bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 5116: Stable Arteriogenesis of Ischemic Hind Limbs by a Conditionally Silenced AAV9 Gene Therapy Vector Expressing Human VEGF
Authors: Boden Jeffrey , Webster Keith A
Published: April 2024

INTRODUCTION: Therapeutic angiogenesis to treat coronary and peripheral artery disease with genes or cells has not been successful.

RATIONALE: Previous protocols focused on angiogenesis with two major flaws

Inadequate delivery vehicles

Use of unregulated genes with no directional cues for new vessel growth.

HYPOTHESIS: VEGF gene delivery in a semipermanent muscle-tropic AAV9 vector with tight hypoxia-regulation supports directional angiogenesis, arteriogenesis and stable reperfusion.

METHODS: The human (h)VEGF gene driven by a PGK promoter containing tandem arrays of HIF-1
α
and NSF (silencer) binding sites was cloned into AAV9. Gene expression and regulation were quantified in myocytes and ischemic hind limbs of normal and atherosclerosis-prone mice. Arteriogenesis was evaluated in ischemic hind limbs with intramuscular delivery.

RESULTS: Systemic delivery of AAV9-HR-VEGF to mice showed high tropism for muscle with a 3-fold greater preference for cardiac over skeletal muscle. VEGF expression from AAV9-HR-hVEGF was negligible in aerobic myocytes or normally perfused hind limb and displayed 50 –100-fold induction by hypoxia. Concurrent atherosclerosis muted transgene expression. Delivery of AAV9-HR-hVEGF to ischemic hind limbs supported a progressive increase of limb perfusion that reached 87% of the contra lateral limb after 12 weeks and was sustained at 40 weeks. Limps treated with PBS or adenoviral-hVEGF auto-amputated at 12 weeks. Immunohistology of hind limbs at 2, 4, 12, 16 and 30 weeks revealed significantly larger vessels containing smooth muscle actin in limbs treated with AAV9-HR-VEGF compared with an unregulated vector. DiI imaging combined with scanning confocal microscopy revealed a progressive longitudinal (hip to foot) growth of vessels with the appearance of arteries at 16-weeks. HVEGF expression peaked at 12 weeks and declined thereafter, confirming regulation by ischemia.

CONCLUSION: Sustained, ischemia-regulated VEGF expression supports early angiogenesis that evolves into arteriogenesis with the generation of stable vessels, muscle perfusion and limb salvage. Down-regulation of the transgene coincident with therapy provides a safety switch that will allow the clinical translation of this strategy.
Abstract 5146: Pre-diabetes is Associated With Masked Hypertension
Authors: Ishikawa Joji , Schwartz Joseph E , Pickering Thomas G
Published: April 2024

Background:
Masked hypertension, a condition where awake ambulatory blood pressure (BP) is elevated (≥135/85 mmHg) while office BP is within the normal range (<140/90 mmHg), is associated with an increased risk of cardiovascular events; however, mechanisms causing masked hypertension remain unclear. We evaluated whether increased glucose within the non-diabetic range (fasting glucose <126 mg/dl) and/or presence of pre-diabetes (fasting glucose 100 –125 mg/dl) is associated with masked hypertension.

Methods:
We performed office BP measurements (3 readings/visit, at 3 visits), a single 24-hr ambulatory BP (ABP) monitoring and a fasting blood glucose test in 475 healthy, employed volunteers who did not have either office hypertension or diabetes.

Results:
The average age was 44.7±11.1 years (male; 37.5%). The subjects with masked hypertension (N=76) were older (47.5±8.9 vs. 44.2±10.2 years, P=0.009), more likely to be male (53.9 vs. 34.3%, P=0.001) and pre-diabetic (32.9 vs. 11.8 %, P<0.001); they had higher fasting glucose (94±12 vs. 88±11 mg/dl, P<0.001), body mass index (29.0±5.7 vs. 27.1±5.2 kg/m
2
, P=0.004), serum creatinine (0.87±0.15 vs. 0.82±0.15 mg/dl, P=0.009), office SBP (123±8 vs. 112±9 mmHg, P<0.001) and office DBP (81±5 vs. 73±7 mmHg, P<0.001) than those with normal ABP (<135/85 mmHg). In a logistic regression analysis, fasting glucose level was significantly associated with masked hypertension [odds ratio (OR) per 10 mg/dl, 1.35, 95% confidence interval (CI) 1.01–1.80, P=0.044], controlling for the confounding factors. In a parallel analysis, presence of pre-diabetes was also associated with masked hypertension (OR=2.68, 95%CI 1.42–5.09, P=0.002). Prevalence of masked hypertension was 51.2% among those with both pre-hypertension and pre-diabetes, 34.5% among those with pre-hypertension alone, 10.3% among those with pre-diabetes alone, and 3.8% among those without either of them.

Conclusion
: Both pre-diabetes status and fasting glucose (measured on a continuum) in non-diabetic individuals are associated with presence of masked hypertension, suggesting that ambulatory BP monitoring can improve CV risk stratification (over and above office BP) in persons with pre-diabetes in addition to pre-hypertension.
Abstract 5168: Adenosine Formed by CD73 on Treg Inhibits T-cell Mediated Immune Response via NFkB
Authors: Romio Michael , Reinbeck Benjamin , Burghoff Sandra , Schrader Jürgen
Published: April 2024

