bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 63: A Regulatory Enhancer Variant at the Hypertensive GRAF3 Locus Increases SMC-Selective Expression of GRAF3 by Promoting SRF Binding
Authors: Mangum Kevin , Taylor Joan , Mack Christopher
Published: March 2022

We have recently shown that the Rho-specific GAP, GRAF3, is specifically expressed in smooth muscle cells (SMC) in mouse and humans and negatively regulates blood pressure by limiting Rho-dependent SMC contractility. Genome wide association studies identified a hypertension-associated locus within the first intron of the GRAF3 gene (currently defined by SNPs rs633185 and rs604723) and demonstrated that the minor allele was significantly correlated with decreased blood pressure in patients. The aims of the current study were to identify transcription mechanisms that regulate SMC-specific GRAF3 expression and to test the hypothesis that SNPs within the GRAF3 hypertension locus affect blood pressure by altering GRAF3 expression and RhoA activity. Using DNaseI Hypersensitivity of chromatin structure, ChIP-seq approaches for previously implicated transcription factors, and information from sequence/regulatory databases, we identified two regions within the human GRAF3 gene that exhibited strong SMC-selective transcription activity in cultured cells. Importantly, the enhancer at +52 Kb contains SNP rs604723 and mutation of the major C allele to the minor T allele significantly increased enhancer activity from 20- to 40-fold over promoterless. The DNA surrounding the minor T allele conforms to the consensus SRF binding element at 8 out of 10 residues. Gel shift and targeted ChIP assays revealed preferential binding of SRF to the minor allele sequence, and the SRF co-factor, myocardin, significantly transactivated the minor but not the major allele enhancer (p<0.05). Allele-specific quantitative RT PCR on RNA isolated from human aortic SMC heterozygous at rs604723 revealed T allele-specific mRNA expression. Finally, data within the Genotype-Tissue Expression (GTEx) database indicated that GRAF3 RNA levels in tibial artery samples were 3-fold higher in patients homozygous for the minor T allele compared to patients homozygous for the major C allele (p<1.5e-10). Taken together, our data indicate that the minor T allele at rs604723 upregulates GRAF3 transcription via an SRF-dependent mechanism and provides the first functional link between this SNP and blood pressure regulation.
Abstract 682: Pharmacological Inhibition of COX and LOX Pathways Leads to Substrate Rediversion: A Novel Plasma Lipidomics Assay to Screen Drugs in Human Blood
Authors: Mazaleuskaya Liudmila L , Ricciotti Emanuela , Lawson John A , Li Xuanwen , Grant Gregory , Grosser Tilo , FitzGerald Garret A
Published: March 2022

Cardiovascular complications from cyclooxygenase (COX)-2-selective nonsteroidal anti-inflammatory drugs (NSAIDs) prompted interest in microsomal prostaglandin synthase 1 (mPGES-1), downstream of COX-2, as an alternative drug target. Unlike COX-2, global deletion of mPGES-1 in mice suppresses prostaglandin E2 (PGE2) and augments prostacyclin PGI2 through substrate rediversion, and does not predispose to thrombogenesis and hypertension. However, after cell-specific deletions of mPGES-1, the predominant substrate rediversion product amongst the prostaglandins varies by cell type. We hypothesized that inhibition of mPGES-1 leads to changes in bioactive lipids beyond the prostaglandin pathway, and that substrate rediversion associated with a more favorable cardiovascular profile after mPGES-1 vs COX-2 inhibition can be translated from mouse to human studies. In this project, we aimed 1) to develop a mass spectrometry-based, broad-spectrum lipidomics approach to measure arachidonate-derived products additional to prostanoids; and 2) to work out an in vitro human whole blood assay (hWBA) to study substrate rediversion upon mPGES-1 vs COX-2 inhibition. We have developed a novel, reproducible analytical method to scan the impact of drugs on the bioactive lipidome. Using an in vitro hWBA and HPLC-MS/MS lipidomics analysis, we observed substrate rediversion in both COX and LOX pathways after mPGES-1 inhibition, which was distinct from inhibiting COX-1/2. We further optimized this assay to screen combinations of inhibitors from COX and lipoxygenase (LOX) cascades. Overall, we have shown that a clear signature of inhibiting diverse targets - mPGES-1, COX and LOX enzymes - can be readily detected from experiments of small samples sizes, and discovered novel aspects of drug effect due to substrate rediversion. Our findings contribute to the comparative cardiovascular safety of inhibiting mPGES-1 vs COX-2 and demonstrate the impact of combining mPGES-1 with LOX inhibitions on the hemostatic system
Abstract 695: Low-density Lipoprotein Receptor-related Protein in the Endothelium Regulates Metabolic Responses
Authors: Lockyer Pamela , Mao Hua , Pi Xinchun
Published: March 2022

