bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 5431: Activation of Myocardin Function by an Ubiquitin E3 Ligase, EDD
Authors: Hu Guoqing , Wang Xiaobo , Saunders Darren , Henderson Michelle , Russell Amanda , Herring Brian , Zhou Jiliang 
Published: April 2024

Fully differentiated mature smooth muscle cells (SMCs) are characterized by the presence of a unique repertoire of contractile proteins. The expression of these proteins is markedly attenuated during the de-differentiation of smooth muscle that occurs under many pathological conditions. SRF is a transcription factor that plays a central role in the expression of these smooth muscle-specific genes through physical association with various cell-restricted or signal dependent accessory factors. Of the SRF-associated proteins identified, myocardin is the most potent for stimulating expression of smooth muscle-specific genes. Although several proteins have been identified that can modify myocardin function, the mechanism of how myocardin regulation is still poorly understood. To identify additional myocardin associated proteins that are required for myocardin-dependent activation smooth muscle-specific genes we performed a yeast 2-hybrid screen of mouse embryo cDNA library using myocardin as bait. From this screen we identified a HECT domain containing protein EDD as a myocardin binding protein. Previous studies have shown that HECT domain containing proteins are ubiquitin E3 ligases that play an important role in protein degradation. EDD has however, also been shown to regulate transcription independent of its E3 ligase activity. In the current study we demonstrated that the amino-terminal domain of myocardin bound to the HECT domain of EDD. We also show that EDD specifically enhanced trans-activation of smooth muscle-specific promoters by myocardin. Furthermore, EDD significantly augmented myocardin’s ability to induce expression of endogenous SMC marker genes, and these effects of EDD were independent on its E3 ligase function. Conversely, depletion of endogenous EDD in fibroblast cells by RNAi attenuated myocardin-induced smooth muscle-specific gene expression, and EDD knockdown in smooth muscle cells resulted in down-regulation of smooth muscle-specific genes. Ongoing experiments are investigating the mechanism by which EDD activates myocardin function. This study reveals an unexpected role for the ubiquitin E3 ligase EDD as an activator of smooth muscle differentiation through its ability to potentiate myocardin function.
This research has received full or partial funding support from the American Heart Association, National Center.
Abstract 5488: Comparison of Primary With Fibrinolytic-based Facilitated PCI and Comparison of Fibrinolytic-based PCI With Fibrinolysis Followed by Standard Care for STEMI: Updated Quantitative Review of Randomized Trials
Authors: Thiele Holger , Eitel Ingo , Desch Steffen , Schuler Gerhard 
Published: April 2024

Background: Facilitated PCI for STEMI is defined as the use of pharmacological substances before a planned immediate intervention (< 3h), to improve coronary patency. We undertook an updated meta-analysis of randomized controlled trials to compare

fibrinolytic-based facilitated and primary PCI and

fibrinolytic-based facilitated PCI and fibrinolysis followed by standard care.

Methods: We identified 12 trials of patients with STEMI assigned to

facilitated (n=2498) or primary (n=2488) PCI and

8 trials with comparison facilitated PCI (n=969) vs fibrinolysis with standard care (n=955).

Outcomes of death, stroke, non-fatal reinfarction, urgent TVR, and major bleeding were assessed.
Results: The facilitated approach resulted in a greater than two-fold increase in the number of patients with initial TIMI-3-flow compared with primary PCI approach (p<0.001); however, final rates did not differ (p=n.s.). Significantly more patients assigned to facilitated PCI than those assigned to primary PCI died (5.3% vs 3.9%; RR 1.36; 95% CI 1.04; 1.77; p=0.02), had higher non-fatal reinfarction rates (3.3% vs 1.7%; RR 1.99; 95% CI 1.36; 2.92; p<0.001), higher urgent TVR rates (3.3% vs 1.3%; RR 2.48; 95% CI 1.57; 3.97; p<0.001), and higher stroke rates (1.3% vs 0.5%; RR 2.76; 95% CI 1.38; 5.64; p=0.002). Facilitated PCI was also associated with higher rates of major bleeding (6.4% vs 3.9%; RR 1.62 95% CI 1.26; 2.09; p<0.001). In the 2nd comparison, facilitated PCI did not result in a mortality difference (5.1% vs 5.7%; RR 0.89; 95% CI 0.60; 1.33; p=0.63) and no difference in non-fatal reinfarction (4.2% vs 5.2%; RR 0.81; 95% CI 0.52; 1.25; p=0.37). However, there was a difference in non-fatal reinfarction in the stenting era (4.0% vs 7.0%; RR 0.57; 95% CI 0.34; 0.95; p=0.03). There was no difference in major bleeding and stroke in the comparison of facilitated PCI vs fibrinolysis with standard care.

