bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 49: A Proposed Role for Transmembrane Activator and Cyclophilin Ligand Interactor (TACI) in Cardiovascular Disease: Adaptive Immunity and Atherosclerosis
Authors: Meelu Omar A
Published: June 2021

Objective:
To elucidate a proposed role of TACI, a tumor necrosis factor (TNF) family member of human B cells, in atherosclerosis. Whereas atherosclerosis has been characterized by inflammation, adaptive immunity, and specifically the role of B cells and their respective cytokine processes have not, and thus we focused on the role of a B cell homeostasis regulator, TACI. We chose TACI because it has been shown, in combination with immunoglobulins, to inhibit cells involved in the innate response, known to be the initial responders to atherosclerosis.

Approach and Results:
As there are two isoforms of TACI in mice, we needed to first confirm whether human B cells had one or two isoforms, before understanding preferential expression. To this end, we examined peripheral blood mononuclear cells (PBMCs), multiple myeloma cell lines and Hodgkin’s disease cells and found that both the longer and the shorter isoform of TACI are expressed. We did this by: stimulation of cell lines with IL-6, western blot analysis and densitometry, polymerase chain reaction. Student’s t-test, with the use of Statistical Analysis System (Cray, NC), for statistical analysis.

Conclusion:
Thus, based on our findings of the existence of both isoforms, functional analyses in cardiomyocytes would shed light on any proposed role of TACI in the context if coronary artery disease.
Abstract 59: Antimir Therapy Ameliorates HIF1-mediated Loss of SERCA2 and Cardiac Contractility
Authors: Williams Allison L , Walton Chad , MacCannell Keith , Avelar Abigail , Shohet Ralph
Published: June 2021

Ischemic heart disease is a major precipitant of heart failure in the developed world. Hypoxia inducible factor 1 (HIF1) is a highly oxygen-sensitive protein and the principal regulator of hypoxia-related signaling including changes in metabolism and angiogenesis. To more fully understand the role of HIF1 in the heart, our lab has developed a transgenic mouse model in which a stable form of HIF-1alpha can be inducibly expressed under normoxic conditions in cardiomyocytes. Upon HIF1 induction, genes associated with calcium signaling are downregulated and metabolism is shifted toward glycolysis. Strikingly, these mice also show a rapid decrease in cardiac contractility and concurrent loss of SERCA2 protein expression (up to 90%) within one week. SERCA2a (sarcoplasmic/endoplasmic reticulum calcium ATPase 2) is the ATP-dependent calcium pump required for calcium re-uptake during excitation-contraction coupling in the heart and its dysfunction has been implicated in heart failure. To extend the physiological significance of these findings, we examined ischemic heart tissue from WT mice subjected to ligation of the left anterior descending (LAD) artery to mimic myocardial infarction. At one day and 3 days post-ligation, we observed an increase in transcripts needed for glycolysis and decreased abundance of transcripts from the genes for proteins that regulate excitation-contraction including the ryanodine receptor, phospholamban and SERCA2. We also observed that while SERCA2 transcript decreased in HIF1 transgenic hearts, it did not fully account for the observed protein loss. Microarray analysis found miR-29c to be substantially upregulated and a potential regulator of SERCA2 expression. Subsequent in vitro analysis confirmed miR-29c reduced SERCA2 expression by 20-30% and could be modulated with an anti-sense inhibitor. In vivo administration of the antimir to miR-29c also improved cardiac contractility and SERCA2 expression in HIF1 transgenic mice. These results suggest that a HIF1 regulated microRNA, miR-29c, contributes to loss of SERCA2 in hypoxia and can be therapeutically targeted to improve cardiac function.
Abstract 71: Model of Induced Ventricular Tachycardia and Cardiac Electrophysiological Mapping
Authors: Chinyere Ike , Weigand Kyle , Moukabary Talal , Witte Russell , Lancaster Jordan , Goldman Steven , Juneman Elizabeth
Published: June 2021

Introduction:
We have developed a computer user interface able to provide prescribed programmed electrical stimulation (PES) to induce sustained-ventricular tachycardia (VT) in rats with chronic heart failure (CHF). We propose this program to examine the cardiac electrophysiology (EP) properties and arrhythmogenic potential in varying disease models and as a method of evaluating drug safety in an intact animal.

Methods:
Using custom MATLAB software developed in our laboratory, we performed monophasic action potential (MAP) recordings and initiated protocols to induce sustained-VT through right ventricular epicardium PES outputs. Studies were performed in adult male Sprague-Dawley rats (N=22) six weeks after left coronary artery ligation under anesthesia and open chest.

Results:
CHF was verified by standard hemodynamic and echocardiographic parameters as is standard in our laboratory. In the CHF group, 71% (10/14) of the rats exhibited sustained-VT in response to PES versus 0% (0/8) of Sham rats. MAP recordings taken prior-to and during VT induction provided examples of localized activity for arrhythmia mechanisms such as delayed afterdepolarizations. Mechanical alternans, electrical alternans, intermittent pulse generations, and pulseless electrical activity were all observed in this model. EP data analysis showed a decreased (p<0.05) electrogram amplitude in border and infarct zones (Healthy (H): 8.7 ±2.1 mV, Border: 5.3±1.6 mV, Infarct (I): 2.3±1.2 mV), a similar trend for MAP amplitudes, and an increased (p<0.05) repolarization heterogeneity in the border zone (H: 8.1±1.5 ms, B: 20.2±3.1 ms).