CD73-derived adenosine (ADO) acts as potent inhibitor of inflammation and we have recently shown that the CD73−/−/ApoE−/− double mutants show enhanced plaque formation with local accumulation of immune cells. Since regulatory T-cells (Treg) have been recently shown to express CD73 as a novel marker enzyme, the present study explored the potential role of CD73-derived adenosine on NFkB activity and cytokine release in CD4+ cells. We asked: does adenosine physiologically formed by Treg inhibit the immune response of the entire Th-cell population? Does the lack of CD73 alter the expression pattern of various enzymes of the ecto-nucleotidase cascade and ATP/adenosine receptors in Tregs? T-cells, isolated from murine spleens by magnetic micro beads, were stimulated with
α
CD3/
α
CD28 which activates the nuclear translocation of transcriptionfactor NF
κ
B, known to be involved in the secretion of pro-inflammatory cytokines such as IL-2. Using EMSA we found activated NF
κ
B in T-cells lacking CD73 to be significantly upregulated (WT: 4.36±0.21, CD73−/−: 6.58±0.75; n=4; p=0.029) which was paralleled by increases in release of the pro-inflammatory cytokines IL-2 and IFN-
γ
(ELISA). Treatment of T-cells with ADO (25
μ
M) caused a massive A2a ADO receptor mediated reduction of IL-2 and IFN-ã release. Similarly, AMP (50
μ
M) decreased the IFN-ã release of WT CD4+ cells. Expression analysis revealed that there were no differences in the enzymes and receptors of the ecto-nucleotidase cascade in WT and CD73−/− Tregs. Upon stimulation with ãCD3/ãCD28, however, mRNA levels of the ATP receptor P2X7 - a ligand-gated ion channel that controls CD4+ activation - was significantly reduced. Expression of the ADO transporter CNT2 was also significantly reduced while the expression of the A2b ADO receptor was increased. In summary, our data show that stimulated CD73-deficient T-cells show a proinflammtory phenotype with increased levels of active NF
κ
B and released proinflammatory cytokines. Thus, CD73-derived ADO is a physiological inhibitor of the immune response relevant to atherogenesis. Downregulation of P2X7 on Tregs may be important to prevent apoptosis and to maintain suppressive function of Tregs by converting extracellular ATP to ADO.
Abstract 5204: Vascular Remodelling in Patients With Psoriatic Arthritis: The Impact of Blood Pressure
Authors: Puato Massimo , Zanardo Marco , Tirrito Carmen , Balbi Giulia , Faggin Elisabetta , Zanon Marta , Ramonda Roberta , Modesti Valentina , Nigro Alessandro Lo , Doria Andrea , Pauletto Paolo
Published: April 2024

Increased cardiovascular mortality have been observed in several immune mediated rheumatic diseases, including psoriatic arthritis (PsA). We evaluated subclinical atherosclerosis in PsA patients according with hypertensive status by studying non invasively structural and functional properties of arteries. We studied 41 consecutive patients with PsA (of whom, 48% hypertensives) attending hospital outpatient clinics. 40 normotensives healthy subjects (NC) and 18 hypertensives (HT-C) served as controls. We evaluated by B-mode ultrasound the carotid intima media thickness (IMT) expressed as mean-IMT (cumulative mean of mean IMT measured in each carotid segment, common, bulb, and internal carotid artery, bilaterally) and as M-MAX (cumulative mean of maximum IMT). Endothelial function was evaluated by post-occlusion flow mediated dilation (FMD) of the brachial artery using ultrasonography. NO-independent vasodilation was evaluated by the response to sublingual glyceril trinitrate (GTN). PsA had a higher mean-IMT compared to NC. Hypertensive PsA displayed higher M-MAX versus both HT-C (p=0.007) and normotensive PsA (p=0.026). FMD was lower in PsA than in NC, whereas there was no difference between hypertensive PsA, normotensive PsA, and HT-C. GTN response was similar in all groups. In conclusion, our data show that subclinical atherosclerosis is enhanced in PsA compared to NC. In PsA, the hypertensive status proved to exert an additional effect on M-MAX, a parameter of advanced pro-atherogenic remodeling. FMD was reduced in PsA irrespective of hypertensives status. Thus, PsA
per se
implies a pro-atherogenic remodeling which is enhanced by the hypertensive status.
Abstract 5216: The Effect of Erythropoietin on Restenosis in Patients With Acute Myocardial Infarction Undergoing PCI
Authors: Ott Ilka , Schulz Stefanie , Mehilli Julinda , Fichtner Stefanie , Hadmamitzky Martin , Ibrahim Tareq , Martinoff Steffan , Dirschinger Josef , Kastrati Adnan , Schömig Albert
Published: April 2024

Background:
In experimental models erythropoietin not only improves myocardial function in acute myocardial infarction but also enhances reendothelialization after vessel wall injury and reduces neointima generation. Aim of this study was to analyze the angiographic restenosis in patients from the randomized, controlled
Regeneration of Vital Myocardium in ST-Segment Elevation Myocardial Infarction by Erythropoietin
(REVIVAL-3) study.

Methods and Results:
Patients with STEMI treated with primary percutaneous coronary intervention (PCI) were randomly assigned to receive epoetin beta (3.33×10
4
U, n=68) or placebo (n=70) immediately, at 24 and 48 hours after PCI. Angiographic follow-up was performed after 6 months. Quantitative coronary angiography at follow-up revealed a binary restenosis rate of 12% in the erythropoietin group and 7% in the placebo group (P=0.298). In segment diameter stenosis was 32±9% in the erythropoietin group versus 26±14% in the placebo group (P=0.046). Accordingly in segment minimal lumen diameter was 2.1±0.7mm after erythropoietin as compared to 2.3±0.6 in the placebo group P=0.070).

Conclusion:
In contrast to experimental findings, erythropoietin use in patients with STEMI treated with primary PCI is associated with a certain risk increase of lumen renarrowing.
Abstract 5223: Dimethylarginine Dimethylaminohydrolase 1Modulates Endothelial Cell Growth and Injury Repair Through No and Akt
Authors: Zhang Ping , Hu Xinli , Xu Xin , Wang Huan , Huo Yuqing , Chen Yingjie , Bache Robert J
Published: April 2024