The endothelium, a biologically active barrier, is the site of metabolic exchange between circulation and underlying tissues. Endothelial dysfunction is a common feature in obesity, dyslipidemia and diabetes. Yet the roles endothelial cells play in systemic metabolic responses are unclear. A recent study showed that PPARγ in the endothelium regulates metabolic responses to high fat diet in mice, suggesting that endothelial cells may play a direct role in regulation of obesity, lipid metabolism and insulin sensitivity. Our studies demonstrate that Low-density lipoprotein receptor-related protein 1 (LRP1), a member of LDL receptor family, influences many biological processes such as lipid metabolism, endocytosis and signal transduction, is required for endothelial functions. More interestingly, we discovered that LRP1 interacts with and positively regulates PPARγ activity. Therefore, we wanted to investigate the role of endothelial cell-specific LRP1 in the regulation of systemic metabolic responses. To do this, we used mice deficient in LRP1 in the endothelium and fed a high-fat diet. After 12 weeks of feeding, we found LRP1EC-KO mice to be more sensitive to insulin following a glucose bolus, demonstrated by decreased glucose level in serum (LRP1EC-KO=114.4±15.59 mg/dL glucose; LRP1EC-WT=157.60±14.64 mg/dL glucose). These LRP1EC-KO mice exhibited a smaller increase in body weight (LRP1EC-KO=34.65±2.996g; LRP1EC-WT=44.55±1.752g) compared to wild-type mice following high fat food feeding. In addition to these findings, The composition of white adipose tissue such as the inguinal fat in LRP1EC-KO mice are only 3.9±1.42% of total body weight compared to 6.37±0.342% of total body weight in LRP1EC-WT mice. The levels of circulating leptin and resistin, factors associated with adipogenesis, were also significantly different, with leptin at 7419.25±2031.197 pg/ml in LRP1EC-KO; 16496.2±1921.146 pg/ml in LRP1EC-WT mice; and resistin at 6033.55±3034.54pg/ml in LRP1EC-KO; 17382.6±2894.95pg/ml in LRP1EC-WT mice. Taken all together, our data demonstrate a previously unrecognized role for endothelial cell-specific LRP1, very similar to PPARγ, in regulating systemic metabolic responses, thus expanding the functional roles of endothelium.
Abstract 704: Glycation of Vitronectin Inhibits VEGF-induced Angiogenesis by Uncoupling VEGF Receptor-2-αvβ3 Integrin Cross-talk
Authors: Wang Liqun , Li Yongjie , Chen Ni , Wu Jianbo
Published: March 2022

Objective:
Glycation of proteins of the vessel wall is thought to play an important role in the pathogenesis of vascular complications in diabetes mellitus. However, no previous study has implicated glycated vitronectin (VN) in controlling vascular endothelial growth factor (VEGF) signaling. Therefore, the goal of this study was to investigate the effects of methylglyoxal (MGO)-glycated VN on VEGF signaling. We tested the hypothesis that glycation of VN down-regulates VEGF receptor-2 (VEGFR-2) activation by uncoupling interaction between VEGFR-2 and αvβ3.

Approach and Results:
MGO-glycated or unmodified vitronectin were used as substrate for human umbilical vein cells (HUVECs). We studied the effects of glycated VN on VEGF signaling in HUVECs. Glycation of VN inhibited VEGF-induced phosphorylation of VEGFR-2 and intracellular signaling pathway downstream of VEGFR-2. Glycated VN inhibited the binding of VEGFR-2 to β3 integrin and phosphorylation of β3 integrin. LM609 (anti-αvβ3 antibody) inhibited VEGF-induced activation of VEGFR-2 in HUVECs grown on VN. Glycation of VN also significantly decreased VEGF-induced HUVECs migration in vitro and vessel outgrowth in ex vivo angiogenesis model.

Conclusions:
Glycation of VN inhibits VEGF-induced VEGFR-2 activation by uncoupling VEGFR-2-αvβ3 integrin cross-talk. Glycation of VN causes a reduction in endothelial cells migration and vessel outgrowth. This may provide a mechanism for the failure of collateral sprouting in diabetic microangiopathy.
Abstract 720: Edaravone Reduces Kidney Damage Induced by Myonephropathic Metabolic Syndrome in Rats
Authors: Yamamura Mitsuhiro , Mitsuno Masataka , Tanaka Hiroe , Ryomoto Masaaki , Fukui Shinya , Kajiyama Tetsuya , Miyamoto Yuji
Published: March 2022

Background:
Free radicals cause myonephropathic metabolic syndrome (MNMS), which damages not only muscles but also kidneys. At ATVB 2014, we previously reported edaravone (Radicut®, Mitsubishi Tanabe Pharma Co., Japan), the free radical scavenger may suppress kidney damage induced by MNMS.

Objective:
In this study, we evaluated whether edaravone suppresses MNMS-induced kidney damage, focusing on active neutrophils and tubular function.

Methods:
Either 3.0 mg/kg of edaravone (edaravone group; n = 4), or saline (MNMS group; n = 6) was intraperitoneally injected into male Lewis male rats (508 ± 31 g). MNMS models were induced, by clamping the bilateral common femoral arteries for 5 hours, followed by de-clamping after more 5 hours. The both kidneys were stained with naphthol AS-D chloroacetate esterase to detect active neutrophils and with Periodic Acid Schiff (PAS) to assess glycogen storage. Normal kidneys were also stained as control (n = 3). Kidney damage was evaluated by counting the number of chloroacetate esterase-positive cells (neutrophil infiltration) and the percentage of PAS-positive area (tubular function).

Results:
The number of chloroacetate esterase-positive cells was significantly increased in the MNMS group than that in normal kidneys (62.4 ± 5.6 vs. 17.5 ± 2.4 cells/mm2, p<0.01). However, the number of chloroacetate esterase-positive cells was significantly reduced in the edaravone group (32.8 ± 5.7 cells/mm2, p<0.01, Figure attached). The percentage of PAS-positive area in the MNMS group was significantly decreased than that in normal kidneys (36.9 ± 2.6 vs. 66.9 ± 1.2%, p<0.01). The percentage of PAS-positive area was maintained in the edaravone group (52.9 ± 0.9 %, p<0.01, Figure attached).

