bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 621: Attenuated Expression of Profilin-1 Confers Protection from Atherosclerosis in the LDL Receptor-null Mouse
Authors: Romeo Giulio R , Moulton Karen S , Kazlauskas Andrius 
Published: July 2021

INTRODUCTION. Atherosclerosis-related events are a major cause of morbidity and death worldwide. The mechanisms underlying atherogenesis are not fully understood. We showed in previous studies that the actin-binding protein profilin-1 (pfn) was upregulated in atheroscle-rotic lesions and in endothelial cells (EC) treated with oxidized LDL (oxLDL).
HYPOTHESIS. In the present study, we tested the hypothesis that pfn levels play a role in early atheroma formation.
RESULTS. Since Pfn −/− mice die during embryonic development, we tested the role of pfn in atherogenesis by crossing adult Pfn +/− mice, which are viable and display a ∼50% reduction in pfn levels, into the Ldlr −/− mouse model of atherosclerosis. Atherosclerotic lesion burden was assessed in Pfn +/−Ldlr −/− (PfnHet) and Ldlr −/−control mice (PfnWT) fed an atherogenic high-cholesterol diet (HCD) for two months. Both males and females PfnHet exhibited a significant reduction in the percent lesion area in the descending aorta, when compared to PfnWT (females: 1.1 Â 27> 0.1 vs PfnWT 4.17 Â27> 0.49%, n=10; males: 1.34 Â 27> 0.22 vs PfnWT 3.4 Â 27> 0.36%, n=6). Similarly, aortic arch lesions were reduced in PfnHet (0.08 Â 27> 0.3 vs PfnWT 0.20 Â 27> 0.05 mm2, n=10). Total cholesterol and triglyceride levels were comparable in the two groups. Relevant atheroprotective changes were identified in PfnHet. When compared to PfnWT, aortas from PfnHet mice showed preserved eNOS activation, NO production and NO-dependent signaling. These changes were observed under HCD but not under a normal diet, thus indicating that pfn levels modified the response of the endothelium to atherogenic injury rather than its basal function. Similarly, knockdown of pfn in cultured aortic EC was protective against endothelial dysfunction triggered by oxLDL. Also, bone marrow-derived macrophages from PfnHet showed reduced internalization of oxLDL and oxLDL-induced inflammatory pathways, including activation of p38.
CONCLUSIONS. These studies demonstrate that pfn levels modulate critical processes for atherosclerotic lesion formation through combined endothelial- and macrophage-dependent mechanisms.
Abstract 740: Statin Prevents Structural and Electrical Atrial Remodeling in Rat Hypertensive Heart Failure Induced by Chronic Inhibition of NO Synthesis
Authors: Okazaki Hidetoshi , Minamino Tetsuo , Wakeno Masakatsu , Tsukamoto Osamu , Kim Jiyoong , Takashima Seiji , Mochizuki Naoki , Kitakaze Masafumi 
Published: July 2021

Background: The prevalence of atrial fibrillation (AF) increases in patients with hypertension and heart failure in which endothelial function is impaired. Oxidative stress in atrial tissues is involved in the development of AF. Statins have pleiotropic effects including antioxidant effects. Thus, we developed hypertensive heart failure in rats by the chronic inhibition of NO synthesis and we investigated the role of the statin in atrial remodeling.
Methods and Results: Ten-week-old male Wistar-Kyoto rats were randomly divided into four groups: the control (no treatment), the L-NAME (received L-NAME, an inhibitor of NO synthase, 1 g/L in drinking water), the L-NAME+atorvastatin (L(a low dose)) (5 mg/kg/day) and the L-NAME+atorvastatin (H(a high dose)) (20 mg/kg/day). Chronic inhibition of NO synthesis increased systolic blood pressure (control group: 138±5 mmHg; L-NAME group: 212±8 mmHg L+atorvastatin (L) group: 210±6 seconds , L+atorvastatin (H): 208±6 seconds) and decreased LV dP/dt max/LVSP (control group: 54±3, L-NAME group: 36±8, L+atorvastatin (L) group: 40±3, L+atorvastatin (H) group: 38±6 1/second), whereas co-administration of atorvastatin did not affect them. The sustained duration of AF was always induced by transesophageal burst pacing (The pacing pulse used for induction of AF was rectanglar in shape, of 60V and 6ms-width. The atrium was paced at a cycle length of 12ms (83Hz) for 30seconds) and it was prolonged by L-NAME, which was prevented by atorvastatin in a dose-dependent manner (control: 8.8 ± 1.0 seconds, L-NAME: 20.7 ± 3.0 seconds, L+atorvastatin (L): 12.8 ± 3.2 seconds , L+atorvastatin (H): 8.8 ± 2.6 seconds). The chronic inhibition of NO synthesis increased the extent of fibrosis and markers of oxidative stresses such as proteins modified with 4-hydroxy-trans-2-nonenal and p47phox and rac1 in left atrial tissue, either of which was prevented by atorvastatin.
Conclusion: Atorvastatin attenuates oxidative stresses in atrium and prevents atrial electrical and morphological remodeling in rat hypertensive failing hearts without hemodynamic changes. Statin may be beneficial in patients with coronary heart disease and atrial fibrillation, and may contribute to the prevention of the onset of heart failure.
Abstract 902: A Beta-2 Adrenergic Receptor Polymorphism Is Associated With Increased Left Ventricular Remodeling Following First ST-Elevation Myocardial Infarction
Authors: McLean Rhondalyn C , Hirsch Glenn A , Gerstenblith Gary , Schulman Steven P 
Published: July 2021

