bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract O.16: Comprehensive Genotyping Of The FCGR2/3 Locus Reveals A Novel Association Of FCGR2C-ORF With Susceptibility To Kawasaki Disease
Authors: Kuijpers Taco W , Tacke Carline E , Nagelkerke Sietse Q , Breunis Willemijn B , Hoang Long T , Png Eileen , Geissler Judy , Tanck Michael W , Ellis Justine A , Ponsonby Anne L , Davila Sonia , Khor Chiea C , de Boer Martin , van der Heijden Joris , Fijnvandraat Karin , van der Schoot E. , van den Berg Timo K , Yeung Rae S , Levin Michael L , Burgner David , Shimizu Chisato , Burns Jane C , Hibberd Martin L 
Published: July 2021

The human FCGR2/3 locus contains highly homologous genes encoding the five major receptors for IgG (Fc-gamma receptors, FcγRs). In two prior GWAS on Kawasaki disease (KD), a SNP in FCGR2A (131H>R; rs1801274) was identified to be associated with disease susceptibility. However, the FCGR2/3 locus contains multiple single nucleotide polymorphisms (SNPs) and copy number variations (CNVs), which were not covered by the detection platforms used in the GWAS. In this study we therefore focused on further fine-mapping of this locus to investigate the association of the different genetic variations with KD susceptibility. A highly accurate and validated multiplex ligation-dependent probe amplification (MLPA) assay was used to analyze all functionally relevant SNPs and CNVs within this locus. In a genetic association study involving case-control and family-based testing with 1028 patients with KD, the previous finding of FCGR2A-131H as a susceptibility marker for KD was confirmed (OR 1.16; 95%CI 1.08-1.32, meta-P = 0.01). In addition, we found a novel significant association of the FCGR2C-ORF haplotype with susceptibility to KD (OR 1.34; 95% confidence interval 1.11-1.62, meta-P = 0.003). FCGR2C-ORF leads to the expression of an extra, functionally activating FcγR (i.e. FcγRIIc) on myeloid cell types and NK cells. Being absent in Asian individuals, the FCGR2C-ORF haplotype only contributed to KD susceptibility in European subjects, independent of the established association with FCGR2A-H131R. We did not find any significant association of CNV of the locus with susceptibility to KD.
Our data point to an important role of the activating FcγRs in KD pathology. We hypothesize that the identified functional SNPs might alter the balance between the activating and inhibitory FcγRs leading to unbalanced inflammation and KD.
Abstract 11722: Excellent Preclinical Results of Honeycomb Microporous Covered Stents for the Treatment of Large and Wide-Necked Cerebral Aneurysms
Authors: Nakayama Yasuhide , Satow Tetsu , Funayama Marina , Moriwaki Takeshi , Tajikawa Tsutomu , Furukoshi Maya , Hamano Eika , Ishii Daizo , Hayashi Masataka , Sugata Sei , Ishibashi-Ueda Hatsue , Takahashi Jun C 
Published: July 2021

Background:
We developed covered stents with a microhoneycomb mesh for the treatment of large or giant wide-necked cerebral aneurysms. These aneurysms are extremely difficult to treat and carry high risks, even with the use of clinically available surgical methods or the endovascular technique using coils with the assistance of bare stents. This preclinical study confirmed the excellent performance of our stents in three animal experimental models.

Methods and Results:
The covered stents were prepared by using balloon-expandable stents (CoCr; diameter, 3.5-5.0 mm; length, 16-28 mm) and by dip coating to cover their struts completely with a polyurethane membrane (thickness, 20 μm), followed by laser microprocessing (pore size, approximately 100 μm; opening ratio, approximately 30%).

Model 1: In prepared tortuous canine arteries (n = 12) that simulated the shape of the human internal carotid artery, all stent-mounted delivery catheters could smoothly navigate with little arterial damage.
Model 2: In a prepared canine model (n = 10) of large (over 7 mm), wide-necked (over 4 mm) outer-sidewall aneurysms, at 3 months after stenting, completely endothelialized neointima was formed at the aneurysm openings and the aneurysm sacs were entirely organized.
Model 3: In rabbit branching abdominal aortas, the vascular flow pattern at all the branching arteries remained unchanged for over 1 year, even after performing overlap stenting (n = 5) in addition to single stenting (n = 5). On all the stented luminal surfaces, complete thin neointimal formation with complete endothelialization was observed.