Conclusions:
Edaravone may reduce neutrophil infiltration around the glomerulus and tubular dysfunction induced by MNMS.
Finalists, The Jeremiah and Rose Stamler Research Award for New Investigators Fatty fish consumption and ischemic heart disease mortality in older adults: The Cardiovascular Health Study
Authors: Mozaffarian Dariush , Lemaitre Rozenn N , Kuller Lewis H , Fried Linda P
Published: March 2022

P01

OBJECTIVE. To assess whether dietary intake of fatty fish is associated with a reduced risk of ischemic heart disease mortality in older adults. BACKGROUND. Dietary intake of fatty fish has been associated with a reduced risk of fatal ischemic heart disease in middle-aged men and women. This association, however, has not been specifically evaluated in older adults. METHODS. In 3968 men and women ≥ age 65 enrolled in the Cardiovascular Health Study and free of known cardiovascular disease at baseline, dietary intake of fish (fried fish, tuna fish, and other fish) was assessed through a detailed picture-sort food frequency questionnaire. In a subset of 348 persons, dietary intake of tuna and other fish combined (“fatty fish”) correlated with plasma phospholipid levels of combined docosahexaenoic acid (DHA, 22:6 n-3)) and eicosapentaenoic acid (EPA, 20:5 n-3) (r = 0.40, p < 0.001); intake of fried fish did not correlate with plasma phospholipid levels of these long-chain n-3 fatty acids (r = 0.05, p = 0.37). Participants were prospectively followed for a mean period of 6.8 years for death from ischemic heart disease (incident fatal myocardial infarction [MI] plus coronary heart disease death) and nonfatal MI. FINDINGS. Compared to intake of less than one serving of fatty fish per week, intake of at least one serving of fatty fish per week was associated with a 35% lower risk of death from ischemic heart disease (hazard ratio [HR] 0.65; 95% confidence interval [CI] 0.45 to 0.94) but was not associated with a lower risk of nonfatal MI (HR 0.86; 95% CI 0.64 to 1.15), after adjustment for age, gender, diabetes, smoking status, education, and total caloric intake. CONCLUSIONS. These findings suggest that consumption of at least one serving of fatty fish per week is associated with a lower risk of death from ischemic heart disease in persons aged 65 years and older.
Changing Congestive Heart Failure Incidence in an Older Population, 1970-74 and 1990-94.
Authors: Barker William H , Getchell William , Mullooly John , Kimes Terry
Published: March 2022

P45

Congestive heart failure (CHF) mortality rates have increased dramatically in recent years, particulary among the elderly, but little is known about CHF incidence changes. This study compares CHF incidence between 1970-74 and 1990-94 in a large well-defined population of over 30,000 persons >65 y/o with access to complete medical records. Using Framingham Heart Study (FHS) criteria for confirming CHF we identified 387 and 1555 confirmed cases in the respective periods. We observed unchanging incidence rates of 7.6 and 7.9, and increasing incidence from 15.1 to 18.7 and 27.8 to 42.4 cases per 1000 person-years among persons 65-74, 75-84, and 85+ years old, respectively. In both time periods, 21% of potential cases were unconfirmed by FHS criteria. Onset of confirmed CHF occured as an outpatient in 16% and 22% of cases in the respective periods. Profiles of FHS major and minor diagnostic criteria were very similar for cases in the two timed periods. This study documents significant increase in CHF incidence among old-old(>75y/o) but not young old (65-74). Possible explanations are under study.
Pulse Pressure and Prevalence of Peripheral Arterial Disease: The Atherosclerosis Risk in Communities (ARIC) Study
Authors: Taylor Herman A. , Brown Andrew C. , Burchfiel Cecil M. , Jones Daniel W. , Skelton Thomas N. , Arnett Donna K. , Clemons Traci L. , Clark Bobby L. , Criqui Michael H.
Published: March 2022

P55

Recent reports have shown that pulse pressure (PP) is a meaningful independent predictor of some types of cardiovascular disease. However, the relationship between PP and peripheral arterial disease (PAD) is largely unexplored. We examine data from a large biracial cohort in order to determine whether PP has an independent association with PAD. A total of 10,592 subjects were examined from the ARIC study (20.5% African American) in 1987-1989. Participants taking anti-hypertension medications were excluded but those with untreated elevated (>=140/90) blood pressure (12.3%) were not. The presence of PAD was indicated by an ankle-brachial index <=0.9. Clinical predictors of PAD and potential confounders were examined individually for associations with PAD, after controlling for age. In a bivariate model, it was found that subjects in the highest quartile of PP (>52 mm Hg) were 2.27 times more likely to have PAD than those in the lowest quartile (<37 mm Hg). Controlling for age, sex, race, mean arterial pressure (MAP), diabetes mellitus, body mass index, high-density lipoprotein, cholesterol level, and smoking status in a multivariate logistic regression model, PP had a significant independent association with PAD. Every 10 mm Hg difference in PP was associated with a 41% increase in prevalence of PAD. PP was also shown to have a stronger association with PAD than either the non-significant systolic blood pressure (SBP) or non-significant diastolic blood pressure (DBP). Each of the three blood pressure components was analyzed independently in the above multivariate model, controlling for MAP. When SBP and DBP were included separately in the multivariate model (excluding MAP), every 10 mm Hg difference in PP was associated with a 73% and 31% increase in prevalence of PAD, respectively, whereas both SBP and DBP were not statistically significant. Conclusion: PP was independently associated with PAD in this population-based sample of subjects who were not being treated for hypertension.
Plasma Lipids, Lipoproteins, and Apolipoproteins and Incident Ischemic Stroke:
Authors: Shahar Eyal , Chambless Lloyd E , Rosamond Wayne D , Boland Lori L , Ballantyne Christie M , McGovern Paul G , Sharrett A. Richey
Published: March 2022

