM profile definitions: 1) Mismatch ratio (MR)≥1.2, penumbral volume (Pen)≥10ml, and (if CTP) core volume defined by CBF<30%; 2) MR≥1.8, Pen≥15ml, CBF<30%; 3) MR≥1.2, Pen≥10ml, CBF<38%; 4) MR≥1.8, Pen≥15ml, CBF<38%. Two definitions of large core were evaluated (≥50ml or ≥70ml; defined as CBF<30% or CBF<38% on CTP and as ADC ≤620x10
-6
mm
2
/s on MRP). We used the Cochran-Armitage Trend Test to evaluate for trends in MM presence over time and McNemar Test to compare MM profiles for differences in prevalence.

Results:
Of 2012 patients included, 49% were male, median age was 75 (IQR 64-84), median baseline NIHSS was 15 (8-20), median time from LKN to imaging was 5.7 hours (1.8-8), 44% had CTP. In patients with cores ≥50ml and ≥70ml (Figures 1&2), MM profile defined by MR≥1.2 and Pen≥10ml was more prevalent than MM defined by MR≥1.8 and Pen≥15ml (P<0.001). MM prevalence decreased over time in three of four MM profiles in the ≥50ml core group but only one MM profile in the ≥70ml group (P<0.001 in all comparisons). In all core sizes (Figure 3), there was a significant trend toward decreased MM prevalence over time in all four profiles (P<0.0001) and between the MR≥1.2 and Pen≥10 vs MR≥1.8 and Pen≥15 profiles (P<0.001 in all comparisons). Similar significant differences between the MR≥1.2 and Pen≥10 vs MR≥18 and Pen≥15 existed in CTP only, MRP only, and witnessed onset only subgroup analyses (P<0.001 for all).

Conclusions:
MM is more prevalent when defined as MR≥1.2 and Pen≥10ml than when defined as MR≥1.8 and Pen≥15ml, especially in large cores. MM prevalence decreases over time in all core sizes, though trends are less clear with increasing infarct size. Understanding real-world prevalence of MM in large core strokes may help contextualize results of recent trials, while clarifying the optimal definition and prevalence of MM in the large core population is of import to future prospective study design. This study is limited by potential bias in favor of higher MM presence due to one site’s patient transfer acceptances contingent on favorable CTP profiles.
Abstract TP346: Investigating Transcriptomic Profiles of Peripheral Blood Mononuclear Cells in Moyamoya Disease Using Single-Cell RNAseq
Authors: DEMIRAG ZEYNEP , Uchino Haruto , Tokairin Kikutaro , Rao Shailaja , Chiang Terrance , Morton Gabriella , Lee Alex , Cheng Michelle , Steinberg Gary
Published: January 2025

Objectives:
Moyamoya Disease (MMD) is a complex cerebrovascular disease marked by progressive narrowing in the cerebral arteries. Previous studies have suggested that dysregulated genes in the peripheral blood of MMD patients are involved in extracellular matrix organization and immune responses. However, the mechanism by which the immune response contributes to MMD progression is unclear. In this study we investigated the immune landscape in the peripheral blood mononuclear cells (PBMCs) of MMD patients using single-cell RNA sequencing (scRNA-seq).

Methods:
Blood samples were collected from 3 bilateral MMD patients and 2 healthy controls. PBMCs were isolated and processed for 10x single-cell RNA sequencing. Immune cell heterogeneity and differentially expressed genes (DEGs) in each cell type were identified. Top biological pathways were determined using Ingenuity Pathway Analysis (IPA). Flow cytometry with an 11-fluorochrome panel was use to validate key immune populations and fluorescence-activated cell sorting (FACS) was used to isolate specific immune populations for transcriptome analysis.

Results:
Preliminary analysis revealed notable increase of cell percentages in certain cell types in MMD, such as CD8+T (9.21% vs 7.11%), B cells (6.94% vs 3.45%) and CD4+NKT (5.78% vs 3.22%), and decrease in CD8+NKT (9.59% vs 15.74%), compared to controls. Higher transcriptome dynamics were observed in certain cell type in MMD, with 2402 DEGs in classical monocytes, 1379 DEGs in CD4+T, 1312 DEGs in CD8+NKT and 997 DEGs in B cells. Interestingly, many cell types, except for classical and non-classical monocytes, displayed a higher number of downregulated DEGs in MMD, particularly in T cells, B cells, and NKT cells. Pathway analysis indicated that CD4+T, CD8+NKT, and B cells show predicted inhibition in several pathways, including T cell receptor and mTOR signaling, interleukins and CXCR4 signaling, as well as cell surface interactions at the vascular wall.

Conclusion:
These preliminary findings point to a distinct immunological profile in MMD, suggesting significant alterations in both immune cell activity and interaction within the vascular environment. Ongoing studies utilize FACS to validate specific immune populations and their transcriptomes in a larger cohort. These findings reveal unique transcriptomic signatures of peripheral blood immune cells in MMD and highlight key immune cell types that may contribute to its pathogenesis.
Abstract TP348: Overload of efferocytic cargoes in lysosome shifts microglia/macrophages towards pro-inflammatory phenotype through activating STING after stroke.
Authors: Liu Zhiruo , Cai Wei
Published: January 2025

Objective:
Incomplete clearance of cell debris in stroke lesion contributes to post-stroke cognitive impairment. Microglia and infiltrated macrophages are the major scavenger of debris in stroke lesion. However, as stroke progresses, their phagocytic capacity is impaired, while secretion of pro-inflammatory factors was enhanced. This study aims to identify the mechanisms driving the phenotypic shift after excessive phagocytosis.