Conclusions:
We have developed a custom computer user interface capable of performing clinically relevant
in-vivo
EP studies in rats with CHF. This rat model reproduces common clinical prognosis factors such as mechanical alternans, electrical alternans, and pulseless electrical activity. These EP studies demonstrate this program’s ability to test the arrhythmogenic potential of pharmaceutic agents, biologics, and implantables in an intact animal model before clinical advancement. We introduce this program to study an animal model’s EP characteristics before, during, and after treatments for CHF, and potentially other disease states.
Abstract 72: The Tumor Suppressor Gene Tip30 Supports Cardiac Compensation During Overload and Inhibits Myocardial Protein Synthesis
Authors: Grund Andrea , Szaroszyk Malgorzata , Korf-Klingebiel Mortimer , Tiedje Christopher , Gaestel Matthias , Batkai Sandor , Thum Thomas , Jungmann Andreas , Bauer Ralf , Yin Xiaoke , Mayr Manuel , Wollert Kai C , Pich Andreas , Xiao Hua , Katus Hugo A , Bauersachs Johann , Müller Oliver J , Heineke Joerg
Published: June 2021

Background:
Here, we examined the currently unknown cardiac expression and function of Tip30, which has previously emerged as a tumor suppressor gene.

Results:
Myocardial TIP30 mRNA and protein were significantly upregulated in response to experimental transverse aortic constriction (TAC). TIP30 contributed to cardiac compensation, since a reduction of cardiac TIP30 in heterozygous Tip30 knock-out mice (HET) led to exaggerated hypertrophy and cardiac dysfunction compared to wild-type mice (WT) after TAC. In turn, TIP30 overexpression by an adenovirus in isolated neonatal cardiomyocytes or by an adeno-associated-virus (AAV9) in mouse hearts led to reduced hypertrophy after pro-hypertrophic stimulation in cells and reduced hypertrophy and improved cardiac function after TAC in mice. Interestingly, cardiac TIP30 levels were strongly diminished in mouse models of genetic cardiomyopathy (mdx mice) and in endstage human cardiomyopathy. Reduced cardiac TIP30 contributed to disease progression, since reconstitution of myocardial TIP30 via AAV9 in mdx mice prevented hypertrophy and improved cardiac function. A protein interaction screen and subsequent characterization showed that TIP30 interacts with the middle domain of the eukaryotic translation factor 1A1 (eEF1A1). As revealed by immunoprecipitation and in situ proximity ligation assay, the cellular interaction of eEF1A1 and its essential cofactor eEF1B was diminished by TIP30 overexpression, but enhanced in cardiomyocytes isolated from HET mice after pro-hypertrophic stimulation. Because eEF1A1 is a crucial mediator of translational elongation, we hypothesized that TIP30 regulates cardiac hypertrophy by interfering with protein synthesis. Indeed, overexpression of TIP30 inhibited cardiac protein synthesis during pro-hypertrophic stimulation, while reduced TIP30 levels in HET (vs. WT) mice triggered enhanced protein synthesis after TAC. Interestingly, administration of the eEF1A1 inhibitor narciclasine ablated the increased hypertrophy in HET mice after TAC.

Conclusion:
TIP30 inhibits protein synthesis by binding to eEF1A1 and inhibiting its interaction with eEF1B. It thereby reduces pathological hypertrophy and supports cardiac compensation during overload.
Abstract 76: Experimentally Increasing Titin’s Compliance in a Mouse Model of Heart Failure with Preserved Ejection Fraction Ameliorates Diastolic Dysfunction
Authors: Methawasin Mei , Strom Joshua G , Slater Rebecca E , Fernandez Vanessa , Saripalli Chandra , Granzier Henk L
Published: June 2021

Increasing titin length in the heart through functional deletion of the RNA recognition motif (RRM) of RBM20 protein leads to a reduction of left ventricular (LV) chamber stiffness in sedentary mice. We tested the therapeutic effect of upregulating compliant titins by creating diastolic dysfunction in a mouse model with inducible RBM20 inhibition (MerCreMer;cRbm20ΔRRM). Under transcriptional control of an MHC6 promoter, RBM20 function of MerCreMer;cRbm20ΔRRM mice was inhibited in time- and cardiac-specific manners.

Adult male MerCreMer;cRbm20ΔRRM mice had undergone transaortic constriction (TAC) surgery with Deoxycorticosterone acetate (DOCA) pellet implantation. Two weeks after TAC+DOCA surgery, mice developed LV hypertrophy and diastolic dysfunction as demonstrated by echocardiography. Four weeks after surgery, pressure volume analysis revealed that LV chamber stiffness was markedly increased in TAC+DOCA mice (140% increased versus sham). However, the TAC+DOCA mice with RBM20 inhibition developed less severe LV chamber stiffness (29% increased versus sham). Passive stiffness measurements showed a reduction of cardiomyocyte stiffness in TAC+DOCA mice with RBM20 inhibition.

Inhibition of the RBM20-based splicing system results in expression of compliant mutant N2BA titin which consequently lead to reduction of cardiomyocyte stiffness and LV chamber stiffness in a mouse model with diastolic dysfunction.
Abstract 78: TNFR2 Stimulation Promotes Mitochondrial Fusion via Stat3 and NF-kB Dependent Activation of OPA1 Expression
Authors: Nan Jing-Liang , Zhu Wei , Wang Ying-Chao , Zhong Zhi-Wei , Zhu Lian-lian , Wu Yan , Zhao Jing , Chen Jing-Hai , Hu Xin-Yang , Yu Hong , Gupta Mahesh , Wang Jian'an
Published: June 2021

Background:
TNFR2 stimulation is known to possess protective effects for the cardiomyocytes, however, the underlying mechanisms remain unknown.