P63

The role of plasma lipids, lipoproteins, and apolipoproteins in the pathogenesis of ischemic stroke is unclear. We examined the association of incident hospitalized ischemic stroke with baseline plasma measurements of LDL-cholesterol, apoB, HDL-cholesterol, apoA-1, and triglycerides in a population-derived sample of 14,175 middle-aged men and women. Over an average follow up of 8.2 years, we documented 233 incident ischemic strokes (120 in men; 113 in women). After multivariable adjustment for stroke risk factors, categorical analysis revealed weak, inconsistent, and statistically non-significant associations between ischemic stroke and the five analytes. For example, the adjusted hazard ratios (95% CI) of ischemic stroke for the top fourth versus the bottom fourth were 1.17 (0.81-1.68) for LDL-cholesterol; 1.29 (0.90-1.87) for apoB; 0.90 (0.57-1.41) for HDL-cholesterol; 0.83 (0.55-1.26) for apoA-1; and 1.25 (0.82-1.92) for triglycerides. Sex-specific analysis (supported by evidence of statistical interaction) indicated lack of any association in men. In women, however, we observed moderate associations of ischemic stroke with LDL-cholesterol, apoB, and triglycerides, primarily restricted to the right tail of their distributions. Hazard ratios of ischemic stroke in women are shown in the

table below. These findings are compatible with the following hypotheses: 1) The effect of LDL-cholesterol, apoB, and triglycerides on ischemic stroke might differ qualitatively between men and women; 2) The pathogenesis of a substantial proportion of ischemic strokes (e.g., lacunar strokes) does not involve classical atherosclerotic mechanisms.

Table 1.
Alcohol Variability and Blood Pressure: Results from the Usual Care Group of the Multiple Risk Factor Intervention Trial (MRFIT)
Authors: Chee Deborah , Stamler Jeremiah
Published: March 2022

P84

Background: Previous studies suggest associations between pattern of alcohol consumption and blood pressure levels. However, there is little information on years-long patterns and blood pressure. Aim: Assess relationship of year-to-year variability in alcohol intake to blood pressure in alcohol users of the Usual Care (UC) group, Multiple Risk Factor Intervention Trial. Methods: 6,438 men from the MRFIT Usual Care group. Data on alcohol variability, blood pressure (BP) and confounding variables from five 24-hour dietary recalls and BP measurements at years 0,1,2,3, and 6. The intra-individual coefficient of variability (CV) in alcohol intake was computed for three sub-groups, defined by mean of reported intake: sub-group 1, less than 2 drinks/d; sub-group 2, 2-3 drinks/d; sub-group 3, more than 3 drinks/d. Multiple linear regression models --dependent variable blood pressure --were adjusted for mean alcohol intake, age, ethnicity, education, BMI, cigarette smoking, serum cholesterol, special diet, mean calories (in per day models), and mean dietary sodium and potassium intake. Results: Alcohol variability was inversely and significantly related to systolic and diastolic blood pressure in the whole group with alcohol expressed both as grams per day (e.g., for SBP: β= -0.026488, z=-2.132) and per cent energy (e.g., for SBP: β= -0.028594, z=-3.371). In sub-group 1 (n=5,128), the association was inverse but not significant; in sub-group 2 (n=1,073), the association was inverse and significant; in sub-group 3 (heavy drinkers) (n=237), the association was direct but not significant. Conclusions: Whereas year-to-year variability in reported alcohol intake was inversely related to blood pressure in men reporting less than 3 drinks/d, in heavy drinkers this evidence of erratic intake was associated with higher blood pressure.
Survival after Treatment during Hospitalization for Acute Myocardial Infarction:
Authors: Rosamond Wayne D. , Sullivan Michael D. , Chambless Lloyd E. , Wang Chin-Hua , Cooper Lawton S. , Folsom Aaron R.
Published: March 2022

0010

Background: The wealth of data from clinical trials has done as much to diversify standard therapy for acute myocardial infarction (MI) as it has to define it. To better characterize the use of treatments during hospitalization for MI in clinical practice, we analyzed one-year post MI survival in relation to common treatments in the Atherosclerosis Risk in Communities (ARIC) Study. Methods: Between 1987 and 1996 potential cases of MI among men and women, ages 35-74 were identified through hospital surveillance in Forsyth Co. NC, Jackson, MS, Minneapolis, MN, and Washington Co. MD. Diagnosis was validated using a standardized algorithm. Information on treatments was obtained from the medical record. Medication use included those given or continued during the hospitalization or prescribed at discharge. Cox proportional hazards models were used to estimate survival associated with treatments. Results: Of the 15,032 MI events, 34% were female and 18% black. One-year case-fatality of hospitalized MI events ranged from a high of 12% in 1987 to a low of 6.5% in 1994. Between 1987 and 1996, use of thrombolytic therapy (TT) increased from 11% to 16% of patients, beta-blockers (BB) 45% to 66%, and aspirin (AS) 49% to 87%. Coronary artery bypass graft (CABG) surgery increased from 13% to 19% and coronary angioplasty (PTCA) from 11% to 26%. After adjusting for age, race, sex, center, history of MI, severity indicators, complications, and the other therapies, AS (hazard ratio (HR) =0.47, 0.40-0.54) and PTCA (HR=0.47, 0.39-0.57) held the strongest association with survival, followed by CABG (HR=0.61, 0.50-0.75), BB (HR=0.62, 0.53-0.72), and TT (HR=0.71, 0.59-0.87). In 1996, the use of PTCA varied by center from 12% to 36% and AS use varied from 79% to 92%. Conclusions: Although not a randomized trial and therefore subject to selection bias, the ARIC data suggest that established treatments may be associated with better MI survival in community practice. Although no other therapy studied was more strongly associated with survival or has fewer contraindications than aspirin, it is underused in some communities.
Risk Factors for Progression of Common Carotid Atherosclerosis:
Authors: Chambless Lloyd E , Folsom Aaron R , Evans Gregory W
Published: March 2022