Dimethylarginine dimethylaminohydrolase-1 (DDAH1) modulates endothelial nitric oxide (NO) production by degrading asymmetric methylarginines that act as endogenous NO synthase (NOS) inhibitors. Since we recently found DDAH1 strongly expressed in vascular endothelial cells, this study examined effects of DDAH1 on endothelial function. Here, by using selective gene silencing of DDAH1 with small interfering RNA (siRNA) and overexpression of DDAH1 in HUVEC, we demonstrate that DDAH1 acts to promote endothelial cell proliferation, migration and tube formation both by the traditional role of degrading asymmetric methylarginines and a novel molecular pathway of increasing Akt activity. Specifically, overexpression of DDAH1 Abstracts From Scientific Sessions 2009 S1073 dose-dependently increased p-Akt
Ser473
, Akt activity and p-eNOS
Ser1177
, while selective gene silencing of DDAH1 with siRNA decreased endothelial p-Akt
Ser473
and p-eNOS
Ser1177
. Importantly, incubation of HUVEC with the NOS inhibitors L-NAME or ADMA, the soluble guanylyl cyclase inhibitor ODQ, or the membrane permeable cGMP analog 8-pCPT-cGMP had no effect on p-Akt
Ser473
, indicating that the increase of p-Akt
Ser473
produced by DDAH1 was NO and cGMP independent. Overexpression of constitutively active Akt rescued HUVEC from DDAH1 knockdown, while the increases of HUVEC tube formation, proliferation and migration after DDAH1 overexpression were abolished by the Akt inhibitor ly294002. Moreover, using endothelial specific DDAH1 gene deficient mice generated in our laboratory, we demonstrated that endothelial spouting from cultured aortic rings and
in vivo
vascular endothelial repair after wire injury were impaired compared to wild type mice. DDAH1 KO also attenuated vascular p-Akt
Ser473
but not total Akt. Together, the data demonstrate that DDAH1 plays an important role in vascular endothelial cell growth and injury repair by regulating both NO production and Akt activity.

This research has received full or partial funding support from the American Heart Association, National Center.
Abstract 5461 In vivo Knockdown of Aortic Nicotinic Acetylcholine Receptor Alpha1 (nAChR
α
1) Diminishes Aortic Atherosclerosis and Valve Calcification
Authors: Zhang Guoqiang , Kernan Kelly A , Thomas Alison , Su Yanyuan , Zheng Yi , LeBoeuf Renee C , Eddy Allison A
Published: April 2024

Urokinase plasminogen activator (uPA) contributes to atherosclerosis, restenosis and vascular remodeling. We have recently identified nAChR
α
1 as a functional cell receptor for uPA in addition to its classic receptor, uPAR. Here, we test the hypothesis that nAChR
α
1 plays a role in the pathogenesis of atherosclerosis. C57BL/6J
ApoE
−/−
mice (male) were initially fed a Western diet for 8 wks. Plasmid DNA encoding scramble RNA (
pScr
) or siRNA (
pSir2
) for nAChR
α
1 was then injected into the mice (n=15) using an aortic hydrodynamic gene transfer protocol. Three mice from each group were sacrificed 7 days after DNA injection to confirm the nAChR
α
1 gene silencing. The rest of the mice continued on the Western diet for an additional 16 wks. Aortas were harvested for paraffin-embedding (aorta root), protein (ascending aorta and aortic arch) and RNA (descending aorta) (n=8). Whole aortas were isolated for oil red staining in 4 mice of each group. The nAChR
α
1 was highly up-regulated in aortic atherosclerotic lesions. Knockdown of the nAChR
α
1 gene was achieved as early as day 7 (76%↓) after DNA injection, and aortic nAChR
α
1 in
pSir2
-treated mice remained lower (78%↓) at 16 wks compared to the
pScr
(n=8,
P
<0.05). The nAChR
α
1 reduction resulted in less severe aortic atheroslcerosis at 16 wks. The ratio of the intimal/medial thickness (I/M ratio) was smaller in the nAChR
α
1-silenced aortas (3.1±1.8 versus 8.4±1.4,
pSir2
versus
pScr
, n=8,
P
<0.05). There were significantly fewer F4/80+ macrophages (60%↓) and
α
SMA+ myofibroblasts (80%↓) in the neointima of the
pSir2
-treated mice compared to the
pScr
(both
P
<0.05). The TGF-
β
1 and
α
SMA mRNA levels were down-regulated by 80% and 72% respectively in the nAChR
α
1-silenced aortas (
pSir2 vs pScr
; n=5, both
P
<0.05). Since nAChR
α
1 is a ligand-gated Ca
2+
channel that may be associated with osteopathic responses in aortic intimal myofibroblasts, aortic calcification was evaluated with the Von Kossa staining. The calcification was most prominent in the aortic valves; a 41% reduction in area stained was observed in the
pSir2
-treated mice (
pSir2 vs pScr
; n=8
P
<0.05). These data suggest that the nAChR
α
1 gene plays a significant role at the artery wall and reducing its expression serves to reduce aortic plaque development and valve calcification.

This research has received full or partial funding support from the American Heart Association, National Center.
Abstract 5509: Association of Pericardial Adipose Tissue With Coronary Atherosclerotic Plaque and Inflammatory Blood Biomarkers
Authors: Schlett Christopher L , Bamberg Fabian , Koenig Wolfgang , Brady Thomas J , Nagurney John T , Hoffmann Udo
Published: April 2024

Background:

Pericardial adipose tissue (PAT) may exert paracrine effects on the pathogenesis of coronary artery disease (CAD). Thus, we assessed the relationship of PAT and different stages of coronary atherosclerosis, and inflammatory biomarkers.

Methods:

We included 355 subjects (mean age: 52±11 years, 60% male) without known CAD who had acute chest pain but no acute coronary syndrome. All subjects underwent contrast-enhanced 64-slice cardiac CT. PAT was defined by any pixel with a CT attenuation of −190 HU to −30 HU within the pericardial sac. Blood samples were collected at the time of CT scan, frozen, and subsequently analyzed for inflammatory biomarkers (E-Selectin, hs-CRP, IL-18, MCP-1, MMP-2, MMP-9, PAI-1, TNF-
α
). We determined the association of PAT to the presence and extent of coronary atherosclerosis, and inflammatory biomarker levels.

