bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 13249: European and American Blood Cholesterol Guidelines Result in Similar Rates of Statin Recommendations when Applied to a Country with High Cardiovascular Disease Risk
Authors: Navar-Boggan Ann M , Zdrojewski, Tomasz , Wyszomirski Adam , Lachacz Mateusz , Opolski Grzegorz , Bandosz Piotr , Rutkowski Marcin , Gaciong Zbigniew , Wyrzykowski Bogdan , Pencina Michael J
Published: July 2021

Introduction:
The American Heart Association and American College of Cardiology (AHA/ACC) recently released updated guidelines for management of blood cholesterol, which differ from current European Society of Cardiology and European Atherosclerosis Society (ESC/EAS) guidelines. How these differences affect the overall number of individuals recommended for statin therapy in a country with high cardiovascular disease (CVD) risk remains unclear.

Hypothesis:
Due to the lower threshold for statin recommendations for primary prevention based on 10-year CVD risk under the AHA/ACC guidelines, more adults overall would be recommended for statin therapy under American compared to European guidelines.

Methods:
Using 2011 data from a nationwide cross-sectional survey in Poland (NATPOL), we estimated the number and characteristics of adults aged 40-65 recommended for lipid lowering therapy under the ESC/EAS and AHA/ACC guidelines. The survey sample of 1060 adults represented 13.5 million adults in Poland aged 40-65.

Results:
Under ESC/EAS guidelines, 47.6% of adults (44.6-50.7%) aged 40-65 were recommended for immediate statin therapy, compared to 49.9% (46.9-52.9%) under AHA/ACC guidelines. Among adults free of cardiovascular disease (CVD), 10.5% had discordant recommendations between guidelines. Individuals recommended for statin therapy under ACC/AHA but not ESC/EAS guidelines had less chronic kidney disease, higher HDL cholesterol, higher 10-year (AHA/ACC calculator) risk, and higher 30-year (Framingham) risk than adults recommended under ESC/EAS but not under ACC/AHA guidelines. Ten-year CVD mortality risk estimated by the SCORE algorithm was similar between the two groups.

Conclusions:
In spite of differences between current European and American cholesterol guidelines, when applied to a nationwide representative sample from a country with high CVD risk, the number of adults aged 40-65 recommended for cholesterol lowering therapy under each guideline was nearly identical. Although more adults met criteria for primary prevention based on 10-year CVD risk under new American guidelines, the impact of this is offset by additional criteria for statin therapy in current European guidelines.
Abstract 133: Electrical Insulating Gloves for Safe Hands-on Defibrillation Do Not Compromise the Quality of Chest Compressions
Authors: Thomsen Jakob E , Harpsø Martin , Petley Graham W , Vittinghus Svend , Deakin Charles D , Løfgren Bo
Published: July 2021

Introduction:
We have recently shown that Class 1 electrical insulating gloves are safe for hands-on defibrillation. Continuous chest compressions during defibrillation reduce the peri-shock pauses and increase the subsequent chance of successful defibrillation. In this study we have investigated the effect of these electrical insulation gloves on the quality of chest compressions, compared with normal clinical examination gloves.

Methods:
Emergency medical technicians trained in 2010 resuscitation guidelines delivered uninterrupted chest compressions for 6 min on a manikin, whilst wearing Class 1 electrical insulating gloves or clinical examination gloves. The order of gloves was randomized and each session of chest compressions was separated by at least 30 min to avoid fatigue. Data were collected from the manikin. Compression depth and compression rate were compared.

Results:
Data from 35 participants are shown in Figure 1. There was no statistically significant difference between Class 1 electrical insulating gloves in chest compression depth (median±range: 45 (28-61) vs 43 (28-61) p=0.69) and chest compression rate (113 (67-150) vs 113(72-145), p=0.87) when compared to clinical examination cloves.

Conclusion:
These preliminary data suggest that the use of Class 1 electrical insulation gloves does not reduce the quality of chest compressions during simulated CPR compared to clinical examination gloves.
Abstract 13347: Injectable Collagen-based Hydrogel Matrix Modulates Micro-RNA Expression and Prevents Cardiac Deterioration Post-myocardial Infarction
Authors: Blackburn Nick J , McNeill Brian , Chiarella-Redfern Helene , Sofrenovic Tanja , Kuraitis Drew , Ahmadi Ali , Ruel Marc , Rayner Katey J , Suuronen Erik J
Published: July 2021

Background:
Injectable hydrogel biomaterials have emerged as promising therapies for treating myocardial infarction (MI). We developed a collagen type I based injectable hydrogel matrix that can prevent the deterioration of cardiac function when delivered soon post-MI and found that the effects may be mediated through a microRNA (miRNA) mechanism.

Methods/Results:
C57BL/6J mice underwent LAD ligation to induce MI. Mice then received myocardial injections of PBS or matrix delivered at 3hours post-MI. Analyses were performed at 2 days, 1 and 3 months post-treatment. At one month post-treatment, mice that received the matrix had superior left ventricular ejection fraction (LVEF; 45.1±2.3%) compared to the PBS group (29.6±2.4%;
p<
0.001). LVEF was maintained in matrix-treated mice at 3 months (42.9±3.8%). Matrix treatment was also associated with reduced infarct sizes and improved ventricular volumes. Matrix-treated mice had more angiogenesis, mitigated apoptosis and reduced inflammation in the infarcted myocardium at both 2 and 28 days post-treatment. To better understand the mechanisms, we performed miRNA microarrays on infarct and peri-infarct tissue. Matrix treatment resulted in 120 miRNAs with differential expression within +/- 0.3 log2 fold change. In particular, we found matrix treatment down-regulated miR-92a (
p<
0.0005), an anti-angiogenic miRNA. Integrins α5 (Itgα5) and αV (ItgαV), involved in angiogenesis and cell-matrix interactions, were identified as putative miR-92a targets and pursued further
in vitro
using circulating angiogenic cells (CACs). CACs cultured on the matrix had increased Itgα5 and ItgαV expression after 4 days (12.4-fold and 13.9-fold, respectively vs. fibronectin;
p<
0.01). When applied in an
in vitro
angiogenesis assay, the number of CACs that incorporated into capillary-like structures was greater (by 4.2-fold) for cells derived from matrix culture (
p
<0.005).

