bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 54: Syndesome-Based Dressings for Enhanced Wound Healing in Diabetic Ulcers
Authors: Das Subhamoy , Singh Gunjan , Monteforte Anthony , Martinez Matthew , Wright Catherine , Martin Patricia , Dunn Andrew , Baker Aaron 
Published: July 2021

Incidence of chronic non-healing wounds has significantly increased over the last decade due to a rising epidemic in type-II diabetes and peripheral arterial disease (PAD). Previous research has attempted to use growth factor proteins or genes to enhance the healing of cutaneous wounds but have achieved only limited success in healing chronic wounds in the long-term. Our previous work has demonstrated a significant reduction of syndecan-4 protein due to long-term diabetic condition and co-delivery of syndesomes (syndecan-4 proteoliposomes) with FGF-2 enhanced angiogenesis. In this study, we tested the efficacy of a novel wound dressing that delivered FGF-2 with syndesomes in ob/ob mice.
To recapitulate the human disease state, we used ob/ob mice and fed them a high fat diet for 15 weeks. We utilized a splinted, excisional wound model and implanted 2% alginate disks containing treatments into the wound, which were fabricated using a custom made high throughput mold. We monitored the perfusion of the wounds over time using laser speckle imaging.
At day 14, wounds treated with syndesomes (S4PL) and FGF-2 healed the wound significantly more than all other groups (A, B). Histological analysis demonstrated increased re-epithelialization of the wounds treated with S4PL with FGF-2 (C, D). Laser speckle imaging of the wound showed increased perfusion in the S4PL with FGF-2 treated group. Furthermore, immunostaining for the M1 macrophage marker (CD86) showed significantly reduced inflammatory macrophages in both S4PL+FGF-2 and S4PL groups (E, F). Staining for an M2 macrophage marker (CD163) revealed enhanced levels when the syndesomes were delivered, compared to control and FGF-2 groups (G, H).
Taken together, our studies support that syndesomes significantly enhance FGF-2 activity in wound healing in diabetic mice. Thus, syndesome-containing wound dressings may be useful in treating chronic wounds and restoring growth factor activity in diseased states.
Abstract 599: Lighting Up P2Y12 Oligomers: Insights into the Role of Oligomerization in P2Y12 Function
Authors: Khan Aasma , Caplan Jeffrey L , Woulfe Donna 
Published: July 2021

Introduction:
Little is known about the role of P2Y12 oligomerization in receptor function and whether P2Y12 receptor mutations associated with human bleeding disorders may be explained by alterations in oligomerization.

Objectives:

1) To determine whether P2Y12 homo- and hetero-oligomers are constitutive or dynamically regulated.
2) To explore whether P2Y12 mutants R256Q and R265W (previously detected in patients with abnormal bleeding, but with unaltered ADP binding) have different oligomerization affinities or kinetics and determine whether differences in P2Y12 oligomerization explain the functional defects.

Methods:
We employed a Venus-based Bimolecular Fluorescence Complementation (BiFC) approach in HEK293T cells transiently co-expressing P2Y12 or its mutant forms (R256Q or R265W) tagged with either the N-terminal (P2Y12-VN) or C terminal fragment (P2Y12-VC) of Venus, to characterize their homomeric interactions, in live cells using confocal microscopy and quantitative flow cytometry assays.

Results:
Agonist-independent formation of P2Y12 receptor homo-oligomers were detected on cell membranes. Time lapse imaging showed movement of P2Y12 receptor pairs from the endoplamic reticulum and Golgi network to the plasma membrane, suggesting that they are constitutive and required for export. Co-expression of P2Y12-VN with increasing amounts of P2Y12-VC demonstrated a dose-dependent increase in the fluorescence intensity of Venus, and reached saturation at a ratio of 1:3. Interestingly, the fluorescence intensities of homomeric P2Y12-R256Q-VN and R256Q-VC and, separately, P2Y12-R265W-VN and P2Y12R265W-VC were almost 4 times stronger than that of the wild type receptor as quantified in flow cytometry-based BiFC. Similar results were obtained in confocal microscopy. This suggests that these P2Y12 mutants form an increased number of dimers or oligomers with increased self-affinities.