Results:

In multivariable analyses, age, gender male, BMI, and current smoking were associated with higher levels of PAT (all p≤0.01). Moreover, the presence of any (
β
: 18.3 cm
3
, 95%-CI: 7.0 –29.6, p=0.002), non-calcified (
β
: 31.5 cm
3
, 95%-CI: 11.3–51.8, p=0.002), and mixed plaque (
β
: 15.6 cm
3
, 95%-CI: 1.7–29.5, p=0.03) was associated with increased PAT independent of age, gender, and current smoking. This association persisted even after adjustment for BMI, except for non-calcified plaque. Hs-CRP, IL-18 and PAI-1 were significantly higher in subjects with increased PAT, independent of the presence of CAD (see Table

; following symbols within the table indicate a significant difference (p<0.0253) for the subgroup comparison: * ‘low PAT/no plaque’ vs. ‘low PAT/yes plaque’; ° ‘high PAT/no plaque’ vs. ‘high PAT/yes plaque’; † ‘low PAT/no plaque’ vs. ‘high PAT/no plaque’; ‡ ‘low PAT/yes plaque’ vs. ‘high PAT/yes plaque’).

Conclusion:

PAT is associated with plaque by CT above and beyond the effect of traditional CVD risk factors. These data may support a central role of PAT in the development of coronary atherosclerosis.

Level of blood biomarkers within the four subgroups as median plus lower and upper quartile
Abstract 5513: Shear Stress Independent Formation of Collaterals in Porcine Coronaries
Authors: van den Wijngaard Jeroen P , Schulten Henny , van Horssen Pepijn , ter Wee Rene D , Siebes Maria , Post Mark J , Spaan Jos A
Published: April 2024

Objectives: To test the hypothesis that intracardiac collateral connections do not result from observable (resolution 40 microns) preexisting vessels and shear stress modulation is not required for collateral formation.

Methods: An LCX ameroid constrictor was placed in 6 pigs and the LCX exposed (sham) in 2. After six weeks the hearts were processed and sliced at 40
μ
m thickness (3000 slices) in a fluorescence imaging cryomicrotome. High resolution images (2000 * 2000 pixels) were captured, yielding a 3D reconstruction (40
μ
m3 voxels) of the vasculature. With custom algorithms, collaterals were detected between (Bridge) and within (Mid) LCX, LAD and RCA.

Results: In shams, collaterals were absent. In stimulated hearts 192 collaterals, median ô111
μ
m (22–1049), were detected, of which 65% endocardial (p= 0.004). Figure 1

shows a typical case (local opacity altered for visibility). Only 64% of all connections were bridging the LCX to LAD or RCA regions. Mid collaterals (n=24), were found in non-LCX regions with no apparent perturbation of shear stress. Mid collaterals (median 76
μ
m, 23–1049) were smaller than bridge (137
μ
m, 22–990, Mann-Whitney U p=0.047).

Conclusion: Since mid collaterals are parallel to normal vasculature, the novel finding of these vessels contra-indicates a dominant role of shear stress in generating and maintaining collaterals. Higher flow rates are to be expected in Bridge versus Mid connections supporting the hypothesis that shear stress plays a role in outward remodeling. Hence, the detection of substantial numbers of collaterals in regions without flow limitation, suggests additional pathways contributing to collateral growth.
Abstract 5592: Role of Afadin in Vascular Endothelial Growth Factor- and Sphingosine 1-Phosphate-Induced Angiogenesis
Authors: Tawa Hideto , Rikitake Yoshiyuki , Takahashi Motonori , Amano Hisayuki , Kinugasa Mitsuo , Nagamatsu Yuichi , Majima Takashi , Ogita Hisakazu , Miyoshi Jun , Takai Yoshimi , Hirata Ken-ichi
Published: April 2024

[Background]
Better understanding of mechanism of angiogenesis and angiogenic signaling of pro-angiogenic factors such as vascular endothelial growth factor (VEGF) and sphingosine 1-phosphate (S1P) will aid in developing therapeutic angiogenesis for ischemic heart disease and peripheral arterial disease. Recently, Rap1 small G protein was identified as a key angiogenic signaling molecule. However, mechanisms involved in the Rap1 regulation of angiogenesis and downstream effectors of Rap1 are not known. Afadin is an adherens junction protein that connects immunoglobulin-like adhesion molecule nectins to the actin cytoskeleton and binds activated Rap1. Recently, we identified that, besides the role in cell-cell adhesion formation in epithelial cells, afadin plays a role in the regulation of directional cell movement of fibroblasts. We therefore attempted to investigate the role of endothelial afadin in angiogenesis and clarify the underlying molecular mechanism.

[Method and Results]
Knockdown of Rap1 or afadin by siRNA inhibited the VEGF- and S1P-induced tubulogenesis, migration and proliferation, and conversely, increased the serum deprivation-induced apoptosis of human umbilical vein endothelial cells (HUVECs). Immunoprecipitation and immunofluorescence studies revealed that Rap1 regulated the interaction between afadin and phosphatidyl-inositol 3-kinase (PI3K) and recruitment of the afadin-PI3K complex to the leading edge, suggesting that Rap1 and afadin regulates the PI3K signaling. Indeed, knockdown of Rap1 or afadin by siRNA inhibited the VEGF- and S1P-induced phosphorylation of Akt. Thus, afadin regulated the PI3K-Akt signaling. To examine the role of afadin in angiogenesis in vivo, we developed endothelial cell-specific afadin-knockout mice using the Cre/loxP system. Majority (96%) of afadin-knockout mice were embryonically lethal, suggesting an essential role of endothelial afadin in embryonic vascular development and angiogenesis. We performed Matrigel plug assay in born adult afadin-knockout mice and observed that VEGF- and S1P-induced angiogenesis was impaired in afadin-knockout mice.

[Conclusion]
These results demonstrate that afadin regulates VEGF- and S1P-induced angiogenesis downstream of Rap1.
Abstract 5623: Early Antithrombotic Therapy Performance Measure in Ischemic Stroke and Clinical Outcomes
Authors: Fonarow Gregg C , Smith Eric E , Reeves Matthew J , Pan Wenqin , Olson DaiWai , Hernandez Adrian F , Peterson Eric D , Schwamm Lee H
Published: April 2024

Background:
Guidelines recommend that patients hospitalized with ischemic stroke should receive antithrombotic therapy within 48 hours of symptom onset to reduce stroke mortality and morbidity. An early antithrombotic process of care measure is being considered by the Center for Medicare and Medicaid Services (CMS). However, it is unknown if this process of care measure is associated with outcomes in routine clinical practice and provides discrimination of the quality of stroke care provided at the hospital level.