Background: Prior studies demonstrate an association between specific β-adrenergic receptor polymorphisms and clinical outcomes in patients with chronic heart failure and following an acute coronary syndrome. The mechanism may relate to an effect on left ventricular (LV) remodeling. We hypothesized that β-adrenergic receptor polymorphisms predict LV remodeling after acute ST elevation myocardial infarction (STEMI).
Methods: We assessed the presence of β-1 and β-2 adrenergic receptor polymorphisms in 122 patients after their first STEMI enrolled in a single-center randomized, double-blind placebo controlled trial of L-arginine vs. placebo, 91% of whom received successful early reperfusion therapy. All patients were treated with a beta-1 receptor antagonist and underwent baseline (mean 5.9 days following STEMI) and 6-month LV volume evaluation using gated blood pool imaging. Univariate and multivariate linear and logistic regressions were used to assess the relationships between the polymorphisms, β1 Arg389Gly, β1 Ser49Gly, β2 Gly16Arg and β2 Gly27Glu and the six-month changes in LV volumes. The top quintiles of LV end-systolic (ESV) and end-diastolic (EDV) 6-month volume increases and LV ejection fraction decrease were compared to the lower quintiles in the logistic regression analyses.
Results: The polymorphisms β1 Arg389Gly, β1 Ser49Gly, β2 Gly16Arg were not associated LV remodeling. However, the 25% of patients homozygous for the β2 Glu27 variant were 5.2 times more likely to have an increase in 6-month ESV than those who had the Gln27 variant (OR 5.2, 95%CI 1.4 –19.0). Multiple linear regression analyses demonstrated that absolute ESV at six months was 19 ml greater (p = 0.02) and EDV was 21 ml greater (p = 0.01) in post STEMI patients with the β2 Glu27 polymorphism compared to the wild type or heterozygous patients.
Conclusions: Increased LV volumes post-STEMI are associated with an increased risk of heart failure and death. The common β2 receptor polymorphism, Glu27Glu, is associated with increased odds of adverse LV remodeling in patients treated with a beta-one receptor antagonist. Whether treatment with a non-specific β-adrenergic receptor blocker guided by this genetic polymorphism ameliorates the effect requires further study.
Abstract 991: The Novel BTB/POZ Protein, RhoBTB3, Regulates Mitochondrial Function Via The Ubiquitin Proteasome System
Authors: Yuasa Shinsuke , Onizuka Takeshi , Kenichiro Shimoji , Ohno Yohei , Katayama Masaharu , Arai Takahide , Hattori Fumiyuki , Murata Mitsushige , Hakuno Daihiko , Kimura Kensuke , Makino Shinji , Sano Motoaki , Ogawa Satoshi , Fukuda Keiichi 
Published: July 2021

Although BTB/POZ family proteins play a critical role in the development of various organs, and are recently known as a substrate adaptor of CUL3 ubiquitin ligases, the involvement of these proteins in heart development remains unknown. We recently isolated the novel protein, RhoBTB3, an 83-kD protein which comprises an N-terminal Rho GTPase resembling domain and two tandem BTB domains. The purpose of this study was to investigate the molecular and functional characteristics of this protein in mice heart.
[Methods and Results]

RhoBTB3 was faintly expressed in the embryonic heart, but was strongly expressed in the adult heart. It was weakly expressed in adult brain and testis.

RhoBTB3-GFP fusion protein cDNA was transfected into primary cultured rat cardiomyocytes using adenovirus. Fluorescence microscopy revealed that RhoBTB3 was located at the cytoplasm of living cardiomyocytes.

