BioMed Research International: Vascular Medicine The latest articles from Hindawi Publishing Corporation © 2016 , Hindawi Publishing Corporation . All rights reserved. The Discovery of Novel Genomic, Transcriptomic, and Proteomic Biomarkers in Cardiovascular and Peripheral Vascular Disease: The State of the Art Thu, 19 May 2016 14:10:28 +0000 Cardiovascular disease (CD) and peripheral vascular disease (PVD) are leading causes of mortality and morbidity in western countries and also responsible of a huge burden in terms of disability, functional decline, and healthcare costs. Biomarkers are measurable biological elements that reflect particular physiological or pathological states or predisposition towards diseases and they are currently widely studied in medicine and especially in CD. In this context, biomarkers can also be used to assess the severity or the evolution of several diseases, as well as the effectiveness of particular therapies. Genomics, transcriptomics, and proteomics have opened new windows on disease phenomena and may permit in the next future an effective development of novel diagnostic and prognostic medicine in order to better prevent or treat CD. This review will consider the current evidence of novel biomarkers with clear implications in the improvement of risk assessment, prevention strategies, and medical decision making in the field of CD. Stefano de Franciscis, Laurent Metzinger, and Raffaele Serra Copyright © 2016 Stefano de Franciscis et al. All rights reserved. Roles and Clinical Applications of OPG and TRAIL as Biomarkers in Cardiovascular Disease Thu, 21 Apr 2016 12:43:55 +0000 Cardiovascular diseases (CVD) remain the major cause of death and premature disability in Western societies. Assessing the risk of CVD is an important aspect in clinical decision-making. Among the growing number of molecules that are studied for their potential utility as CVD biomarkers, a lot of attention has been focused on osteoprotegerin (OPG) and its ligands, which are receptor activator of nuclear factor κB ligand (RANKL) and TNF-related apoptosis-inducing ligand. Based on the existing literature and on our experience in this field, here we review what the possible roles of OPG and TRAIL in CVD are and their potential utility as CVD biomarkers. Stella Bernardi, Fleur Bossi, Barbara Toffoli, and Bruno Fabris Copyright © 2016 Stella Bernardi et al. All rights reserved. Genetic Variants of CD40 Gene Are Associated with Coronary Artery Disease and Blood Lipid Levels Wed, 20 Apr 2016 08:09:51 +0000 Objectives. The present study aimed to evaluate the effect of CD40 and CXCR4 genes polymorphisms on CAD susceptibility and the blood lipid levels and history of cardiovascular risk factors in a Chinese Han population. Materials and Methods. A total of 583 unrelated patients with CAD and 540 controls were recruited. Two tag SNPs (rs4239702 and rs1535045) at the CD40 locus and one tag SNP (rs2228014) at the CXCR4 locus were genotyped using the SEQUENOM Mass-ARRAY system. Results. After adjusting the risk factors, the frequency of rs1535045-T allele was also higher in patients than controls. Haplotype analysis showed that the rs4239702(C)-rs1535045(T) haplotype was associated with CAD. People with rs4239702-TT genotype had higher blood lipid levels in case group while it was not in the control group. History of cardiovascular risk factors showed no association for the three SNPs in case group and control group. Conclusions. rs1535045 in CD40 gene is likely to be associated with CAD in the Chinese Han population. rs4239702(C)-rs1535045(T) haplotype was associated with CAD. Only in CAD patients, the blood lipid level of patients with rs4239702-TT genotype was higher than other patients. CXCR4 gene may not relate to CAD. Liting Zhou, Lin Xie, Dongchun Zheng, Na Li, Jian Zhu, Shuyue Wang, Bo Li, and Lin Ye Copyright © 2016 Liting Zhou et al. All rights reserved. Pressure Drop in Tortuosity/Kinking of the Internal Carotid Artery: Simulation and Clinical Investigation Mon, 18 Apr 2016 07:24:22 +0000 Background. Whether carotid tortuosity/kinking of the internal carotid artery leads to cerebral ischemia remains unclear. There is very little research about the hemodynamic variation induced by carotid tortuosity/kinking in the literature. The objective of this study was to research the blood pressure changes induced by carotid tortuosity/kinking. Methods. We first created a geometric model of carotid tortuosity/kinking. Based on hemodynamic boundary conditions, the hemodynamics of carotid tortuosity and kinking were studied via a finite element simulation. Then, an in vitro system was built to validate the numerical simulation results. The mean arterial pressure changes before and after carotid kinking were measured using pressure sensors in 12 patients with carotid kinking. Results. Numerical simulation revealed that the pressure drops increased with increases in the kinking angles. Clinical tests and in vitro experiments confirmed the numerical simulation results. Conclusions. Carotid kinking leads to blood pressure reduction. In certain conditions, kinking may affect the cerebral blood supply and be associated with cerebral ischemia. Lijun Wang, Feng Zhao, Daming Wang, Shen Hu, Jiachun Liu, Zhilun Zhou, Jun Lu, Peng Qi, and Shiying Song Copyright © 2016 Lijun Wang et al. All rights reserved. Analyzing Dynamic Changes of Laboratory Indexes in Patients with Acute Heart Failure Based on Retrospective Study Wed, 06 Apr 2016 06:49:55 +0000 Background. Changes of N-terminal probrain natriuretic peptide (NT-proBNP) have been studied whether in the long term or the short term in patients of acute heart failure (AHF); however, changes of NT-proBNP in the first five days and their association with other factors have not been investigated. Aims. To describe the dynamic changes of relevant laboratory indexes in the first five days between different outcomes of AHF patients and their associations. Methods and Results. 