Conclusion:
Our newly developed novel covered stents exhibited excellent preclinical performance in three animal experimental models in terms of stent delivery, aneurysm embolization, and flow maintenance in branching arteries. We highly expect that the stents will be useful as endovascular embolization devices for large and wide-necked aneurysms in humans.
Abstract 12566: Novel Genes Associated With Platelet Function Discovered Through Whole Genome Sequencing
Authors: Yanek Lisa R , Taub Margaret A , Iyer Kruthika R , Ruczinski Ingo , Vaidya Dhananjay , Kral Brian G , Faraday Nauder , Becker Diane M , Becker Lewis C , Mathias Rasika A 
Published: July 2021

Platelet aggregation on ruptured or eroded atherosclerotic plaques can initiate arterial thromboses. GWAS have identified common loci associated with platelet aggregation, but cumulatively they account for only a small portion of the variability in platelet aggregation. We used whole genome sequencing (WGS) in 286 healthy subjects from families with early onset coronary artery disease (GeneSTAR Study) to identify additional genes harboring rare/novel functional variants that determine platelet aggregation at baseline and post-aspirin therapy (ASA, 81mg/day for 14 days). Deep (>30X) WGS was performed on the Illumina Hiseq platform with an average of ~3.7M variants per individual and analyses presented here were limited to coding functional (i.e., missense, stop-gained and stop-lost) variants that were either rare (MAF < 5%) or novel (not observed in public catalogs of variation).
Using a 2-Step process, we: (1) tested for perfect co-segregation between these variants in each gene and a variety of dichotomized platelet aggregation phenotypes in n=34 subjects from 5 African American (AA) families and n=38 subjects from 7 European American (EA) families; (2) any gene with evidence across multiple families was then replicated in a set of independent samples (n=95 AA, n=119 EA) using Skat-O. To dichotomize phenotypes, ‘hyper-aggregators’ were defined as subjects in the upper 10th percentile while ‘normals’ were those between the 25th-75th percentile for each trait.
We identified three novel genes harboring coding functional variants not previously reported to be associated with platelet aggregation in both races: (1) HMG20B, associated with baseline aggregation to ADP; (2) PGA5, associated with baseline aggregation to collagen; and (3) PLAC4, associated with post-ASA aggregation to collagen. HMG20B is involved in megakaryocyte development and platelet production and interacts with KIF4, a microtubule based motor protein, while the role of PGA5 and PLAC4 is less clear. Follow-up studies may identify new platelet biological pathways that can be targeted for clinical purposes.
Abstract 13458: Caspase Recruitment Domain-Containing Protein 9 Deficiency Mitigates Obesity-Induced Cardiac Dysfunction
Authors: Cao Li , Qin Xing , Zheng Tiantian , Nair Sreejayan , Ren Jun , He Guanglong 
Published: July 2021

Obesity has become an epidemic and is associated with cardiovascular anomalies. Obesity-induced low-grade chronic inflammation and macrophage infiltration into the heart plays a critical role in the development of cardiac dysfunction. Because the adaptor protein caspase recruitment domain-containing protein 9 (CARD9) regulates the innate immunity in macrophages via activation of pro-inflammatory cytokines, we hypothesized that CARD9 mediates chronic inflammation and plays a detrimental role in obesity-associated cardiac dysfunction.
Male C57BL/6 wild-type (WT) and CARD9 knockout (CARD9-/-) mice were fed normal chow (ND, 12% fat) or a Western diet (WD, 45% fat) for 5 months starting at 4 weeks of age. At the end of 5-month WD feeding, cardiac geometry and function were evaluated using echocardiography. Immunofluorescence assay was performed to detect macrophage infiltration in the heart. Heart tissue homogenates, plasma, and supernatants from cultured macrophages were collected to measure the activities of pro-inflammatory cytokines by ELISA kits. Western immunoblotting analyses were performed on isolated peritoneal macrophages and heart tissue to dissect the pro-inflammatory signaling pathways. WD feeding induced insulin resistance and glucose intolerance in the WT mice and the effects were attenuated in the CARD9-/- mice. Myocardial fractional shortening and ejection fraction were significantly compromised in the WD-fed WT mice and CARD9 knockout preserved cardiac function. CARD9 knockout also significantly decreased the number of infiltrated macrophages in the heart. In addition, myocardium-, plasma- and macrophage-derived cytokines IL-6, IL-1β and TNFα were attenuated in the CARD9-/- mice compared to the WT mice. WD feeding resulted in increased protein phosphorylation levels of p38 MAPK in the heart and macrophages which were suppressed in the CARD9-/- mice.
Taken together, these results suggest that CARD9 knockout ameliorates obesity-induced cardiac dysfunction, potentially through reduction of macrophage infiltration and suppression of p38 MAPK phosphorylation in the heart.
Abstract 13709: High-Density Lipoprotein is a Novel Activator of Cardiac Glucose Metabolism in both Healthy and Insulin Resistant Myocardium
Authors: Heywood Sarah E , Henstridge Darren C , Richart Adele L , Alt Karen , Carey Andrew L , Kammoun Helene L , Delbridge Lea M , Hagemeyer Christoph E , Febbraio Mark A , Kingwell Bronwyn A , Siebel Andrew L 
Published: July 2021