HA-tagged RhoBTB3 and MYC-tagged CUL family proteins (1 – 5) were co-transfected into COS7 cells. RhoBTB3 was co-immunoprecipitated with CUL3
in vitro
, but not with other CUL proteins.

The various mutant RhoBTB3 and wild type CUL3 protein were examined by immunoprecipitation assay. Both the first BTB domain and second BTB domain can bind to CUL3 protein independently.

It was difficult to detect RhoBTB3 in protein analysis. To determine whether or not RhoBTB3 itself could be the Cul3 ubiquitin target, RhoBTB3 protein was measured under the presence of proteasome inhibitor MG132 and protein synthesis inhibitor cycloheximide. The RhoBTB3 protein was immediately degraded and dropped below the detection level within 2 hours.

The transgenic mice which overexpressed the heart-restricted RhoBTB3 were healthy at a young age.

But after 3 months, they gradually showed heart failure and finally fell into sudden death. Interestingly before heart failure, electron microscopy demonstrated that mitochondria dramatically increased in number and showed deformity in RhoBTB3 transgenic hearts.
[Conclusions] RhoBTB3 is mainly expressed in the adult heart, and its protein was rapidly degraded with the CUL3-mediated ubiquitin proteasome system in the heart. The overexpression of RhoBTB3 caused a marked increase in abnormal mitochondria, followed by severe heart failure.
Abstract 1042: Viral Chemokine Modulating Protein, M-T7, Interrupts Chemokine : Glycosaminoglycan (GAG) Interaction: M-T7 Inhibitory Activity is Dependent upon Y46 and V210 aminoacid residues
Authors: Dai Erbin , McIvor Dana , Liu Liying , Munaswamy-Ramanujam Ganesh , Sun Yunming , Esko Jeffrey , Charo Israel F , McFadden Grant , Lucas Alexandra 
Published: July 2021

Background: Chemokines bind to glycosaminoglycans (GAGs) forming gradients that direct inflammatory cell invasion. The viral chemokine modulating protein (CMP), MT-7 binds the C terminal, GAG-binding domain of chemokines and has been previously reported to significantly reduce cell invasion and plaque growth in rat aortic and renal transplant models. Two other viral CMPs, M-T1 and M3 CMPs bind the N terminal domain of chemokines that bind to cell surface receptors. To determine the role of CC chemokine receptor 2 (CCR2) and GAGs for M-T7 anti-inflammatory activity, effects of M-T7 on plaque growth were assessed after mouse CCR2 deficient (CCR2−/−) or GAG deficient (NDST1−/−) aortic allograft transplant. Mononuclear cell migration in response to MCP-1 or RANTES into mouse ascites was also tested. Active sites necessary for M-T7 inhibition of chemokine function and monocyte activation, were assessed by infusion of in the mouse cell migration and human monocyte membrane fluidity assays.
Results: M-T7 significantly reduced cell migration and intimal hyperplasia in wild type CCR2+/+ (p<0.009), and CCR2−/− aortic transplants (p<0.026). M-T1 and M3 inhibited cell invasion and plaque in CCR2+/+, but not CCR2−/− mice. M-T7 inhibited plaque growth and CC chemokine (MCP-1 and RANTES)-induced cell migration in wild type mice (P<0.01), but not in NDST1−/− mice (P=0.34). Selected M-T7 point mutations Ty (Y)46A, and Val (V) 210A no longer block chemokine-induced cell migration nor monocyte activation, whereas Asn (N) 40, Asn (N) 63 and Val (V)129 retain inhibitory activity.
Conclusions:

M-T7 but not M-T1 nor M3, blocks cell migration and plaque growth in CCR2 deficient (CCR2−/−) mouse aortic transplant models.

M-T7 loses the ability to block cell migration and plaque growth in NDST1−/−, GAG (heparan sulfate) deficient mice.

Point mutations Tyr46 and Val 210 lack inflammatory for mouse and human inflammatory monocyte responses indicating that these amino acid residues on the M-T7 CMP protein are required for inhibitory activity.
Abstract 1046: The ApoA-I Mimetic Peptide, 4F, Inhibits LPS-induced Inflammatory Reactions Through Dysregulation Of The NFkB Activation Pathway.
Authors: Datta Geeta , White C. R , Chaddha Manjula , Palgunachari M. N , Anantharamaiah G. M , Smythies Lesley 
Published: July 2021

Background: The apoA-I mimetic peptide 4F possesses potent anti-inflammatory properties. We previously reported that 4F inhibits lipopolysaccharide (LPS)-induced cytokine, chemokine and adhesion molecule expression
in vivo and in vitro
. Other data show that 4F reduces macrophage content in atheroma in susceptible mice.