Methods and Results:
Using cultured neonatal cardiomyocytes that were infected with lentivirus containing shRNA targeting TNFR1, we showed that TNFR2 activation by TNFα (5nmol/L) resulted in increased mitochondrial fusion, mitochondrial membrane potential which were associated with both elevated intracellular ATP levels and oxygen consumption rate. Intriguingly, these changes were associated with increased protein levels of OPA1, with no changes in the expression levels of Drp1, Mfn1, Mfn2. We went further and reproduced previously reported data that NF-kB acetylation (Lys310) was increased with TNFR2 activation. Interestingly, however, we also observed dose-dependent effects on increase in Stat3 acetylation. Using shRNA approach, we then demonstrated that either Stat3 or NF-kB knockdown can attenuate TNFR2 induced OPA1 expression. The close interaction between these two signalings was validated by co-IP assay and confocal immunofluorescence staining. Aided by bio-informatics searching, we then performed ChIP assay to show that the binding sites of OPA1 promoter regions for STAT3 (-156 to -167) and NFkB (-192 to -203) were adjacent. We further validated that p300 induced Stat3 acetylation was indispensable for complex formation by the interaction between Stat3-DBD and NF-kB -ΔRHD, which in turn was a key event for OPA1 transcription activation. And silence of p300 can abolish OPA1 upregulation upon TNFR2 activation. Computerized data analysis based on zdock and zrank score followed by molecular dynamic simulation model for the whole Stat3 structure revealed higher value of the exterior dielectric constant (obtained from MM/PBSA calculation) for the two sites, K370 and K383, of Stat3, suggesting the essential roles for these two sites for Stat3-NFkB interaction, which were confirmed by co-IP with Stat3-DBD mutants (K370Q,K370R,K383Q,K383R) approach.

Conclusions:
Our data suggested that p300 mediated Stat3 acetylation cooperates with NF-kB to modulate TNFR2 activation induced OPA1 upregulation, leading to improved mitochondrial morphology and function.
Abstract 79: Sarcospan Has a Protective Effect During Development of Cardiac Disease
Authors: Parvatiyar Michelle S , Nguyen Reginald T , Jordan Maria C , Roos Kenneth P , Crosbie-Watson Rachelle H
Published: June 2021

Sarcospan (SSPN) has an important role in stabilizing sarcolemmal dystrophin- and utrophin-glycoprotein adhesion complexes at the cell membrane. Loss of cell adhesion leads to contraction-induced muscle damage, causing muscle dysfunction and cell death. Recently we have shown a specific role for SSPN in modulating cardiac cell adhesion and physiological function. After transthoracic aortic constriction (TAC), SSPN-null mice transitioned toward failure faster than wild-type (WT) mice. Muscle histology revealed large focal areas of collagen deposition in SSPN-null hearts after TAC compared to WT hearts, suggesting that increased membrane fragility affected cardiomyocyte survival. Our laboratory has shown that SSPN loss reduces sarcolemmal dystrophin levels and associated adhesion complexes in the heart. Whereas, the complete loss of dystrophin leads to development of Duchenne muscular dystrophy (DMD), causing cardiac dysfunction and early mortality. Overexpression of SSPN in DMD mice increased cell adhesion and laminin binding in hearts, leading to improvements in tissue histopathology and increased expression of utrophin, a functional homologue of dystrophin. Examining the restorative potential of SSPN in dystrophic cardiac tissue, led us to query whether compensatory upregulation of SSPN occurs in failing hearts of TAC-treated WT mice. In failing non-DMD hearts, we found that SSPN expression is increased. We have evidence of a chaperone role for SSPN, and its increased expression in the failing heart may contribute to the increased localization of dystrophin and associated glycoprotein complexes at the sarcolemma, which we observed in failing WT hearts compared to untreated controls. The upregulation of cell-stabilizing cell adhesion complexes may compensate for increased wall stress and counter pathological processes that culminate in cardiomyocyte demise, and we are exploring whether naturally increased expression or transgenic overexpression of SSPN in the heart may protect against damage. In summary, we have found that SSPN promotes cardiac function by maintaining cell adhesion and promoting cell survival during disease conditions.
Abstract 90: Modulation of Cardiac Fibroblast to Myofibroblast Transition by Rho-associated Kinases ROCK1 and ROCK2
Authors: Hartmann Svenja , Jatho Aline , Tiburcy Malte , Zimmermann Wolfram H , Ridley Anne J , Lutz Susanne
Published: June 2021

Background:
Rho-associated kinases ROCK1 and ROCK2 play a critical role in the pathogenesis of myocardial fibrosis; however, their specific function in cardiac fibroblasts (CF) remains unclear. Remodelling processes in diseased hearts propels the transition of CF to a myofibroblast phenotype exemplified by increased proliferation, migration and synthesis of extracellular matrix (ECM) proteins. Therefore, we sought to investigate whether ROCK1/2 have an impact on CF characteristics in isolated cells and engineered cardiac tissue.

Methods:
Neonatal wild type (WT) rat CF and cardiomyocytes (CM) were isolated and lentivirally transduced/transfected resulting in downregulation of ROCK1 and ROCK2 by 75%. In addition, WT CF were treated with 10 μM Fasudil or 3 μM H1152P for non-specific ROCK1/2 inhibition. CF gene and protein expression, morphology, proliferation, and migration were assessed. Subsequently, CF and CM were mixed within a hydrogel to engineer heart constructs with assessments of contractile function/rigidity performed by isometric force and rheological measurements, respectively.

Results:
Knockdown of ROCK1 and ROCK2 and inhibition of ROCK1/2 activity altered CF morphology, disrupted cytoskeletal structures, and increased adhesion velocity. Moreover, decreased migration velocity and distance was detected, and the double knockdown and inhibition of ROCK1/2 attenuated proliferation of CF. In contraction measurements, engineered heart muscle (EHM) treated with ROCK inhibitors developed a significantly higher force of contraction per cross sectional area than control EHM. Destructive tensile strength measurement of engineered connective tissue (ECT) treated with ROCK inhibitors showed that rigidity was significantly reduced compared to control, suggesting that ROCK1/2 influence the regulation and turnover of the ECM and thus viscoelastic properties of engineered tissues. Indeed, RNA sequencing of ROCK inhibitor treated ECT showed that ROCK1/2 are involved in the regulation of ECM proteins, such as collagens, biglycan, decorin, elastin and its degrading enzyme MMP12.