0005

Intima-media thickness (IMT) of the carotid arteries, as measured by B-mode ultrasound, is a marker of atherosclerosis, and as such has been shown to be positively associated with both prevalent and incident coronary heart disease (CHD) and incident stroke, and with risk factors for CHD. We examined the association of baseline risk factors or change in those risk factors with change in carotid IMT over an average nine years’ follow-up (1987-98) in the bi-ethnic population-based cohort of the Atherosclerosis Risk in Communities (ARIC) study (45-64 years old at baseline, n = 15,792). We found common carotid IMT to progress at 6 - 10 micrometers/ year, which is similar to other studies. We found, similar to other large epidemiologic studies, sizeable measurement error in IMT, which can cause large bias in estimating risk factor associations with change in IMT if adjusting for baseline IMT using standard methods. We concluded that adjustment for the potential confounding of baseline IMT, with proper correction for measurement error, generally had only a small effect on estimates of the risk factor associations. Either not adjusting for baseline IMT at all, or if doing so also correcting for measurement error by regression calibration, we found statistically significant (p < 0.05) associations of IMT change with baseline diabetes, current smoking, HDL cholesterol, Lp(a), pulse pressure, white blood cell count, and fibrinogen, all in the expected direction except Lp(a). The associations were of the same order of magnitude as anticipated from our cross-sectional findings. The association with baseline triglycerides was marginally significant, while those for systolic and diastolic blood pressure, hypertension, BMI, and LDL cholesterol were not significant. We examined the associations of IMT change with change in HDL cholesterol, LDL cholesterol, triglycerides, and BMI and with onset of diabetes and hypertension, and found statistically significant associations with all but HDL cholesterol. Modifiable, established CHD risk factors are associated with the rate of change of subclinical atherosclerosis, as indexed by carotid IMT.
Abstract 101: Serum Pro Brain Natriuretic Peptide (pro BNP) Levels In Children With Kawasaki Disease
Authors: Rawat Amit , Reddy Mounika V , Suri Deepti , Gupta Anju , Singh Surjit
Published: March 2022

Introduction:
Brain natriuretic peptide (BNP) is a useful biomarker for diagnosing heart failure in adults and children. ProBNP, the precursor form of BNP has been shown to predict myocardial involvement in acute Kawasaki disease (KD) especially in cases of incomplete KD. As the phenotype of KD in North India is different from that in Japan and North America, this study was carried out to validate the utility of proBNP in North Indian children with KD.

Objectives:
To assay the levels of proBNP in North Indian children with KD.

Methodology:
Serum pro brain natriuretic peptide (pro BNP and NT-proBNP) levels were estimated in 25 children with Kawasaki disease, both during acute phase and later in convalescence. The levels were similarly assayed in an equal number of age and sex matched controls. The proBNP levels were estimated by an enzyme linked immunosorbent assay (ELISA) using the RayBio Human proBNP ELISA kit which detects both pro BNP and NT-pro BNP.

Results:
Pro BNP levels were significantly elevated in children during acute phase of KD (4259±8015 pg/ml) when compared to febrile controls (407±542pg/ml) or to children in convalescent phase of KD (509±1066 pg/ml). Pro BNP levels were also higher in children with coronary abnormalities in acute phase.

Conclusions:
In our experience, pro BNP is a useful biomarker for KD especially with regard to myocardial involvement. ProBNP level is markedly elevated during the acute phase in all children with KD. In children with persistent coronary artery abnormalities, the proBNP level remains elevated even during convalescence.

Future impact:
ProBNP may be useful marker for children with incomplete forms of KD
Abstract 111: Identification of the agent causing Kawasaki Disease via throat swab cultures
Authors: Menikou Stephanie , Sancho-Shimizu Vanessa , Levin Zoe , M. Hamilton Shea , Langford Paul , Levin Mike
Published: March 2022

Kawasaki disease (KD) is a paediatric inflammatory disease and is the most common cause of acquired heart disease in children in developed countries. The cause of KD is unknown but epidemiological data indicate that it is most likely to be an infectious agent. The clinical features of KD are consistent with either bacterial or fungal infection rather than a virus, as KD is associated with an intense inflammatory process. It has been suggested that the oral and respiratory mucosa are the primary site of infection as the initial infection in KD patients is likely to be in the mucosal surfaces of the respiratory track with prominent lymphadenopathy and oral mucous membrane inflammation. This study examines whether the agent causing KD induces disease through production of novel toxins, or inflammation inducing antigens in the supernatants of cultured bacteria isolated from KD patients and healthy controls. Throat swabs were collected from children with KD (n=14), other diseases (n=10) and healthy children (n=8), and were inoculated into growth medium, incubated and then centrifuged. The supernatants were collected and toxicity was measured by microscopic observation as well as by resazurin dye reduction of cells after growth in the presence of the KD or control supernatant. An effect of the toxins in activating inflammation was also tested by the addition of the supernatants to healthy donor peripheral blood mononuclear cells (PBMCs) and measurement of interleukin 6 production by Enzyme linked immune assay (ELISA). There was no significant difference in the reduction of cells after growth between control and KD supernatants. No difference in the production of IL-6 was observed between the three groups. Further analysis and larger sample size are required to establish whether novel toxins are present in cultured supernatants of KD patients.
Abstract 128: Efficacy Of Plasma Exchange For Refractory Kawasaki Disease: The Earlier, The Better
Authors: Hashimoto Kunio , Motomura Hideki , Moriuchi Hiroyuki
Published: March 2022

Background:
Amelioration of systemic inflammatory changes during the acute phase of Kawasaki Disease (KD) is critical for preventing coronary artery lesions (CALs). Plasma exchange (PE) therapy has been applied to patients with severe KD in several medical facilities in Japan and been covered by health insurance since 2012.