Conclusions:
Fibrinolytic-based facilitated PCI has substantial harm in comparison to primary PCI. In comparison to fibrinolysis with standard care facilitated PCI leads to a small reduction in reinfarction but has no beneficial effect on mortality. A strategy of primary PCI should therefore be implemented for all STEMI patients.
Abstract 5504: Transient Decrease in Circulating Dendritic Cell Precursors After Acute Stroke - Potential Recruitment Into the Brain
Authors: Yilmaz Atilla , Fuchs Tanja , Dietel Barbara , Altendorf Regina , Cicha Iwona , Stumpf Christian , Schellinger Peter , Blümcke Ingmar , Schwab Stefan , Daniel Werner G. , Garlichs Christoph D. , Kollmar Rainer 
Published: April 2024

Background: The role of dendritic cells (DC) as potent mediators of inflammation has not been sufficiently investigated in stroke.
Methods: Circulating myeloid (mDCP), plasmacytoid (pDCP), and total DCP (tDCP) were flow cytometrically analyzed in

healthy controls (n=29), patients with

asymptomatic A. carotis interna stenosis (ACI-S, n=46),

transient ischemic attack (TIA, n=39),

acute ischemic stroke (AIS, n=73), and

acute hemorrhagic stroke (AHS, n=31).

The National Institutes of Health Stroke Scale (NIHSS) and the infarction size in CT scan were evaluated after stroke. In a patient subgroup, postmortem immunohistochemical brain analyses were performed to detect mDC (CD209), pDC (CD123), T cells (CD3), and HLA-DR.
Results: In AIS and AHS, circulating mDCP, pDCP, and tDCP (each p<0.005) were significantly reduced. A significant inverse correlation was found between the NIHSS and circulating DCP (p<0.02), as well as between hsCRP and circulating DCP (p<0.001). Patients with large stroke size in CT scan had significantly lower mDCP, pDCP, and tDCP (each p<0.01) than those with smaller stroke. Follow-up analysis showed a significant recovery of circulating DCP in the first days after stroke. In the infarcted brain, a dense infiltration of mDC colocalized with T cells, single pDC, and high HLA-DR expression were observed.
Conclusions: Acute stroke leads to a decrease in circulating DCP. Potentially, circulating DCP are recruited from the blood into the infarcted brain, and probably trigger cerebral immune reactions there.
Abstract 5536: Strain-dependent Susceptibility for Hypertension in Mice Resides in the Natural Killer Gene Complex
Authors: Taherzadeh Zhila , VanBavel Ed , de Vos Judith , Matlung Hanke , van Montfrans Gert , Brewster Lizzy , Seghers Leonard , Quax Paul H.A. , Bakker Erik N.T.P. 
Published: April 2024

Hypertension is accompanied by vascular remodeling and inflammation. We tested the hypothesis that the sensitivity to develop hypertension and vascular remodeling depends on the immunological background.
Methods: Blood pressure, vascular remodeling and expression of inflammatory cytokines were determined in mice with a genetically determined difference in immune background: C57BL/6 mice and BALB/c mice; and additionally BALB.B6-Cmv1r mice, a congenic strain where the natural killer (NK) gene complex of C57BL/6 mice is introduced in BALB/c background. Mice were treated for 4 weeks with L-NAME (NG-nitro-L-arginine methyl ester). Systemic and local inflammatory responses were quantified in spleen and mesenteric arteries respectively.

Results: L-NAME treatment induced a significant increase in mean arterial blood pressure in C57BL/6 mice, from 101±4 mmHg (n=7) to 154±5 mmHg (n=5, P<0.001). This was accompanied by a highly significant increase in vessel wall-to-lumen ratio, from 0.104±0.002 (n= 6) to 0.126±0.003 in L-NAME treated mice (n=6, P<0.001). In BALB/c mice the blood pressure (117±3 mmHg for untreated vs. 126±6 mmHg for L-NAME treated mice) and wall to lumen ratio (0.104±0.003 in untreated vs. 0.113±0.003 in L-NAME treated mice) were not altered after 4 weeks of L-NAME treatment. The congenic BALB.B6-Cmv1r mice showed a similar sensitivity to develop hypertension and vascular remodeling as C57Bl/6 mice: blood pressure increased from 111±11 mmHg to 155±7 mmHg (P<0.01). The wall to lumen ratio increased from 0.107±0.003 to 0.125±0.008 (P<0.05). Expression of inflammatory markers such as IFN
γ
, IL1, IL10 and CD68 (macrophages) was similar in C57BL/6 and BALB/c, with exception of CD11b, which increased in vessels of C57BL/6 mice only. Confocal microscopy showed that CD11b co-localized with NK1.1, and reflected the recruitment of natural killer cells in C57BL/6 mice.