Conclusion:
This study demonstrates that ROCK signalling controls myofibroblast characteristics of CF via remodelling of the cytoskeleton and the ECM.
Abstract 91: Genome-wide Identification and Characterization of Cardiac Hypertrophy-related Long Noncoding RNAs in Mice
Authors: Huang Zhanpeng , Wu Gengze , Yang Jian-Hua , Ding Jian , Chen Jinghai , Lin Zhiqiang , Drakos Stavros G , Selzman Craig H , Kyselovic Jan , Pu William T , Qu Liang-Hu , Zeng Chunyu , Wang Da-Zhi
Published: June 2021

Long noncoding RNAs (LncRNAs) are RNA transcripts longer than 200 nucleotides that lack protein-coding potential. Although thousands of lncRNAs have been identified, only a few have been linked to cardiac gene expression and function. In this study, we identified, from genome-scale RNA-seq data, 12 candidate lncRNAs associated with cardiac hypertrophy (CH-lncRNAs). The expression of these lncRNAs was altered in mouse models of cardiac hypertrophy induced by transverse aortic constriction (TAC)- or CnA transgene. To determine the function of these lncRNAs, we developed an adeno-associated virus serotype 9 (AAV9)-based functional screening in postnatal mice. An AAV9:cTNT vector, in which the cardiac troponin T (cTNT) promoter was used to direct cardiac-specific expression of target genes, was utilized to overexpress or knockdown candidate lncRNAs in mouse hearts. Postnatal day 1 wild type or CnA transgenic pups were injected with AAV9 viruses and cardiac function was measured one and two months later. Thus far, we have tested 15 candidate lncRNAs for both gain- and loss-of-function studies. Among them, two lncRNAs were demonstrated regulating hypertrophy growth when knocked down. Finally, we identified the human homologues of CH-lncRNA through analyzing the conservation of the promoter regions of lncRNA genes. We showed that the expression of these human CH-lncRNA was dysregulated in human diseased hearts, suggesting the functional conservation of these lncRNAs in cardiac disease. Our study therefore demonstrated that lncRNAs are important regulator of cardiac hypertrophy and disease.
Abstract 100: A Simple Bedside Grading Scale Can Effectively Predict Severe Post-stroke Upper-extremity Spasticity
Authors: Bayona Hernan F , Chhatbar Pratik Y , Schlaug Gottfried , Feng Wayne
Published: June 2021

Introduction:
Upper-limb spasticity is a very disabling complication after stroke. There has been no simple clinical standard scale to predict spasticity immediately after stroke. This study aims to develop a simple bedside grading scale with the information collected during the acute phase to predict spasticity at 3 month post-stroke

Methods:
This is a prospective cohort study (Prediction and Imaging Biomarker of Post-stroke Motor Recovery) of patients with first-ever acute ischemic stroke with various degrees of motor impairment. NIH stroke scale (NIHSS) was assessed 2-7 days after onset of stroke symptoms. Modified Ashworth Spasticity Scale was used as an assessment tool in biceps, wrist flexors and finger flexors at 90 days (± 15 days) and score ≥2 at any muscle was considered as severe spasticity. Infarction volume was measured based on the lesion on MRI/DWI. Independent predictors of upper-limb spasticity at 90 days were identified by multivariate logistic regression. A risk stratification scale was developed with weighting independent predictors based on beta coefficient.

Results:
One hundred twenty three patients were recruited for this study. Covariates associated with upper-limb spasticity are NIHSS arm score (p<0.0001), sub-cortical location (p=0.004) and lesion volume >65 cc (p=0.025). The proposed grading scale is summation of individual points as followed: NIHSS Arm Score: =4 (2 point), <4 (0 point); infarct location: sub-cortical (1 point), non sub-cortical (0 point); infarct volume: ≥65 cc (1 point), <65cc (0 point). The rates of severe upper limb spasticity for the bedside spasticity scale, in order 0-4, are 8.9%, 29.2%, 65%, 88.7%, 96.2%. In other words, the likelihood of developing severe spasticity increases steadily using the score.

Conclusion:
A simple bedside grading scale can effectively predict severe post-stroke upper-limb spasticity at 90 days. Validation with an independent external dataset is a planned next step.
Abstract 101: Predicting Motor Recovery at One Week After Ischemic Stroke: a Combined Transcranial Magnetic Stimulation and MRI Study
Authors: Takahashi Yukako , Oishi Naoya , Mima Tatsuya , Fukuyama Hidenao , Takahashi Ryosuke , Toyoda Kazunori , Nagatsuka Kazuyuki , Ihara Masafumi
Published: June 2021

Introduction:
The failure of numerous stroke clinical trials partially lies in the difficulty in predicting motor recovery in the acute phase. Because of the interindividual variability in subsequent recovery, better prediction of motor prognosis and earlier patient stratification are required to design a promising protocol for clinical trials. Recently, an algorithm to predict motor recovery at 2 weeks after stroke has been reported by combining diffusion-weighted MRI and transcranial magnetic stimulation (TMS); however, 2 weeks may be too late for therapeutic intervention. The aim of this study was therefore to explore how to predict motor recovery even earlier at 1 week after ischemic stroke.

Subjects and Methods:
Twenty-five patients with acute ischemic stroke (67.9±10.5 years old) who showed supratentorial lesions and hemiparesis of the upper extremity were prospectively enrolled. Integrity of the corticospinal tract was assessed structurally with MRI and functionally with TMS within 7 days after onset (acute phase), at 10-20 days (subacute), and at 6 months (chronic). The fractional anisotropy (FA) asymmetry index (FAcontra-FAipsi)/(FAcontra+FAipsi) at the level of the cerebral peduncle was calculated on MRI, while motor evoked potential (MEP) was recorded on TMS. The Fugl-Meyer scale was used to evaluate upper limb impairment in the subacute and chronic phase.

Results:
Patients with detectable MEP at 1 week after onset showed significantly higher Fugl-Meyer score at 6 months compared to those without MEP (65.4 versus 33.6, p=0.011). Those with FA asymmetry index of less than 0.024 at 1 week showed significantly higher Fugl-Meyer score at 6 months compared to those with the index above 0.024 (64.2 versus 40.1, p=0.037).