284 AHF with dynamic values recorded were analyzed. Changes of NT-proBNP, troponin T, and C-reactive protein were different between patients with different outcomes, with higher values in adverse group than in control group at the same time points (). Then, prognostic use and risk stratification of NT-proBNP were assessed by receiver-operating characteristic curve and logistic regression. NT-proBNP levels at day 3 showed the best prognostic power (area under the curve = 0.730, 95% confidence interval (CI): 0.657 to 0.794) and was an independent risk factor for adverse outcome (odds ratio, OR: 2.185, 95% CI: 1.584–3.015). Classified changes of NT-proBNP may be predictive for adverse outcomes in AHF patients. Conclusions. Sequential monitoring of laboratory indexes within the first 5 days may be helpful for management of AHF patients. Yurong Wang, Lei Fu, Qian Jia, Hao Yu, Pengjun Zhang, Chunyan Zhang, Xueliang Huang, Kunlun He, and Yaping Tian Copyright © 2016 Yurong Wang et al. All rights reserved. A Nested Case-Control Study of Association between Metabolome and Hypertension Risk Tue, 29 Mar 2016 12:24:57 +0000 We aimed to explore novel small metabolites that associated with hypertension risk in a population-based nested case-control study. Among 460 individuals with optimal blood pressure (<120/80 mmHg) at baseline, 55 progressed to hypertension during 5 years of follow-up. Twenty-nine cases of incident hypertension and 29 controls, matched for age, sex, and baseline systolic blood pressure, were included in this study. Serum metabolites were measured by gas chromatography-tandem mass spectrometry. -test and logistic regression analysis were applied to investigate the association between metabolites and incident hypertension. Among the 241 metabolites identified in this study, baseline levels of 26 metabolites were significantly different between hypertension and control groups. After adjusting for body mass index, smoking, and drinking, 16 out of the 26 metabolites were still associated with hypertension risk including four amino acids. Amino acids were negatively associated with risk of future hypertension, with odds ratio (OR) ranging from 0.33 to 0.53. Two of these amino acids were essential amino acids including threonine and phenylalanine. Higher level of lyxose, a fermentation product of gut microbes, was associated with higher risk of hypertension. Our study identified multiple metabolites that associated with hypertension risk. These findings implied that low amino acid levels and gut microbiome might play an important role in the pathogenesis of hypertension. Yongchen Hao, Ying Wang, Lu Xi, Guoqi Li, Fan Zhao, Yue Qi, Jing Liu, and Dong Zhao Copyright © 2016 Yongchen Hao et al. All rights reserved. Ramipril and Losartan Exert a Similar Long-Term Effect upon Markers of Heart Failure, Endogenous Fibrinolysis, and Platelet Aggregation in Survivors of ST-Elevation Myocardial Infarction: A Single Centre Randomized Trial Tue, 15 Mar 2016 17:03:42 +0000 Introduction. Blocking the renin-angiotensin-aldosterone system in ST-elevation myocardial infarction (STEMI) patients prevents heart failure and recurrent thrombosis. Our aim was to compare the effects of ramipril and losartan upon the markers of heart failure, endogenous fibrinolysis, and platelet aggregation in STEMI patients over the long term. Methods. After primary percutaneous coronary intervention (PPCI), 28 STEMI patients were randomly assigned ramipril and 27 losartan, receiving therapy for six months with dual antiplatelet therapy (DAPT). We measured N-terminal proBNP (NT-proBNP), ejection fraction (EF), plasminogen-activator-inhibitor type 1 (PAI-1), and platelet aggregation by closure times (CT) at the baseline and after six months. Results. Baseline NT-proBNP ≥ 200 pmol/mL was observed in 48.1% of the patients, EF < 55% in 49.1%, and PAI-1 ≥ 3.5 U/mL in 32.7%. Six-month treatment with ramipril or losartan resulted in a similar effect upon PAI-1, NT-proBNP, EF, and CT levels in survivors of STEMI, but in comparison to control group, receiving DAPT alone, ramipril or losartan treatment with DAPT significantly increased mean CT (226.7 ± 80.3 sec versus 158.1 ± 80.3 sec, ). Conclusions. Ramipril and losartan exert a similar effect upon markers of heart failure and endogenous fibrinolysis, and, with DAPT, a more efficient antiplatelet effect in long term than DAPT alone. Martin Marinšek and Andreja Sinkovič Copyright © 2016 Martin Marinšek and Andreja Sinkovič. All rights reserved. The Diagnostic Role of Adiponectin in Pulmonary Embolism Sun, 06 Mar 2016 08:57:05 +0000 Background and Aims. Pulmonary thromboembolism (PTE) is a frequent disease with difficult diagnosis and high mortality. Misdiagnosis occurs in 2/3 patients and mortality rates reach up to 30%. The aim of our study was to investigate the role of adiponectin used in emergency service in diagnosis of PTE. Materials and Methods. 