Conclusions: These data show that vascular remodeling and hypertension are strain dependent. We have identified the NK gene complex as important determinant in the genetically determined sensitivity to develop hypertension in mice.
Abstract 5785: A Novel Function of Regulator of G-protein Signaling 2: Stabilization of Signal Transducer and Activator of Transcription 3 Phosphorylation
Authors: Yue Hong , Desnoyer Russell , Li Wei , Karnik Sadashiva 
Published: April 2024

Background: Regulator of G protein signaling proteins (RGS) are known to regulate G protein coupled receptor (GPCR) signaling at the plasma membrane through their GTPase-activating protein activity. We have previously identified RGS2 as a downstream target of unphosphorylated Signal Transducer and Activator of Transcription 3 (U-STAT3). U-STAT3 mediated Angiotensin II (AngII) type 1 receptor (AT1R), a GPCR, induced cardiac hypertrophy but the mechanism is unclear.
Hypothesis: RGS2 is the mediator of U-STAT3 regulated Ang II/AT1R signaling.
Methods and Results: In AT1R overexpressing HEK293 cells and HL-1 cells (a mouse cardiomyocyte cell line), Ang II stimulation induced RGS2 expression in parallel with U-STAT3. The localization, distribution and expression of RGS2 were changed upon Ang II stimulation. RGS2 was mainly localized in the nucleus, which was also confirmed in human vascular smooth muscle cells. Gene transfection of RGS2 dose-dependently increased phosphorylated STAT3 (P-STAT3), and maintained a high level of P-STAT3 up to 24 hrs. Co-immunoprecipitation assay identified a complex formed between RGS2 and P-STAT3, suggesting that RGS2 plays a role in maintaining P-STAT3 stability. Treatment with AT1R antagonist, losartan, decreased RGS2/P-STAT3 interaction, suggesting that this interaction is dependent on AT1R activation. On the other hand, in cells overexpressing AT1R and Y705F mutant STAT3, which cannot be phosphorylated on residue 705 (U-STAT3), chromatin immunoprecipitation assay demonstrated that Y705F mutant STAT3 binds to a consensus GAS motif located in the RGS2 promoter, providing direct evidence that RGS2 is a downstream target of U-STAT3. siRNA-mediated gene knockdown of STAT3 consistently caused decreased RGS2 expression in AT1R overexpressing cells.
Conclusions: Chronic activation of AT1R induced U-STAT3 overexpression and subsequently induced overexpression of RGS2, which interacted with and stabilized P-STAT3. P-STAT3 further increases the transcription of stat3 gene, forming a positive feedback loop. We reported for the first time a novel function of RGS2, stabilization of P-STAT3, in mediating U-STAT3 regulated Ang II/AT1R signaling. Further studies are now underway utilizing engineered mice model.
This research has received full or partial funding support from the American Heart Association, Great Rivers Affiliate (Delaware, Kentucky, Ohio, Pennsylvania & West Virginia).
Abstract 5804: Embryonic Stem Cell-Derived Endothelial Cells Engraft Into the Ischemic Hindlimb and Restore Perfusion
Authors: Huang Ngan F , Niiyama Hiroshi , Peter Christoph , De Abhiiit , Natkunam Yasodha , Fleissner Felix , Li Zongjin , Rollins Mark D , Wu Joseph C , Gambhir Sanjiv S , Cooke John P 
Published: April 2024