Conclusion:
Presence of MEP and smaller asymmetry of FA at 1 week after ischemic stroke could be a useful biomarker for predicting better motor recovery. This finding can be useful in earlier patient stratification in future clinical trials.
Abstract 102: NIHSS Lacks Validity and Sensitivity in Postacute Stroke
Authors: Basobas Brittani , Hand Brittany , Peters Heather , Page Stephen
Published: June 2021

Stroke remains a leading cause of disability, with those affected usually experiencing long-term motor deficits. The National Institutes of Health Stroke Scale (NIHSS) is commonly used to measure acute neurological status, but recently is being deployed in at least 29 chronic stroke trials as a secondary UE motor outcome despite a lack of evidence surrounding its validity in chronic stroke. The purpose of this secondary, retrospective analysis of randomized controlled, multicenter, trial data was to (a) determine the NIHSS’s association with the Stroke Impact Scale (SIS), a poststroke health status measurement, the upper extremity Fugl Meyer (UEFM) and the Arm Motor Ability Test (AMAT), measures of upper extremity (UE) impairment and functional limitation and (b) determine the NIHSS’s sensitivity to UE motor change over time
.
There was no association between total NIHSS scores and SIS physical dimension, overall perception of recovery, activity of daily living scores (
P = -.036, P = .782; P = -.039, P = .640; P = -.054, P 5.520,
respectively) or functional limitation (
P; = –0.141; P; = 0.089
). NIHSS and UE impairment associations were significant (
P; = –0.204; P; = 0.014
) but explained less than 4% of the variance among UEFM scores. Subjects scoring a “zero” on the NIHSS exhibited discernible UE motor deficits and varied scores on the UEFM, AMAT, and SIS. A statistically significant sensitivity to change was found for the UEFM (

χ
2
(3) = 10.783, p = 0.013

), but not for NIHSS (

X
2
(3) = 1.250, p = 0.741

) or either AMAT functional ability or quality of movement scores (

χ
2
(3) = 3.953, p = 0.267; χ
2
(3) = 4.199, p = 0.241, respectively

). There was a significant weak, negative correlation between the NIHSS and UEFM (

r
s
(47) = -.325, p = .026

), but nonsignificant weak, negative correlations between the NIHSS and either AMAT scores (

r
s
(47) = -.051, p = .740; r
s
(47) = -.103, p = .489, respectively

). While being increasingly used in postacute trials, the NIHSS’s weak correlation with measures of motor impairment, functional limitation, and health status measurements as well as its insensitivity to change suggest that its use be reserved to acute bedside status. Its utility as a predictor of real world UE motor outcomes has been purported and is currently under investigation as a next step.
Abstract 103: A Phase I Current Escalation Study for Transcranial Direct Current Stimulation in Ischemic Stroke Patients
Authors: Chhatbar Pratik Y , Chen Rong , DeVries William , Lee Ha M , George Mark S , Adams Robert J , Kautz Steven A , Feng Wuwei
Published: June 2021

Introduction:
Positive dose-response relationship with the use of Transcranial direct current stimulation (tDCS) in post-stroke upper limb motor recovery demands investigating the efficacy of higher tDCS doses. Safety and tolerability of tDCS current higher than typically used 2 mA has not been investigated in stroke patients.

Hypothesis:
tDCS application up to 4 mA for 30 minutes is safe and tolerable in stroke patients

Methods:
A 3+3 study design with current escalation schedule of 1, 2, 2.5, 3, 3.5 and 4 mA was adopted for this tDCS safety study (
Fig. A
). Patients with first-ever ischemic stroke and unilateral hemiparesis were recruited. Single session of tDCS and customatory occupational therapy for 30 minutes using a device (Chattanooga Ionto) with 5х7 cm
2
sponge electrodes (Soterix Medical) on a bihemispheric montage (C3/C4, anode on lesional side) were administered. Stopping rules were (1) second degree scalp burn, or (2) seizure, or (3) new lesion(s) on DWI sequence of MRI or decreased ADC. Tolerability was assessed by tDCS questionnaire administered before and after tDCS. Body resistance and skin temperature at electrode contact site were monitored in real time.

Results:
Eighteen patients were enrolled and completed the study. The current was escalated to 4 mA without any major safety concerns (stopping rule). 50% of patients revealed transient skin redness at anodal site and 17% at cathodal side. No patient had any persistent skin perception issues at the end of tDCS (
Fig. B
). We also present real-time monitoring of body resistance and skin temperature to ensure safety during tDCS administration (
Fig. C
). Skin barrier remained intact through tDCS, as demonstrated by absence of abrupt decrease in body resistance (<400 Ω). Skin temperatures remained well below body temperature (range 26°C-35°C)

Conclusion:
Our data support that tDCS current up to 4 mA is safe and tolerable in ischemic stroke patients. Efficacy and safety should be further tested in a phase II study.
Abstract 113: Streamlined Workflows for Triage and Transfer Reduce Door-to-Puncture Time in Mechanical Thrombectomy
Authors: Kansagra Akash P , Wallace Adam N , Curfman David , McEachern James D , Moran Christopher J , Cross DeWitte T , Williams Jennifer , Ford Andria L , Goyal Manu S , Panagos Peter D , Lee Jin-Moo , Derdeyn Colin P
Published: June 2021

Introduction:
Rapid thrombectomy produces improved neurological outcomes in select patients with acute ischemic stroke. Here, we detail process improvements to reduce door-to-puncture (DTP) times in a multi-hospital network organized around an endovascular-capable center (ECC).

Methods:
A lean workflow was adopted to minimize time door-to-imaging and imaging-to-puncture time intervals. Key features of this workflow included minimalist and parallelized clinical and radiological evaluation with single-session CT and CT angiography (CTA) and point-of-care image interpretation, IV fibrinolysis in the CT suite for patients presenting to the ECC, pre-transfer IV fibrinolysis and CTA for patients transferred to the ECC, immediate transport from the CT suite to the angiography suite (potentially before neurointerventional team arrival), and minimalist setup in the angiography suite. DTP time was tracked between January 2015 and December 2015. Trends over time were characterized using a linear regression model.