Conclusion:
We demonstrate pronounced benefits associated with our hydrogel matrix when delivered at 3h post-MI. The matrix effects may be mediated, at least in part, through its ability to regulate miR-92a and integrin-mechanisms. Overall, the matrix may provide a promising therapeutic approach for protecting the myocardium post-MI.
Abstract 13354: Event-Free Survival in African American Patients with Heart Failure
Authors: Adams Kelly L , Dekker Rebecca L , Lennie Terry A , Chung Misook L , Dracup Kathleen , Moser Debra K
Published: July 2021

Introduction:
Health outcomes such as event-free survival (cumulative end-point in time to first health event) in heart failure (HF) patients is worse in African American than Caucasians. While the direct impact of traditional risk factors on outcomes are recognized, it is unknown how sociodemographic and psychosocial variables, disease, and treatment factors may alter the relationship between race and event-free survival.

Hypothesis:
Sociodemographics (age, gender, economic status), psychosocial factors (anxiety, depression), disease factors (smoking, functional status, diabetes) and treatments (beta blockers, ACE inhibitors) moderate the relationship between race and shorter event-free survival among patients with HF.

Methods:
Data were analyzed from 993 outpatients in a multicenter HF registry who were followed for a median of 1.9 years (37% female, 11.3% African American, 64±13 years, 44% NYHA Class III/IV). Data were collected via chart review and interview. Potential proposed moderators were analyzed with race as the predictor and the outcome event-free survival. Regressions were conducted on event-free survival using race and each proposed moderator, and the product of race and each moderator.

Results:
A primary analysis showed that African American patients are 1.54 times more likely to experience a cardiac event within this data set (p=.003). Further regression analyses indicate event-free survival in African American patients with HF is not moderated by the proposed moderators (all p>.05). Although an incomplete moderation, interactions with medication and race demonstrated better outcomes in African Americans than Caucasians not on ACE inhibitors, but Caucasians on prescribed ACE inhibitors have better comparative outcomes.

Conclusions:
Although many modifiable and non-modifiable risk factors may be associated with event free survival in African American HF patients, sociodemographic, psychosocial, disease, and treatment factors do not moderate the relationship between race and event-free survival. Future research is needed to better understand what factors contribute to and moderate evident disparities in the event-free survival of African American patients with HF.
Abstract 13372: Diagnostic Value of a Novel Coronary Computed Tomography Angiography-based Approach for Assessing Fractional Flow Reserve: Comparison with Invasive Measurement
Authors: Baumann Stefan , Renker Matthias , Wang Rui , Meinel Felix G , Rier Jeremy D , Bayer Richard R , Akin Ibrahim , Borggrefe Martin , Steinberg Daniel H , Schoepf U J
Published: July 2021

Background:
Noninvasive fractional flow reserve (FFR) from cCTA correlates well with invasive FFR and substantially improves the detection of obstructive CAD. However with current algorithms, CT-based FFR is derived off-site in an involved, time-consuming manner. We sought to investigate the diagnostic performance of a novel CT-based FFR algorithm (Siemens, Germany), developed for time-efficient in-hospital evaluation of hemodynamically indeterminate coronary lesions.

Methods and Results:
In a blinded fashion, CT-based FFR was assessed in 67 coronary lesions of 53 patients. Pressure guidewire-based FFR<0.80 served as the reference standard to define hemodynamically significant stenosis and assess the diagnostic performance of CT-based FFR compared to standard evaluation of cCTA (stenosis ≥50%). On a per-lesion and per-patient basis, CT-based FFR resulted in a sensitivity of 85% and 94%, specificity of 85% and 84%, positive predictive value of 71% and 71%, and negative predictive value of 93% and 97%. The area under the ROC curve on a per-lesion basis was significantly greater for CT-based FFR compared with standard evaluation of cCTA (0.92 vs. 0.72, p=0.0049). A similar trend, albeit not statistically significant, was observed on per-patient analysis (0.91 vs. 0.78, p=0.078). Mean total time for CT-based FFR was 37.5±13.8 min.

Conclusions:
The CT-based FFR algorithm evaluated here outperforms standard evaluation of cCTA for the detection of hemodynamically significant stenoses while allowing on-site application within clinically viable timeframes.
Abstract 13378: Atypical Mechanosensitive MicroRNA-712 Derived From Pre-ribosomal RNA Induces Endothelial Inflammation and Atherosclerosis
Authors: Kumar Sandeep , Son Dong J , Takabe Wakako , Kim Chanwoo , Jang Inhwan , Ni Chih W , Alberts-Grill Noah , Jo Hanjoong
Published: July 2021

Atherosclerosis is the underlying cause of cardiovascular events, such as heart attack and stroke, and preferentially occurs in arterial regions exposed to disturbed flow (
d-flow
) by mechanisms involving broad changes in gene expression. While microRNAs (miRNAs) are known to regulate various aspects of cardiovascular biology, their role in atherosclerosis has not been directly demonstrated.

We used partial carotid ligation surgery to induce d-flow in the Left carotid artery (LCA) of mouse while the contralateral right carotid artery (RCA) continued to experience stable flow (s-flow). We devised a technique to quickly extract endothelial-enriched RNA from these carotids and performed miRNA array and subsequent validations. Also, using a cone-and-plate shear device, we exposed the immortalized mouse aortic endothelial cells to either laminar or oscillatory shear stress. Use used pre-miR-712 to induce and anti-miR-712 to knockdown the expression of miR-712 in our system.