Objective: In this study, we explored anti-inflammatory mechanisms of 4F action under basal conditions and in the presence of LPS using THP1 monocytes/macrophages and primary human blood monocytes.
Methods and Results: Blood monocytes isolated by Ficoll and CD14+ MACS separation were analyzed by FACS for CD14, TLR-4 and p-NFκB expression. Cytokines, chemokines and adhesion molecules were measured by the Searchlight multi-analyte analysis (Pierce, MA, USA). 4F treatment (3 days) of both THP-1 and blood monocytes enhanced the secretion of GM-CSF (25-fold), a potent inducer of monocyte to dendritic cell differentiation. 4F also inhibited (by 80%) the PMA-induced phagocytosis of FITC-labeled beads by THP-1 cells. In subsequent studies, effects of 4F on LPS-induced inflammatory responses in monocytes were tested. Pre-treatment with 4F significantly inhibited LPS-induced MCP-1 expression (by 50%) in both THP-1 cells and blood monocytes. Under these conditions, the expression of the anti-inflammatory cytokine IL-10 was increased by 200%. The effects of 4F on the activation of NFκB, a downstream mediator of LPS action, were studied using flow cytometry. 4F pre-treatment significantly inhibited LPS-induced NFκB phosphorylation (NFκBp65) compared to vehicle treatment. Protective effects of 4F in LPS-treated monocytes were associated with a 50% reduction in the expression of TLR-4 and CD14, cell surface receptors for LPS.
Conclusions: These data provide evidence that 4F modulates functional properties of monocytes and show for the first time that 4F inhibits LPS-induced inflammatory responses by inhibiting the expression of CD14, TLR-4 and phosphorylation of NFκB. Enhanced GM-CSF expression and inhibition of monocyte phagocytic activity also suggest that 4F may favor the differentiation of monocytes to a dendritic cell rather than a macrophage phenotype.
Abstract 1102: Cardiac-Specific Myosin Light Chain Kinase (MLCK), a Third Member of MLCK Family, Potentiates Sarcomere Formation and Cardiac Contraction
Authors: Chan Jason Y , Takeda Morihiko , Briggs Laura E , Lu Jonathan T , Horikoshi Nobuo , Weinberg Ellen O , Aoki Hiroki , Chien Kenneth R , Kasahara Hideko 
Published: July 2021

Background: Two myosin light chain kinase (MLCK) proteins, skeletal (encoded by mylk2 gene) and smooth muscle MLCK (encoded by mylk1 gene) have been shown to be expressed in mammals. Human mylk2 has been mapped as a disease locus for familial cardiac hypertrophy (OMIM
606566
), suggesting that abnormal function of skeletal MLCK stimulates cardiac hypertrophy. While phosphorylation of the putative substrate of skeletal MLCK, myosin light chain 2 (MLC2), is recognized as a key regulator of cardiac contraction, the abundance of skeletal MLCK in the heart is controversial, suggesting the existence of an additional MLCK that is preferentially expressed in cardiac muscle.

Methods and Results: We characterized a new kinase named cardiac MLCK that is encoded by a gene homologous to mylk1 and 2 and is specifically expressed in the heart in both atrium and ventricle. Expression of cardiac MLCK was highly regulated by the cardiac homeobox transcription factor, Nkx2.5, in neonatal cardiomyocytes. The overall structure of cardiac MLCK protein is conserved with skeletal and smooth muscle MLCK including putative catalytic and adjacent Ca2+/calmodulin binding domains at the carboxyl-terminus. The amino-terminus is unique without significant homology to other known proteins. Cardiac MLCK phosphorylated MLC2v with a catalytic value of Km=4.3 micro M (Lineweaver-Burk analysis) indicating high affinity of cardiac MLCK to MLC2v, similar to the affinity of skeletal muscle MLCK to skeletal muscle MLC2 and smooth muscle MLCK to smooth muscle MLC2. Adenoviral-mediated overexpression of cardiac MLCK and knockdown of cardiac MLCK using RNAi in cultured cardiomyocytes revealed that cardiac MLCK regulates MLC2v phosphorylation, sarcomere organization and cardiac myocyte contraction. Expression of cardiac MLCK protein was significantly decreased in severe heart failure in vivo (post-myocardial infarction heart failure mouse model).
Conclusion: Cardiac MLCK is a new key regulator of cardiac contraction and sarcomere organization. Reduction of cardiac MLCK function leading to decreased phosphorylation of MLC2v may contribute to compromised contractile function in the failing heart.
Abstract 1124: CD40L Induces Inflammation in Adipose Tissue - a Potential Link Between the Metabolic Syndrome and Atherosclerosis
Authors: Missiou Anna , Platzer Isabel , Ernst Sandra , Schonbeck Uwe , Libby Peter , Bode Christoph , Zirlik Andreas 
Published: July 2021