95 patients with suspected PTE included in the study. Plasma adiponectin and D-dimer levels were measured and chest X-ray and multidetector row computed tomography scan obtained. Diagnosis was supported by vascular filling defect on tomography. Control group consisted of patients with suspected PTE and normal chest computed tomography findings. Results. Mean D-dimer level was  ng/mL in patients and  ng/mL in the control group (). Mean adiponectin level was  μg/mL in patients and  μg/mL in the control group (). Wells and Geneva scores were higher in patients compared to the control group. Conclusions. As a result, we conclude that lower adiponectin levels have an important role in the diagnosis of PTE. Evrim Gul, Yeliz Gul, Ersin Yıldırım, Mustafa Safa Pepele, Mustafa Yıldız, Mehmet Nuri Bozdemir, Mehmet Ruhi Onur, Bengü Mutlu, Feti Yıldız, Ömer Doğan Alataş, and Necip İlhan Copyright © 2016 Evrim Gul et al. All rights reserved. TBS Predict Coronary Artery Calcification in Adults Sun, 06 Mar 2016 08:18:55 +0000 Purpose. This study analyzes the association between the bony microarchitecture score (trabecular bone score, TBS) and coronary artery calcification (CAC) in adults undergoing health exams. Materials and Methods. We retrospectively collected subjects () who underwent coronary computed tomography and bone mineral density studies simultaneously. CAC was categorized to three levels (Group 0, G0, no CAC, score = 0, ; Group 1, G1, moderate CAC, score = 1–100, ; Group 2, G2, high CAC, score 101, ). Multinomial logistic regression was used to study the association between TBS and CAC levels. Results. CAC is present in 44.4% of the population. Mean TBS ± SD was . Per 1 SD increase in TBS, the unadjusted odds ratio (2.393) of moderate CAC compared with no CAC was significantly increased (95% CI, 1.219–4.696, ). However, there has been no association of TBS with high CAC (OR: 1.026, 95% CI: 0.586–1.797, ). These relationships also existed when individually adjusted for age, sex, and multiple other covariates. Conclusions. Higher TBS was related to moderate CAC, but not high CAC; a possible explanation may be that bone microarchitecture remodeling becomes more active when early coronary artery calcification occurs. However, further researches are needed to clarify this pathophysiology. Tzyy-Ling Chuang, Fu-Tsung Hsiao, Yi-Da Li, and Yuh-Feng Wang Copyright © 2016 Tzyy-Ling Chuang et al. All rights reserved. Circulating Long Noncoding RNA UCA1 as a Novel Biomarker of Acute Myocardial Infarction Mon, 01 Feb 2016 13:02:41 +0000 Acute myocardial infarction (AMI) is the most serious cardiovascular disease with high morbidity and mortality. Recent studies have showed that long noncoding RNAs (lnc RNA) play important roles in pathophysiology of cardiovascular diseases, but the investigations are still in their infancy. An lnc RNA named urothelial carcinoma-associated 1 (UCA1) is found in tumors such as bladder cancers and lung cancer. And the UCA1 could be as a predictive biomarker for bladder cancer in urine samples or lung cancer in plasma, respectively. In normal states, UCA1 is specifically expressed in heart of adult, indicating that UCA1 might be as a biomarker for heart diseases such as AMI. To test the speculation, we detect the level of UCA1 in plasma of AMI patients and health control using quantitative reverse transcription-polymerase chain reaction (qRT-PCR). In addition, we also test the level of miR-1 as it is reported to regulate the expression of UCA1. The results show that the level of plasma UCA1 is decreased at the early state of AMI patients and increased at day 3 after AMI. In addition, the UCA1 alteration is inversely associated with the expression of miR-1. These findings indicate that the circulating UCA1 could be used as a promising novel biomarker for the diagnosis and/or prognosis of AMI. Youyou Yan, Bin Zhang, Ning Liu, Chao Qi, Yanlong Xiao, Xin Tian, Tianyi Li, and Bin Liu Copyright © 2016 Youyou Yan et al. All rights reserved. Gender-Specific Association of ATP2B1 Variants with Susceptibility to Essential Hypertension in the Han Chinese Population Mon, 11 Jan 2016 07:43:51 +0000 Previous genome-wide association studies (GWASs) found that several ATP2B1 variants are associated with essential hypertension (EHT). But the “genome-wide significant” ATP2B1 SNPs (rs2681472, rs2681492, rs17249754, and rs1105378) are in strong linkage disequilibrium (LD) and are located in the same LD block in Chinese populations. We asked whether there are other SNPs within the ATP2B1 gene associated with susceptibility to EHT in the Han Chinese population. Therefore, we performed a case-control study to investigate the association of seven tagSNPs within the ATP2B1 gene and EHT in the Han Chinese population, and we then analyzed the interaction among different SNPs and nongenetic risk factors for EHT. A total of 902 essential hypertensive cases and 902 normotensive controls were involved in the study. All 7 tagSNPs within the ATP2B1 gene were retrieved from HapMap, and genotyping was performed using the Tm-shift genotyping method. Chi-squared test, logistic regression, and propensity score analysis showed that rs17249754 was associated with EHT, particularly in females. The MDR analysis demonstrated that the interaction of rs2070759, rs17249754, TC, TG, and BMI increased the susceptibility to hypertension. Crossover analysis and stratified analysis indicated that BMI has a major effect on the development of hypertension, while ATP2B1 variants have a minor effect. Jin Xu, Hai-xia Qian, Su-pei Hu, Li-ya Liu, Mi Zhou, Mei Feng, Jia Su, and Lin-dan Ji Copyright © 2016 Jin Xu et al. All rights reserved. Autocrine Human Urotensin II Enhances Macrophage-Derived Foam Cell Formation in Transgenic Rabbits Thu, 12 Nov 2015 10:25:55 +0000 Circulating urotensin II (UII) is involved in the development of atherosclerosis. However, the role of autocrine UII in the development of atherosclerosis remains unclear. Here, we tested the hypothesis that autocrine UII would promote atherosclerosis. Transgenic rabbits were created as a model to study macrophage-specific expressing human UII (hUII) and used to investigate the