Methods:
Brain cells from 8-12 week-old male C57BL/6J mice were collected for single-cell RNA sequencing at 1, 5, and 14 days after transient middle cerebral artery occlusion (tMCAO), as well as from sham-operated control group. Key transcription factors mediating the phenotypic changes of microglia and macrophages were identified through bioinformatics analysis, and further validated
in vivo
and
in vitro
using immunofluorescence staining, flow cytometry, and Western blot. Additionally, tMCAO models were treated with the key factor inhibitor to evaluate microglia and macrophage phenotypic shifts and the progression of stroke.

Results:
Through single-cell RNA sequencing analysis, we identified a concurrent activation pattern between the stimulator of interferon genes (STING) mediated Type I interferon signaling and the pro-inflammatory phenotype of microglia/macrophages. In the first 5 days after stroke, microglia and macrophages displayed an inflammatory-resolving phenotype with enhanced phagocytic activity, but from days 5 to 14, they transitioned to a pro-inflammatory phenotype with sustained upregulation of STING-mediated Type I interferon pathway molecules. We validated the phenomenon observed in bioinformatics using tMCAO models.
In vitro
and
in vivo,
we found that continuous phagocytosis by macrophages and microglia led to STING activation, triggering Type I interferon signaling and driving the phenotypic shift towards a pro-inflammatory state. Treatment with STING inhibitor H-151 reversed these detrimental shifts..

Conclusions:
These findings suggest that continuous post-stroke phagocytosis by macrophages and microglia led to the activation of STING, thereby triggering type I interferon signaling and driving the phenotypic shift towards a pro-inflammatory state. STING inhibition may serve as a therapeutic strategy for mitigating neuroinflammatory responses following stroke.
Abstract TP362: Estrogen Receptor-Beta Activation Reduces Cognitive Deficits After Stroke in Middle-Aged Female Rats
Authors: Cabeda Diaz Indy , Dave Kunjan , Perez-Pinzon Miguel , Raval Ami
Published: January 2025

Introduction:
Menopause increases the risk and severity of ischemic stroke (IS), while endogenous 17β-estradiol (E2) naturally protects premenopausal women against IS. The female sex hormone E2 is a potent neuro- and cognitive-protective agent. Studies have shown that periodic E2 or estrogen receptor subtype-beta (ER-β) agonist pre-treatments every 48 hours before an ischemic episode ameliorated ischemic brain damage in young ovariectomized or reproductively senescent (RS) aged female rats. The current study investigates the underlying mechanism of ER-β agonist-mediated neuroprotection.

Methods:
Retired breeder (9–10 months) Sprague–Dawley female rats were considered RS after remaining in constant diestrus phase for more than a month. The RS rats were exposed to transient middle cerebral artery occlusion (tMCAO; 90 mins) and treated with either ER-β agonist (beta 2, 3-bis(4-hydroxyphenyl) propionitrile; DPN; 1 mg/kg; s.c.) or DMSO vehicle at 4.5 hours after induction of tMCAO. Subsequently, rats were treated with either ER-β agonist or DMSO vehicle every 48 hours (48-h) for ten injections. Forty-eight hours after the last treatment, animals were tested for cognitive deficits via the Morris water maze. At 1-month post-tMCAO, brains were collected for histopathological analysis. A second cohort of RS rats underwent DPN or DMSO treatment for a month; 48-h after last injection, brains were collected for unbiased global metabolomic analysis (conducted by Metabolon Inc.). The metabolomic study was complemented with western blot analysis and enzyme activity measurements of key altered pathways.

Results:
Data showed significant (p<0.05) decreases in glucose-6-phosphate and increases in 5-phosphyribosyl diphosphate, UDP-galactose, and N-acetylglucoseamine-6-phosphate in the brain of DPN-treated RS female rats as compared to DMSO-treated controls. DPN treatment also changes the enzymatic activity of glycolytic rate limiting enzyme hexokinase. Metabolomics data also showed significant increase in choline and arginine levels in the brain of DPN-treated RS female rats as compared to DMSO-treated controls.

Conclusion:
The observed changes in glycolytic and amino acid pathway metabolites could enhance brain energy metabolism and increase choline availability, potentially contributing to the improved post-stroke cognitive outcomes in RS rats.
Abstract TP366: Impact of Neutrophil Extracellular Traps on Stroke Clot Mechanics, Radiomics, and Histology
Authors: Santo Briana , Jenkins TaJania , SHAH JAY PURNAKANT , Siddiqui Adnan , Tutino Vincent
Published: January 2025

Introduction:
In stroke clots, neutrophil extracellular traps (NETs) promote thrombosis, enhance clot stability, and decrease clot amenability to thrombectomy and thrombolysis.

Objective:
We examine the relationship between NET enrichment and clot mechanical characteristics, radiomics, and histological microstructure.

Methods:
White blood cells (WBCs) were concentrated from human donor whole blood. Platelet-rich plasma and red blood cells (RBCs) were isolated by centrifugation and mixed with WBCs to produce 0%, 20%, and 40% RBC clot analogs. Lipopolysaccharide (LPS) was added (0μL, 2μL, 6μL into 1mL dH2O) to enrich clots with varying NET amounts. CaCl2 induced clotting. Clots were incubated while rotating at 37°C. Clot analogs were assessed mechanically by uniaxial stretch tester and stress-strain curves, radiomically through high-resolution microCT and radiomic feature analysis, and histologically through whole-slide imaging of H&E-stained tissue and immunofluorescence (Citrullinated Histone Antibody and DAPI counterstain) to quantify NETs. Statistical analysis assessed what mechanical, radiomic, and histological features were significantly different among clots of various NET enrichment. unsupervised clustering was performed on radiomic and histological features to identify signatures unique to NET-enriched clots.

