bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 319: Small Molecule Gβγ Inhibition Attenuates Cardiac Fibroblast Inflammatory and Pro-Fibrotic Signaling
Authors: Travers Joshua G , Kamal Fadia A , Burhans Michael S , Blaxall Burns C 
Published: July 2021

Heart failure (HF) is a devastating disease characterized by chamber remodeling, interstitial fibrosis and reduced ventricular compliance. Prolonged sympathetic overstimulation promotes excess signaling through G-protein Gβγ subunits and ultimately results in pathologic GRK2-mediated β-adrenergic receptor (β-AR) downregulation. We have recently demonstrated the therapeutic potential of the small molecule Gβγ-GRK2 inhibitor Gallein in limiting HF progression. Pathologic activation of the cardiac fibroblast (CF) induces the transition to a myofibroblast phenotype, which plays a fundamental role in myocardial fibrosis and remodeling. We hypothesized that Gβγ-GRK2 inhibition plays an important functional role in the CF to attenuate pathologic CF activation, inflammation and interstitial fibrosis.
To explore the effect of Gβγ-GRK2 inhibition on inflammation and pro-fibrotic signaling, mice were subjected to 7 days of transverse aortic constriction, a pressure-overload model of HF. In addition to the attenuation in overall cardiac hypertrophy, animals treated with Gallein displayed reduced expression of pro-inflammatory cytokines, including macrophage inflammatory protein 1 alpha (MIP-1α) and MIP-1β, along with Interleukin-6, as assessed by qPCR. Gallein-treated animals also exhibited diminished pro-fibrotic signaling, as evidenced by a reduction in the expression of TGFβ, a major driver of myocardial fibrosis, and decreased expression of collagen. To recapitulate these findings in vitro, primary adult mouse ventricular fibroblasts were pathologically stimulated using Isoproterenol (ISO, β-AR agonist) or Angiotensin II and treated +/- Gallein for 24 hours. CFs treated with Gallein displayed an analogous reduction in the expression of these pro-inflammatory cytokines and collagen.
In summary, these data suggest a potential therapeutic role for small molecule Gβγ-GRK2 inhibition in limiting pathologic myofibroblast activation, inflammation and interstitial fibrosis. We believe this fibroblast-targeted approach will lead to the refinement of existing targets and compounds, and possibly the generation of novel therapeutics for the treatment of HF.
Abstract 10: MicroRNA 17 in Angiogenesis: Lessons Learned From Immobilized Vascular Endothelial Growth Factor
Authors: Aday Sezin , Besnier Marie , Zoldan Janet , Saif Jaimy , Santos Tiago , Carreto Laura , Bernardino Liliana , Langer Robert , Emanueli Costanza , Ferreira Lino 
Published: July 2021

Several pre-clinical and clinical studies are exploring the therapeutic effect of cell-based therapies in ischemic diseases, including Peripheral Artery Disease (PAD). Unfortunately, most of the cells (more than 80%) die few days after delivery. We postulate that better understanding of VEGF Biology might be important for the design of more effective pro-survival strategies. Release of VEGF from ECM activates a carcinogenic program since it triggers vasculogenesis, tumor growth and metastasis. Contrarily, immobilized VEGF might have all the properties of soluble VEGF without inducing a carcinogenic program. Thus, identification of downstream players such as miRNAs mediating this process might be important. Herein, we evaluated therapeutic effect of miR-17 downregulation, which mediates the effect of immobilized VEGF both in vitro and in vivo.
We recently showed that conjugated VEGF modulates cell activity by decreasing the expression miR-17 both in vitro and in vivo. In the present study, cell survival and angiogenesis were evaluated firstly in vitro using endothelial cells (ECs) transfected with antagomiR-17 to mimic the down-regulation of miR-17 by conjugated VEGF. AntagomiR-17 increased EC survival at least 1.5 times (n=6) compared to pro-angiogenic miRNAs reported in the literature (e.g. miR-424 and miR-132) and sprout formation on Matrigel at least 2 times (n=5) compared to all groups. The effect of antagomiR-17 was more pronounced under hypoxia conditions. In vivo, antagomiR-17 accelerated hemodynamic recovery of the whole limb (n=12) in unilateral limb ischemia obtained by occlusion of the left femoral artery. Blood flow recovery evaluated by Laser Doppler analysis was significantly higher 21 days after surgery in antagomiR-17 group compared to all other groups. Immunohistochemical analyses showed an increase in the capillary density of skeletal muscle in antagomiR-17 condition. In order to determine the gene target and potential pathway involved in the biological effect of antagomiR-17, next generation mRNA sequencing was performed.
In conclusion, here we show the potential and underlying molecular mechanism of antagomiR-17 treatment in endothelial cell survival and angiogenesis both in vitro and in vivo.
Abstract 58: Dicarbonyl Stress and Diabetic Heart Failure: The Role of Endothelial Cells
Authors: Vulesevic Branka , McNeill Brian , Giacco Ferdinando , Brownlee Michael , Milne Ross , Suuronen Erik 
Published: July 2021