role of autocrine UII in the development of atherosclerosis. Transgenic rabbits and their nontransgenic littermates were fed a high cholesterol diet to induce atherosclerosis. Comparing the transgenic rabbits with their nontransgenic littermates, it was observed that hUII expression increased the macrophage-positive area in the atherosclerotic lesions by 45% and the positive area ratio by 56% in the transgenic rabbits. Autocrine hUII significantly decreased the smooth muscle cell-positive area ratio in transgenic rabbits (by 54%), without affecting the plasma levels of total cholesterol, triglycerides, low-density lipoprotein cholesterol, high-density lipoprotein cholesterol, and glucose and adipose tissue contents. These results elucidated for the first time that autocrine UII plays an important role in the development of atherosclerosis by increasing the accumulation of macrophage-derived foam cell. Sihai Zhao, Yafeng Li, Shoucui Gao, Xiaojing Wang, Lijing Sun, Daxing Cheng, Liang Bai, Hua Guan, Rong Wang, Jianglin Fan, and Enqi Liu Copyright © 2015 Sihai Zhao et al. All rights reserved. Puerarin Inhibits oxLDL-Induced Macrophage Activation and Foam Cell Formation in Human THP1 Macrophage Wed, 21 Oct 2015 12:13:41 +0000 Puerarin, an isoflavone derived from Kudzu roots, has been widely used for treatment of cardiovascular and cerebral vascular diseases in China and other Asian countries. However, the underlying mechanisms are largely unknown. The present study investigated whether puerarin inhibited atherogenic lipid oxLDL-mediated macrophage activation and foam cell formation in human THP1 macrophage. Treatment with oxLDL significantly increased the mRNA expression of proinflammatory cytokines tumor necrosis factor α (TNFα, 160%) and interleukin (IL) 1β (13 fold) accompanied by upregulation of toll-like receptor 4 (TLR4, 165%) and the ratio of phospho-IκBα/IκBα in THP1 macrophage. Puerarin dose-dependently prevented an increase in oxLDL-induced proinflammatory gene expression with downregulation of TLR4 and the ratio of phospho-IκBα/IκBα. Furthermore, puerarin prevented oxLDL-mediated lipid deposition and foam cell formation associated with downregulation of scavenger receptor CD36. Flow cytometry analysis showed that puerarin reduced the number of early apoptotic cells of macrophages induced by oxLDL. Our results show that puerarin has anti-inflammatory and antiatherogenic effects in vitro; the underlying mechanisms may involve the inhibition of TLR4/NFκB pathway and downregulation of CD36 expression. The results from the present study provide scientific evidence and may expand our armamentarium to use puerarin for prevention and treatment of cardiovascular and atherosclerotic diseases. Heng Zhang, Zhenhua Zhai, Hongyu Zhou, Yao Li, Xiaojie Li, Yuhan Lin, Weihong Li, Yueping Shi, and Ming-Sheng Zhou Copyright © 2015 Heng Zhang et al. All rights reserved. Shear Stress Induces Differentiation of Endothelial Lineage Cells to Protect Neonatal Brain from Hypoxic-Ischemic Injury through NRP1 and VEGFR2 Signaling Mon, 05 Oct 2015 07:45:17 +0000 Neonatal hypoxic-ischemic (HI) brain injuries disrupt the integrity of neurovascular structure and lead to lifelong neurological deficit. The devastating damage can be ameliorated by preserving the endothelial network, but the source for therapeutic cells is limited. We aim to evaluate the beneficial effect of mechanical shear stress in the differentiation of endothelial lineage cells (ELCs) from adipose-derived stem cells (ASCs) and the possible intracellular signals to protect HI injury using cell-based therapy in the neonatal rats. The ASCs expressed early endothelial markers after biochemical stimulation of endothelial growth medium. The ELCs with full endothelial characteristics were accomplished after a subsequential shear stress application for 24 hours. When comparing the therapeutic potential of ASCs and ELCs, the ELCs treatment significantly reduced the infarction area and preserved neurovascular architecture in HI injured brain. The transplanted ELCs can migrate and engraft into the brain tissue, especially in vessels, where they promoted the angiogenesis. The activation of Akt by neuropilin 1 (NRP1) and vascular endothelial growth factor receptor 2 (VEGFR2) was important for ELC migration and following in vivo therapeutic outcomes. Therefore, the current study demonstrated importance of mechanical factor in stem cell differentiation and showed promising protection of brain from HI injury using ELCs treatment. Chia-Wei Huang, Chao-Ching Huang, Yuh-Ling Chen, Shih-Chen Fan, Yuan-Yu Hsueh, Chien-Jung Ho, and Chia-Ching Wu Copyright © 2015 Chia-Wei Huang et al. All rights reserved. Pigment Epithelium-Derived Factor Induces Endothelial Barrier Dysfunction via p38/MAPK Phosphorylation Sun, 04 Oct 2015 13:44:56 +0000 Endothelial barrier dysfunction, which is a serious problem that occurs in various inflammatory conditions, permits extravasation of serum components into the surrounding tissues, leading to edema formation and organ failure. Pigment epithelium-derived factor (PEDF), which is a major endogenous antagonist, has been implicated in diverse biological process, but its role in endothelial barrier dysfunction has not been defined. To assess the role of PEDF in the vasculature, we evaluated the effects of exogenous PEDF using human umbilical vein endothelial cells (HUVECs) in vitro. Our results demonstrated that exogenous PEDF activated p38/MAPK signalling pathway in a dose- and time-dependent manner and induced vascular hyperpermeability as measured by the markedly increased FITC-dextran leakage and the decreased transendothelial electrical resistance (TER) across