Results:
LPS addition produced clots with increasing NET enrichment and microstructural complexity. For fibrin-platelet rich clots (0% RBC), NET enrichment produced significant increase in mechanical stiffness, measured by breaking strength (max load) and Young’s Modulus. For each percent composition, radiomic and histomic profiling clustered clot analogs well by NET-enrichment, with NET-enriched clots demonstrating distinct radiomic and histological feature trends. See Figure 1.

Conclusion:
NET enrichment produces mechanically stiffer clot analogs with distinct microstructure, radiomic, and histological profiles.
Abstract TP367: Role of STAT1 and type I interferon signaling in ischemic preconditioning-induced microglial responses
Authors: McDonough Ashley , Weinstein Jonathan
Published: January 2025

Background:
Ischemic preconditioning (IPC) is a robust protective phenomenon in which a brief ischemic exposure confers tolerance against a subsequent prolonged ischemic challenge. Elucidating the mechanisms of IPC is a critical challenge in the stroke field. Innate immune pathways, particularly type I interferon (IFN) signaling, in microglia are critical in establishing protection in both grey and white matter. Our previous studies demonstrated that type I IFN receptor (IFNAR1) in microglia is required for both IPC-induced axonal protection and interferon stimulated gene (ISG) expression. We also showed that exposure of primary microglia to either IFN-beta or ischemia/reperfusion-like conditions results in phosphorylation of STAT1, a key signaling kinase downstream of IFNAR1. Therefore, we hypothesized that STAT1 is a critical mediator of the microglial response to IPC. Here we report the impact of systemic STAT1 knock-out on microglial responses after IPC.

Methods:
We performed a transient (15 min) middle cerebral artery occlusion on 4 wild-type (WT) and 4

Stat1
-/-

mice and collected tissue 72 hours later. We processed tissue for single nucleus RNA-sequencing using 10X Genomics kits. We captured ~10,000 nuclei per sample and sequenced at a depth of 25,000 reads. We performed comparative analysis between ipsilateral and contralateral hemispheres to identify focal changes in microglial subpopulations and gene expression due to IPC. We performed immunofluorescent microscopy and quantitative stereology to validate IPC-induced changes in microglial number/morphology and expression of ISG protein products such as IFITM3.

Results:
We describe global in vivo transcriptomic changes in microglial subpopulations after IPC and found that STAT1-deficiency alters both the distribution of microglial subpopulations and cluster-specific transcriptional profiles. We identify and define an interferon responsive microglial cluster induced by IPC that is dependent on STAT1. We also provide data demonstrating how ischemia/reperfusion and innate immune signaling affect STAT1 phosphorylation signaling dynamics in microglia in vitro. We show that phosphorylation is dependent on both Toll-like receptor 4 (TLR4) and IFNAR1.

Conclusions:
Our results support a central role for STAT1 in mediating microglial phenotype in vivo following IPC. Our findings indicate that STAT1 is an important regulator of microglial type I IFN signaling in the context of ischemia.
Abstract TP370: Deep Learning Applied Analysis of Post-Stroke Mice During Corner Test Provides Quantitative Assessment of Locomotion
Authors: Varghese Jeremiah , DiPersio James , Schappell Laurel , Tang Meiyi , Madeira Miguel , Nadkarni Neil
Published: January 2025

Introduction:
Despite efforts to improve stroke outcomes in patients, a translational gap exists between preclinical and clinical studies. Due to this gap, the Stroke Preclinical Assessment Network has incorporated the corner test (CoT) for behavioral outcome as a primary measure for evaluating whether a treatment is successful or not. Standard behavioral analysis for CoT uses the laterality index to detect if there is mouse turning preference on a scale from -1 to 1. We sought to determine if a deep learning approach using “DeepLabCut” could be applied towards enriching our CoT data to better evaluate aspects of mouse locomotion.

Methods:
Six C57/Bl6 mice were subjected to an 1 hr transient middle cerebral artery occlusion in the right hemisphere of the brain. CoT were recorded with an isometric view and performed at both baseline (BL) prior to the stroke and one day post stroke (D1). The same set of six mice performed 10 turns per CoT, totalling to 60 turns for BL and 60 turns for D1. The pose estimation model was made using a ResNet-101 neural network trained on 1064 manually-labeled frames, with the assistance of “DeepLabCut” software packages. Videos were analyzed by the pose estimation model and sequentially processed through a newly developed R script and DLC Analyzer R script. Turns were defined as in SPAN with a 90 degree head turn upon vibrissae contact on both sides of the corner boards. Turn latency was defined as the time lapsed during a turn, and head turn speed as the average speed during a turning event.

Results:
The average laterality index showed clear preference towards ipsilateral turning in all D1 mice (-1.0 ± 0.04, N = 6). Furthermore, a comparison between BL data and respective D1 data showed significantly longer turn latencies and slower head turn speeds (p < 0.05) for D1 mice. The average turn latency for BL mice was 3.58 ± 0.57 s, which was 4.6 times shorter than that of D1 mice (16.45 ± 3.11 s). The average speed for BL mice was 2.01 cm/s ±0.21, which was 2.3 times faster than that of D1 mice (0.86 ± 0.14 cm/s).