Conclusion:
We demonstrate that P2Y12 forms constitutive homo-oligomers. Two mutations associated with bleeding disorders in patients have altered receptor-receptor interactions. Future investigation will explore the effect of mutations and receptor oligomers on G protein coupling and receptor: G protein stoichiometry.
Abstract 862: Ser517:-phosphorylation Of Sirt1 Is Required For Its Pi3k/akt-mediated Nuclear Translocation And Cardiomyocyte Protection Against Oxidant Stress In Failing Hearts.
Authors: Tanno Masaya , Miura Tetsuji , Miki Takayuki , Yano Toshiyuki , Horio Yoshiyuki , Shimamoto Kazuaki 
Published: July 2021

[Purpose] We recently found that SIRT1, a protein deacetylase, shuttles between the nucleus and cytoplasm. In this study, we examined the role of nuclear SIRT1 in cardiomyocyte protection against oxidant stress and involvement of PI3K/Akt in the nuclear translocation.
[Methods and Results] First, the critical intracellular location of SIRT1 for its anti-apoptotic function was examined. C2C12 cells were transfected with wild-type SIRT1 (WT) or SIRT1 with site-directed mutations in the nuclear localizing signal (mtNLS) and exposed to antimycin A (AA), an oxidative stressor. AA-induced apoptosis was suppressed in WT-transfected cells expressing SIRT1 in the nuclei compared with that in mtNLS-transfected cells expressing SIRT1 in the cytoplasm (TUNEL-positive cells = 4.4±0.7% vs. 34.6±8.0%). AA-induced apoptosis and also angiotensin II-(angII)-induced apoptosis in neonatal rat cardiomyocytes (NRCM) were suppressed by resveratrol, a SIRT1 activator. This protective effect of resveratrol was attenuated by transfection of SIRT1-siRNA but not by transfection of control siRNA. Next, we assessed the role of PI3K/Akt in nuclear translocation of SIRT1. SIRT1 in NRCM was localized in both the nucleus and cytoplasm under baseline conditions, and IGF-1 induced its nuclear translocation. This effect of IGF-1 was suppressed by LY294002 (LY), a PI3K inhibitor. Deletion mutagenesis study showed that LY-induced nuclear exclusion was observed for SIRT1[223–540] but not for SIRT1[223– 489]. Replacement of serine517 with alanine (S517A) increased cytoplasmic SIRT1, and S517A showed attenuated nuclear translocation in response to IGF-1, indicating that serine517 is the target site of PI3K/Akt. Finally, to confirm heart failure-associated SIRT1 translocation in vivo, myocardial infarction was induced in WKY rats. The number of ventricular cardiomyocytes with nuclear SIRT1 at 4 weeks after infarction was significantly larger than that in sham-operated hearts (10.2±2.9% vs. 0.7±0.2%).
[Conclusion] The results suggest that phosphorylation of SIRT1 at Ser517 by PI3K/Akt is involved in nuclear translocation of SIRT1, which contributes to cardiomyocyte protection from oxidant stress-mediated injury in failing hearts.
Abstract 194: Rosuvastatin Reduces Progression of Carotid Atherosclerosis within 12 Months of Treatment: The METEOR Trial
Authors: Bots Michiel L , Raichlen Joel S , Evans Gregory W , Palmer Mike K , O’Leary Daniel H , Grobbee Diederick E , Crouse John R 
Published: July 2021

Background: In several statin trials, vascular event rates for treatment groups begin to separate 1 year after commencement of treatment. For atherosclerosis progression, the temporal sequence of the effect has not been defined. We sought to determine the earliest time point at which significant differences in atherosclerosis progression rates were detectable after initiation of statin therapy using data from the METEOR trial (Measuring Effects on intima media Thickness: an Evaluation Of Rosuvastatin).
Methods: METEOR was a double-blind, randomized, placebo-controlled trial among 984 low risk subjects, which studied the effect of LDL lowering with 40 mg rosuvastatin on the rate of change in carotid intima media thickness (CIMT) over time. Ultrasound assessments were made at 12 carotid artery sites at baseline and every 6 months up to two years. In these analyses, the data were cut at 6 months, 1 year, and 18 months, and compared with analysis of all data at 2 years, using the same statistical method.
Results: The difference in rate of maximum CIMT progression for all carotid artery sites (primary endpoint - near and far walls of the left and right common carotid artery [CCA], carotid bulb and internal carotid artery) between the rosuvastatin and placebo groups was apparent 6 months after baseline (0.0023 mm/yr and 0.0106 mm/yr, respectively p =0.36). After 12 months CIMT progression rates were significantly different between groups: 0.0032 mm/yr and 0.0133 mm/yr (p=0.049). This divergence grew with further follow-up: − 0.0009 mm/yr and 0.0131 mm/yr after 18 months (p<0.0001), and − 0.0014 mm/yr and 0.0131 mm/yr after 24 months of treatment (p<<178>0.0001). For the individual carotid artery segments, significant differences were seen at 12 months for the mean CIMT of the CCA, and at 18 months for the maximum CIMT of the bulb and the CCA.
Conclusion: Aggressive LDL lowering with rosuvastatin exerts its beneficial effect on atherosclerosis during the first year of treatment, which parallels the timing of event rate reduction seen in clinical trials. These findings suggest that, in trials examining the effects of treatment on CIMT progression, a duration of 12 months may be adequate, given sufficient sample size, high precision of measurements, and treatment effect.
Abstract 883: Elevated Epidermal Growth factor Receptor Phosphorylation Induces Resistance Artery Dysfunction in Type 2 Diabetes
Authors: Matrougui Khalid , Palen Desiree I 
Published: July 2021