Methods:
We reviewed medical records of five KD patients, aged 3 to 44 months, who were treated with PE in Nagasaki University Hospital from December 2012 to August 2015.

Results:
All patient were treated with two courses of intravenous immunoglobulin (IVIG). After IVIG that had offered little or no improvement, two of the five patients used infliximab and another used cyclosporine A before PE. PE was performed for the consecutive 3 or 5 days until defervescence and clinical improvement were obtained. Acute inflammatory changes were successfully ameliorated upon PE therapy in all five patients, although two patients who had already developed CALs before PE on days 20 and 21, respectively, residual CALs were confirmed one year after the treatment. No CALs had developed in the remaining three patients who started PE therapy within nine days after onset (days 8, 8 and 9, respectively). PE therapy was carefully performed in the intensive care unit, and no severe adverse events occurred.

Discussion:
PE therapy is able to ameliorate acute inflammation in most cases of severe KD refractory to standard IVIG therapy; however, it is critical to start PE therapy earlier than day-10 in order to prevent CALs. Thus, pediatricians have to make prompt decision on referral of their KD patient who did not respond IVIG to the institutions where PE is applicable in Japan.
Abstract 130: Clinical Efficacy Of Modified RAISE Protocol In Acute Kawasaki Disease
Authors: Ikehara Satoshi , Takatsuki Shinichi , Naoi Kazuyuki , Nakayama Tomotaka , Matsuura Hiroyuki , Kobayashi Tohru , Saji Tsutomu
Published: March 2022

Backgrounds and Objectives:
RAISE protocol demonstrated that IVIG plus prednisolone (PSL) therapy reduced the incidence of coronary artery abnormalities (CAAs) in children with high risk of refractory Kawasaki disease (KD). However, this protocol has longer hospital stay because PSL is tapered over the next 15 days when CRP was under 5mg/L. We proposed modified RAISE protocol which PSL is tapered over 9 days. The aim of this study was to evaluate clinical utility of modified RAISE protocol.

Methods:
This is a retrospective study to evaluate clinical data from KD between May, 2012 and August, 2014. Composite outcomes included incidence of relapse and coronary arterial abnormalities. Using Kobayashi score, children were divided into high and low risk group.

Results:
We enrolled 36 KD patients with high risk group in total 147 patients with KD. Median of age was 33 months (range 2-83 months), 13 males. Median days of illness were 4.0 (range 2-7) and Kobayashi score was 6.0 (range 5-9). Overall, 31 children (86.1%) were responders, whereas 5 children were non-responders (resistance; 1 (2.8%) relapse; 4 (11.1%)). Among 5 non-responders, 4 patients received methylprednisolone pulse therapy (mPSL) and the remaining one patient administrated two times mPSL and infliximab. During follow-up (1-29 months), 3 children (8.3%) had CAAs (dilatation: 3 patients). Modified RAISE protocol was not inferior to conventional RAISE protocol (CAAs 3.3%, p=0.68, responder 84.3%, p=1.0, resistance 5.0%, p=0.92, relapse 10.7%, p= 0.66).

Conclusions:
Clinical safety and efficacy of modified RAISE protocol (3days х3) were similar to conventional RASIE protocol(5days х3).
Abstract 142: Plasma Exchange Therapy in a Patient with Refractory Kawasaki Disease Initially Treated with Intravenous Immunoglobulin and Prednisolone
Authors: Okunushi Kentaro , Ebata Ryota , Hattori Noriyuki , Kobayashi Hironobu , Saito Naoki , Oda Shigeto , Shimojo Naoki
Published: March 2022

Prednisolone combined with intravenous immunoglobulin (IVIG) is the standard option for the initial treatment of Kawasaki disease (KD) in Japan, as described in the 2012 Japanese guidelines for KD. However, there is still debate regarding treatment in patients with refractory KD initially treated with IVIG plus prednisolone, especially in those with comorbid coronary artery aneurysm (CAA). Here, we present the case of a 3 year-old-girl initially treated with IVIG plus prednisolone, whose clinical and laboratory parameters improved after plasma exchange therapy (PE). She was admitted to a local hospital on day 4 of illness and diagnosed as having severe KD (Kobayashi score, 7). Therefore, prednisolone plus IVIG was administered as initial treatment. The initial course was ineffective. She was treated with methylprednisolone pulse therapy, and a second and third round of additional IVIG, but low-grade fever persisted. After these treatments the patient’s general condition had improved and major KD symptoms improved by day 13 of illness, despite a high serum C-reactive protein level (19.9 mg/dl) and hypoalbuminemia (1.8 g/dl). Expanding medium-sized aneurysm was detected on the same day, so she was immediately referred to our hospital for further treatment. The CAA continued to expand, so PE was performed for 3 consecutive days and prednisolone was gradually reduced from day 15 of illness. After that, she became afebrile and CAA progression stopped. Laboratory findings such as serum C-reactive protein and albumin levels were dramatically improved after PE. This case suggests that PE might be effective for refractory KD initially treated with IVIG plus glucocorticoids and may stop the CAA development. Doctors must remain aware that patients treated with glucocorticoids may have masked clinical conditions, and take care not to underestimate the signs of refractory KD.
Abstract 148: The Optimal Duration Of Asa Treatment After Acute Phase Of Kawasaki Disease Based On Inflammatory Markers
Authors: Yoo Jae-won , kim ji mok , Kil Hong Ryang
Published: March 2022

Purpose:
Outcomes of Kawasaki disease (KD) are determined by the development of cardiovascular complications. Low dose aspirin(ASA) treatment is used as antithrombotic effect after acute phase of KD for 6-8 weeks.There are no standardized inflammatory markers used as guidance for completing low dose ASA treatment. The optimal duration of low dose ASA treatment is not determined. The aim of this study is to decide the duration of low dose ASA treatment based on inflammatory markers.