We identified mechanosensitive miRNAs using a mouse partial carotid ligation model and endothelial miRNA array. Of those mechanosensitive miRNAs identified, miR-712 was the most shear-sensitive miRNA upregulated by
d-flow
both
in vivo
and
in vitro
. We found that miR-712 is derived from the internal transcribed spacer 2 (ITS2) region of pre-ribosomal RNA (
RN45S gene)
in a XRN1 exonuclease-dependent, but DGCR8-independent manner, suggesting that it is an atypical miRNA derived from an unexpected source. Using gain-of-function (pre-miR-712) and loss-of-function (anti-miR-712) approaches and target-binding assays showed that miR-712 directly downregulated the tissue inhibitor of metalloproteinase 3 (TIMP3) expression. This in turn activated downstream metalloproteinases (MMPs and ADAM family) and stimulated pro-atherogenic responses, endothelial tubule formation and sprouting, in a flow-dependent manner.

Treatment with anti-miR-712 prevented atherosclerosis in two different models of murine atherosclerosis using ApoE
-/-
mice: a chronic conventional western-diet or an acute carotid partial ligation model on a high-fat diet. Targeting mechanosensitive "athero-miRs" with anti-miR-based approaches may provide a new treatment paradigm in atherosclerosis.
Abstract 13390: Metabolomic Changes during Short-term Water-only Fasting: the FEELGOOD Trial
Authors: Horne Benjamin D , Cox James E , Miao Ren , Muhlestein Joseph B , Butler Amy R , May Heidi T , Carlquist John F , Anderson Jeffrey L
Published: July 2021

Background:
Routine, periodic fasting is associated with a lower risk of coronary artery disease and diabetes in coronary angiography patients, and may reduce weight and alter metabolic risk in obese people. Fasting reportedly extends longevity in animals. This study evaluated the effect of fasting on metabolic profiles among apparently healthy people.

Methods:
Individuals (N=30) with no fasting history underwent a 24-hour water-only fast and 24 hours of ad libitum eating in a randomized cross-over trial (clinicaltrials.gov NCT01059760). All participants were free from cardiovascular diseases. Study endpoints included 75 metabolic analytes measured by gas chromatography-mass spectrometry. With the Bonferroni correction, p≤0.000667 was needed for statistical significance (p≤0.05 was suggestively significant).

Results:
Participants averaged 43.6±13.5 years of age and 66.7% were female. The 24-hour fasting intervention reduced plasma levels of proline (p=0.00002), tyrosine (p=0.00033), urea (p=0.00034), and galactitol (p=0.00058) compared to the eating day. Overall, 29 factors had p≤0.05 (the expected type I error rate at p≤0.05 for 75 hypothesis tests is 4 false positives), including 16 amino acids (all were reduced by fasting) and 6 fatty acids (all were increased by fasting). Among participants randomized to fast the first 24 hours (n=16), 48-hour levels had returned to baseline for all 29 factors except for suggestively significant lower final values of pyruvate (p=0.014), glutamic acid (p=0.025), and tryptophan (p=0.028).

Conclusion:
A 24-hour fasting intervention reduced plasma levels of two amino acids (proline, tyrosine), and metabolomics literature connects lower proline and tyrosine with lower insulin resistance, better cognitive function, and less depression. The other findings confirm the known utilization of urea in the natriuresis of fasting (via excretion of nitrogen) that may lower blood pressure and that, during fasting, glucose is stored as galactose such that galactitol production is limited and fatty acids are used for energy. Changes in these four plus potentially 25 other metabolites suggest a beneficial health impact, which along with the potential to reduce insulin resistance warrants further study of fasting.
Abstract 13435: Gender Differences in Impact of CYP2C19 Polymorphism on Risk of Coronary Artery Disease
Authors: Hokimoto Seiji , Akasaka Tomonori , Nakagawa Kazuko , Ogawa Hisao
Published: July 2021

Background:
Several cytochrome P450 (CYP) enzyme families have been identified in extra hepatic tissues such as heart, vasculature, kidney, and lung. Specific CYPs, such as CYP2C9 and 2C19, localized in vascular smooth muscle and endothelium may contribute to the pathophysiology of cardiovascular diseases. The aim was to clarify whether CYP2C19 polymorphism would be associated with the development of coronary artery disease (CAD) as a disease-susceptibility gene independently of the conventional coronary risk factors.

Methods:
This study enrolled 723 patients with CAD (male 71%, 70yrs), and healthy subjects undergoing medical checkup (n=453) as control (male 69%, 53yrs). CAD was defined as definitive stable angina and acute coronary syndrome. We analyzed the incidence of CYP2C19 polymorphism for presence of CAD in all subjects and in the absence of diabetes, dyslipidemia, and chronic kidney disease (CKD) to minimize the influence of acquired and environmental coronary risk factors. CYP2C19 genotype was classified into 3 genotypes: (1) EM having normal function alleles, (2) IM having one loss-of-function (LOF) allele, and (3) PM having two LOF alleles.

Results:
The distribution of CYP2C19 genotype in total subjects between both groups was not different (EM, IM, PM: CAD; 34, 47, 19%: control; 32, 49, 19%). In the analysis of non-diabetes, non-dyslipidemia, and non-CKD status, there was no difference in incidence of CYP2C19 genotype between CAD (n=115) and control (n=194) groups (EM, IM, PM: non-risk CAD, 33, 46, 21%; non-risk control, 31, 52, 17%). When patients were stratified by gender, there was significant difference in distribution of CYP2C19 genotype between male and female in non-risk CAD patients (EM, IM, PM: male; 35, 50, 15%: female; 27, 36, 36%). In short, the PM frequency was significantly higher in female than in male (15% vs. 36%, P=0.011). Moreover, in females between CAD and control, incidence of PM was significantly higher in non-risk CAD patients than in non-risk control (36 vs. 14%, P=0.006), but not in male (16 vs. 19%, P=0.546).