Conclusion:
This deep learning approach enriches current stroke behavioral analysis methods by offering additional quantitative information upon which behavior can be assessed. Future studies can use these behavioral metrics for stratification or correlation with variables of interest (e.g. infarct size) to provide a more refined assessment of preclinical stroke behavior.
Abstract TP378: Malondialdehyde as a Clinical Indicator for Oxidative Stress: Associations with Age, BMI, and Cognitive Impairment
Authors: Petway Ana , Eagen Patrick , Baskin Lynae , Cahuiche Ana , Couture Lucy , Delevati Colpo Gabriela
Published: January 2025

Introduction:
The molecular and metabolic changes that occur after acute ischemic stroke (AIS) are not fully understood. One mechanism known to trigger systemic inflammatory responses and neuronal death during ischemic stroke cascades the rapid increase in Reactive Oxygen Species (ROS). Accumulation of oxidative stress has been shown to trigger the initiation and progression of cognitive deficits, including mild cognitive impairment (MCI) and Alzheimer’s Dementia (AD). One emerging biomarker able to reliably measure oxidative stress is Malondialdehyde (MDA), a reactive carbonyl compound originating from polyunsaturated fatty acid oxidation and lipid peroxidation. Due to its composition, MDA readily reacts with lipid membranes, making it a sensitive oxidative stress biomarker. This study assessed MDA levels in the plasma of AIS patients to evaluate its ability to predict cognitive impairment and long-term functional outcomes.

Hypothesis:
We hypothesized that oxidative stress correlates with long-term functional outcomes in AIS patients and varies based on non-modifiable risk factors such as sex and race.

Methods:
In this study, we used peripheral blood plasma from healthy volunteers (HV, N=24), and from ischemic stroke patients (N=27) at 3d and 7d post-stroke to capture the temporal profile of MDA after injury. Cognitive impairment was assessed during hospitalization with the Brief Neurocognitive Screening Test (BNST), with a score of 8 or below denoting cognitive impairment.

Results:
AIS patients had an increase in MDA levels compared to the control group, as seen in prior literature. There was a significant correlation with increase age of stroke patients and higher levels of MDA (p<0.05). Higher levels of oxidative stress was also associated with increased levels of cognitive impairment among the stroke population (p<0.05). There was no relationship between levels of MDA during hospitalization and functional outcome at 3 months post discharge.

Conclusion:
MDA appears to be a promising biomarker for oxidative stress in AIS patients, with notable associations with age and cognitive impairment. These findings underscore the possible mechanisms behind the rapid development of cognitive impairment seen in patients post stroke.
Abstract TP380: TMEM97 and Lipid Metabolism: Unraveling Post-Stroke Lipid Regulation
Authors: Arkelius Kajsa , Won Seok Joon , Gupta Lavanya , Zhang Mengqi , Swanson Raymond , SINGHAL NEEL
Published: January 2025

Recent research on neurodegenerative diseases has highlighted a pathophysiological cascade characterized by abnormal lipid accumulation within the brain. Although progress has been made, the role of lipid metabolism following stroke remains unclear. This study aims to further explore changes in lipid metabolism after stroke, with a specific focus on the endoplasmic reticulum transmembrane protein 97 (TMEM97), which has emerged as a promising therapeutic target due to its critical role in cholesterol metabolism.

Spatial transcriptomic analyses following experimental stroke induction in mice revealed neuronal upregulation of crucial genes involved in lipid metabolism pathways, including LDLR, acetyl-CoA acetyltransferase 1, and elongation of very long-chain fatty acids protein 6, indicating dynamic adaptations post-stroke or under cellular stressors.

Initial findings demonstrate TMEM97 expression predominantly within neurons in vivo (mouse model). In an in vitro model of lipid starvation using mouse neural progenitor cells (MNPCs), pharmacological inhibition of lysosomal lipid release leads to increased expression of TMEM97. Interestingly, when TMEM97 was knocked down in MNPCs using lentiviral shRNA transfection, these cells exhibited no significant difference in neutral lipid accumulation compared to control cells. However, upon treatment with the lysosomal lipid release inhibitor, a significant increase in neutral lipid accumulation was observed exclusively in the TMEM97 knockdown cells.

Furthermore, Thapsigargin-induced endoplasmic reticulum (ER) stress in the MNPCs reduced TMEM97 expression, while also increasing low-density lipoprotein (LDL) uptake and the mRNA levels of the LDL receptor (LDLR). The observed elevation of neuronal LDLR expression in vitro closely aligns with the heightened expression of LDLR in vivo, suggesting a coordinated modulation of lipid metabolism pathways triggered by ER stress in response to stroke or metabolic stress induction. This synchronous upregulation of LDLR underscores the crucial role of ER stress as a potential trigger for this regulatory response. Additionally, considering the known involvement of TMEM97 in lipid homeostasis within the ER, it further highlights the intricate interplay between these proteins in post-stroke lipid metabolism regulation.