Aims:
High-density lipoprotein (HDL) is known to increase skeletal muscle glucose uptake, but whether this action extends to the myocardium and has relevance in the setting of ischemia is unknown. This study aimed to determine the effect of HDL on cardiac glucose metabolism both in vitro and in vivo, including during ischemia/reperfusion injury.

Methods and Results:
HDL treatment (50μg/mL) increased glucose uptake (147±10%), glycolysis (154±11%) and glucose oxidation (130±6%) in neonatal ventricular cardiomyocytes compared to saline control. HDL rapidly activated the Akt pathway and an Akt inhibitor (Akti 1/2) abolished HDL-mediated glucose uptake, suggesting an Akt-dependent mechanism. To determine the upstream receptor mechanism HDL binding to sphingosine-1-phosphate (S1P) receptors was blocked with either 1uM VPC23019 (S1P1&3 antagonist) or 1uM CAY10444 (S1P3 antagonist). Both similarly inhibited HDL-mediated glucose uptake. S1P is a known activator of the Akt pathway, thus our data suggests the S1P component of HDL increases glucose uptake via S1P3 activation of the Akt pathway.

The effect of a single intravenous bolus of reconstituted HDL (CSL-111; 80mg/kg) in chow diet (CD) and high fat diet (HFD) insulin resistant C57Bl6 mice was investigated during an oral glucose tolerance test (OGTT). CSL-111 treatment increased cardiac muscle glucose clearance in both CD (168±16%) and HFD (138±11%) mice compared to saline infusion during the OGTT.
To assess whether CSL-111 could increase cardiac glucose uptake in an ischemic setting, a single bolus of CSL-111 (80mg/kg) was given at the beginning of reperfusion in mice following left anterior descending artery (LAD) ligation. CSL-111 trended to increase cardiac glucose uptake ([18F]Fluorodeoxyglucose tracer and PET-CT imaging) by 66% (chow) and 81% (HFD) (treatment effect p=0.07) 1 hour post-reperfusion following LAD surgery. Furthermore, CSL-111 improved long term cardiac functional recovery.

Conclusion:
These data show that HDL increases glucose uptake and utilisation in the heart suggesting a role for HDL therapies in preserving functional myocardium subsequent to acute coronary ischemic syndromes.
Abstract 15194: Multiple Co-morbidities Produce Left Ventricular Diastolic Dysfunction in a Large Animal Model
Authors: Sorop Oana , Heinonen Ilkka , van Kranenburg Matthijs , de Beer Vincent J , Octavia Yanti , van Geuns Robert-Jan M , van den Meiracker Anton H , van der Velden Jolanda , Danser Jan A , Merkus Daphne , Duncker Dirk J 
Published: July 2021