Conclusions:
CYP2C19 PM status might be associated with the development of CAD. CYP2C19 polymorphism seems to have an important role in pathophysiology of atherosclerotic disease as a disease-susceptibility gene.
Abstract 13476: Selective Estrogen Receptor Modulator Retards Arterial Senescence and Atherosclerosis Through Restoration of Down-regulated Sirt-1 in Post-menopause Model Mouse
Authors: Miyauchi Takahiro , Ikeda Yoshiyuki , Akasaki Yuichi , Iwabayashi Masaaki , Sasaki Yuichi , Miyata Masaaki , Ohishi Mitsuru
Published: July 2021

Background:
Incidence of cardiovascular disease (CVD) increases with development of arterial senescence and atherosclerosis, which are accelerated after menopause in women. This study aims 1) to clarify the mechanism of which post-menopose facilitates arterial senescence and atherosclerosis, and 2) to develop optical treatment.

Methods and Results:
Female Apolipoprotein E deficient (ApoE KO) mice underwent ovariectomy at 8 weeks old were used as post-menopausal model mice (PM). Age-matched female mice with sham operation were used as control (Ctr). We confirmed that serum estrogen concentration was significantly lower in PM than in Ctr. Twelve weeks after surgery, aortas were harvested to examine molecular biological analysis. Sirt-1 and the ratio of p-eNOS/eNOS protein expressions were significantly lower in PM than in Ctr. Ratio of senescence associated β galactosidase (SA-β Gal) positive cells and expressions of acetyl (Ac)-p53, p21, p16, and PAI-1 were significantly higher in PM than in Ctr. Aortic atherosclerotic lesions assessed by Oil Red O staining was significantly greater in PM than in Ctr. Sirt-1 overexpression retarded ovariectomy-induced arterial senescence. These results suggest that ovariectomy accelerates arterial senescence and atherosclerotic development through downregulation of Sirt-1. Next, we administrated selective estrogen receptor modulator (SERM) to PM mice. Sirt-1 and the ratio of p-eNOS/eNOS protein expression were significantly higher in SERM-treated PM mice than in untreated mice. Ratio of SA-β Gal positive cells, protein expressions of Ac-p53, p21, p16, and PAI-1, and aortic atherosclerotic lesions were significantly lower in SERM-treated PM mice than in untreated mice. The effect of SERM on arterial senescence and atherosclerosis was attenuated by administration of either L-NAME, eNOS inhibitor, or sirtinol, Sirt-1 inhibitor. In addition, L-NAME administration decreased Sirt-1 expression in SERM-treated PM mice, suggesting that SERM-induced eNOS activation upregulates Sirt-1.

Conclusion:
SERM retards development of arterial senescence and atherosclerosis by activation of eNOS and Sirt-1 in postmenopause mice.
Abstract 13485: Prevalence and Treatment of “Balloon Uncrossable” Coronary Chronic Total Occlusions
Authors: Patel Siddharth , Pokala Nagendra , Menon Rohan , Kotsia Anna , Raja Vijay , Christopoulos George , Michael Tesfaldet , Rangan Bavana , Sherbet Daniel , Patel Vishal , Abdullah Shuaib , Hastings Jeffrey , Grodin Jerrold , Banerjee Subhash , Brilakis Emmanouil
Published: July 2021

Background:
In percutaneous coronary intervention (PCI) of coronary chronic total occlusions (CTOs), inability to cross the lesion with a balloon is the second most common cause of technical failure following inability to cross the lesion with a guidewire. While various techniques have been reported for such "balloon uncrossable" CTOs, the frequency and outcomes of these techniques have received limited study.

Methods:
We retrospectively examined 373 consecutive CTO PCIs performed at our institution between 2005 and 2013 to determine the frequency and outcomes of “balloon uncrossable” CTOs.

Results:
Mean age was 63.7 ± 8.3 years and 98.9% of the patients were men. Twenty four patients (6.4%) were found to have a "balloon uncrossable" CTO. These patients had similar clinical and angiographic characteristics compared to the other CTO PCI patients (Table). Successful crossing of the lesion was achieved in 22 of 24 patients (91.7%) using a variety of techniques, such as successive balloon inflations (43.5%), microcatheter advancement (21.7%), laser (8.7%), increased guide catheter support (13.0%), and subintimal lesion crossing (13.0%) (Figure). Patients with “balloon uncrossable” CTOs had longer procedure time (184.5 ± 77.9 vs. 134.0 ± 69.0 min, p<0.01), fluoroscopy time (55.2 ± 24.9 vs. 37.9 ± 20.8 min, p<0.01), and received higher contrast volume (404.4 ± 137.9 vs. 351.7 ± 138.5 ml, p=0.085), but had similar incidence of major complications (8.3% vs. 3.2%, p=0.25) compared to patients without “balloon uncrossable” CTOs.

Conclusion:
“Balloon uncrossable” CTOs occur in 6.4% of contemporary CTO PCIs and can be successfully treated in most patients using a variety of techniques.
Abstract 13493: Complete Disruption of All Nitric Oxide Synthase Genes Markedly Reduces Cerebral Infarct Size after Middle Cerebral Artery Occlusion in Mice
Authors: Tsutsui Masato , Kubota Haruaki , Noguchi Katsuhiko , Matsuzaki Toshihiro , Hattori Fumiya , Sakanashi Mayuko , Kina-Tanada Mika , Uchida Taro , Nakasone Junko , Shimokawa Hiroaki , Ohya Yusuke , Sugahara Kazuhiro , Kakinohana Manabu
Published: July 2021

Background:
The role of each nitric oxide synthase (NOS) isoform in the pathogenesis of cerebral infarction has been studied in individual NOS isoform-deficient mice. It has been reported that, in a model of middle cerebral artery occlusion (MCAO), neuronal and inducible NOSs exacerbate cerebral infarction, whereas endothelial NOS conversely alleviates cerebral infarction. Although the role of the whole NOSs system in cerebral infarction has been examined in pharmacological studies with non-selective NOS inhibitors, the results are quit inconsistent, possibly because of non-specificity of the agents. In this study, we addressed this point in mice in which all three NOS genes are completely disrupted.