Dicarbonyl stress (DS) caused by the accumulation of α-oxoaldehyde metabolites, like methylglyoxal (MG), leads to detrimental DNA and protein modifications. Under normal conditions, MG is detoxified by glyoxalase-1 (GLO1) and -2 enzymes, but this system fails in diabetes. While the role of DS in diabetic cardiomyopathy through changes in cardiomyocyte function has been well described, this study aimed to link DS with the development of endothelial dysfunction (ED) and early heart failure in diabetes.
Transgenic mice that over-express GLO1 in endothelial cells (ECs) but not in cardiomyocytes, and their wild-type (WT) littermates were treated with STZ to induce hyperglycemia (WT-diabetic and GLO1-diabetic mice) or vehicle (non-diabetic controls). Hyperglycemia increased the circulating levels of ED markers in WT-diabetic (E-selectin 1.5-fold, ICAM 1.4-fold, and VCAM 1.1-fold), but not GLO1-diabetic mice. The number of vWF+ ECs in WT-diabetic hearts was reduced 2-fold compared to other groups, whereas GLO1 over-expression preserved capillary density. Cell death, determined by TUNEL staining, was greater in the hearts of WT-diabetic mice compared to all other groups. GLO1 over-expression resulted in reduced inflammation: TNF-α protein expression was increased in both diabetic groups (≥2-fold), but significantly less so in GLO1-diabetic mice (p=0.03). The preservation of ECs in GLO1-diabetic mice was associated with delayed signs of heart failure. At 4wk of hyperglycemia, WT-diabetic mice had reduced heart function compared to all other groups (p=0.04). At 8wk, cardiac function in GLO-diabetic mice was greater than in WT-diabetic mice, but both were reduced compared to non-diabetic controls (p=0.02; p=0.4). A possible mechanism for EC survival in GLO1 mice despite the presence of inflammation was examined in vitro using human aortic ECs. ECs exposed to high glucose or MG for 24h had increased apoptosis induced by TNF-α compared to cells treated only with TNF-α (by 2- and 3-fold, respectively), suggesting that reduced MG protects ECs from TNF-α mediated death.
Taken together, these results suggest that DS in diabetes increases inflammation and ED, leading to the loss of ECs in the heart, which contributes to the development of heart failure.
Abstract 6: Novel Tools for Generation, Purification, and Analysis of Pluripotent Stem Cell Derived Cardiomyocytes
Authors: Eckardt Dominik , Noack Kristin , Bosio Andreas 
Published: July 2021

Pure and well-characterized cardiac cells derived from human pluripotent stem cells (hPSCs) are of high interest for cardiovascular disease modeling, drug safety studies and development of cell replacement strategies. Although several protocols for cardiac differentiation of hPSCs have been developed, major limitations are clone-to-clone variations in differentiation efficacy as well as heterogeneity of generated cardiomyocyte populations. Therefore, we have developed novel tools for cardiomyocyte differentiation, magnetic cell sorting-based purification and cell analysis using new antibody-conjugates enabling for flow cytometry- and immunofluorescence-based identification of cardiomyocytes and subtypes.
hPSCs were maintained under xeno-free conditions in our recently developed StemMACS iPS-Brew XF medium to keep pluripotency for more than >20 passages and enable for efficient cardiac differentiation. We chose a monolayer differentiation protocol based on the timely regulated activation and inhibition of Wnt signaling by small molecules.
In order to identify antibodies suitable for cardiomyocyte enrichment or depletion of non-myocytes, we performed a surface marker screen with more than 400 antibodies between days 10-20 of differentiation. Besides identification of new surface markers, our screen confirmed expression of recently published markers like CD172a and CD106. In order to evaluate kinetics of their expression and correlation with intracellular markers of cardiomyocytes and subtypes, we performed a flow cytometry-based analysis of marker expression. Our data indicate a dynamic expression pattern for both CD172a and CD106, either completely or partially overlapping with intracellular cardiomyocyte marker expression. Based on these data we developed magnetic cell separation procedures for the isolation of cardiomyocytes. Magnetically enriched cardiomyocytes initiated contractions after replating and could be stably maintained in culture.
Taken together, we have developed novel tools supporting the workflow for efficient generation of PSC-derived cardiomyocytes, magnetic purification and flow cytometry or immunofluorecence-based characterization of cardiomyocytes.
Abstract 312: Visualization of Cardiomyocyte cGMP Dynamics in Hypoxia/Reoxygenation
Authors: Bork Nadja I , Nikolaev Viacheslav O 
Published: July 2021