Peripheral arterial disease (PAD), which affects about 10 million Americans, is typically due to atherosclerotic arterial occlusive disease of the limbs and is associated with dysfunctional vascular endothelium. Cell-based therapy using endothelial cells derived from embryonic stem cells (ESC-ECs) is a promising approach for regenerating the endothelium. However, studies that characterize the localization and engraftment of embryonic stem cells (ESCs) and ESC-ECs into the ischemic limb are limited. In this study, we examined the survival, localization, and functional effects of exogenously administered ESC-ECs for repair of hindlimb ischemia, which is a murine model of PAD. Murine ESCs or ESC-ECs were stably transduced with a construct for bioluminescence imaging (BLI) and fluorescent detection. In a syngenic murine model of hindlimb ischemia, ESCs or ESC-ECs were delivered by intramuscular (IM), intra-femoral artery (IA), or intra-femoral vein (IV) injections (n=5 in each group). For 2 weeks, cell survival and localization were tracked by BLI and confirmed by gene expression, and functional improvement was assessed by laser Doppler blood perfusion. BLI results showed ESC persisting in the ischemic limb after IM or IA, but not after IV administration. Regardless of the route of administration, ESCs were detected outside of the hindlimb circulation in the spleen or lungs. ESCs did not improve limb perfusion, and induced teratomas. In contrast, ESC-ECs localized to the ischemic limb in all cases as assessed by BLI. Immunohistochemical studies confirmed the engraftment of ESC-ECs into the limb vasculature after 2 weeks. Intriguingly, ESC-ECs were not detected in the spleen or lungs after 2 weeks, even after IV administration. Furthermore, ESC-ECs improved limb perfusion. In comparison to parental ESCs, ESC-ECs preferentially localized in the ischemic hindlimb, even after IV delivery. ESC-EC localization was temporally related to the improvement in hindlimb perfusion, which was associated with engraftment of ESC-ECs into the ischemic microvasculature.
This research has received full or partial funding support from the American Heart Association, Western States Affiliate (California, Nevada & Utah).
Abstract 16986: A New Cardiac Muscle Post-Translational Modification: Detection of Protein Arginine Deiminases and Protein Citrullination in Heart Tissue
Authors: Fert-Bober Justyna , Uhrigshardt Helge , Halushka Marc , Andrade Felipe , Giles Jon , Bathon Joan , Van Eyk Jennifer 
Published: April 2024

ABSTRACT:
Protein citrullination originates from enzymatic deimination of amino acid arginine in a protein and is involved in various biological processes during health and disease. However, it has not been investigated in heart. Therefore, our goal was to identify novel substrates for peptidylarginine deiminase (PAD), enzyme responsible for this modification and associate it with changes in protein citrullination during heart failure (HF).

METHODS:
Proteomics methods were used to identify citrullinated proteins and the site of modification in control mouse and control vs. HF human heart tissue (5 per group). The characterization of deiminated proteins was facilitated by modification of peptide-bound citrullineted Arg residues with antipyrine and butanedione, allowing for the unambiguous identification by mass spectrometry. Verification of citrullinated proteins was carried out by 2DE (modification causes pI shift) and western blot with anti-citrullinated antibody. PAD enzyme expression was determined using RT-PCR reaction and immunohistochemistry.

RESULTS:
We detected up to 20 proteins markedly modified by PAD in vitro and in vivo in mouse heart including myofilament and mitochondria proteins. Similar proteins were identified to be citrullinated in control human heart tissue including tropomyosin (R220), myosin heavy and light chain (R1176, 1434 and 1479) and myosin binding protein C (R696). Importantly, we showed a specific residues deimination associated with the HF, e.g. tropomyosin from HF samples has 3 different citrullinated sites (R 125, 133, 238) compared to control. The PAD enzyme is present in mouse isolated cardiomyocytes and cardiac fibroblast (predominantly PAD2 vs. PAD2/PAD4, respectively). Immunohistochemistry of control human hearts had positive PADs staining in cardiomyocytes.

CONCLUSIONS:
Our study demonstrates, for the first time that protein citrullination is a viable post-translational modification in heart tissue and corresponds to the cardiac expression of PAD. Furthermore we showed that citrullinatated residues are altered HF suggesting that this novel PTM could have a regulatory role.
Abstract 17551: The Impact of Ordering Provider Specialty on Appropriateness Classification
Authors: Carryer Damita J , Askew John W , Hodge David O , Miller Todd D , Gibbons Raymond J 
Published: April 2024

The Impact of Ordering Provider Specialty on Appropriateness Classification In prior studies, our laboratory applied the ACCF/ASNC AUC for SPECT MPI to several cohorts of patients in Mayo Clinic Rochester. The purpose of this study was to re-examine our data to test the hypothesis that the specialty of the ordering provider affected the appropriateness.
We compared the ordering providers specialty with appropriateness determined in prior studies. The study group consisted of 281 patients who underwent stress SPECT MPI studies in May 2005 and 282 patients in October 2006 (before any quality improvement initiatives were begun at Mayo). Appropriateness categories were assigned by 2 experienced nurses from the CV Research Studies Unit not affiliated with the nuclear laboratory. Patients who were studied for pre-operative evaluation (n = 20) or post myocardial infarction (n = 7) were excluded because they comprised less than 5 percent of the patients. Pre-operative testing in our institution is done primarily by stress echocardiography. The final study group consisted of 526 patients.
Excluding the 64 unclassified patients, AUE categories were appropriate (75 %), uncertain (13 %), inappropriate (12%). Results by ordering provider are shown in the figure, where CV = Cardiovascular MDs, ED = Emergency Department MDs, IM = General and Internal Medicine MDs, RN = Registered Nurses, and SP = other Specialty MDs. There was no significant association (p = 0.19); Chi-square analysis between appropriateness and specialty groups.
In a clinical practice where pre-operative testing using SPECT is infrequent, the rate of inappropriate studies was similar for all ordering providers. Quality improvement efforts in similar practices will likely require a broad focus on all ordering physicians/nurses.
MicroRNA Regulation of the Pathological Angiogenic Response
Authors: Cheresh David 
Published: April 2024