Our findings provide a compelling starting point for further exploring TMEM97's significance in post-ischemic lipid homeostasis.
Abstract TP385: Vitamin D3 prevent blood-brain barrier dysfunction after ischemic stroke
Authors: Mushtaq Zyad , Zheng Yuting , Tyagi Neetu
Published: January 2025

Emerging research suggests that Vitamin D3 might help protect against brain damage caused by ischemic stroke, as a deficiency in Vitamin D3 can worsen the severity and outcomes of a stroke. To investigate this, we studied the impact of Vitamin D3 on maintaining the integrity of the blood-brain barrier (BBB) using a mouse model of stroke. The BBB is severely disrupted during all kinds of strokes, which can lead to further injury and functional deficits. We measured several biomarkers related to BBB disruption to see if Vitamin D3 treatment could protect the BBB after a stroke. Mice were randomly divided into two diet groups: one receiving a Vitamin D3-sufficient diet (VDHsuf) and the other a Vitamin D3-deficient diet (VDHdef). The VDHsuf group ate standard chow, while the VDHdef group consumed a diet lacking Vitamin D3 for 8 weeks. Both groups underwent middle cerebral artery occlusion (MCAO) surgery for 40 minutes followed by reperfusion for 72 h. We assessed BBB disruption by measuring even blue leakage, matrix metalloproteinase-9 (MMP-9/MMP-2) activity, and changes in tight junction (TJ) proteins. We also evaluated the expression of OPG, RANKL, osteopontin (OPN), and the Vitamin D receptor (VDR) in both diet groups. At the time of MCAO, mice on the Vitamin D3-deficient diet had low levels of Vitamin D3. After MCAO, Evans blue leakage, indicating more severe BBB injury, was significantly greater in the VDHdef group compared to the VDHsuf group. Additionally, the expression of MMP-9, GLUT1, VDR, OPG, and OPN increased, while TJ proteins occluding, claudin-5 and RANKL decreased in the VDHdef group. These findings suggest that Vitamin D3 deficiency can impair the BBB, potentially worsening stroke severity and its long-term effects.
Abstract TP395: Alzheimer's Disease model APP/PS1 mice exhibit motor deficits following 15-minute focal ischemia.
Authors: Waits Paxton , Otero Anthony , Zimmerly Madison , Vedati Vidhaath , Orfila James , Herson Paco
Published: January 2025

Introduction:
Ischemic stroke is one of the leading causes of death in the United States and is a known risk factor for Alzheimer’s Disease (AD) development. One of the characterizations of AD is the accumulation of β-amyloid peptide due to the proteolysis of Amyloid Precursor Protein (APP) by the protein Presenilin 1 (PS1) among others. In APP/PS1 mice, which contain an additional human copy of APP and PS1, a 15-minute Middle Cerebral Artery Occlusion (MCAO) model was developed. Here we investigate the effects of increased β-amyloid peptide on motor coordination when subjected to local ischemia.

Methods:
APP/PS1or Wt male mice are initially subjected to either a 15-minute MCAO or Sham surgery. Injury volume using MRI is assessed at 3-days using T2 imaging. To test motor coordination the mice went through a tapered beam analysis at the 7-day time point. Following the tapered beam test, Cresyl Violet was used to stain brain slices. All mice were 8-12 weeks old at the time of surgery. Differences between groups were determined by Welch’s T-Test. Significance was determined as p < 0.05.

Results:
No significant difference in infarct volume was observed between the APP/PS1-MCAO and Wt-MCAO groups. In the hind legs, it was observed that there is a significant difference in the number of slips off the tapered beam in the APP/PS1-MCAO group when compared to the Wt-MCAO group (9.4 ± 3.356, n=7, p < 0.05 and 2.5 ± 0.289, n=4, p < 0.05 respectively). No significant difference was found in the Cresyl Violet staining.

Conclusions:
Our study shows motor deficit in the APP/PS1-MCAO experimental group when compared to the Wt-MCAO group as measured on hind-limb coordination. Therefore, further studies are warranted to assess the interaction between ischemia and β-amyloid peptide on histological injury and functional recovery.
Abstract TP397: Effect of Intracranial Pressure Spikes Modeling Subarachnoid Aneurysm Rupture on the Brain and Dura Barrier Immune System
Authors: Mattke Jordan , Shah Vaidehi , Mastorakos Panagiotis
Published: January 2025

Steep intracranial pressure (ICP) rise constitutes a hallmark of aneurysmal subarachnoid hemorrhage (SAH). While during aneurysm rupture there are multiple simultaneous triggers that can initiate an inflammatory response, in this study we isolate the impact of a single ICP spike on the brain and meningeal barriers and the resulting secondary injury.

Using a rodent model, we induced an ICP spike through cisterna magna injection of artificial cerebrospinal fluid or autologous blood, with simultaneous ICP monitoring. We then assessed the innate immune response in the brain parenchyma and dura using immunohistochemistry and multiparameter flow cytometry. Glia limitans integrity was evaluated via intrathecal injection of the SR101 tracer, and microthrombi presence was assessed using intravenous fluorescent fibrin. Upregulated pathways were identified through bulk RNA sequencing.

Our findings revealed that a single ICP spike exerts global effects on brain and meningeal barrier immunology, potentially leading to secondary injury. Histological analysis and flow cytometry of dura whole mounts showed extensive cell death of resident CD206+ macrophages, essential for steady-state immune surveillance, alongside infiltration of Ly6C+ pro-inflammatory monocytes and Ly6G+ neutrophils, indicating a robust acute response to the ICP spike. In the brain parenchyma, we observed minimal infiltration of circulating immune cells, consistent with the effects of aneurysm rupture. However, diffuse microglia clustering suggested multifocal parenchymal injury. Additionally, there was evidence of widespread glia limitans disruption and microthrombi formation in the brain parenchyma. Bulk RNA sequencing supported these findings, demonstrating upregulation of MyD88 and TNFα pathways in the dura, which were absent in the brain.