Objective: We previously showed that epidermal growth factor receptor (EGFR) transactivation is a key mechanism in the regulation of resistance artery myogenic tone. Type 2 diabetes is associated with microvascular complications. We hypothesized that elevated EGFR phosphorylation contributes to resistance artery dysfunction in type 2 diabetes.
Methods and results: Type 2 diabetic db-/db- (diabetic) and non-diabetic db-/db+ (control) mice were treated with EGFR inhibitor (AG1478, 1 microM) for 2 weeks. Isolated coronary artery (CA) and mesenteric resistance artery (MRA) were mounted in an arteriograph. Pressure-induced myogenic tone (MT) was increased in MRA and CA from diabetic mice and normalized by AG1478 treatment. Phenylephrine-induced contraction and nitric oxide donor-induced relaxation were similar in all groups. Relaxation endothelium-dependent in response to shear-stress and acetylcholine of MRA and CA from diabetic mice were altered and associated with reduced eNOS expression and phosphorylation. The treatment of diabetic mice with AG1478 improved CA and MRA endothelial function and restored eNOS expression. Immunostaining and western blot analysis showed increased endothelial and SMC EGFR phosphorylation of MRA and coronary from diabetic mouse, which was reduced by AG1478 treatment. Primary cultured endothelial cells (EC), from resistance arteries, treated with high glucose for 48hrs showed an increase of EGFR phosphorylation associated with a decrease of eNOS expression and phosphorylation in response to calcium ionophore. Pretreatment of endothelial cells with AG1478 prevented the effect of high glucose.
Conclusion: This study provides evidence of the role of elevated EGFR phosphorylation in CA and MRA dysfunction in type 2 diabetes. Therefore, EGFR should be a potential target for overcoming diabetic small arteries complications.
Abstract 997: Innate Immune Adaptor MyD88 Mediates Myocardial Inflammation And Infarction In A Mouse Model Of Ischemia-reperfusion Injury
Authors: Zhao Huailong , Xu Xinhua , Schmidt Ulrich , Chao Wei 
Published: July 2021

INTRODUCTION: The innate immune response to ischemia-reperfusion (I/R) is, by far, the most common cause of myocardial inflammation and contributes to ischemic myocardial injury. MyD88 is an adaptor protein critical for transducing signals from most innate immune receptors and plays an important role in host defense against microbial infection; but its role in non-infectious myocardial inflammation and injury following I/R is unclear.
METHODS: I/R injury was studied in wild-type (WT) and MyD88-deficient (MyD88−/−) mice. Mice were anesthetized and mechanically ventilated. The heart was exposed via intercostal thoracotomy. The left coronary artery was ligated for 30 min followed by reperfusion. The coronary artery was re-ligated at 24 hours and fluorescent microsphere was injected into LV to detect area-at-risk (AAR). The hearts were harvested, sectioned, and examined for inflammation or infarct size. Myocardial neutrophil infiltration [GR-1 immunohistochemistry and myeloperoxidase activity] and NF-kB signaling (I-kBa degradation and NF-kB binding activity) were examined to assess myocardial inflammation. In ex vivo studies, isolated mouse hearts were exposed to global ischemia for 20 min followed by reperfusion for 30 min.
RESULTS: In vivo I/R induced significant neutrophil infiltration and NF-kB activation in myocardium in WT mice compared with the sham-operated mice. However, in MyD88−/− mice subjected to the same I/R, the level of neutrophil recruitment and NF-kB signaling was significantly lower compared with WT mice. The ratio of AAR to LV was similar in WT and MyD88−/− mice (34 +/− 3% vs. 41 +/− 3%). However, the ratio of MI to AAR was 58% less in MyD88−/− than in WT mice (14 +/− 2% vs. 33 +/− 6% (p < 0.014, n = 11–12). In a Langendorff model of I/R injury, a system devoid of circulating blood components, global I/R induced similar MI sizes, as a fraction of LV size (MI/LV), in MyD88−/− and WT mice (34 +/− 6% and 25 +/− 7%, respectively, p = 0.3, n = 8).
CONCLUSIONS: These studies demonstrate that systemic MyD88 deficiency decreases myocardial inflammation and infarction after ischemia-reperfusion in vivo and support the hypothesis that MyD88 mediates ischemic myocardial injury via mechanisms that are dependent upon circulating blood components.
Abstract 1190: The Complement Component C5a is Present in Human Unstable Coronary Lesions in Vivo and Induces the Expression of MMP-9 in Human Macrophages via Nf-kappaB Activation In Vitro
Authors: Speidl Walter S , Kastl Stefan P , Hutter Randolp , Kaun Christoph , Bauriedel Gerhard , Fuster Valentin , Maurer Gerald , Badimon Juan J , Wojta Johann 
Published: July 2021