the monolayer cells, which was accompanied by microtubules (MTs) disassembly and F-actin rearrangement. However, the aforementioned alterations can be arrested by the application of low concentration of p38/MAPK inhibitor SB203580. These results reveal a novel role for PEDF as a potential vasoactive substance in inducing hyperpermeability. Furthermore, our results suggest that PEDF and p38/MAPK may serve as therapeutic targets for maintaining vascular integrity. Ting He, Liping Zhao, Dongxia Zhang, Qiong Zhang, Jiezhi Jia, Jiongyu Hu, and Yuesheng Huang Copyright © 2015 Ting He et al. All rights reserved. Extracellular Calcium-Dependent Modulation of Endothelium Relaxation in Rat Mesenteric Small Artery: The Role of Potassium Signaling Sun, 04 Oct 2015 13:40:18 +0000 The nature of NO- and COX-independent endothelial hyperpolarization (EDH) is not fully understood but activation of small- and intermittent-conductance Ca2+-activated K+ channels ( and ) is important. Previous studies have suggested that the significance of depends on . Also it has been suggested that K+ is important through localized signaling causing activation of the Na+,K+-ATPase and inward-rectifying K+ channels (). Here we tested the hypothesis that the modulating effect of on the EDH-like response depends on . We addressed this possibility using isometric myography of rat mesenteric small arteries. When was 4.2 mM, relaxation to acetylcholine (ACh) was stronger at 2.5 mM than at 1 mM . Inhibition of with TRAM34 suppressed the relaxations but did not change the relation between the relaxations at the low and high . This -dependence disappeared at 5.9 mM and in the presence of ouabain or BaCl2. Our results suggest that are involved in the localized signaling which acts through the Na+,K+-ATPase and channels and that the significance of this endothelium-dependent pathway is modulated by . Lise Hangaard, Peter B. Jessen, Dmitrii Kamaev, Christian Aalkjaer, and Vladimir V. Matchkov Copyright © 2015 Lise Hangaard et al. All rights reserved. Endothelial Expression of Scavenger Receptor Class B, Type I Protects against Development of Atherosclerosis in Mice Sun, 04 Oct 2015 10:38:25 +0000 The role of scavenger receptor class B, type I (SR-BI) in endothelial cells (EC) was examined in several novel transgenic mouse models expressing SR-BI in endothelium of mice with normal C57Bl6/N, apoE-KO, or Scarb1-KO backgrounds. Mice were also created expressing SR-BI exclusively in endothelium and liver. Endothelial expression of the Tie2-Scarb1 transgene had no significant effect on plasma lipoprotein levels in mice on a normal chow diet but on an atherogenic diet, significantly decreased plasma cholesterol levels, increased plasma HDL cholesterol (HDL-C) levels, and protected mice against atherosclerosis. In 8-month-old apoE-KO mice fed a normal chow diet, the Tie2-Scarb1 transgene decreased aortic lesions by 24%. Mice expressing SR-BI only in EC and liver had a 1.5 ± 0.1-fold increase in plasma cholesterol compared to mice synthesizing SR-BI only in liver. This elevation was due mostly to increased HDL-C. In EC culture studies, SR-BI was found to be present in both basolateral and apical membranes but greater cellular uptake of cholesterol from HDL was found in the basolateral compartment. In summary, enhanced expression of SR-BI in EC resulted in a less atherogenic lipoprotein profile and decreased atherosclerosis, suggesting a possible role for endothelial SR-BI in the flux of cholesterol across EC. Boris L. Vaisman, Tatyana G. Vishnyakova, Lita A. Freeman, Marcelo J. Amar, Stephen J. Demosky, Chengyu Liu, John A. Stonik, Maureen L. Sampson, Milton Pryor, Alexander V. Bocharov, Thomas L. Eggerman, Amy P. Patterson, and Alan T. Remaley Copyright © 2015 Boris L. Vaisman et al. All rights reserved. Dipeptidyl Peptidase 4: A New Link between Diabetes Mellitus and Atherosclerosis? Thu, 04 Jun 2015 09:55:18 +0000 Type 2 diabetes mellitus (T2DM) has become one of the most prevalent noncommunicable diseases in the past years. It is undoubtedly associated with atherosclerosis and increased risk for cardiovascular diseases. Incretins, which are intestinal peptides secreted during digestion, are able to increase insulin secretion and its impaired function and/or secretion is involved in the pathophysiology of T2DM. Dipeptidyl peptidase 4 (DPP4) is an ubiquitous enzyme that regulates incretins and consequently is related to the pathophysiology of T2DM. DPP4 is mainly secreted by endothelial cells and acts as a regulatory protease for cytokines, chemokines, and neuropeptides involved in inflammation, immunity, and vascular function. In T2DM, the activity of DPP4 seems to be increased and there are a growing number of in vitro and in vivo studies suggesting that this enzyme could be a new link between T2DM and atherosclerosis. Gliptins are a new class of pharmaceutical agents that acts by inhibiting DPP4. Thus, it is expected that gliptin represents a new pharmacological approach not only for reducing glycemic levels in T2DM, but also for the prevention and treatment of atherosclerotic cardiovascular disease in diabetic subjects. We aimed to review the evidences that reinforce the associations between DPP4, atherosclerosis, and T2DM. Wellington Santana da Silva Júnior, Amélio Fernando de Godoy-Matos, and Luiz Guilherme Kraemer-Aguiar Copyright © 2015 Wellington Santana da Silva Júnior et al. All rights reserved. Vibration Training Triggers Brown Adipocyte Relative Protein Expression in Rat White Adipose Tissue Mon, 01 Jun 2015 11:19:19 +0000 Recently, vibration training is considered as a novel strategy of weight loss; however, its mechanisms are still unclear. In this