In conclusion, this study demonstrates that a single ICP spike triggers significant meningeal inflammation, disrupts steady-state dura immune surveillance, and causes diffuse parenchymal injury with glia limitans disruption. These results highlight a novel blood-independent pathway of injury following aneurysm rupture, offering new insights into potential therapeutic targets.
Abstract WMP99: Circulating Plasma Biomarkers Associated with Familial Cerebral Cavernous Malformation, Hereditary Hemorrhagic Telangiectasia and Sturge-Weber Syndrome
Authors: Weinsheimer Shantel , Nixon Andrew , Nelson Jeffrey , Brady Chris , McCulloch Charles , Marchuk Douglas , Awad Issam , Faughnan Marie , Loeb Jeffrey , Lawton Michael , Kim Helen
Published: January 2025

Introduction:
Circulating plasma protein profiling in individuals with brain vascular disorders may aid in the identification of robust diagnostic biomarkers, stratification of high-risk patients for treatment, and monitoring of disease progression or treatment response. This Brain Vascular Malformation Consortium (BVMC) study aimed to identify circulating inflammatory and angiogenic proteins that associate with familial Cerebral Cavernous Malformation (FCCM), Hereditary Hemorrhagic Telangiectasia (HHT), or Sturge-Weber Syndrome (SWS).

Methods:
We used the Angiome multiplex ELISA biomarker panel to assess the circulating plasma levels of 22 proteins related to inflammation and angiogenesis in 234 individuals enrolled in the BVMC, including 114 FCCM, 101 HHT and 19 SWS cases. Protein levels were measured in duplicate and absolute measurements obtained. We tested for biomarker associations between disease states using linear regression models adjusting for age at blood collection and sex. We calculated the intraclass correlation coefficient (ICC) to describe the similarity in replicates, and report proportional increase (PI) for protein levels for statistically significant results with Bonferroni-corrected P-values <0.05 (adjusted for 22 markers).

Results:
All 22 proteins were successfully measured in the 114 FCCM (39% male, median age 52 years), 101 HHT (49% male, median age 42 years, 36% with brain arteriovenous malformation), and 19 SWS (53% male, median age 15 years). The ICC was high for all markers (range 0.904-0.990). As expected, HHT cases with endoglin mutations had ~50% lower endoglin. We observed increased levels of endoglin (PI=1.38, 95% CI:1.27-1.50, P=6.50E-13) and IL10 (PI=1.48, 95% CI:1.16-1.91, P=0.04) in CCM compared to HHT cases, and decreased IL8 (PI=0.60, 95% CI:0.43-0.83, P=0.04) and OPN (PI=0.63, 95% CI:0.47-0.84, P=0.04) in CCM compared to SWS cases. We also observed higher levels of endoglin (PI=1.45, 95% CI:1.24-1.71, P=1.47E-04), GP130 (PI=1.24, 95% CI:1.11-1.38, P=2.91E-03), and OPN (PI=1.80, 95% CI:1.36-2.40, P=1.13E-03), and lower levels of IL1β (PI=0.30, 95% CI:0.15-0.59, P=0.01) in SWS compared to HHT cases.

Conclusions:
We identified circulating plasma biomarkers associated with FCCM, HHT and SWS. Larger ongoing work will confirm whether these inflammatory/angiogenic markers are relevant as potential diagnostic or therapeutic targets or have broader implications as clinical biomarkers in other brain vascular diseases.
Abstract WP164: Creating an Acute Sleep Medicine Service for Veterans Hospitalized with an Acute Ischemic Stroke or Transient Ischemic Attack – A Comparative Case Study from the Addressing Sleep Apnea Post-Stroke/TIA Hybrid Type I Stepped-Wedge Cluster Randomized Trial
Authors: Sico Jason , Rattray Nicholas , Burrone Laura , Sexson Ali , Miech Edward , Story Kristin , KOO BRIAN , Taylor Stanley , Myers Laura , Perkins Anthony , Daggy Joanne , Bravata Dawn
Published: January 2025

Introduction:
Rates of guideline-concordant obstructive sleep apnea (OSA) testing among those with a recent cerebrovascular event are exceedingly low. Understanding the role contextual factors play is necessary to inform successful implementation of quality improvement (QI) initiatives designed to address this gap in stroke/transient ischemic attack (TIA) care.

Methods:
Longitudinal data was collected via questionnaires and semi-structured interviews to evaluate the implementation of QI initiatives conducted at six diverse VA Medical Centers (VAMCs) participating in Addressing Sleep Apnea Post-Stroke/TIA (ASAP), a Hybrid Type I, stepped-wedge cluster-randomized trial. Intervention components included a Systems Redesign Virtual Collaborative and data monitoring (Figure 1). Implementation strategies included external facilitation and audit and feedback. Provider- (e.g., clinical training) and systems-level contextual elements (e.g., Champion Team members and their roles) were collected. Select Consolidated Framework for Implementation Research (CFIR) constructs were rated in terms of magnitude and valence. The primary outcome of successful implementation was defined at the end of 21 months of active implementation as obtaining a Group Organizational (GO) score of ≥6 a measure of programmatic development and maturation. Comparisons of sites were conducted across contextual elements and stratified by those achieving a GO score of ≥6 (Figure 2).