Results:
The number of thrombectomies performed each quarter increased by 173% (
p
=0.0017), and there was an increase in the proportion of transferred patients that underwent pre-transfer CTA (
p
=0.0438) (Fig. 1). Quarterly median DTP time decreased by 74% (147 minutes to 39 minutes,
p
<0.0001); this decrease was greatest in transferred patients with pre-transfer CTA, in whom DTP time decreased 81% (129 minutes to 25 minutes,
p
=0.0008) (Fig. 2).

Conclusions:
Workflow improvements to streamline in-hospital evaluation at an ECC and perform critical workup such as CTA at referring hospitals before transfer to an ECC can dramatically improve DTP time.

Fig. 1

Fig. 2
Abstract 114: 24/7 Stroke Nurse Team Coverage Reduced Door-to-needle Time and Improved Tissue Plasminogen Activator Utilization Rates in a Stroke Level 1 Community Hospital
Authors: Haque Zohaib , Enchelmeier Marsha , Bern Stephanie , Malik Maheen
Published: June 2021

Background:
Although the American Heart Association has placed particular emphasis on the importance of improving door-to-imaging (DTI) and door-to-needle (DTN) times, treatment with intravenous tissue Plasminogen Activator (IV tPA) remains nationally low in Acute Ischemic Stroke (AIS) patients, especially for community hospitals due to a lack of in-house neurovascular physician coverage. We sought to remedy this problem by implementing round-the-clock coverage of the Emergency Department by Stroke-trained nurses for patients presenting with AIS who may be appropriate to receive IV tPA.

Methods:
AIS patients treated with IV tPA at a Community Hospital and Primary Stroke Center in St. Louis, Missouri were studied between 2013 and 2016. A 24/7 Rapid Response Team of nurses was trained as specialized stroke responders for which the DTI time, DTN time, and IV tPA utilization rates were then compared between pre-implementation (12 months) and post-implementation (30 months).

Results:
We studied 189 Stroke Code patients who were treated with IV tPA (40 pre-implementation and 149 post). The median DTI time was reduced from 16 minutes (interquartile range [IQR]10-21) to 11 minutes (IQR 6-12) (P < .05), and the median DTN time was reduced from 64 minutes (IQR 44-79) to 51 minutes (IQR 39-65) (P<.05). Compliance within the 60-minute benchmark DTN time improved from 55% (27 of 44 patients) to 76.1% (113 of 149 patients) treated in less than 60 minutes with 53.5% (80 of 149 patients) being treated in less than 45 minutes (P<.05). The tPA treatment rates also increased pre and post-implementation by 77.5% (40 to 71 patients) (P<.05) while IV tPA complication rates decreased from 7.3% to 2.7% (P<.10).

Conclusions:
Implementation of round-the-clock on-site stroke nurse coverage for Acute Stroke Code significantly reduced the DTI and DTN time while increasing treatment rates and decreasing complications amongst patients with AIS treated with IV tPA.
Abstract 121: Transcriptome Profiling of Laser-Capture Microdissected Endothelial Cells From Human Cerebral Cavernous Malformations
Authors: Zeineddine Hussein , Girard Romuald , Li Yan , Lightle Rhonda , Shenkar Robert , Saadat Laleh , Shen Le , Moore Thomas , Fam Maged , Kotipatruni Rama , Shi Changbin , Cao Ying , Polster Sean , Faber Pieter , Ginsberg Mark , Andrade Jorge , Awad Issam
Published: June 2021

Introduction:
Cerebral cavernous malformations (CCMs) are common lesions causing stroke and epilepsy. Mechanistic studies in cultured endothelial cells (ECs) and in murine models have correlated CCM gene loss with signaling aberrations related to MEKK3-KLF2/4, Rho/ROCK and angiogenesis activity, and disruption of junctional proteins and EC-mesenchymal transition. Other studies demonstrated a robust antigen driven clonally expanded B-cell immune response, and pathogenetic B-T cell interactions. We hypothesized that the transcriptome of endothelial ECs from human CCM lesions reflects these postulated mechanisms.

Methods:
Resected human CCMs were harvested at the time of surgical resection, snap frozen, embedded in OCT and stored at -80 °C. Normal brain tissue was acquired at autopsy from patients free of neurological disease. Five human CCMs and 3 normal brain tissues were included in the study. ECs from CCMs and normal capillaries were collected using laser-captured microdissection. RNA was extracted using the TRIzol protocol. RNA libraries, generated using low-input strand specific RNA-Seq kit (Clontech), were multiplexed and sequenced with 50 basepair (bp) single end reads (SR 50 bp), and mapped using STAR v2.4.0g1 alignment. Differentially expressed (DE) genes between CCM and normal ECs were identified with DESeq2. QIAGEN’s Ingenuity® was used to conduct functional enrichment analysis on the DE genes, defining canonical pathways associated with DE genes.

Results:
There were 774 DE genes (
P
<0.05; Fold-change>4). Most significant enriched canonical pathways were inflammation related (the highest enriched were signaling in T-helper cells, complement system and leukocyte extravasation signaling). Other pathways related to extracellular matrix and actin signaling (Rac, Paxillin, actin cytoskeleton and integrin signaling), angiogenesis, MMPs and the coagulation cascade.

Conclusion:
Differential transciptome analysis is feasible in CCM ECs in situ. It validates previously postulated disease mechanisms, and identifies other candidates for future study. This approach can categorize disease-related gene activity in lesions with different genotypes, clinical severity, and therapeutic interventions.
Abstract 124: Plasma Biomarkers of inflammation Reflect Seizures and Hemorrhagic Activity of Cerebral Cavernous Malformations
Authors: GIRARD Romuald , Zeineddine Hussein A. , Fam Maged D. , Mayampurath Anoop , Cao Ying , Shi Changbin , Shenkar Robert , Jesselson Michael , Duggan Ryan , Tan Huan , Mikati Abdul-Ghani , Andrade Jorge , Whitehead Kevin J. , Li Dean Y. , Awad Issam A.
Published: June 2021

Introduction:
The clinical course cerebral of cavernous malformations (CCMs) is highly variable, with a limited number of recent studies querying factors associated with disease severity. We hereby explore a panel of peripheral plasma biomarkers implied with inflammation and angiogenesis in relation to CCM clinical activity.