study, normal or high-fat diet-induced rats were trained by whole body vibration for 8 weeks. We observed that the body weight and fat metabolism index, blood glucose, triglyceride, cholesterol, and free fatty acid in obesity rats decreased significantly compared with nonvibration group . Although intrascapular BAT weight did not change significantly, vibration enhanced ATP reduction and increased protein level of the key molecule of brown adipose tissue (BAT), PGC-1α, and UCP1 in BAT. Interestingly, the adipocytes in retroperitoneal white adipose tissue (WAT) became smaller due to vibration exercise and had higher protein level of the key molecule of brown adipose tissue (BAT), PGC-1α, and UCP1 and inflammatory relative proteins, IL-6 and TNFα. Simultaneously, ATP content and PPARγ protein level in WAT became less in rats compared with nonvibration group. The results indicated that vibration training changed lipid metabolism in rats and promoted brown fat-like change in white adipose tissues through triggering BAT associated gene expression, inflammatory reflect, and reducing energy reserve. Chao Sun, Ruixia Zeng, Ge Cao, Zhibang Song, Yibo Zhang, and Chang Liu Copyright © 2015 Chao Sun et al. All rights reserved. Heart Failure: Advanced Development in Genetics and Epigenetics Thu, 09 Apr 2015 10:16:53 +0000 Heart failure (HF) is a complex pathophysiological syndrome that arises from a primary defect in the ability of the heart to take in and/or eject sufficient blood. Genetic mutations associated with familial dilated cardiomyopathy, hypertrophic cardiomyopathy, and arrhythmogenic right ventricular cardiomyopathy can contribute to the various pathologies of HF. Therefore, genetic screening could be an approach for guiding individualized therapies and surveillance. In addition, epigenetic regulation occurs via key mechanisms, including ATP-dependent chromatin remodeling, DNA methylation, histone modification, and RNA-based mechanisms. MicroRNA is also a hot spot in HF research. This review gives an overview of genetic mutations associated with cardiomyopathy and the roles of some epigenetic mechanisms in HF. Jian Yang, Wei-wei Xu, and Shen-jiang Hu Copyright © 2015 Jian Yang et al. All rights reserved. Angiogenesis Sun, 29 Mar 2015 13:18:54 +0000 Qiang Zhao and Zongjin Li Copyright © 2015 Qiang Zhao and Zongjin Li. All rights reserved. Skin-Derived Precursor Cells Promote Angiogenesis and Stimulate Proliferation of Endogenous Neural Stem Cells after Cerebral Infarction Thu, 26 Mar 2015 06:25:47 +0000 Stroke is one of the most common diseases that caused high mortality and has become burden to the health care systems. Stem cell transplantation has shown therapeutic effect in ameliorating ischemic damage after cerebral artery occlusion mainly due to their neurogenesis, immune regulation, or effects on the plasticity, proliferation, and survival of host cells. Recent studies demonstrated that skin-derived precursor cells (SKPs) could promote central nervous system regeneration in spinal cord injury model or the neonatal peripheral neuron. Here, we investigated the therapeutic potential of SKPs in a rat model of cerebral ischemia. SKPs were isolated, expanded, and transplanted into rat cortex and striatum after transient middle cerebral artery occlusion. Our results revealed that SKPs transplantation could improve the behavioral measures of neurological deficit. Moreover, immunohistology confirmed that SKPs could secrete basic FGF and VEGF in the ischemic region and further markedly increase the proliferation of endogenous nestin+ and βIII-tubulin+ neural stem cells. Furthermore, increased angiogenesis induced by SKPs was observed by vWF and α-SMA staining. These data suggest that SKPs induced endogenous neurogenesis and angiogenesis and protected neuron from hypoxic-ischemic environment. In conclusion, SKPs transplantation may be a promising approach in treatment of stroke. Duo Mao, Xinpeng Yao, Guowei Feng, Xiaoqing Yang, Lina Mao, Xiaomin Wang, Tingyu Ke, Yongzhe Che, and Deling Kong Copyright © 2015 Duo Mao et al. All rights reserved. Activating Transcription Factor 4 Promotes Angiogenesis of Breast Cancer through Enhanced Macrophage Recruitment Thu, 26 Mar 2015 05:57:34 +0000 Angiogenesis plays an important role in the progression of tumor. Besides being regulated by tumor cells per se, tumor angiogenesis is also influenced by stromal cells in tumor microenvironment (TME), for example, tumor associated macrophages (TAMs). Activating transcription factor 4 (ATF4), a member of the ATF/CREB family, has been reported to be related to tumor angiogenesis. In this study, we found that exogenous overexpression of ATF4 in mouse breast cancer cells promotes tumor growth via increasing tumor microvascular density. However, ATF4 overexpression failed to increase the expression level of a series of proangiogenic factors including vascular endothelial growth factor A (VEGFA) in tumor cells in this model. Thus, we further investigated the infiltration of proangiogenic macrophages in tumor tissues and found that ATF4-overexpressing tumors could recruit more macrophages via secretion of macrophage colony stimulating factor (M-CSF). Overall, we concluded that exogenous overexpression of ATF4 in breast cancer cells may facilitate the recruitment of macrophages into tumor tissues and promote tumor angiogenesis and tumor growth indirectly. Chen Liu, Zongjin Li, Lina Wang, Lingling Tong, Ningning He, Yanan Chen, Yanhua Liu, Zhongjun Wu, Peiqing Sun, Rong Xiang, Guosheng Ren, and Weijun Su Copyright © 2015 Chen Liu et al. All rights reserved. Corrigendum to “SIRT1 Inhibition Affects Angiogenic Properties of Human MSCs” Wed, 25 Mar 2015 