The second messenger cyclic guanosine 3′5′-monophosphate (cGMP) plays an important role in the regulation of multiple physiologic processes in the cardiovascular system for example it is known to inhibit hypertrophy and to protect against ischemia-reperfusion injury. As a consequence of hypoxia in the heart, maladaptive signaling cascades are activated that can result in cardiac damage and finally lead to heart failure. However, cellular responses to hypoxia/reoxygenation (H/R) are still incompletely understood and almost nothing is known about cGMP dynamics in the context of H/R.
The aim of this project was to investigate the effects of H/R on cGMP dynamics in mammalian cardiomyocytes. Therefore transgenic mice with cardiomyocyte-specific expression of the cytosolic Förster resonance energy transfer (FRET)-based cGMP sensor red cGES-DE5 were used to study cGMP dynamics in single adult cardiomyocytes exposed to H/R. In addition, a Langendorff system was used for FRET measurements in whole-heart.
Basal cGMP levels in single adult cardiomyocytes exposed to H/R were increased. This increase was generated already during the hypoxic period and was maintained during reoxygenation. PDE3 protein expression and activity were significantly downregulated after H/R, however RNA expression level of PDE3 was not significantly changed. At the same time, protein levels of the soluble guanylyl cyclase β-subunit showed a tendency towards downregulation during H/R.
In whole heart measurements, we could show that cGMP levels increase during anoxia and decrease during reoxygenation. This indicates that there is also an influence of other cell-types on cGMP dynamics or on cGMP cell-cell transfer.
In conclusion, we developed FRET-based cGMP measurements in single cardiomyocytes and whole hearts in context of H/R and found an increase of intracellular cGMP in hypoxia. This offers great opportunities to dissect the molecular mechanism of cGMP signaling regulation during ischemic injury and should help to distinguish between direct protective effects on cardiomyocytes and indirect mechanisms such as cell-cell interactions in cGMP signaling during and after H/R.
Abstract NS 7: Code 1 (Acute Stroke Response Team): Improving Patient Outcomes Utilizing Simulation Training
Authors: Poole Karen , Filomena Patricia 
Published: July 2021

The use of innovative blended learning for stroke program training has enabled the Washington Hospital Center’s Stroke Program in Washington, D.C. to decrease their patient “door presentation time” to Tissue Plasminogen Activator (tPA) “needle time” by 30minutes. Stroke programs always seem to be in a constant state of performance improvement and looking for way s to create seamless care from admission to treatment. A review of stroke response times and needs assessment in 2008 led the MedStar Health Washington Hospital Center Stroke Coordinator to collaborate with MedStar Health Simulation and Training Environmental Lab (SiTEL) to develop team training and individual stroke nurse responder training that includes both online modules and clinical simulation scenarios. Over the past three years this program has evolved and now includes online modules on stroke diagnosis and treatment, didactic instruction, preceptorship with an experienced stroke nurse responder and the use of clinical scenarios with live actors as patients to give nurses and the rest of the team practice with running a stroke code.
Initial objectives/goals included testing the team that had been trained and developing practice and validation scenarios for the nurse stroke responders so they could practice how to assess potential stroke victims, carrying out the stroke protocol and administer tPA. A second scenario is conducted quarterly for the entire team and is used to evaluate the entire process. Throughout this immersive simulation the team of physicians, nurses and radiology technicians must make crucial assessments, critical treatment decisions, follow a stroke protocol, transport the patient, administer tPA, monitor the patient and escalate care as complications arise. This scenario is timed and videotaped to allow the team to debrief after the scenario and review decisions made and actions taken thus increasing their efficiency and effectiveness of critical decision making regarding patient care. The overall success of this education program can be seen in the shortened door to tPA administration time for the acute stroke patient.
Abstract WP241: Value Stream Analysis Reduces Door-To-Needle Times in Acute Stroke
Authors: Bentaleb Abdeljalil , Golding Larisa , Southee Susan , Eberly Lewis B 
Published: July 2021

Introduction:
An important goal in treating acute stroke is minimizing door-to-needle times, consistent with evidence based practice and quality goals. Utilizing methods to streamline the process of patient and information flow in the emergency department (ED) is necessary to reduce treatment times.