Methods
: We enrolled 80 KD children admitted to the Chungnam National University Hospital,South Korea between Sep 2012 and May 2014. We reviewed the medical records for clinical characteristics, duration of fever, cardiovascular complications and laboratory data on admission. Inflammatory markers were measured every 7-14 days at OPD after immunoglobulin(IVIG) and high dose ASA treatment in acute phase. Low dose ASA was stopped when the level of inflammatory markers were down to the normal without any cardiovascular complications. Echocardiograms were performed at admission, discharge and prior to completion of ASA treatment when the inflammatory marker turned to normal.

Results
: 80 KD patients,(Complete, n=44; Incomplete, n=36). The incidence of cardiovascular complications was similar both two groups. No significant differences in the laboratory studies either complete or incomplete KD. The levels of inflammatory marker were normalize after the acute phase as 13.0±13.6 days for platelets, 13.8± 8.3 days for CRP, 14.4±9.6 days for d-dimer, and 24.1 ± 10.5 days for ESR. The levels of platelets and CRP were first normalized within 2 weeks, and that of ESR was last within 3-4 weeks. The duration of raised CRP levels showed significantly shorter in incomplete group compared to complete groups (p=0.02). No new coronary lesions were developed among patients who didn’t have the coronary lesions at subacute phase.

Conclusion:
The majority of the inflammatory marker levels were normalized within 2-3 weeks after acute phase of KD. There were no new cardiovascular complications when the duration of low dose ASA treatment was guided by level of the inflammatory markers. So, the optimal duration of low dose ASA treatment can be reduced to 3-4 weeks without development of coronary lesions.
Abstract 154: CMR Feature Tracking in Kawasaki Disease convalescence
Authors: Bratis Konstantinos , Hackmann Pauline , Child Nicholas , Mavrogeni Sophie , Krasemann Thomas , Tarique Hussain , Botnar Rene , Razavi Reza , Greil Gerald
Published: March 2022

Objective:
To determine whether left ventricular (LV) myocardial deformation indices can detect subclinical myocardial abnormalities in Kawasaki Disease (KD) convalescence. We hypothesized that subclinical myocardial abnormalities due to inflammation represent an early manifestation of the disease that persist in convalescence.

Background:
Myocardial inflammation has been described as a global finding in the acute phase of Kawasaki Disease. Despite normal LV systolic function by routine functional measurements, reduced longitudinal strain (S) and strain rate (SR) have been detected by echocardiography in the acute phase, which may potentially predict late onset heart failure.

Method and results:
Peak systolic LV myocardial longitudinal, radial and circumferential S and SR were examined in 29 KD convalescent patients (15 males; mean (SD) age 11 (6.6) years, range 3- 27 years; median interval from KD onset 5.8 (5.4) years) and 10 healthy volunteers (5 males; mean age 14 (3.8) years, range 6- 19 years) with the use of cardiac magnetic resonance feature tracking (CMR-FT). Routine indices of LV systolic function were normal in both groups.

Comparisons were made between normal controls and (i) the entire KD group, (ii) KD group subdivided by coronary artery involvement. Compared to controls, KD patients had lower longitudinal S. Average longitudinal and circumferential S at all levels was lower in KD patients compared normal controls. In subgroup analysis, both KD patients with and without any history of CAD had similar longitudinal and circumferential S at all levels and lower when compared against controls. There was a non-significant trend for lower circumferential and longitudinal values in KD patients with persisting CAD when compared against those with regressed CAD.

Conclusion:
In this CMR-FT study in KD convalescent patients with preserved conventional functional indices, we observed a trend for lower circumferential and longitudinal strain in KD patients compared to normal controls, irrespective of their coronary artery status.
Abstract 161: Investigating the Added Value Of Repeated ECHO Imaging In Patients With Kawasaki Disease and Always-Normal Coronary Arteries
Authors: de Ferranti Sarah D , Gauvreau Kimberlee K , Berry Erika , Tang Alexander , Baker Annette , Fulton David , Friedman Kevin , Tremoulet Adriana , Burns Jane C , Newburger Jane W
Published: March 2022

Background:
Echocardiography (echo) is recommended in patients (pts) with KD at baseline, 2 wks, and 6 wks after fever onset to detect coronary artery (CA) abnormalities. However, echo imaging is expensive and in infants and toddlers often requires sedation, which is burdensome and carries some risk.

Objective:
To assess the benefit of additional echo imaging in uncomplicated KD pts with normal CAs at baseline and 2 wks.

Methods:
In this retrospective two-center study, eligibility criteria were: 1) admission and treatment at center for acute KD; 2) no significant congenital heart disease; 3) available echo measurements of both the right (R) CA and left anterior descending (LAD) at ≤ Day 10 (baseline), 2 (±1) wks and 6 (±3) wks; and 4) normal CAs = RCA and LAD z scores [zMax] ≤2.0 and no aneurysms.