13:45:31 +0000 Chiara Botti, Ilaria Caiafa, Antonietta Coppola, Francesca Cuomo, Marco Miceli, Lucia Altucci, and Gilda Cobellis Copyright © 2015 Chiara Botti et al. All rights reserved. Actin-Tethered Junctional Complexes in Angiogenesis and Lymphangiogenesis in Association with Vascular Endothelial Growth Factor Wed, 25 Mar 2015 13:35:45 +0000 Vasculature is present in all tissues and therefore is indispensable for development, biology, and pathology of multicellular organisms. Endothelial cells guarantee proper function of the vessels and are the original component in angiogenesis. Morphogenesis of the vascular system utilizes processes like cell adhesion, motility, proliferation, and survival that are closely related to the dynamics of actin filaments and actin-tethered adhesion complexes. Here we review involvement of actin cytoskeleton-associated junctional molecules of endothelial cells in angiogenesis and lymphangiogenesis. Particularly, we focus on F-actin binding protein afadin, an adaptor protein involved in broad range of signaling mechanisms. Afadin mediates the pathways of vascular endothelial growth factor- (VEGF-) and sphingosine 1-phosphate-triggered angiogenesis and is essential for embryonic development of lymph vessels in mice. We propose that targeting actin-tethered junctional molecules, including afadin, may present a new approach to angiogenic therapy that in combination with today used medications like VEGF inhibitors will benefit against development of pathological angiogenesis. Dimitar P. Zankov and Hisakazu Ogita Copyright © 2015 Dimitar P. Zankov and Hisakazu Ogita. All rights reserved. Enhanced Vascularization in Hybrid PCL/Gelatin Fibrous Scaffolds with Sustained Release of VEGF Wed, 25 Mar 2015 13:19:25 +0000 Creating a long-lasting and functional vasculature represents one of the most fundamental challenges in tissue engineering. VEGF has been widely accepted as a potent angiogenic factor involved in the early stages of blood vessel formation. In this study, fibrous scaffolds that consist of PCL and gelatin fibers were fabricated. The gelatin fibers were further functionalized by heparin immobilization, which provides binding sites for VEGF and thus enables the sustained release of VEGF. In vitro release test confirms the sustained releasing profile of VEGF, and stable release was observed over a time period of 25 days. In vitro cell assay indicates that VEGF release significantly promoted the proliferation of endothelial cells. More importantly, in vivo subcutaneous implantation reflects that vascularization has been effectively enhanced in the PCL/gelatin scaffolds compared with the PCL counterpart due to the sustained release of VEGF. Therefore, the heparinized PCL/gelatin scaffolds developed in this study may be a promising candidate for regeneration of complex tissues with sufficient vascularization. Kai Wang, Xuejiao Chen, Yiwa Pan, Yun Cui, Xin Zhou, Deling Kong, and Qiang Zhao Copyright © 2015 Kai Wang et al. All rights reserved. PGCCs Generating Erythrocytes to Form VM Structure Contributes to Tumor Blood Supply Wed, 25 Mar 2015 13:18:47 +0000 Shiwu Zhang, Xiaochun Xu, Siwei Zhu, and Jun Liu Copyright © 2015 Shiwu Zhang et al. All rights reserved. Attenuating Tumour Angiogenesis: A Preventive Role of Metformin against Breast Cancer Wed, 25 Mar 2015 12:57:55 +0000 Metformin is one of the most widely prescribed antidiabetics for type 2 diabetes. A critical role of metformin against tumorigenesis has recently been implicated, although several studies also reported the lack of anticancer property of the antidiabetics. Given the controversies regarding the potential role of metformin against tumour progression, the effect of metformin against breast, cervical, and ovarian tumour cell lines was examined followed by in vivo assessment of metformin on tumour growth using xenograft breast cancer models. Significant inhibitory impact of metformin was observed in MCF-7, HeLa, and SKOV-3 cells, suggesting an antiproliferative property of metformin against breast, cervical, and ovarian tumour cells, respectively, with the breast tumour cells, MCF-7, being the most responsive. In vivo assessment was subsequently carried out, where mice with breast tumours were treated with metformin (20 mg/kg body weight) or sterile PBS solution for 15 consecutive days. No inhibition of breast tumour progression was detected. However, tumour necrosis was significantly increased in the metformin-treated group, accompanied by decreased capillary formation within the tumours. Thus, despite the lack of short-term benefit of metformin against tumour progression, a preventive role of metformin against breast cancer was implicated, which is at partially attributable to the attenuation of tumour angiogenesis. Shan Gao, Jingcheng Jiang, Pan Li, Huijuan Song, Weiwei Wang, Chen Li, and Deling Kong Copyright © 2015 Shan Gao et al. All rights reserved. Diffuse Calcifications Protect Carotid Plaques regardless of the Amount of Neoangiogenesis and Related Histological Complications Wed, 25 Mar 2015 11:20:59 +0000 Background. Neoangiogenesis is crucial in plaque progression and instability. Previous data from our group showed that Nestin-positive intraplaque neovessels correlated with histological complications. The aim of the present work is to evaluate the relationship between neoangiogenesis, plaque morphology, and clinical instability of the plaque. Materials and Methods. Seventy-three patients (53 males and 20 females, mean age 71 years) were consecutively enrolled. Clinical data and 14 histological variables, including intraplaque hemorrhage and calcifications, were collected. Immunohistochemistry for CD34 and Nestin