Hypothesis:
Use of value stream analysis to improve efficiency in processing acute stroke patients in the ED should reduce door-to-needle times.

Methods:
Value stream is the movement of patient and information to deliver the desired outcome. The process from door to treatment was divided into individual steps. We defined process time (the time each step takes) and wait time (the time between each step). The mission was to identify which steps were absolutely necessary before a patient could receive IV-tPA, and to streamline those steps to reduce the process time. All patients who received IV-tPA in 2011 and thus far in 2012 are shown in the graph. Each bar represents an individual patient.

Results:
Prior to stream analysis, our average door-to-needle time was 122.2 minutes. After the process was rolled-out, the average door-to-needle time was down to 73.8 minutes. Before stream analysis we treated 7.6% of our IV-tPA patients in less than 60 minutes. After analysis we treated 69.2% of patients in less than 60 minutes. The number of patients treated rose from 11 in 2011 (5%) to 17 thus far in 2012 (mid-year, projected 15%). We also found that the number of thrombectomy treated patients was reduced from 39% prior to analysis to 23% following analysis, perhaps partly due to reduced processing time and more patients evaluated within the window for IV-tPA.

Conclusion:
Streamlining the process of patient flow in the ED by value stream analysis resulted in several outcome improvements in acute stroke patients:

1) Reduced door-to-needle times.
2) Increased number of patients treated within the golden hour.
3) Increased number of patients treated with IV-tPA.
Abstract WP269: Increased Sensitivity to recombinant Tissue Plasminogen Activator (rtPA)-induced Lysis in vitro of Thrombi Produced in Human Platelet Rich Plasma containing Dabigatran
Authors: van Ryn Joanne , Kink-Eiband Monika , Fischer Davina , Schurer Johanna , Clemens Andreas 
Published: July 2021

Dabigatran is a direct thrombin inhibitor approved for the prevention of stroke in patients with atrial fibrillation. It significantly reduced ischemic and hemorrhagic stroke in these patients vs warfarin. However, in patients treated with dabigtran who developed a stroke, there remain unresolved issues regarding thrombolysis therapy, both in terms of timing of lytic treatment to minimize hemorrhage risk and also lytic dose. It is hypothesized that if the fibrin network in a thrombus formed in patients on dabigatran treatment is altered and is less compact, then it may be more sensitive to lysis at lower doses of rtPA.

Thrombi were produced in human platelet rich plasma (PRP) in the presence of low conc. of dabigatran (0, 30, 75 ng/ml). PRP was supplemented with 594-Alexa fluorescent labelled human fibrinogen. Thromboplastin (Thromborel
®
) and 200mM CaCl
2
were added and this was incubated at 37°C for 60 min to allow clot formation. Thrombi were removed from PRP and washed twice in saline. They were then added to human plasma from the same subject containing increasing conc. (0-250 IU/ml) of rtPA (Actilyse
®
) and incubated for 2 hrs under continual stirring conditions at 37°C. Lysis was measured as amount of fluorescence released into plasma from the thrombus over 2hrs.

Preformed thrombi added to plasma containing only dabigatran underwent no lysis. Lysis of preformed control thrombi (in the absence of dabigatran) occurred in a conc-dependent manner when incubated with rtPA and maximal lysis was achieved with 300 IU/mL. Thus further experiments were performed with ≤250 IU/ml rtPA to look for potential synergistic effects with dabigatran. Clots preformed in PRP containing either 30 or 75 ng/mL dabigatran were more susceptible to rtPA-induced lysis, with maximal lysis achieved with 100 IU/mL rtPA, and ~ 90% lysis was achieved with 50 IU/mL rtPA.
These data provide evidence that thrombi preformed in low concentrations of dabigatran appear to be more susceptible to thrombolysis with rtPA than those preformed in the absence of dabigatran, thus lower concentrations of rtPA achieve a similar degree of thrombolysis under these experimental conditions. This may imply that in patients that suffer a stroke while on dabigatran therapy, lower doses of tPA may also be effective.
Abstract WP4: Conscious Sedation With Dexmedetomidine For Acute Stroke Patients Undergoing Intrarterial Therapy.
Authors: Jumaa Mouhammad A , Lin Ridwan , Zaidi Syed F , Linares Guillermo , Jadhav Ashutosh , Jankowitz Brian , Jovin Tudor G 
Published: July 2021