Results:
ASAP Sites 1 through 4 obtained a GO score ≥6 (range: 7-9); across these sites, the Clinical Champion had: (1) field staff engaged in activities such as care coordination; (2) full to partial support of their local sleep personnel, and; (3) consistently positive CFIR scores values. These sites also received a greater amount of external facilitation and used a quality dashboard more often. All 4 sites created a change in health care personnel and medical center culture that stressed the importance of OSA testing soon after a cerebrovascular event occurred.

Conclusions:
Developing strong and consistent Champion teams who meaningfully engaged with local VAMC personnel within and across sleep medicine and stroke service lines was important for implementation success. Other key contextual factors for changing culture and creating a healthcare system wide approach to improving OSA testing for stroke/TIA patients included external facilitators and using performance data.

Clinical Trials registration:
NCT04322162
Abstract WP299: Anticoagulant therapy for cerebral venous sinus thrombosis: A propensity score matching study
Authors: Liu Kai , Wei Zhao , Meng Zhang , Qin Dai , Song Bo , Xu Yuming
Published: January 2025

Background:
Newer Oral Anticoagulants (NOACs) offer potential advantages for patients with cerebral venous sinus thrombosis (CVST). However, there is a lack of evidence to evaluate the efficacy and safety of NOACs in CVST. The purpose of this study was to compare the benefit and safety between NOACs and warfarin in CVST patients based on propensity score matching.

Methods:
We performed a single-center prospective analysis including CVST patients from the First Affiliated Hospital of Zhengzhou University between January 1, 2018 and December 31, 2021. The primary outcome was recurrent thrombotic events during 6-month follow-up. Secondary outcomes included modified Rankin Scale (mRS), bleeding events, death and cerebral venous recanalization during anticoagulant therapy. Propensity score matching (PSM) was used to balance baseline bias.

Results:
A total of 650 patients were identified. NOACs were treated in 184 patients and 466 were used warfarin following hospitalization. After 1:2 and 1:3 PSM, there were no statistically significant differences between the two groups in recurrent CVST (odd ratio [OR], 0.543; 95% CI, 0.258-1.143; P=0.108), bleeding events (OR, 0.823; 95% CI, 0.074-9.143; P=0.874) and partial/complete recanalization (OR, 0.980; 95% CI, 0.546-1.760; P=0.946) in all patients. Similarly, there were no significant differences in patients received anticoagulation therapy group and patients who were performed endovascular therapy plus anticoagulation therapy regarding any of the clinical outcomes. However, there were a statistically significant differences between the two groups in death (2.2% vs 8.0%, P=0.014) and mRS≤2 (95.1% vs 88.7%, P=0.020) in all patients.

Conclusions:
Our study demonstrates that the use of NOACs in CVST has similar efficacy and safety compared to warfarin treatment. NOACs treatment may reduced mortality at six months and good functional outcome compared with warfarin.

Keywords:
new oral anticoagulants; cerebral venous sinus thrombosis; warfarin; propensity score matching
Abstract WP3: Large Vessel Recanalization with Thrombolysis Prior to Thrombectomy: Incidence, Outcomes, and Associations With Collateral Status
Authors: Skorseth Paige , Chen Huanwen , Rewinkel Scott , Kim Daniel , Amin Sonesh , Shakal Scott , Priest Ryan , Nesbit Gary , Clark Wayne , Colasurdo Marco
Published: January 2025

Introduction:
Large vessel recanalization (LVR) is a well described phenomenon following intravenous thrombolysis (IVT). However, predictors of LVR are elusive, and whether early LVR for large vessel occlusion (LVO) is beneficial for patients who cannot undergo immediate endovascular thrombectomy (EVT) is unclear. Moreover, the interplay between collateral circulation and the likelihood of LVR is poorly understood and even less studied.

Methods:
This was a retrospective study at a comprehensive stroke center that centralizes patient transfers for EVT from a network of 15 hospitals. Adult patients with an internal carotid artery or middle cerebral artery LVO who received IVT and did not undergo EVT within one hour of LVO diagnosis were included. Clinical and radiographic information at the time of LVO diagnosis were collected. Collateral status was graded using the Tan score (Tan CS). LVR was defined as vessel recanalization identified on repeat vessel imaging leading to a decision to defer EVT. The primary endpoint was infarct progression defined as decrease in ASPECTS of two or more. The secondary endpoint was 90-day modified Rankin Scale (mRS) outcomes. Multivariable logistic regression models were used to adjust for various confounders.

Results:
Of the 150 patients included, 42 (28.0%) experienced LVR prior to EVT. LVR patients had lower NIH Stroke Scale (p=0.006) and higher Tan CS (p=0.019). In multivariable analyses, LVR was associated with significantly lower odds of infarct growth (OR 0.15 [95%CI 0.07-0.36], p<0.001), and compared with patients who underwent successful EVT recanalization, lower 90-day mRS (common OR 0.44 [95%CI 0.22-0.90], p=0.024). Higher Tan CS was significantly associated with higher odds of LVR (2.95 [95%CI 1.60-5.45], p<0.001), and both collateral status and LVR were independently associated with lower odds of infarct growth (0.34 [95%CI 0.18-0.63], p<0.001 and OR 0.21 [95%CI 0.09-0.52], p<0.001, respectively).