Background: CD40L figures prominently as marker and mediator of atherosclerosis and its clinical complications. Recent data also demonstrate an association between CD40L and the metabolic syndrome. In the light of these data we hypothesized a functional pro-inflammatory role of CD40L in adipose tissue that provokes systemic pro-inflammatory responses and potentially mediates some of the high cardiovascular risk associated with the metabolic syndrome.
Methods and Results: Adipocytes and preadipocytes isolated from adipose tissue obtained from bariatic surgery did not express CD40L but its receptor CD40 as assessed by PCR and immunohistochemistry. Furthermore, sections of human adipose tissue from obese donors contained more macrophages and T cells colocalizing with CD40L than those from lean controls. Stimulation of both adipocytes and preadipocytes with recombinant CD40L resulted in a concentration- and time-dependent release of the pro-inflammatory cytokine IL-6, the chemokines MCP -1 and IL-8 as well as the inhibitor of fibrinolysis PAI-1 into the supernatant as quantified by ELISA. Membranes isolated from a Murine cell line overexpressing human CD40L reproduced these pro-inflammatory effects compared to respective controls. Interestingly, fenofibrate but neither rosiglitazone, metformin, nor atorvastatin attenuated the CD40L-inducible expression of these pro-atherogenic mediators. Finally, supernatants from adipocytes and preadipocytes stimulated with CD40L activated endothelial cells and macrophages, typical cell types resident in atherosclerotic lesions, as assessed by FACS for tissue factor, Mac-1, and ICAM-1, respectively.
Conclusions: Our data suggest that CD40L induces inflammatory cytokine production in adipose tissue resulting in activation of endothelial cells and macrophages. This new mechanism may contribute to the high cardiovascular risk of patients suffering from the metabolic syndrome.
Abstract 225: Role Of PEDF In Regulation Of Skeletal Blood Flow And Neovascularization In Ischemic Limb Via Interaction With VEGF
Authors: Arima Ken , Katsuda Yousuke , Saito Yutaka , Katsuki Yoshio , Sasaki Kenichiro , Yamagishi Shoichi , Ikeda Hisao , Kai Hisashi , Imaizumi Tsutomu 
Published: July 2021

[Introduction] We reported that pigment epithelium-derived factor (PEDF) plays an important role in the angiogenesis, vascular permeability and regulation of blood flow in the eye and brain via interaction with VEGF.
[Objective] We examined the role of PEDF in ischemia or VEGF-related angiogenesis or regulation of blood flow in severe limb ischemia.
[Methods] Unilateral hindlimb ischemia was performed in klotho-mice by binding the unilateral femoral artery. After hindlimb ischemia was surgically induced, we injected saline (control group) or VEGF (3 μg/day × 7, VEGF group) to ischemic limb from day 1 to 7. The expression level of PEDF in the ischemic tissue was examined by RT-PCR method. Furthermore, we injected recombinant mouse PEDF (4 μg/day × 7) or anti-PEDF antibody (60 μg/day × 7) in addition to VEGF (VEGF + PEDF group and VEGF + anti-PEDF group, respectively) to ischemic limb from day 1 to 7. We evaluated limb blood flow by laser doppler blood-flow (LDBF) at day 0, 3, 7, and 14, and number of CD34 positive cells in the ischemic tissue by histology as an index of ischemic angiogenesis at day 7 in all groups.
[Results] Expression levels of PEDF in ischemic tissue were significantly elevated in VEGF group. In VEGF group, the ischemic/normal limb blood flow ratio by LDBF analysis and number of CD34 positive cells were significantly increased compared with control group (p < 0.05). The effects of VEGF on blood flow or number of CD34 positive cells were completely abolished by administration of PEDF. Furthermore, anti-PEDF group, accelerated recover of blood flow in ischemic limb, but not number of CD34 positive cells compared in VEGF administration.
[Conclusion] VEGF augmented the expression level of PEDF in the ischemic limb. VEGF-induced ischemic angiogenesis was abolished by exogenous administration of PEDF. Furthermore, anti-PEDF antibody augmented VEGF-induced increase of blood flow. These results suggest that PEDF may play an important role in regulation of skeletal blood flow and neovascularization in ischemic limb via interaction with VEGF.