Background. The complement component C5a formed during activation of the complement cascade exerts chemotactic and proinflammatory effects. We have recently shown that serum levels of C5a predict cardiovascular risk in patients with advanced atherosclerosis. However, it is not known whether C5a may play an active role in the onset of acute coronary events.
Methods and Results. Immunohistochemical analysis of human coronary plaques obtained by atherectomy demonstrated the presence of C5a in macrophage rich areas and the necrotic core. Unstable plaques (n=9) showed significantly higher staining for C5a than stable coronary lesions (n=10; p<0.05). Double-labeling studies demonstrated colocalization of C5a and MMP-9. When human monocyte derived macrophages were stimulated in vitro with rhC5a, MMP-9 mRNA levels were increase 6-fold. In addition, MMP-9 antigen as detected by ELISA and MMP-9 activity as detected by a specific activity assay significantly increased after stimulation with rhC5a. Nuclear shift assay revealed that rhC5a stimulated NF-κB DNA binding and the NF-κB inhibitor 6-amino-4-phenoxyphenethylaminoquinazoline blocked the up-regulation of MMP-9 by rhC5a.
Conclusion. C5a is increased in unstable coronary plaques, colocalizes with MMP-9 and induces the expression of MMP-9 in human macrophages via NF-κB activation. Taken this observations together, activation of the complement cascade and formation of C5a may be involved in the onset of acute coronary events.
Abstract 1407: Interferon Regulatory Factor-9 (irf-9) Mediates Short Term Host Protection, But Promotes Long Term Immune Injury In Evolution Of Myocarditis Leading To Dilated Cardiomyopathy
Authors: Konviser Michael , Liu Youan , Chen Manyin , Shi Yu , Sun Mei , Gramolini Anthony , Fish Eleanor , Opavsky Anne , Liu Peter 
Published: July 2021

INTRODUCTION: Evolution of viral myocarditis to dilated cardiomyopathy (DCM) represents a delicate balance between host innate immunity and T-cell acquired immunity. IRF-9 is a key member of a transcription factor family that regulates type I interferon (IFN) production, critical for innate antiviral protection. However, the influence of innate immunity in general and IRF-9 in particular on acquired immunity and DCM is unknown.
HYPOTHESIS: IRF-9 signaling provides immediate host protection through interferon production, but stimulates acquired immunity leading to DCM in a coxsackievirus murine myocarditis model.

METHODS: Interferon-regulatory factor 9 (IRF-9) homozygous knockout mice were generated, and show impaired type I interferon production. Wild-type (WT, n=51) and IRF-9
−/−
(n=124) littermates were inoculated with 10^2 p.f.u. of coxsackievirus B3 as previously described. Survival, viral titers, cardiac hypertrophy, inflammation and fibrosis were evaluated on days 0, 4, 7, 10, 14, 28 and 42. Splenocyte subpopulations were cell sorted and quantitated by FACS.

RESULTS: IRF9−/− mice showed dramatically increased mortality compared to the wild-type littermates (0% WT vs 72% IRF-9
−/−
on day 14, P<0.0001). On day 42, there was less cardiac hypertrophy and inflammation in IRF-9
−/−
mice compared to WT controls (p<0.05). There was no difference in fibrosis. The mature T-lymphocyte population, defined as CD4 or CD8 single positive, was statistically identical in the two populations up until and including day 28 post-infection. However on day 42 there was a dramatic increase in the number of cytotoxic and helper T-Cells in the wild-type mice that was not observed in the IRF-9
−/−
spleens (p<0.05).