Conclusions:
LVR with IVT prior to EVT was significantly associated with lower odds of infarct progression and better clinical outcomes. Better collateral circulation was independently associated with both higher odds of LVR, as well as lower odds of infarct progression. In conclusion, these findings suggest the clinical benefit of LVR for patients who cannot receive timely EVT and highlight the potential for modulating collaterals with the goal of limiting infarct growth and maximizing the odds of LVR.
Detecting glioblastoma infiltration beyond conventional imaging
tumour margins using MTE-NODDI
Authors: Karamched Saketh R. , Gorup Dunja , Tolomeo Daniele , Brooks Lucy J. , Gong Ting , Jørgensen Andreas Christ Sølvsten , Hill Ciaran Scott , Clements Melanie , Kalber Tammy L. , Wells Jack A. , Stuckey Daniel J. , Thorne Lewis , Shahrezaei Vahid , Marguerat Samuel , Zhang Hui , Parrinello Simona , Lythgoe Mark F.
Published: January 2025

Glioblastoma (GBM) is the most common and aggressive brain tumour with stark resistance to available therapies, leading to relapse and a median survival of <15 months. A key cause of therapy resistance is diffuse infiltration of tumour cells into brain regions surrounding the tumour, which presents a major clinical challenge as existing imaging techniques offer limited detection of the resectable margin. Here, we use diffusion weighted imaging (DWI) and apply the multiple echo time neurite orientation dispersion and density imaging (MTE-NODDI) model as a tool to detect tumour cells in the hard-to-distinguish margin. We used the G144 patient-derived xenograft model, with characteristic invasion along white matter tracts, in combination with MTE-NODDI. Tumour development was monitored, and magnetic resonance imaging (MRI) data were acquired over a 4-week period, starting at 4 weeks after stereotactic injection of tumour cells. MTE-NODDI demonstrated sensitivity to the developing tumour in the invading margin, and changes in measured parameters were apparent from 6 weeks after injection. In comparison to standard DWI, MTE-NODDI showed increased sensitivity to the tumour-associated changes in the margin. Furthermore, extraneurite volume fraction (fen) and neurite density index (NDI) measured from MTE-NODDI correlated with immunohistological measurement of tumour cells. These findings suggest that MTE-NODDI may non-invasively detect infiltrating cells and tumour-induced pathology in margin regions without T2 or DWI changes in a patient-derived mouse model of GBM. MTE-NODDI is clinically translatable and could be a powerful tool for neurosurgeons to maximise surgical resection, resulting in better survival outcomes for patients with GBM.
Fast-Forwarding the Future: Takarazuka Revue’s Recalibration of Youth Cultures
Authors: Grajdian Maria
Published: January 2025

Within a mixed-methodological framework, this paper analyzes Takarazuka Revue’s groundbreaking performances City Hunter: The Stolen xyz (2021, snow troupe) and High & Low: The Prequel (2022, cosmos troupe): famously rigid in its business model targeted at a clearly defined socioeconomic demographic (upper-middle and upper class female citizens aged from 35 up to 80s or even 90s), these two performances manifest a subtle paradigm shift in Takarazuka Revue’s performance strategy towards a consideration of younger audiences of both genders through the employment of a diverse narrative and musical material and the immersion into the problematic of global concerns such as family dysfunctionality, rampant loneliness and loss of valid role-models.
A new species of hedgehog-lizard of the genus Echinosaura (Squamata: Gymnophthalmidae) from Colombia and Panama with a reevaluation of the conservation status of the genus
Authors: VÁSQUEZ-RESTREPO JUAN D. , DAZA JUAN M.
Published: January 2025

The gymnophthalmid semi-aquatic lizards of the genus Echinosaura have been subject to taxonomic scrutiny in recent years. Here, we combine previously available and new morphological and phylogenetic evidence to describe a new species formerly known under the name E. palmeri. Our results are consistent with previous phylogenetic hypotheses and recover an overlooked evolutionary lineage which is both morphologically and genetically different from the other ones currently recognized within the genus. This species is diagnosable based on external and hemipenial morphology and is phylogenetically related to E. centralis, E. palmeri, and E. panamensis. The new species inhabits the northern portion of the Pacific region in Colombia and the Darien zone between Colombia and Panama. We also re-evaluate and propose a new IUCN extinction risk categorization for the nine species in the genus, assessing three uncategorized species, removing two species from a threatened category, and including a previously considered Least Concern species into a threatened status.
2. Conservation as a marathon vs. a sprint
Authors: Sherman Krista Danielle
Published: January 2025

Krista D. Sherman, describes the rapidly emerging and persistent challenges to save the critically endangered Nassau Grouper in The Bahamas. This effort has been characterized by short-term gains and setbacks, which requires cutting edge science and high-level engagement with local fishers, coastal communities, and government. Her high level of engagement in the community, as well as the science, seems to be the key to success.
5. Climate change impacts on the fisheries and aquaculture sectors with a focus on Malaysia
Authors: Alam Lubna , Mokhtar Mazlin , Sumaila U. Rashid
Published: January 2025

Alam Lubna, Mazlin Bin Mokhtar, and Ussif Rashid Sumaila detail the effects of climate change on fisheries and aquaculture. This case study, focused on Malaysia, characterizes the shift from constraining human actors to harmonizing human needs with maintaining functional marine ecosystems. Small-scale fisheries provide 90% of the jobs in fisheries and half the protein for human consumption. As wild-capture fisheries have continued to decline, aquaculture has been on the rise to meet the nutritional demands of a growing human population. This combination of enterprises is occurring globally, but as this case study shows, the way forward must be shaped by the history, culture, and governance approaches appropriate for each country.