Methods:
Blood samples of 85 CCM patients (49 with solitary/sporadic lesions and 36 with multifocal/familial CCMs) were collected at the time of the clinical visit, concurrently with advanced MRI sequences. Twenty
a priori
chosen plasma biomarkers were quantified and analyzed in relation to established parameters of disease categorization and severity, including genotype, lesion burden, age at symptomatic presentation, CCM-related seizures and the number and timing of prior symptomatic hemorrhages. We first tested classic univariate correlations of each biomarker with disease features, including an FDR correction, and we then applied a multivariate hierarchical clustering approach. We further correlated the peripheral plasma biomarkers with measures of lesional permeability and iron deposition using previously validated MRI protocols.

Results:
MMP2 and ICAM1 levels were significantly higher (p=0.02 and p=0.04 respectively) in patients with seizure activity while MMP9 was lower (p=0.04). VEGF and endoglin/CD105 (p=0.04 for both) plasma levels were both lower in patients who had suffered a symptomatic bleed in the prior 3 months. The hierarchical clustering analysis revealed a cluster of 4 plasma inflammatory cytokines (TNFα, IL1β, IL2 and IFNγ) separating patients into high and low inflammatory states. The high inflammatory state was associated with more CCM hemorrhagic events during a patient’s lifetime (p=0.04) but not recent bleeding. CCM lesion iron concentrations were inversely correlated with IL-10 (r=-0.61, p=0.02), CCL2/MCP1 (r=-0.60, p=0.02) and ROBO4 (r=-0.53, p=0.05) in CCM lesions that recently bled.

Conclusion:
Peripheral plasma biomarkers reflect seizure and recent hemorrhagic activity from CCM. And clusters of pro-inflammatory biomarkers correlate with cumulative chronic disease aggressiveness. Other biomarkers may reflect the clearance of lesional iron after recent hemorrhage.
Abstract 130: Is Intravenous Thrombolysis Far From Obsolete in the Era of Endovascular Thrombectomy? A Hint From a Meta-analysis of Randomized Clinical Trials
Authors: Tsivgoulis Georgios , Katsanos Aristeidis H , Mavridis Dimitris , Alexandrov Anne W , Magoufis Georgios , Arhur Adam , Schellinger Peter D , Alexandrov Andrei V
Published: June 2021

Background & Purpose:
Current AHA recommendations advocate that pretreatment with intravenous thrombolysis (IVT) should first be offered to all eligible acute ischemic stroke (AIS) patients with emergent large vessel occlusion (ELVO) before an endovascular thrombectomy (ET) procedure. Nevertheless, recent single-center observational studies question the utility of IVT pretreatment in ELVO patients eligible for systemic thrombolysis and advocate ET monotherapy. We sought to evaluate the comparative efficacy between ET and bridging therapy (IVT&ET) in AIS due to ELVO.

Methods:
We performed mixed-effects subgroup analyses, according to IVT pretreatment status of both functional independence [modified Rankin Scale (mRS) of 0-2)] and death/dependency (mRS of 5-6) at 90 days using available RCTs that evaluated the safety and efficacy of ET with stent-retrievers in comparison to standard therapy. We additionally performed an ordinal logistic regression analysis of individual patient data on the distribution of 3-month mRS scores (shift analysis) between the two groups.

Results:
We identified 7 eligible RCTs including 1764 ELVO patients (53% men), and 108 patients randomized to ET without IVT pretreatment. Patients receiving bridging therapy (BT) had lower rates (p=0.041) of 90-day death/dependency (19%, 95%CI: 14%-25%) compared to patients receiving only ET (31%, 95%CI: 21%-43%; Figure). Similarly, shift-analysis favored a trend for BT over ET in terms of better 90-day functional outcome (common OR=0.78, 95%CI: 0.53-1.10; p=0.155).

Conclusion:
Our findings indicate that pretreatment with IVT prior ET in ELVO patients may be associated with lower rates of three-month death/dependency and a trend towards more favorable functional outcomes. Until the results from head-to-head RCTs comparing BT to ET monotherapy become available, our findings support AHA guidelines recommending delivery of tPA to appropriate ELVO candidates undergoing ET.
Abstract 136: Genetics of White Matter Hyperintensity Burden in Patients With Ischemic Stroke: The MRI-GENIE Study
Authors: Giese Anne-Katrin , Xu Huichun , Ryan Kathleen , Schirmer Markus D , Dalca Adrian V , Dave Tushar , Cole John W , McArdle Patrick F , Broderick Joseph P , Jimenez-Conde Jordi , Jern Christina , Kissela Brett M , Kleindorfer Dawn O , Lemmens Robin , Lindgren Arne , Meschia James F , Rundek Tatjana , Sacco Ralph L , Schmidt Reinhold , Sharma Pankaj , Slowik Agnieszka , Thijs Vincent , Woo Daniel , Worrall Bradford B , Wu Ona , Kittner Steven J , Golland Polina , Rosand Jonathan , Mitchell Braxton D , Rost Natalia S
Published: June 2021

Introduction:
The MRI-Genetics Interface Exploration (MRI-GENIE) study is the first international collaboration that aims to facilitate genetic discoveries in clinical cohorts of patients with acute ischemic stroke (AIS). We have amassed the largest-to-date collection of AIS cases with brain MRI scans and genome-wide genotyping to test the role of genetic susceptibility in MRI-based cerebrovascular traits.

Objective/Hypothesis:
To elucidate the genetic architecture of white matter hyperintensity (WMH) burden in AIS patients.