Method and Results:
We newly generated triple NOSs-deficient mice and wild-type littermates by crossbreeding single NOS
-/-
mice. Transient (1 hour) and permanent MCAO was performed in male triple NOSs
-/-
and wild-type mice at 8-12 weeks of age (n=9-11). Cerebral infarct size was evaluated by triphenyltetrazolium chloride (TTC) staining. There was no anatomical difference in the structure of cerebral arteries between the triple NOSs
-/-
and wild-type mice (n=3 each). Reductions in cerebral blood flow during MCAO were identical between the two mice. However, cerebral infarct size at 24 hours after transient MCAO was markedly smaller in the triple NOSs
-/-
than in the wild-type mice (
P
<0.05, n=8-12). Cerebral infarct size at 24 hours after permanent MCAO was also markedly decreased in the triple NOSs
-/-
mice as compared with the wild-type mice (
P
<0.05, n=8-10). In addition, neurological deficit was significantly less and the survival rate was significantly better in the triple NOSs
-/-
mice compared with the wild-type mice (both
P
<0.05, n=8-12).

Conclusions:
These results provide the first evidence that complete disruption of all NOS genes markedly reduces cerebral infarct size after MCAO in mice, demonstrating a novel injurious role of the entire NOSs system in cerebral infarction. Inhibition of the NOSs system may be a novel therapeutic option in the treatment of cerebral infarction.
Abstract 13545: MicroRNA-33b Knock-in Mice for an Intron of Sterol Regulatory Element-Binding Factor 1 (Srebf1) Exhibit Reduced HDL-C in vivo
Authors: Horie Takahiro , Nishino Tomohiro , Baba Osamu , Kuwabara Yasuhide , Nakao Tetsushi , Nishiga Masataka , Usami Shunsuke , Izuhara Masayasu , Nakazeki Fumiko , Ide Yuya , Koyama Satoshi , Yokode Masayuki , Kita Toru , Kimura Takeshi , Ono Koh
Published: July 2021

Background:
MicroRNAs (miRs) are small non-protein-coding RNAs that bind to specific mRNAs and inhibit translation or promote mRNA degradation. Recent reports, including ours, indicated that miR-33 (miR-33a) located within the intron of sterol regulatory element-binding protein (SREBP) 2 controls cholesterol homeostasis and can be a possible therapeutic target for treating atherosclerosis. Primates, but not rodents, express a second miR-33 gene (miR-33b) from an intron of SREBF1. To address miR-33b function in vivo, we developed humanized mice, in which a miR-33b transgene is inserted within a Srebf1 intron.

Methods and Results:
The human miR-33b sequence was introduced into intron 16 of mouse Srebf1 by conventional gene targeting methods, because miR-33b is located in intron 16 of human SREBF1 and there are high homologies in exons 16 and 17 between human and mouse. We successfully established miR-33b knock-in (KI) mice with C57BL/6 background and this miR-33b KI strategy did not alter Srebf1 intron 16 splicing, which was confirmed by RT-PCR and sequencing. The expression of miR-33b in miR-33b KI hetero mice were almost half of those in miR-33b KI homo mice. An LXR agonist T0901317, which induces Srebf1 expression, enhanced miR-33b expression in primary hepatocytes and the liver of miR-33 KI homo mice. The protein levels of known miR-33a target genes, such as ABCA1, ABCG1, and SREBP-1, were reduced compared with those in wild-type mice. Peritoneal macrophages from the miR-33b KI mice had a reduced cholesterol efflux capacity via apoA-I and HDL-C. Serum HDL-C levels were reduced by almost 35% even in miR-33b KI hetero mice compared with wild-type mice. HPLC elution analysis showed that the decreased HDL levels were mainly composed of very large-, large-, medium sized HDL, which was compatible with the previous results of miR-33a deficient mice.

Conclusions:
miR-33b KI mice for an intron of Srebf1 showed reduced HDL-C levels. These results indicate that miR-33b can be a potential target for raising HDL-C in humans and may account for lower HDL-C levels in humans than those in mice. These mice will aid in elucidating the roles of miR-33s in different disease models and in screening of the drugs that can alter miR-33a and miR-33b levels and activities.
Abstract 13593: Lower Plasma Omega-6 Polyunsaturated Fatty Acid Level and Malnutrition in Acute Heart Failure Patients and Its Association With Worse Clinical Outcome
Authors: Nagai Toshiyuki , Yamamoto Yoshiya , Sugano Yasuo , Yamane Takafumi , Shibata Tatsuhiro , Nakamura Kenji , Iwakami Naotsugu , Chinen Daigo , Asaumi Yasuhide , Aiba Takeshi , Noguchi Teruo , Ishihara Masaharu , Kusano Kengo , Ogawa Hisao , Yasuda Satoshi , Anzai Toshihisa
Published: July 2021

Backgrounds:
Malnutrition was reported to be associated with poor clinical outcome in acute heart failure (AHF) patients. Plasma omega-6 (n-6) polyunsaturated fatty acids (PUFA) were parts of the essential fatty acids absorbed with intestine, and its absorption impairment could occur in right side of heart failure (HF). Thus, we aim to investigate whether the n-6 PUFA level could be a marker for malnutrition and a determinant of prognosis in patients with AHF.

Methods:
We examined 320 consecutive AHF patients who admitted to our institution between January 2013 and January 2014 from prospective registry. Patients with acute coronary syndrome and without complete data-set at admission were excluded. Finally, 284 patients were divided into two groups according to the plasma n-6 PUFA level as follows; Group A [lower plasma n-6 PUFA (dihomo-gamma-linolenic acid plus arachidonic acid) (below 163 μg/ml, the median), Group B [higher plasma n-6 PUFA (above 163 μg/ml) level] at admission. Adverse events were defined as worsening HF and death.