Results:
Of 2,600 pts treated for KD from 1976 - 2014, 457 had normal CAs at baseline and 2 wks and met other inclusion criteria. Of these, 57% were male; median age was 3.3 yrs [IQR 1.8 to 5.4 yrs]; 77% had complete KD; and 14% received IVIG retreatment. At 6 wks, 450 (98.5%) had normal CAs. Of the remaining 7 pts, zMax was 2.0-2.4 in 5 (1.1%); their z scores subsequently normalized at 1 yr. The zMax was 2.5-2.9 in 1 (0.2%) pt who remained on low dose aspirin until 6 mos, when ECHO normalized. The zMax was ≥3.0 in 1 pt (0.2%, 95% CI [0.0%, 1.2%]) with a dilated LAD, which was normal 3 wks later. Sensitivity analysis using zMax of <2.5 was similar (98.0% with nl CAs at 6 wks).

Conclusion:
New CA abnormalities are rarely detected at 6 wks in KD pts with normal CAs at baseline and 2 wks. The 6 wk echo may be unnecessary for uncomplicated KD pts with normal CAs at baseline and 2 wks.

Figure - CA zMax shown as mean (+), median, 25th to 75th %tile.
Abstract 172: Tissue Doppler Imaging as a Predictor of Immunoglobulin Resistance in Kawasaki Disease
Authors: Phadke Daniel R , Patel Sonali S , Dominguez Samuel R , Heizer Heather R , Anderson Marsha , Glode Mary P , Jone Pei-Ni
Published: March 2022

Background:
Kawasaki disease (KD) causes myocarditis and left ventricular dysfunction during the acute phase of the illness. Despite treatment with intravenous immunoglobulin (IVIG), a significant number of patients are IVIG resistant (fail to defervesce or experience recrudescent fever). We evaluated KD patients in the acute phase using Tissue Doppler Imaging (TDI) to assess if myocardial dysfunction may predict IVIG resistance.

Methods:
All patients with acute KD presenting to Children’s Hospital Colorado from February, 2007 to March, 2014 were included in this study. All patients underwent echocardiograms with TDI (pulsed wave sample Doppler were placed at the tricuspid, septal, and mitral annuli) upon diagnosis with KD. Patients were divided into two groups: IVIG Responder group and IVIG Resistant group. Group differences were assessed using Wilcoxon-Mann-Whitney and Chi-square testing. Receiver Operating Characteristic (ROC) Curve analysis was utilized to determine threshold values of TDI measurements associated with IVIG resistance.

Results:
Fifty-one age matched (+/- 6 months) IVIG Responder patients were compared to 51 IVIG Resistant group [33.49 (17.30 - 62.89) months vs 44.57 (20.13 - 77.07) months, p <0.44]. There were 34 males in Responder group versus 33 males in Resistant group (p< 0.83). There were significant differences in the tricuspid, septal, and mitral early diastolic velocities (E’) (p< 0.05, p< 0.001 and p<0.01) respectively. ROC analysis demonstrated that tricuspid E’ < 0.15 cm/s, septal E’ < 0.12 cm/s, and mitral E’ < 0.16 cm/s were good predictors of IVIG unresponsiveness (AUC = 0.66, 0.66, and 0.70 respectively). There were no differences between the systolic velocities (S’) or late diastolic velocities (A’).

Conclusions:
IVIG resistant KD patients present with significantly greater diastolic dysfunction compared to responders in patients with KD. TDI may be a useful tool to differentiate KD patients who may be IVIG resistant.
Abstract 195: Kawasaki Disease in a four-year-old girl with History of Acute Myeloid Leukemia; Report of a Case and Literature Review
Authors: Kawai Reiko , Takatsuki Shinichi , Nakayama Tomotaka , Sawa Tomoka , Yanai Shun , Naoi Kazuyuki , Ikehara Satoshi , Matsuura Hiroyuki , Ohara Akira , Saji Tsutomu , Saji Tsutomu
Published: March 2022

Background:
Kawasaki disease (KD) and acute myeloid leukemia (AML) both occur predominantly in young children. However, considering that the incidence rate of cancer during 0-14 years of age in Japan is 10.1 per a hundred thousand and that of KD during 0-4 years of age is 260 per a hundred thousand, there is little possibility of the same individual developing 2 diseases. Both inside and outside Japan we found at least 19 case reports of KD some relevant to cancer. Herein, we report a case of KD in a four-year-old Japanese girl with history of AML.

Case report:
A four-year-old Japanese girl was diagnosed with KD at 3 days of illness. She had the medical history of AML-M7 at her age of 1. She had no chimeric genes and was classed as intermediate risk. She had been treated following JPLSG AML05 protocol including etoposide, cytarabine (Ara-C), mitoxantrone, idarubicin, methotrexate (MTX), hydrocortisone (HDC) and triple intrathecal therapy (Ara-C, MTX and HDC). More than 2 years had passed without recurrence since her last chemotherapy. When we diagnosed her as KD, laboratory findings showed WBC 15.5 х103/μl with 85% neutrophils, hematocrit 33.7%, platelet count 19.1x10
6
/μl, CRP 10.5mg/dl, serum albumin 3.8g/dl and BNP 12.2pg/ml. She responded well to IVIG (2g/kg) plus prednisolone (2mg/kg/day) and ASA (30mg/kg/day). Although BNP transiently had increased to 151.8pg/ml at 5 days after onset, neither coronary arterial abnormalities nor myocardial complications were detected at 1 year of illness.

Conclusions:
We experienced the rare case of KD in a four-year-old Japanese girl with history of AML. She responded well to IVIG and any cardiovascular complications were not detected at 1 year of illness. Referring to only this case, it is unclear whether the history of AML could influence developing and progress of KD or not. The possibility of the same individual developing KD during 0-4 years of age and cancer during 0-14 years of age would be 2.6 per 10 million in Japan. However, among three hundred thousand cumulative patients of KD in Japan, 14 cases have been reported that they had developed cancer during 0-14 years of age. To discuss the relation to character of cancer, chemotherapy and severity or compilations of KD, more case should be evaluated.