Methods:
We analyzed data from 91,184 Medicare fee-for-service beneficiaries enrolled from 625 Get With The Guidelines-Stroke hospitals between April 2003 and December 2006, which were linked to outcome data from CMS. The process measure assessed was the % of patients with ischemic stroke who received antithrombotic (antiplatelet or anticoagulant) therapy by the end of hospital day two in the absence of IV tPA and as long as no contraindications existed. Cox proportional hazards models were used to estimate the unadjusted and adjusted association of this process measure with 30-day mortality.

Results:
Mean age was 79.3 years, 58% were female, and 32% had prior stroke/TIA. The median hospital conformity rate was 93.8% (25
th
-75
th
, 90.3–96.7%) for this measure. The 30-day mortality rate (measured from hospital day 2) in this cohort was 13.1%. The adjusted odds ratio for 30-day mortality with a 10% increase in adherence at the hospital level was 0.91, 95% CI 0.87– 0.96, P=0.0005.

Conclusions:
An early antithrombotic therapy performance measure in eligible ischemic stroke patients is associated with improved 30-day outcomes, discriminates quality of care for ischemic stroke, and may be considered a useful clinical performance measure.

This research has received full or partial funding support from the American Heart Association, National Center.
Abstract 5673: Engagement of the Sphingosine 1 Phosphate Receptor 1 Induces Macrophage Egression From Atherosclerotic Lesions in Apolipoprotein E Deficient Mice
Authors: Larmann Jan , Westermann Anik , Schmitz Martina , Brinkmann Volker , Bremer Christoph , Nofer Jerzy-Roch , Theilmeier Gregor
Published: April 2024

Background:
The regulation of macrophage (MØ) and T-cell content is crucial to atherosclerotic plaque stability. MØ have the potential to rapidly egress from plaques. Sphingosine 1-Phosphate (S1P) - an integral component of high density lipoproteins (HDL) - reduces leukocyte counts at sites of inflammation. We here tested if short-term treatment with the synthetic S1P analogue FTY720 or the selective S1P receptor 1 (S1P1) agonist AUY954 affects MØ and T-cells in plaques of ApoE deficient (ApoE
−/−
) mice.

Methods:
ApoE
−/−
and wild type (WT) mice were fed a 16wk western diet. For 3D in vivo Fluorescence-Mediated-Tomography (FMT), 5×10
6
peritoneal eGFP-MØ labeled with near infrared fluorescent dye (DiR) were given IV 2d prior to 2 or 4d FTY720 (1
μ
g/gBW/d), AUY954 (10
μ
g/gBW twice/d), or vehicle (CTR) treatment. Baseline MØ content was assessed by FMT in a region of interest covering aortic-valve and -arch. On d2 and 4 MØ influx and egress from plaques into the blood were quantified. MØ egress into aortic lymphnodes (LN) was quantified by flow cytometry. T-cell (Thy1.2), MØ (CD68) and lipid content (Oil Red O) were assessed in plaques.

Results:
FMT revealed net MØ recruitment during 2 days in CTR (487±151%, d2 vs. d0, p<0.05, n=9) but in neither treatment group (FTY720: 67±11.9%, AUY954: 103±26.5%, p=n.s., n=9/5). After 4 days MØ related signal was reduced consistent with MØ egress from plaques in both treatment groups (FTY720: 40±9.5%, AUY954: 38±10.5%, d4 vs. d0, p<0.05, n=5) but not in CTR (14922.9%, p<n.s., n=5). CD68- and Oil-Red O-pos lesion area decreased by 33% and 26% (FTY720 vs. CTR, n=8, p<0.05). FTY720 but not AUY954 promoted MØ relocation into LN (FTY720: 7.21.9, AUY954: 1.270.73, CTR: 2.20.7, eGFP-MØ/10
5
cells, FTY720 vs. CTR & AUY954, n=5, p<0.05;). 2d treatment with FTY720 reduced plaque T-cell content (10±2.6 vs. 19±4.4 cells/mm
2
, FTY720 vs. CTR, p<0.05, n=5).

Conclusions:
Both FTY720 and AUY954 rapidly reduce macrophage content in atherosclerotic lesions indicating S1P1 involvement in this process. However, only FTY720 induces MØ relocation into regional LN and reduces plaque T-cell content. Reducing MØ and T-cells in the plaque through engagement of various S1P receptors (FTY720) vs. S1P1 represents a valid strategy for plaque stabilization.
Abstract 5789: Prevention of Peripheral Arterial Disease With Calcium Channel Blockers in Patients With Hypertension: A Meta-analysis of Randomized Trials
Authors: Feringa Harm H , Emani Usha R , Ardestani Afrooz , Shetty Shilpa , Pearson William N
Published: April 2024

Background:
Peripheral arterial disease (PAD) is a manifestation of atherosclerosis and associated with increased morbidity and mortality. Several preclinical studies suggest that calcium channel blockers (CCB) may prevent the development of atherosclerosis in high-risk patients.

Aim
: This study sought to evaluate the effect of CCB treatment for the primary prevention of PAD in patients with hypertension.

Methods
: We searched the published literature (MEDLINE, the Cochrane Central Register of Controlled Trials and EMBASE) for trials published after 1990 reporting the effect of CCB on the development of PAD. Criteria for inclusion included a study duration >6 months, use of a randomized control group not receiving CCB, and availability of outcome data on PAD. We conducted a meta-analysis with random and fixed effects models to combine the results of these studies.