CONCLUSIONS: These data suggest a novel dual role of IRF-9 in not only regulating interferon in acute stage of viral infection in myocarditis, but also late acquired immunity activation, including CD4/8 populations, contributing to the development of chronic cardiomyopathy.
Abstract 1413: Interleukin 13 Exerts Anti-appoptotic Effects On Hypoxic Cardiomyocytes Through Pi3 Kinase In The Absence Of Hypertrophic Effects
Authors: Poeling Jochen , Kubin Thomas , Rees Wolfgang , Hein Stefan , Shim Gil-Jin , Warnecke Henning , Braun Thomas 
Published: July 2021

Objectives: Recent research has focussed on anti-inflammatory cytokines (a-ICs) as cardio-protective agents. A-ICs were mainly considered in the context of inhibition of secretion of pro-ICs such as tumour necrosis factor-α and with the exception of transforming growth factor β little is known about direct effects of a-ICs on cardiomyocytes (CMC).

Methods: To clarify a direct functional relationship of a-ICs we evaluated effects of interleukin-4, -10 and -13 on cultured CMC by planimetry, confocal microscopy (CF), protein synthesis determination (PS), 2DE-analysis (2DE), mass spectrometry and Western blotting. Intracellular pathways were analysed with specific chemical inhibitors. Anti-appoptotic effects were evaluated by determinating the cell number, cleaved caspase 3 (cC3) and PARP (cP) under 2% O
2
. Untreated cell lines (unCO) or leukemia inhibitory factor (LIF) treated CMC served as controls.

Findings: In contrast to Il-13 we did not detect effects of Il-4 and Il-10 on CMC. Il-13 treated CMC showed a 40% (p<0.01) increased viability over unCO under normoxic and 65% (p<0.01) under hypoxic conditions after 4d and 12h, respectively. Under hypoxia antiapoptotic effects were visible in 60% reduction of cC3 (60%; p<0.05) and cP (40%; p<0.05). Using a battery of more than 300 phospho-specific antibodies we found that phosphorylation of Akt (5-fold, p<0.01) was the predominant activated pathway but we found also mild increases in P-MEK1/2 (1.7-fold). The antiapoptotic effects and Akt activation were abolished in the presence of the PI3 kinase inhibitor (20μM) LY294002 (p<0.01) but not in the presence of the MEK1/2 inhibitor (5μM) UO126. In contrast we did not find any significant morphological changes of CMC in the presence of Il-13 neither in the protein composition of structural proteins (2DE, CF), nor increases in PS nor changes in cell surface area, width and length opposing hypertrophic responses visible after LIF treatment.
Conclusions: These findings indicate that Il-13 can directly support myocyte function in addition to its inactivation of immune cells. Our results and the existence of a Il-13 receptor in the heart altogether throws light on Il-13 as a promising therapeutical target.
Abstract 1534: Diabetes Mellitus Is An Independent Predictor Of Mortality In Defibrillator Recipients
Authors: Chen-Scarabelli Carol , Scarabelli Tiziano M 
Published: July 2021

Background: Previous studies have shown an increased morbidity and mortality risk in patients with diabetes mellitus (DM) undergoing percutaneous coronary interventions and cardiac surgery. However, the impact of DM on mortality in patients with implantable cardioverter defibrillators (ICDs) is unknown.
Aim: to investigate whether DM was associated with increased mortality risk in patients with ICDs
Methods: A retrospective review of all ICD patients followed within a consecutive 2.5 year period was performed. Baseline characteristics and mortality data were compared between DM and non-DM patients. Mann Whitney test was used to analyze the differences between the means, and multivariate logistic regression analysis was performed to identify independent predictors of mortality.

Results: A total of 336 patients with ICDs (including 141 patients with DM and 195 without DM) were studied. The mean age was 66.8 ± 9.6 years in the overall population, with no significant difference in baseline characteristics between the 2 groups, except for beta-blocker therapy, creatinine, and estimated glomerular infiltration rate (EGFR). Use of beta-blockers was greater in the non-DM group (92.8% vs 84.4%, p=0.02). Renal function was worse in the DM group, as documented by a higher serum creatinine (1.6 ± 0.8 mg/dl in DM vs 1.38 ± 0.74 mg/dl in non-DM, p = 0.003) and a lower EGFR (55.9 ± 22.9 ml/min/1.73m
2
vs 63.5 ± 19.9 ml/min/1.73m
2
, p=0.0005). Although the incidence of VT was not significantly different between the two groups, there was a significantly higher mortality in the DM group (27.7% vs 11.3% in non-DM, p=0.0002). On multivariate stepwise logistic regression analysis, DM was a significant independent predictor of mortality (OR 2.38, 95% CI 1.3– 4.36, p =0.005).

Conclusion: DM is an independent predictor of mortality in ICD patients. Prevention of DM, as well as early diagnosis and intervention, should be emphasized in order to reduce the growing epidemic of type 2 DM and its associated complications.