Results:
During a mean period of 231 days, adverse events were occurred in 78 patients (27 %). Group A had higher incidence of adverse events compared with group B (35% vs 20%, P<0.01). Group A had lower blood pressure, hemoglobin, plasma omega-3 (n-3) PUFA levels, higher age and serum creatinine level on admission than group B. Regarding right HF sign, group A had higher prevalence of jugular vein distention (P=0.04), hepatomegaly (P<0.01) and higher bilirubin level (P<0.01) than group B. Furthermore, group A had lower serum total cholesterol (P<0.01), triglyceride (P=0.02), and albumin (P<0.01) levels as malnutrition than group B. In multivariate Cox proportional hazards model, lower plasma n-6 PUFA level (HR 1.62, 95% CI 1.01-2.65), higher serum creatinine (HR 1.83, 95% CI 1.02-3.59), and higher plasma BNP levels (HR 1.60, 95% CI 1.02-2.57) were independent determinants for adverse events, among variables including age, blood pressure, hemoglobin, plasma n-3 PUFA, serum albumin levels.

Conclusions:
Lower plasma n-6 PUFA level was an independent determinant for adverse events in association with right HF sign and malnutrition, suggesting plasma n-6 PUFA could be a potential surrogate marker for clinical outcome in AHF patients.
Abstract 13599: Combined Aerobic and Resistance Exercise Training Decrease Oxidative Stress by Augmenting Antioxidant Potential in Patients with Cardiovascular Disease
Authors: Yoshida Noriko , Shiotani Emiko , Niiyama Hiroshi , Harada Haruhito , Kato Atsushi , Nishiyama Yasuhiro , Ikeda Hisao
Published: July 2021

Introduction:
Oxidative stress plays important roles in pathophysiology of atherosclerotic disease. However, little is known about the effects of aerobic exercise and resistance exercise on the oxidant offense and antioxidant defense systems in cardiovascular disease (CVD).

Hypothesis:
We hypothesized that combined aerobic and resistance exercise training decrease oxidative stress in patient with CVD.

Methods:
Thirty-two patients with CVD refer to in-hospital cardiac rehabilitation were divided to aerobic exercise (AE) group (n=15) and combined aerobic and resistance exercise (AE+RE) group (n=17). AE sessions consisted of 50 minutes of exercise (anaerobic threshold level), 5 days/week during training period (mean 27 days). Resistance exercise (30% of 1-repetition maximum) was performed 3 days/week before AE sessions. Serum reactive oxygen metabolites (d-ROMs) and biological antioxidant potential (BAP) were measured using the FRAS4 (Diacron International, Italy) before and after training period. BAP/d-ROMs ratio was used as an indicator of oxidative stress.

Results:
No significant differences in baseline values of age, body mass index, exercise tolerance and BAP/d-ROMs ratio were observed between the AE+RE group and the AE group. After training period, exercise tolerance (the AE group: 5.1 ± 1.0 to 5.6 ± 1.0 METs, p<0.01, the AE+RE group: 5.1 ± 1.1 to 5.7 ± 1.0 METs, p<0.01) were significantly increased in both groups. In the AE+RE group, d-ROMs decreased and BAP significantly increased (2173 ± 268 to 2338 ± 182, p<0.05). Consequently BAP/d-ROMs ratio significantly increased (7.3 ± 1.5 to 8.9 ± 3.1, p<0.05) after training period. In the AE group, d-ROMs significantly decreased (374 ± 61 to 347 ± 61, p<0.05), and BAP trends to decreased (2389 ± 201 to 2279 ± 189, p<0.1). Consequently BAP/d-ROMs ratio did not significantly change after training period. BAP/d-ROMs ratio in the AE+RE group were significantly higher than those in the AE group (8.9 ± 3.1 vs 6.8 ± 1.3, p<0.05) after training period.

Conclusions:
The present findings suggest that combined aerobic and resistance exercise training decrease oxidative stress by augmenting antioxidant potential in patients with CVD.
Abstract 13607: Differential Response of Local Coagulation after Implantation of New Generation Drug-Eluting Stents Compared with Sirolimus-Eluting Stent
Authors: Yamaguchi Koji , Wakatsuki Tetsuzo , Takashima Akira , Bando Mika , Bando Sachiko , Matsuura Tomomi , Ise Takayuki , Hotchi Jyunko , Tobiume Takeshi , Yagi Shusuke , Iwase Takashi , Yamada Hirotsugu , Soeki Takeshi , Sata Masataka
Published: July 2021

Purpose:
We have previously demonstrated local persistent hypercoagulation after sirolimus-eluting stent (SES) implantation by measuring plasma prothrombin fragment F1+2(frF1+2) levels. The aim of this study is to examine local coagulation response after biolimus-eluting stent (BES) and everolimus-eluting stent (EES) implantation.

Methods:
Sixty-eight patients who were treated with stents about eight months earlier to the mid-segment of the left anterior descending coronary artery, with no evidence of restenosis, were studied (BES: 12pts, EES: 15pts, SES: 23pts, and BMS: 18pts). We measured plasma levels of frF1+2 sampled in coronary sinus (CS) and sinus of Valsalva (V). The transcardiac frF1+2 gradients (ΔfrF1+2) were defined as CS level minus V level. The patients were divided into 2 groups according to total stent length [longer stent group (≥24mm) or shorter stent group (<24mm)].

Results:
A larger percent diameter stenosis was observed in the BMS group than in the SES, EES, and BES groups (25.1±15.6 vs 7.1±16.5, 12.5±12.8, 10.3±14.2%, p<0.05, respectively). The ΔfrF1+2 was greater in the SES group than in the BMS, EES, and BES groups, and no significant differences were observed among the BMS, EES, and BES groups (23.4±21.1 vs 4.7±13.4, 3.2±11.7, 1.5±11.1 pmol/l, p<0.05, respectively). The ΔfrF1+2 in longer stent group (n=11) was significantly greater than in shorter stent group (n=12) (27.4±12.1 vs 19.2±10.1 pmol/l, p<0.05) only in the SES group, however, no significant differences were observed according to total stent length in other groups.