During development, pro- and anti-angiogenic factors orchestrate a complex, dynamic process to allow initial sprouting and invasion, subsequent pruning and remodeling, and finally maturation and survival of blood vessels. However, in pathological conditions such as cancer, diabetic retinopathy, macular degeneration or inflammation the same angiogenic signaling pathways are misregulated and exploited, typically resulting in poorly organized vessels with leaky and tortuous properties. In the last decade, a new class of small non-coding RNA molecules termed microRNAs (miRs) have emerged as key regulators of several cellular processes including angiogenesis.
We recently profiled microRNAs that were regulated upon growth factor activation of human endothelial cells and identified miR-132 as a regulator of pathological neovascularization. We found that miR-132 was highly expressed in activated blood vessels, but was undetectable in normal endothelium. Ectopic expression of miR-132 in endothelial cells increased their proliferation and tube-forming capacity, whereas intraocular injection of an antagomir targeting miR-132, reduced postnatal retinal vascular development in mice. We identified p120RasGAP as a miR-132 target and showed that miR-132 and p120RasGAP are expressed reciprocally during pathological angiogenesis. Our observations in several in vitro and in vivo models suggest that miR-132 acts as an angiogenic switch by suppressing endothelial p120RasGAP expression, leading to Ras activation and the induction of neovascularization, whereas the application of anti–miR-132 inhibits neovascularization by maintaining vessels in the resting state.
In contrast to miR-132, a highly upregulated miR during angiogenesis, we have identified miR-34 as one of the most downregulated microRNAs in response to growth factors. Conversely, miR-34a is highly upregulated during endothelial stress responses. Our observations suggest that miR-34a functions as an ‘apoptomiR’ mediating endothelial cell death during stress by regulating the Notch pathway. Taken together, our data suggests a broad regulatory network of pro and anti-angiogenic microRNAs that target defined pathways to co-ordinate vascular responses in development and disease.
Abstract 9443: Long-Term Clinical Outcomes of Infarct-Related Artery versus Multivessel Revascularization in Acute ST-Segment Elevation Myocardial Infarction with Multivessel Disease: An Analysis from Korea Acute Myocardial Infarction Registry
Authors: Lee Han Cheol , Choi Jin Hee , Kim Bo Won , Lee Hye Won , Yang Mi Jin , An Sung Gyu , Oh Jun-Hyok , Choi Junghyun , Cha Kwang Soo , Hong Taek Jong 
Published: April 2024

Many STEMI patients have multivessel disease. Guidelines recommend multiple revascularization only in cardiogenic shock. We compared clinical outcomes of multivessel revascularization with infarct-related artery(IRA) revascularization in ST-segment myocardial infarction(STEMI) patients.
The 1,283 STEMI patients with multivessel disease[975 in IRA group(group I), 308 in multivessel group(group II)] received primary PCI and were analyzed from a nationwide Korea Acute Myocardial Infarction Registry. Primary outcomes were in-hospital mortality and 12-month major adverse cardiac events(MACE, defined as death, myocardial infarction, and repeated revascularization). Secondary outcomes were in-hospital complications and components of the MACE. There were more hemodynamic unstable patients in group I. In-hospital mortality was higher in group I with marginal significance(6.6% vs. 3.6%, p = 0.051). 12-month MACE occurred in 84(14.6%) in group I, 20(11.5%) in group II(p = 0.294). Each components of MACE were occurred similarly in two groups but non-target vessel revascularization(6.4% vs. 1.7%, p = 0.012). Adjusting for confounding factors, multivessel revascularization did not reduce in-hospital mortality(OR 0.935, 95% CI 0.403-2.167, p=0.875). It was same for 12-month MACE(OR 0.953, 95% CI 0.406-2.233, p = 0.911) and secondary outcomes. But multivessel revascularization reduced non-target vessel revascularization(OR 0.279, 95% CI 0.085-0.922, p = 0.036).
There was no significant difference of clinical outcomes except for low risk of non-target vessel revascularization in multivessel revascularization group. Therefore, results of our study support current guidelines.