Results:
Of 2,006 potentially relevant studies, a total of 7 studies (65,925 patients) met our inclusion criteria. Patients randomized to the control group received either placebo or active treatment with ACE-inhibitors, beta-blockers or diuretics. In patients receiving CCB, 535 of 24,678 (2.2%) developed PAD versus 1,249 of 39,833 (3.1%) in the control group. CCB reduced the relative risk for PAD development by 25% (95% confidence interval 33% to 17%, p<0.001). In the subset of 6 studies with 64,511 participants taking dihydropyridines vs control, dihydropyridines were associated with a significant risk reduction in PAD of 26% (95% CI 33% to 18%, p<0.001).

Conclusion:
Based on the results of this study, calcium-channel blockers appear to be effective in the prevention of PAD in patients with hypertension.
Abstract 5796: Unique Atrial Fibrillation Subpopulations: Thromboembolic Destination Predicts Recurrence Rates and Survival
Authors: Gosk-Bierska Izabela , McBane Robert , McLeod Thomas G , Wysokinski Waldemar E
Published: April 2024

Aim.
Thromboembolic presentation (stroke, mesenteric or limb ischemia) in non-valvular atrial fibrillation may occur randomly or may identify unique subpopulations of patients with this dysrhythmia. The aim of this study was to determine whether future major adverse cardiovascular event rates (myocardial infarction, stroke, peripheral arterial embolism, death) differ between patients with peripheral embolism (AFPE) compared to those suffering stroke (AFS).

Methods.
An inception cohort (1995–2005) with AFPE (n=216) were compared to randomly selected AFS patients (n=241).

Results.
Patients with AFPE were older than AFS patients (79±10 vs 73±13 years; p<0.001) with a greater percentage of women (71% vs 45%; p<0.001). The most common presentation of AFPE patients was limb ischemia (90%). Except for prior stroke, all CHADS-2 variables were more prevalent in AFPE (Table

). During 3.3±3.6 years of follow up, survival was lower in AFPE compared to either AFS (p<0.001) or age/gender matched controls (p<0.001; Figure

). AFPE patients had lower survival free from recurrent embolization compared to AFS (82% vs 100%; p=0.005). Recurrence free survival for MI and stroke were similar.

Conclusions.
Patients with peripheral embolization represent a unique subset of non-valvular atrial fibrillation with an increased risk of recurrence and high mortality rates compared to those suffering stroke.

Comparison of clinical and echocardiographic characteristics
Abstract 16675: Assessment of Initial U.S. Dabigatran Utilization and Short-Term Persistence
Authors: Stanek Eric J , Agatep Barnabie C , Herrera Vivian , Hawk Gosia S , Schrader Bruce J , Pittman Donald G , Frueh Felix W , Charland Scott L
Published: April 2024

Background:
The U.S. Food and Drug Administration approved dabigatran in October 2010 to reduce the risk of stroke and systemic embolism in patients with non-valvular atrial fibrillation, and it became available in November 2010. The purpose of this analysis is to assess early US dabigatran utilization patterns and short-term persistence.

Methods:
A retrospective analysis of a de-identified, integrated pharmacy-medical claims database is described. Patients with a prescription claim for dabigatran Nov 1-Dec 31, 2010, and with continuous eligibility of ≥12 months prior to and up to 4 months after initial dabigatran claim date were included. Demographic and clinical characteristics of patients, prescriber specialty, and 4-month persistence to dabigatran therapy were assessed. Medical diagnoses were determined by the presence of at least 2 ICD-9 or CPT codes in medical claims on 2 separate dates in the prior 12 months. Persistence at 120 days was calculated allowing a 45-day supply gap. Categorical and continuous variables were analyzed using chi-squared and t-tests, respectively (significance at p<0.05).

Results:
A total of 1,143 patients received dabigatran mainly through retail pharmacy distribution (Table). Of these, 152 (13%) did not have a claims history for atrial fibrillation. Overall, cardiologists were the highest prescribers (62%), however primary care physicians prescribed in 23%. Warfarin therapy was prescribed in 71% of patients during the prior 6 months.

Conclusion:
The 13% of patients with an unclear dabigatran indication appear distinct from those with atrial fibrillation. Overall, 17% do not persist on dabigatran therapy for 120 days, which may adversely affect stroke outcomes. Ongoing review and action on dabigatran patient selection and therapy maintenance to optimize clinical outcomes, as well as real-world comparative effectiveness with optimized warfarin therapy, are needed.
Abstract 16677: Allogeneic Cardiospheres Boost Cardiac Function and Attenuate Adverse Remodeling Without Eliciting Immune Reactions Post-Myocardial Infarction in Immunologically-Mismatched Rat Strains
Authors: Tseliou Eleni , Malliaras Konstantinos , Terrovitis John , Luthringer Daniel , Marban Linda , Marban Eduardo
Published: April 2024

BACKGROUND:
Autologous cardiospheres, 3-dimensional heart-derived multicellular structures that form spontaneously in suspension culture, have been shown to ameliorate remodeling and improve global function after intramyocardial (IM) injection in infarcted pig hearts. However, autologous tissue processing is fraught with considerable expense, delays to treatment and potential complications.The possible role of allogeneic cardiospheres in restoring ischemic heart tissue has not been studied.

OBJECTIVES:
We evaluated the immunologic profile of syngeneic and allogeneic cardiospheres in vitro and their in vivo efficacy after IM injection in a rat model of acute myocardial infarction.

METHODS:
Cardiospheres expanded from male Wistar Kyoto (WKY) rat hearts were injected in the peri-infarct zone of WKY female (syngeneic group, n=18) or Brown Norway female rats (allogeneic group, n=20). Brown Norway female rats (n=12) injected with normal saline served as controls.

RESULTS:
In vitro, cardiospheres expressed only MHC I antigens on their outer layer, while MHC II were only gradually expressed after stimulation with γ-interferon over a course of 7 days. In vivo, cell engraftment was equivalent in the two treated groups 1 week (syngeneic 4.5% vs allogeneic 3%, p>0.05) and 3 weeks after injection (<0.5% in both groups). At the same time points, infarct size was significantly and equally reduced in the two cardiosphere-treated groups compared with the vehicle-treated group by quantitative morphometry. Echocardiography revealed similar functional improvement in both cardiosphere-treated groups at one week (EF syngeneic 45.5±2.9, allogeneic 46.4 ± 1.7 vs control 33.4±1.7, p<0.05 ) and three weeks of follow up (EF syngeneic 51.5±1.9, allogeneic 50.5±2.2 vs control 31.9±0.7, p<0.05). Histopathologic evaluation of the infarct site and the peri-infarct zone, where the allogeneic cardiospheres were injected, showed no signs of rejection.