Conclusions:
A lower response of local coagulation after BES and EES implantation was shown, and significant correlation between local hypercoagulation and total stent length was observed only after SES implantation. These findings might be associated with lower strut thickness and unique characteristics of polymer in second-generation drug-eluting stents.
Abstract 13636: Derivation and Validation of a Clinical Prediction Rule for Sleep Disordered Breathing in Patients With Cardiovascular Disease
Authors: Miyazaki Takashi , Kojima Sunao , Matsui Kunihiko , Kamijikkoku Shuichi , Hokimoto Seiji , Honda Takashi , Ogawa Hisao
Published: July 2021

Introduction:
There is a growing demand for polysomnography (PSG) indispensable for the diagnosis of sleep apnea which is associated with the development of cardiovascular disease.

Hypothesis:
We assessed the hypothesis that a prediction rule without PSG could identify obstructive sleep apnea (OSA) patients in which the continuous positive airway pressure (CPAP) treatment was recommended.

Methods:
Consecutive 326 cardiovascular disease patients simultaneously underwent a full-night PSG and a digital-installed built-in memory pulse-oximeter. A logistic regression-based prediction rule was developed in patients at university hospital (n=278) and validated in patients at community hospital (n=48).

Results:
In the derivation group, apnea-hypopnea index (AHI) was correlated with 3% oxygen desaturation index (ODI) (r=0.94, P<0.0001) and 223 patients were diagnosed as sleep disordered breathing (SDB) (AHI >=5). Four independent parameters were identified to distinguish between OSA (n=161) and central sleep apnea patients (n=62): pulse pressure, left atrial diameter, minimum oxygen saturation and B-type natriuretic peptide levels. The observed probability of OSA, by calculated score, was comparable between derivation and validation groups (Figure 1). The model-based rule was well calibrated (Hosmer-Lemeshow χ2=4.23) and demonstrated good discrimination in the derivation group. Discriminatory ability was also reasonable in the validation group and the difference between these groups was limited (Figure 2). Provided that cutoff levels of 3%ODI and 4 independent parameters were entirely satisfied, positive predictive value was 100% in both groups for the diagnosis of OSA necessary for CPAP treatment.

Conclusions:
A prediction rule for the type of sleep apnea without PSG can quantify the difference of pathophysiological mechanism in SDB patients. This index may be useful in detecting eligible candidates for CPAP in cardiovascular disease patients.
Abstract 13672: High Frequency Echocardiography and Speckle Tracking Based Strain Analysis Revealed Delayed Functional Recovery After Myocardial Cryoinjury in Adult Zebrafish
Authors: Hein Selina , Lehmann Lorenz , Katus Hugo A , Hassel David
Published: July 2021

Zebrafish is a widespread used model in cardiovascular research, and recently adult zebrafish, as a more human relevant model compared to larval stages, became particularly valuable to study heart regeneration. While cardiac performance in larval stages can be easily assessed using transillumination microscopy, methods to reliably assess cardiac function in adult zebrafish are largely missing.

Here, we developed the first standardized protocol to reliably assess cardiac function in adult zebrafish using high frequency echocardiography and speckle-tracking algorithms. Respecting distinct anatomic characteristics in zebrafish, we defined three examination planes according to standards in human echocardiography. Further, by combining conventional echocardiographic measurements with modern speckle tracking based strain analysis we are able to measure myocardial performance in extremely high spatial and temporal resolution. Thereby we were able to attain high quality B-Mode imaging and PW-Doppler signals allowing distinct detection of changes in chronotropy, inotropy and, for the first time in adult zebrafish, in dromotropy in response to treatment with Atenolol and Isoproterenol. Additionally, we employed zebrafish that underwent cardiac cryoinjury to longitudinally follow functional cardiac regeneration. High-resolution speckle-tracking strain analysis allowed us to demarcate injured myocardial regions and demonstrate that functional healing extends beyond 30-45 days post injury (dpi) as previously implied by conventional methodologies. Specifically, we found delayed restitution of cardiac displacement and re-synchronization of injured and non-injured myocardium beyond 60 dpi until 120 dpi. In summary, our protocol enables highly reproducible and high throughput measurements of cardiac performance in adult zebrafish in a high spatio-temporal resolution. Thereby, our protocol represents a valuable novel tool for functional cardiac assessment and to detect changes in cardiac function in response to gene mutation or myocardial injury and might lead to further insights into cardiac physiology and disease.
Abstract 13688: Long Chain Fatty Acid Composition Modulated by Elovl6 Plays a Critical Role in Vascular Smooth Muscle Cell Proliferation and Neointimal Formation
Authors: Matsui Hiroki , Sunaga Hiroaki , Anjo Saki , Syamsunarno Mas Rizky A , Iso Tatsuya , Matsuzaka Takashi , Shimano Hitoshi , Yokoyama Tomoyuki , Kurabayashi Masahiko
Published: July 2021

Introduction:
Elovl6, the elongase of long chain fatty acids 6, is a rate-limiting enzyme catalyzing the elongation of saturated and monounsaturated fatty acids with 12, 14 and 16 carbons. Our recent study showed that Elovl6 is abundantly expressed in vascular smooth muscle cells (VSMC) and is dramatically induced in neointima in rat.

Hypothesis:
In this study, we tested the hypothesis that changes of fatty acid (FA) composition by Elovl6 affects the proliferation of VSMC and contributes to neointimal formation in vivo.

Methods and Results:
Abundant Elovl6 expression was observed in mice femoral artery at 2 weeks after wire-injury and in intimal thickening lesion of human coronary artery. Furthermore, Elovl6 mRNA expression in cultured human aortic SMC (HASMC) was significantly increased by platelet-derived growth factor-BB (2.4-fold, p<0.05) or hypoxic stress (6.7-fold, p<0.01) in a dose- or time-dependent manner. Furthermore, knockdown of Elovl6 expression in HASMC markedly suppressed cell proliferation (16%, relative to control, p<0.01) and migration, concomitantly induced the expression of p21 and phospholyration of AMP-activated protein kinase (p-AMPK) and suppressed mTOR expression. Consistent with in vitro data, Elovl6 deficient (Elovl6 -/-) mice at 2 weeks after injury showed markedly suppressed neointimal formation compared with wild-type (WT) mice (intima/media ratio: WT, 1.4 ± 0.6; Elovl6 -/-, 0.5 ± 0.2; Ki67-positive cells: 0.2 fold relative to WT mice; N=6-7, p<0.05). Of an importance, analysis of FA composition in SMC isolated from Elovl6 -/- mice showed that high levels of palmitic acid and low levels of oleic acid were detected as compared with that from WT mice. In accordance with these results, exogenous treatment of palmitic acid in SMC substantially suppressed cell proliferation (42%, relative to control, p<0.01) and migration, induced p21 and p-AMPK expressions. Conversely, these effects were blunted by adenovirus-mediated Elovl6-overexpression or exogenous oleic acid treatment.