Methods:
Using a novel automated algorithm, we extracted WMH volume (WMHv) from clinical MRI scans of 2704 AIS patients (age 63.1 ± 14.7 years, 60.6% male) of European ancestry. Quality control (QC) measures were undertaken per subject and per SNP, excluding subjects with non-European ancestry and poor genotyping, as well as SNPs deviating from Hardy-Weinberg equilibrium and high levels of missingness. Imputation to the Haplotype Reference Consortium (HRC version r1.1) was conducted for 1712 remaining subjects with 2.8 million SNPs on the Michigan Imputation Server. After exclusion of poorly imputed SNPs (R
2
<0.5) and SNPs with minor allele frequency < 1%, 7.7 million SNPs remained for further analysis. Genome-wide association testing of natural log-transformed WMHv on the allelic dosage per SNP was adjusted for age, sex and principal components 1-10.

Results:
Genome-wide association testing has identified a novel locus on chromosome 2 (T allele at rs72856504) near the LDL Receptor related Protein 1B gene (LRP1B) that was significantly associated with WMHv burden in AIS (β=0.54, SE=0.098, p=3.65*10
-8
).

Conclusion:
We have identified a novel locus (T allele rs72856504) on chromosome 2 near the LRP1B gene, which is specific for WMH in AIS and has not been previosuly described in stroke-free WMH cohorts. A replication effort involving additional independent cohorts of AIS patients with brain MRI and genome-wide genotyping is ongoing.
Abstract 144: A Novel Multi-function Small Molecule for the Treatment of Acute Ischemic Stroke in Rodent and Non-human Primate
Authors: Yeh Sheng Wen , Jiang Yinghua , Yeh Tzulin , Chern Wendy
Published: June 2021

Background and rationale:
DC007 is a synthetic small molecule with multiple pharmacological activities. It has been shown to have thrombolytic, anti-platelet and free radical scavenging activities in vitro. To explore its potential in treating acute ischemic stroke, we evaluated its safety and efficacy using a thromboembolic stroke model in rattus and in Cynomolgus macaque.

Methods:
Effects of DC007 (10mg/kg) were compared to intravenous tPA (10mg/kg in rat; 0.9mg/kg in monkey) using a model of focal embolic cerebral ischemia in rats and monkeys. Pre-formed blood clots were introduced into middle cerebral artery through catheter cannulated into internal carotid artery of the animals. Animals (12 rats per group; 4 monkeys per group) were randomized to receive intravenous infusion of saline, tPA or DC007 at 3 hours after embolization was confirmed.

Results:
DC007 reduced brain infarction and hemispheric swelling in the stroke rats more significantly than saline or tPA did, when treatments were given at 3 hours after stroke onset. At 24 hours after stroke, infraction volumes were 49.36%, 31.98%, and 40.66% for saline, DC007 and tPA group, respectively; while brain swelling rates were 17.67%, 13.20% and 22.58% for saline, DC007 and tPA group, respectively. In the thromboembolic stroke model in monkey, DC007 also showed a stronger beneficial effect on vessel patency (DC007: 46.3±10.3% vs. tPA: 22.2±12.8% in the 8 hours time period after stroke), brain infarction (DC007: 3361±354 mm
3
vs. tPA: 9072±2096 mm
3
measured by MRI at 24 hours after stroke) and neurobehavioral score measured at 72 hours after stroke (DC007: 46.3±10.3 vs. tPA: 60±9.8, where score 0 indicated normal, while score 77 indicated death).

Conclusions:
DC007 appears to be a more potent thrombolytic compound with superior safety profile than IV rt-PA when given at 3 hours after the stroke onset in rats and monkeys. Further investigation is warranted for its potential in the treatment of AIS clinically.
Abstract 145: Development of Novel S100A9 Peptide Vaccine for Prevention of Thrombus Formation in Ischemic Stroke in Mice
Authors: Kawano Tomohiro , Shimamura Munehisa , Nakagami Hironori , Koriyama Hiroshi , Sasaki Tsutomu , Sakaguchi Manabu , Morishita Ryuichi , Mochizuki Hideki
Published: June 2021

Background and purpose:
Nonadherence of antithrombotic medicine is one of major risk factors for recurrent strokes. Immunotherapy, such as antithrombotic vaccination, is expected to solve such a problem due to its long-lasting effects. Here, we focused on platelet-derived S100A9, which has directly regulate thrombosis without influence on hemostasis, and examined the antithrombotic effects of newly developed S100A9 vaccine in mice.

Methods:
S100A9 vaccine was designed to select the short peptide as an antigen of S100A9 conjugated to keyhole limpet hemocyanin (KLH). Male C57BL/6J mice were immunized with S100A9 vaccine three times at 2 weeks interval. Middle cerebral arteries (MCA) was exposed to topical application of 20% ferric chloride. The time to occlude the MCA was examined measuring cerebral blood flow with a laser doppler flowmeter. Bleeding time was assessed by cutting the tail arteries. The thrombus formation was also assessed using the laminar flow chamber. The influences of vaccine for T-cell activation were assed using an IgG subclass ELISA and an enzyme-linked immunosorbent spot (ELISPOT) assay.

Results:
S100A9 vaccine successfully produced antibodies specific for S100A9 in serum in a dose-dependent manner. After the third immunization, the time to thrombus formation in MCA was significantly prolonged in the mice vaccinated with S100A9 compared to the non-immunized mice (23.3 ± 4.3 min vs 6.9 ± 0.8 min, p<0.01), but bleeding time was same in both groups. There was no significant difference in platelet count in both groups. The formation of platelet thrombus under ex vivo was significantly reduced in S100A9 vaccine group compared to control group (14.8 ± 0.7 x10
4
/μm
2
vs 17.8 ± 0.9 x10
4
/μm
2
, p<0.05). In the IgG subclass ELISA, the IgG1:IgG2b ratio was greater than 1.0 in the S100A9 vaccine group. In the ELISPOT assay using splenocytes isolated from S100A9 immunized mice, stimulation with S100A9 partial peptide did not induce the production of IFN-gamma and IL-4 in comparison with KLH.

Conclusion:
The S100A9 vaccine could successfully prevent thrombus formation in MCA without influence on hemostasis nor eliciting an autoimmune response. S100A9 vaccine might shed light on the development of safe antithrombotic vaccine in ischemic stroke.