CONCLUSION:
Allogeneic cardiospheres engraft in the infarcted heart, improve cardiac function and limit infarct size, to the same extent as syngeneic ones, without evident immune reaction. Cardiospheres may serve as an effective off-the-shelf cell product for regeneration after acute myocardial infarction.
Abstract 16742: Determinants of Mortality And Neurological Outcome Following Treatment of Acute Type A Dissections Using Different Cerebral Protection Methods
Authors: Wiedemann Dominik , Dorfmeister Marion , Sodeck Gottfried , Herbst Claudia , Kocher Alfred , Mahr Stephane , Laufer Günther , Ehrlich Marek
Published: April 2024

Objective:
To assess the efficacy and midterm results of different cerebral protection techniques in the treatment of acute type A aortic dissection.

Methods and results:
Between 1993 and 2010, 329 patients(220 male, median age 59)undergoing replacement of the ascending aorta/arch with an open distal anastomosis were analyzed. Either hypothermic circulatory arrest(HCA) alone at 18 Degrees(n=116;35%) or in combination with retrograde cerebral perfusion(RCP) (n=120;36%) or antegrade cerebral perfusion(ACP) at 25 Degrees(n=93;29%) were used. Median circulatory arrest time was 29 minutes(4-145). Overall in-hospital mortality was 19% (63/329); 28%(32 pts) in the HCA group(25pts), vs.17% in the RCP group(21pts) vs. 11%(10pts) in the ACP group(p=0.02). Permanent neurological dysfunction(PND) occurred in 53 patients(16%), with no statistically significantly differences between the 3 groups.(25% in HCA vs.13% in the RCP vs.12% in the ACP group;p=0.13) Transient neurological dysfunctions(TND) were observed in 14 of the other 266 patients(5 %) with a trend towards improved clinical outcome with the use of ACP(10% in HCA group, vs.6 % in the RCP vs.2% in the ACP group;p=0.14). Multivariate analysis revealed that preoperative hemodynamic instability was the only statistically significant independent predictor of in-hospital mortality(odds ratio [OR] 5.24; 95% CI 2.6 - 10.5;p<0.001) and the only significant independent predictor of PND was the use of HCA alone(OR 2.52; 95% CI 1.35- 4.69;p=0.004). Overall long-term survival(mean follow up 65 months) was 76% at one year,68% at 5 years and 56% at 10 years with statistically significant differences between the three groups(p=0.03). ACP patients had 1,5 and 10 year survival rates of 86%, 81% and 74% respectively,in comparison to 74%, 66% and 52% in the RCP group and 68%, 65% and 55% in the HCA alone group. (Figure1)

Conclusion:
The use of ACP confers mostly a survival benefit in the treatment of acute type A aortic dissections.
Abstract 16754: Bone Mineral Density is Abnormal in Children with Protein Losing Enteropathy Following the Fontan Procedure
Authors: Goldberg David J , Dodds Kathryn , Avitabile Catharine M , Brodsky Jill L , Glatz Andrew C , Rand Elizabeth B , Mancilla Edna E , Rychik Jack
Published: April 2024

BACKGROUND:
Protein losing enteropathy (PLE) is a rare but potentially devastating complication of single ventricle physiology following the Fontan operation. Pathological spinal compression fractures have been observed clinically in children with PLE but there are no reports of bone mineral density (BMD), vitamin D [25(OH)D)], or calcium metabolism in this population.

OBJECTIVE:
To describe abnormalities in BMD in a cross section of children with single ventricle physiology complicated by PLE following the Fontan operation.

METHODS:
Cross-sectional sample of children and young adults with a confirmed diagnosis of PLE. Serum levels of 25(OH)D, calcium, total protein, and albumin were recorded from the first outpatient encounter of each subject at The Single Ventricle Survivorship Program at The Children's Hospital of Philadelphia. Corrected calcium (cCa) was calculated from the serum calcium and albumin levels. BMD was measured using dual energy X-ray absorptiometry (DXA), and Z-scores were generated using appropriate software.

RESULTS:
DXA results were available for evaluation in 12 patients (8M/4F). Age at DXA ranged from 7.2 - 25.2 years. The mean Z-score was -1.73 SD for the entire cohort, 42% below -2 SDs. Serum 25(OH)D levels were abnormal in 58%. Mean serum calcium level was 8.6 ± 0.6 mg/dL, mean serum total protein was 5.1 ± 1.1 g/dL, mean serum albumin was 2.9 ± 0.8 g/dL, and mean cCa was 9.4 ± 0.4 g/dL. Of those on chronic corticosteroid therapy, the mean DXA Z-score was -3.15 SD, 67% below -2 SDs. There was a positive correlation between cCa and DXA Z-score and a negative correlation between total protein and DXA Z-score. Patients on corticosteroid therapy had a significantly lower DXA Z-score than those not on corticosteroids (-3.15 vs. -0.31, p=0.02).

CONCLUSION:
Children with PLE are at risk for abnormal BMD compared to age- and sex-matched controls. There are a number of factors that may contribute to abnormal BMD including corticosteroid use, chronic illness, and protein loss through the enteric mucosa. In this cohort, corticosteroid therapy, a marker of chronic disease, appeared to be associated with decreased BMD. Routine bone health screening is warranted for children with PLE, particularly those receiving corticosteroid therapy.
Abstract 16772: Quality of Life in Patients After LAD Revascularization by MIDCAB or by PCI
Authors: Martin Breuer , Georgi Björn , Sandhaus Tim , Ferrari Michael , Doenst Thorsten
Published: April 2024

Purpose:
Outcome comparisons of minimally invasive direct coronary artery bypass (MIDCAB) and percutaneous coronary intervention (PCI) for LAD-revascularization demonstrated equal survival. We assessed quality of life (QOL) in propensity matched MIDCAB and PCI patients where revascularization of the LAD was required.