Conclusions:
Collectively, our study demonstrates that proliferation of VSMC is tightly regulated by FA composition modulated by Elvlo6, offering a novel therapeutic target for arterial proliferative disease in which VSMC plays a key role.
Abstract 13760: Dipeptidyl-Peptidase-4 Inhibitor, Des-Fluoro-Sitagliptin, Prevented the Progression of Established Atheromatous Plaques in ApoE Deficient Mice - Possible Involvement of AMP-Activated Protein Kinase Pathway -
Authors: Kurokawa Hirofumi , Sugiyama Seigo , Sugamura Koichi , Matsubara Junichi , Fujisue Koichiro , Ohba Keisuke , Maeda Hirofumi , Yamamoto Eiichirou , Tsujita Kenichi , Kaikita Koichi , Hokimoto Seiji , Ogawa Hisao
Published: July 2021

Background:
We demonstrated that dipeptidyl-peptidase-4 inhibitor (DPP4i) Des-Fluoro-Sitagliptin (DFS) exhibited anti-atherosclerotic effects on the newly development process of atherosclerosis, however it is unknown whether DPP4i also have anti-atherosclerotic effects on the established atheroma (preventing progression or regression). In the practical situation, it is clinically important to investigate whether DPP4i have beneficial effects on the established atherosclerosis.

Methods and Results:
8 weeks old ApoE deficient mice were fed by high fat diet (HFD) for 12 weeks lead to development of the established atherosclerotic plaques, followed by randomized into 4 groups; continued HFD arm [HFD alone, HFD+DFS (200mg/kg/day)] and lipid-lowering arm [low fat diet (LFD) alone, LFD+DFS], for additional 8 weeks. DFS significantly reduced the progression of established plaque compared to HFD alone [plaque area per aorta; HFD (n=11): 53.3±4.9% vs. HFD+DFS (n=15): 43.8±9.0%, p<0.05]. During the lipid-lowering therapy by diet, DFS supported the diet-induced regression of atherosclerosis [baseline 20 weeks old (n=10): 34.6±11.4%, LFD (n=17): 32.7±7.8% vs. LFD+DFS (n=18): 30.2±9.1%]. In vitro, DFS with glucagon-like peptide-1 (GLP-1) increased the phosphorylation of AMPK in human coronary artery endothelial cells (HCAEC) (2.71±0.31 fold, p<0.01) and in THP-1-drived human macrophages (1.29±0.07 fold, p<0.05). DFS with GLP-1 reduced the expression of adhesion molecules induced by tissue necrosis factor-α (intercellular adhesion molecule-1: -18.1±2.6%, p<0.05, vascular cell adhesion molecule-1: -22.4±4.5%, p<0.05), and these effects were abolished by AMPK inhibitor (Compound-C; CC) in HCAEC. In macrophages, DFS with GLP-1 reduced expression of the M1 pro-inflammatory factor (interleukin (IL)-1β; 0.74±0.04 fold, p<0.05) but not the M2 anti-inflammatory factor (IL-10; 1.00±0.06 fold, p=0.99) and the effect on M1 was abolished by CC.

Conclution: DFS exhibited beneficial effects on the established atherosclerosis with attenuating progression and supporting regression by the lipid-lowering therapy. AMPK activation by DFS might partly play a role in these processes. DFS could also be effective for the established atherosclerosis.
Abstract 13770: DWS as an Index of Myocardial Stiffness and a Predictor of Heart Failure Outcome in Patients with Chronic Beta-blockade Therapy
Authors: Soyama Yuko , Mano Toshiaki , Hirotani Shinichi , Masaki Mitsuru , Fukui Miho , Fujiwara Shohei , Sugahara Masataka , Komamura Kazuo , Kawabata Masaaki L , Tsujino Takeshi , Masuyama Tohru
Published: July 2021

Background:
Diastolic dysfunction determines symptoms and prognosis in patients with left ventricular (LV) dysfunction. Diastolic wall strain (DWS) is associated with poor outcomes in heart failure with preserved ejection fraction. However, the utility of DWS is still unknown in heart failure with reduced ejection fraction (HFrEF). Our aim is to determine whether DWS is predictive of the outcome in HFrEF.

Methods:
We studied 54 HFrEF patients (LVEF<50%) and followed DWS as an index of myocardial stiffness for 6 months after the induction of beta blockade (Bisoprolol 2.5-10 mg / day). DWS was determined in the LV M-mode echocardiogram using the following equation: DWS = {(LV posterior wall thickness at end-systole - LV posterior wall thickness at end-diastole) / LV posterior wall thickness at end-systole}. We followed for 7years after the induction of beta-blockade.

Results:
DWS increased after the induction of beta-blockade (0.32±0.11 vs 0.25±0.12,p<0.05). DcT, EF and E’ also increased after the induction of beta-blockade. HR at rest and log BNP decreased following beta blockade. Patients with DWS ≤ median (0.25) before the induction of beta-blockade had higher rate of HF hospitalization than those with DWS >median during 7 years (Log-rank p =0.025). DcT, EF, E’, HR at rest and log BNP before the induction of beta blockade were not significant predictors of HF outcome (Log-rank p=0.263, 0.504, 0.0796, 0.289 and 0.877) respectively.

Conclusions:
Induction of beta-blockade provided an improvement in DWS. DWS might be useful as an index of myocardial stiffness to predict the outcomes in HFrEF patients with chronic beta-blockade therapy.