More than 50% of patients with heart failure have preserved ejection fraction (HFpEF) characterized by diastolic dysfunction. HFpEF usually occurs in female patients with multiple comorbidities such as obesity, hypercholesterolemia, diabetes mellitus (DM) and hypertension. Here we studied the effects of DM, high fat diet (HFD), and chronic kidney disease (CKD) on diastolic dysfunction in swine.
In 11 female swine, DM type 2 (3x50mg/kg iv streptozotocin), hypercholesterolemia (HFD 1% cholesterol) and CKD by renal artery embolization (38-42μm polyethylene beads), were induced (DM+HFD+CKD). Swine were followed for 6 months. Eight female healthy swine on normal pig-chow, matched for age were used as controls (CON).
The DM+HFD+CKD showed hyperglycemia (22.7±0.9mmol/l in DM+HFD+CKD vs 6.1±0.67mmol/l in CON), hypercholesterolemia (16.9±3.4 vs 2.2±0.1mmol/l), hypertriglyceridemia (1.20±0.36 vs 0.28±0.05mmol/l) and hypertension (awake mean arterial pressure: 108±6 vs 84±3mmHg; all P≤0.05). These co-morbidities resulted in chronic systemic inflammation (TNF-α: 231±64 vs 74±24pg/ml), impaired coronary small artery endothelium-dependent vasodilation to bradykinin, capillary rarefaction (cap./fiber ratio 0.58 vs 0.99) and increased myocardial superoxide production (14.8±0.9 vs 6.4±0.3 RLU/sec/g), due to increased NOX activity (all P≤0.05). The coronary microvascular abnormalities were associated with increased myocardial collagen content (4.7±0.3 vs 2.4±0.2%) and elevated passive cardiomyocyte force, (both P≤0.05), resulting in reduced E/A ratio (0.99±0.14 vs 1.53±0.23), measured with cardiac MRI, and increased LV end-diastolic stiffness (0.79±0.19 vs 0.29±0.03 mmHg/ml/mg) derived from LV pressure-volume relations obtained with a conductance catheter (both P≤0.05). In contrast, ejection fraction was maintained, (45±4 vs 54±3%, P=NS).
In summary, multiple comorbidities, including DM, hypercholesterolemia, CKD and hypertension in female swine trigger a series of events, starting with systemic inflammation, increased oxidative stress and coronary microvascular dysfunction, resulting in myocardial stiffening, and leading to diastolic dysfunction.
Abstract 15259: Genetically Engineered eNOS Dimer Destabilization Impairs Blood Pressure Reducing Activity of eNOS in Mice
Authors: Suvorava Tatsiana , Pick Stephanie , Kojda Georg 
Published: July 2021

Endothelial dysfunction and oxidative stress are associated with hypertension but whether endothelial superoxide plays a role in the early development of essential hypertension remains uncertain. In this study we investigated whether eNOS-derived endothelial oxidative stress is involved in the regulation of blood pressure (BP). Normal bovine eNOS (eNOS-Tg) or a novel dimer-destabilized eNOS-mutant harboring a partially disrupted zinc-finger (C101A-eNOS-Tg) was introduced in C57BL/6 mice using the endothelium-specific tie-2 promoter. Mice were monitored for aortic endothelium-dependent relaxation, systolic BP, levels of superoxide and several post-translational protein modifications indicating activity and/or increased vascular oxidative stress. Some groups of mice underwent voluntary exercise training or treatment with SOD mimetic Tempol (1mM).
C101A-eNOS-Tg (aorta, skeletal muscle, left ventricular myocardium and lung) showed significantly increased superoxide generation, protein- and eNOS-tyrosine-nitration, eNOS-S-glutathionylation, eNOS1176/79 phosphorylation and Thr172-AMP kinase (AMPKα) phosphorylation as compared to eNOS-Tg and wildtype (WT)-controls. The localization of eNOS-Tg was restricted to endothelium as evidenced by immunohistochemical staining for eNOS and an endothelial-specific marker CD31. Exercise training increased phosphorylation of eNOS and AMPKα in WT while these physiologic adaptations were absent in C101A-eNOS-Tg. Aortic endothelium-dependent relaxation was similar in all strains. In striking contrast, C101A-eNOS-Tg displayed normal BP despite higher levels of eNOS, while eNOS-Tg showed significant hypotension. Tempol completely reversed the occurring protein modifications and significantly reduced BP in C101A-eNOS-Tg but not in WT.
We observed that oxidative stress generated by endothelial-specific expression of genetically destabilized C101A- eNOS selectively prevents BP reducing activity of vascular eNOS, while having no effect on aortic endothelial dependent relaxation. Our data suggest that oxidative stress generated by eNOS dimer destabilization contributes to the regulation of blood pressure.
Abstract 15568: Selective Deletion of Smad4 in Smooth Muscle Cells Causes Thoracic Aortic Aneurysm in Mice
Authors: Carnevale Daniela , Carnevale Raimondo , Da Ros Francesco , Cifelli Giuseppe , Bizzotto Dario , Braghetta Paola , Bressan Giorgio , Lembo Giuseppe 
Published: July 2021