Methods:
We used the SF-36 Health Survey to assess post-treatment health related QOL (HRQOL) in 330 patients (172 MIDCAB, 158 PCI). MIDCAB patients were all patients having undergone surgery between 11/1999 and 12/2006 in our department. The PCI patients were selected by propensity matching out of 1186 patients having undergone PCI of the LAD in this time period. HRQOL analysis consisted of 8 subscales including general health perception, and physical and emotional role functioning. We further subdivided the patients into two groups (less than 80 years of age and 80 years or older)

Results:
Follow up was up to 84 months, the average follow-up was 38 months. There was no significant difference in survival at latest follow-up ( 94% versus 92% in PCI patients, n.s.). Analyzing all patients, HRQOL was better in MIDCAB patients with significant improvements in general health perception (p<0.05). However, subdividing the groups by age showed continuous improvement of HRQOL by patients younger than 80 years but inferior HRQOL in patients older than 80 years of age. In the older patients specifically physical and emotional role function was improved (p<0.05).

Conclusions:
MIDCAB revascularization of the LAD results in improved HRQOL compared to PCI only in patients under the age of 80 years. Since survival is equal HRQOL aspects maybe helpful in guiding treatment selection for patients with isolated LAD-stenosis.
Abstract 16791: Elevated Post-Prandial Triglyceride Concentrations Are Associated with an Unfavorable Cardiometabolic Risk Factor Profile in Patients with Coronary Artery Disease
Authors: Werner Christian , Groenewold Stephanie , Fritsch Marco , Filmer Anja , Böhm Michael , Laufs Ulrich
Published: April 2024

Background:
The role of the post-prandial triglyceride (ppTG) metabolism for cardiovascular risk has not been studied prospectively.

Study design:
The cohort of this prospective study consisted of N=514 consecutive patients with documented stable coronary artery disease (CAD) (83% male, 17% female, age 66±0.4 years), who were subjected to a novel sequential oral triglyceride and glucose tolerance test (OTT / OGT). The test protocol (75g cream fat given 3 hours prior to 75g glucose) was previously characterized and validated.

Results:
Patients with normal glucose tolerance (N=126) had the lowest fasting (fTG) (108±4mg/dl) and ppTG (195±7mg/dl) concentrations, whereas patients with pathologic glucose tolerance (n=150; fTG 167±10mg/dl vs. ppTG 293±14mg/dl) and patients with diabetes (n=238; fTG 175±11mg/dl vs. ppTG 297±12mg/dl) were characterized by elevated fTG and ppTG. The mean relative TG increase (% vs. baseline) was comparable between subgroups. However, patients showed a large inter-individual variability with respect to ppTG metabolism within each group. Patients in the upper compared to the lower ppTG tertile were younger and had a more severe cardiovascular risk profile (e.g. more active smokers, higher blood pressure, higher body mass index and waist-to-hip-ratio). There were no differences in the angiographic extent of CAD, heart rate or medication. Patients in the upper ppTG tertile had higher concentrations of fasting glucose and insulin, HbA1c, total and LDL cholesterol, whereas HDL was lower. Markers of ppTG metabolism including area under the curve and maximal TG increase per hour were significantly elevated in these patients (p<0.01, Chi
2
or ANOVA where indicated). Underscoring these findings, 53% of patients in ppTG tertile 3 vs. 38% in tertile 1 had diabetes and 76% vs. 32% in tertile 1 met the WHO definition criteria of the metabolic syndrome.

Conclusions:
Elevated ppTG concentrations characterize CAD patients with a high burden of cardiometabolic risk factors. Interestingly, patients differed strongly in the individual ppTG response. Multivariable analyses of cardiovascular events after the 12-months follow-up show that assessment of ppTG metabolism improves risk stratification in CAD patients.
Abstract 16792: KLF6 Modulates Recruitment and Polarization of Inflammatory Cells Through Cardiomyocytes in Initiation of Cardiac Fibrosis
Authors: Sawaki Daigo , Suzuki Toru , Aizawa Kenichi , Matsumura Takayoshi , Munemasa Yoshiko , Ishida Junichi , Fridman Scott L , Nagai Ryozo
Published: April 2024

Introduction:
Previous studies suggest that Kråppel-like factors (KLFs) play important roles in the cardiovasculature. We previously identified and cloned KLF6, whose homozygous mutation in mice results in embryonic lethality with markedly reduced yolk sac vascularization suggesting a role in cardiovascular development. However, it has yet to be addressed how KLF6 contributes to cardiovascular disease.

Methods & Results:
Preliminary investigation showed significantly diminished levels of cardiac fibrosis in response to continuous angiotensin II (Ang II) infusion (fibrotic area; wild-type: klf6
+/-
mice=5.24%:1.6%) in Klf6
+/-
mice. Immunohistochemical analysis revealed remarkably reduced infiltration of Mac3 positive cells in klf6
+/-
mice as soon as 1 to 5 days after initiation of Ang II stimulation and induction of KLF6 in the nucleus of cardiomyocytes. We further investigated infiltrating cells by flow cytometric analysis which revealed marked difference in polarization of macrophages infiltrating cardiac tissue (M1: M2= wild-type mice, 21: 64%, klf6
+/-
mice, 44: 42%). Microarray analysis confirmed reduced expression levels of thrombospondins in the stimulated klf6
+/-
mouse heart (thrombospondin-1 38.9%, thrombospondin-4 27.7%). These biologically active substances subsequently recruited monocytes and modulated their polarization, which in turn shifted activation of residual cardiac fibroblasts or transformation into myofibroblasts thus participating in the early stage of Ang II induced cardiac fibrosis.

Conclusion:
KLF6 plays a functional role in the initiation of cardiac fibrosis in response to pathological stimuli mainly through modulation of recruitment and polarization of infiltrating monocytic cells by biologically active molecules like thrombospondins.