Background:
The aim of this pilot study was to prospectively determine safety and feasibility of Conscious sedation (CS) with dexmedetomidine in stroke patients undergoing intra-arterial therapy (IAT).

Methods:
This industry sponsored prospective single arm study was conducted with institutional IRB approval. The following data points were collected: demographics, pre-procedure NIHSS, mean time to treatment, treatment modalities, sedation and procedure times and the use of adjunctive sedation medications. Outcome measures included intraprocedural pain scores and Richmond agitation sedation scale (RASS), recanalization score, intraprocedural complications, conversion to GA, modified Rankin scale at 30 days as well as a personal account of the patients experience during the procedure.

Results:
Fifty- one patients were enrolled in this study of which 41 patients underwent acute stroke therapy and were included in this analysis.

Baseline characteristics are included in table 1. Site of occlusion was the MCA in 27 patients, ICA-terminus in 10 patients and BA in 4 patients. IAT was performed using the MERCI device in 30 patients, Penumbra device in 3, manual aspiration in 5, and angioplasty and stenting in 3 patients. Adjunctive IA tPA was administered in 6 patients.
All patients tolerated treatment under CS. The mean procedural RASS was 0 and the mean pain score was 1. Thrombolysis in Cerebral Ischemia (TICI) 2B/3 recanalization was achieved in 32 patients (78%). Two intra-procedural wire perforations (5%) were recorded. Ten patients (24%) had a mRs of < 2 at 30 days and 6 patients died (14%). Thirty-one patients described their experience. Only 6 patients were able to remember having the procedure of which 1 had a negative experience.

Conclusion:
To our knowledge, this is the first prospective study to evaluate the safety and efficacy of CS in stroke patients who undergo IAT. In our cohort, CS was safe, feasible and well tolerated.
Abstract T P318: Comparative Effectiveness of tPA on 6 Month, Patient-Centered Outcomes: A Pilot Analysis
Authors: Lang Catherine E , Bland Marghuretta D , Corbetta Maurizio , Lee Jin-Moo 
Published: July 2021

We examined the real-world benefit of tPA, delivered as part of usual care, on patient-centered outcomes. All persons with a diagnosis of stroke or TIA from our hospital are contacted for telephone follow-up at 6 mos. after the event. The survey includes the Stroke Impact Scale (SIS), Patient Health Questionnare-9 (PHQ9), and other questions about aspects of daily life.
This study retrospectively compared outcomes in patients who received tPA as part of usual care with patients who would have received tPA had they arrived at the hospital within the time window, i.e. criteria other than time were met. From hospital records, we acquired demographic and medical data. Patients in each group with completed surveys were matched on initial NIHSS score, age, race, gender, and date of stroke. Staff conducting surveys were independent from staff determining matches and were blinded to the others data. Matching was done with 1:2 ratio of tPA to controls. Results were compared between groups with 1-tailed tests due to directionally-specific hypothesis in favor of the tPA group.
Persons receiving tPA and controls completed the surveys at the same rate. Sample size was 67 for the tPA group and 137 for the control group, with the groups matched across the above variables as intended. On the SIS, the tPA group reported better physical function (p=.001), communication (p=.048), cognition (p=.029), and quality of life (p=.04) compared to the control group. Depressive symptomatology, as indicated by PHQ9 scores, was not different between groups (p=.48). Fewer people in the tPA group (27%) reported 1 or more emergency room visits in the 6 months after their stroke compared to controls (44%, p=.03). Of those driving prior to their stroke, 72% had returned to driving, vs. 58% of controls (p=.045). Return to work rates were not different (52% tPA vs. 39% controls, p=.19) within the smaller subset of people working prior to their stroke.
While it is known that tPA reduces long-term disability, this is the first study to demonstrate the effectiveness of tPA in improving meaningful, patient-centered outcomes, such as physical function and communication. These data indicate that use of tPA provides a large benefit to the daily lives of people with ischemic stroke.