TGFβ signaling have been implicated in the onset of thoracic aortic aneurysms (TAAs). Recently, it has been described that mutations in SMAD4, an intracellular transducer required in the TGFβ canonical pathway, present aortic disease in humans. Although transgenic models recapitulating mutations of TGFβ pathway have been studied, the role of Smad4 in the onset of TAA has never been explored.
Smad4 homozygotes mice are embryonically lethal, thus, we postnatally targeted the gene, using a tamoxifen (TAM)-inducible Cre recombinase under the smooth muscle myosin heavy chain promoter. Activation by TAM suppressed Smad4 in aorta SMCs of Smadf/f;Myh11-CreERT2 mice. Most of these mice died 4/6 months later from hemothorax due to rupture of thoracic aorta, while untreated animals survived. Echocardiography showed a progressive dilatation of aorta and TAA formation. Histological analysis revealed progressive fragmentation of elastic lamellae with increased inflammatory infiltrates at sites of disarrangement, rich in macrophages. At the molecular level, we found that lack of Smad4 was followed by increased levels of P-Smad2 and P-Smad1/5/8 early after TAM. This finding suggested an enhancement of TGFβ receptors activity, a condition that could increase non-canonical TGFβ signaling, the canonical one being certainly depressed. We actually found a significant increase in one of the non-canonical TGFβ signaling, i.e. the p65 subunit of NFkB, a protein complex involved in regulating responses to different stress stimuli, including cytokines. Given the particular relevance that the latter molecules may have in TAA, we tested expression of some of them, finding a selective IL-1β upregulation, produced and stored as an inactive precursor and released in its active form by caspase-1 cleavage. In turn, caspase-1 requires the function of NLRP3 inflammasome, a multiprotein platform that assembles in response to infection and tissue damage and is responsible for activation of inflammatory caspases. Therefore, we also tested NLRP3 mRNA levels and found them to be significantly upregulated. Overall, we demonstrated that selective Smad4 deletion in the SMC of adult mice is sufficient to induce ultrastructural damage and immune reaction culminating in the onset of TAA.
Abstract 15822: Phosphorus Magnetic Resonance Spectroscopy is More Precise at 7 Tesla Field Strength Than 3 Tesla in Patients With Dilated Cardiomyopathy
Authors: Stoll Victoria , Clarke William , Levelt Eylem , Myerson Saul , Robson Matthew , Rodgers Christopher , Neubauer Stefan 
Published: July 2021

Introduction:
Phosphorus magnetic resonance spectroscopy (
31
P-MRS) allows insight into cardiac energetics
in vivo
, but is a technique with an intrinsically low signal to noise ratio (SNR). As ultra-high magnetic field MRI scanners become more widely available this limitation will lessen, allowing detection of smaller changes in metabolite concentrations or measurement of changes in smaller patient groups, which will further the understanding of cardiac energetics.

Hypothesis:
We hypothesised that the 2.4x increase in SNR, predicted by theory at 7T compared to 3T, will allow more precise metabolite concentration measurements in dilated cardiomyopathy (DCM) patients.

Methods:
26 idiopathic DCM patients (69% male, mean age 55±2 yr, mean LVEF 35±2%) underwent
31
P-MRS at 3T and 7T field strengths. Patients were NYHA Class I (46%), II (50%) or III (4%).

Results:
The mean SNR increased from 6.5 ± 0.5 to 16.2 ± 1.5 for PCr (p <0.0001, paired t-test) and from 3.4 ± 0.2 to 11.4 ± 0.8 for γATP (p <0.0001) as the field strength increased from 3T to 7T.

The PCr/ATP ratio was consistent between the 2 field strengths; at 3T PCr/ATP was 1.5 ± 0.1 and at 7T 1.5 ± 0.1; there was as expected no statistically significant difference (p=0.67) (fig a).
The Cramér-Rao lower bounds (CRLB) quantify the minimum precision of fitted parameters. A desirable decrease in the PCr/ATP CRLB was seen from 3T to 7T (47 ± 3% vs 25 ± 2% p<0.0001)(fig b), i.e.-metabolites are indeed quantified more precisely at 7T compared to 3T.

Conclusion:
This study represents the first cardiac
31
P-MRS patient data acquired at a 7T field strength. Compared to a 2.8x SNR increase from 3T to 7T previously reported in normal volunteers, we observed a 2.5x increase in this patient population. There is no significant bias at 7T as the PCr/ATP ratios are similar.

In conclusion 7T
31
P-MRS is feasible in DCM patients and more accurate than at 3T, which in future should allow more accurate quantification and understanding of myocardial energetics.