was performed. RT-PCR was performed to evaluate Nestin mRNA (including 5 healthy arteries as controls). Results. Diffusely calcified plaques (13/73) were found predominantly in females , with a significantly lower incidence of symptoms (TIA/stroke than noncalcified plaques but with the same incidence of histological complications ). Accordingly, calcified and noncalcified plaques showed similar mean densities of positivity for CD34 and Nestin. Nestin density, but not CD34, correlated with the occurrence of intraplaque hemorrhage. Conclusions. Plaques with massive calcifications show the same incidence of histological complications but without influencing symptomatology, especially in female patients, and regardless of the amount of neoangiogenesis. These results can be applied in a future presurgical identification of patients at major risk of developing symptoms. Francesco Vasuri, Silvia Fittipaldi, Rodolfo Pini, Alessio Degiovanni, Raffaella Mauro, Antonia D’Errico-Grigioni, Gianluca Faggioli, Andrea Stella, and Gianandrea Pasquinelli Copyright © 2015 Francesco Vasuri et al. All rights reserved. Gemcitabine, Navelbine, and Doxorubicin as Treatment for Patients with Refractory or Relapsed T-Cell Lymphoma Thu, 19 Mar 2015 12:12:30 +0000 T-cell lymphoma (TCL) is resistant to conventional chemotherapy. We retrospectively evaluated the therapeutic efficiency and toxicity of gemcitabine, navelbine, and doxorubicin (GND) in patients with refractory or relapsed TCL. From 2002 to 2012, 69 patients with refractory or relapsed TCL received GND treatment in our hospital. The treatment protocol comprised gemcitabine (800 mg/m2, group 1; 1000 mg/m2, group 2) on days 1 and 8, navelbine (25 mg/m2) on day 1, and doxorubicin (20 mg/m2) on day 1, repeated every 3 weeks. The overall response rate (ORR) was 65.2%. The median overall survival (OS) was 36 months. The 5-year estimated OS rate was 32.4%. The GND regimen was well tolerated. Subgroup analysis demonstrated that the ORR and CR for group 1 were similar. A longer median OS was observed for group 1. Significant difference in grades 3-4 toxicities was observed between groups 1 and 2 (). Our study indicated that gemcitabine (800 mg/m2) on days 1 and 8 every 21 days was favorable for pretreated TCL patients. Zhengzi Qian, Zheng Song, Huilai Zhang, Xianhuo Wang, Jing Zhao, and Huaqing Wang Copyright © 2015 Zhengzi Qian et al. All rights reserved. Construction of a CXCL12-KDEL Fusion Gene to Inhibit Head and Neck Squamous Cell Carcinoma Metastasis by Intracellular Sequestration of CXCR4 Thu, 19 Mar 2015 11:38:15 +0000 The CXCL12-CXCR4 biological axis consisting of the chemotactic factor CXCL12 and its specific receptor CXCR4 plays an important role in oral cancer metastasis. High expression of CXCR4 may help oral squamous cancer cells invade local tissues and metastasize to lymph nodes. No obvious association was observed between CXCL12 expression and lymph node metastasis, suggesting that CXCL12 chemotaxis may only be related to CXCR4 expression on the tumor cell membrane. KDEL can be retained by receptors on the surface of the intracellular endoplasmic reticulum (ER) and also be called an ER retention signal sequence. So we adopted the KDEL sequence in this study to generate a CXCL12-KDEL fusion protein in combination with a traceable E-tag label. As such, CXCL12 was retained in the ER. Specific receptor CXCR4 binds to the CXCL12-KDEL, was also retained in the ER, and was thus prevented from reaching the oral squamous cancer cell surface. We reduced the cell surface level of CXCR4 and called the technique “intracellular sequestration.” By this way, we have finished blocking of CXCL12-CXCR4 biological axis and inhibiting lymph node metastasis of oral carcinoma. Wenchao Zhang, Xudong Wang, Kai Yue, Su Liu, and Xiaonan Liu Copyright © 2015 Wenchao Zhang et al. All rights reserved. A Phase I Trial to Evaluate the Multiple-Dose Safety and Antitumor Activity of Ursolic Acid Liposomes in Subjects with Advanced Solid Tumors Thu, 19 Mar 2015 09:51:36 +0000 Ursolic acid liposome (UAL), a new antitumor drug, has potential therapeutic value. However, limited clinical data exists regarding multiple-dose safety, antitumor activity, and the recommended dose (RD) of UAL for subjects with advanced solid tumors. All subjects were intravenously administered UAL for 14 consecutive days of a 21-day treatment cycle. Twenty-one subjects were enrolled in 1 of 3 sequential cohorts (56, 74, and 98 mg/m2) to evaluate multiple-dose tolerability and efficacy. Eight additional subjects were treated with UAL (74 mg/m2) to evaluate multiple-dose pharmacokinetics. No ≥grade 3 adverse events (NCI-CTC) were observed. Sixty percent subjects achieved stable disease after 2 treatment cycles. Multiple-dose pharmacokinetic analysis suggested UAL does not accumulate in the body. This trial demonstrates that UAL was tolerable, had manageable toxicity, and could potentially improve patient remission rates. A large phase II study is recommended to confirm these results (i.e., RD of 98 mg/m2). Zhengzi Qian, Xianhuo Wang, Zheng Song, Huilai Zhang, Shiyong Zhou, Jing Zhao, and Huaqing Wang Copyright © 2015 Zhengzi Qian et al. All rights reserved. The Effect of Iloprost and N-Acetylcysteine on Skeletal Muscle Injury in an Acute Aortic Ischemia-Reperfusion Model: An Experimental Study Thu, 05 Mar 2015 10:33:00 +0000 Objective. The objective of this study was to examine the effects of iloprost and N-acetylcysteine (NAC) on ischemia-reperfusion (IR) injuries to the gastrocnemius muscle, following the occlusion-reperfusion period in the abdominal aorta of rats. Materials and Methods. Forty male Sprague-Dawley rats were randomly divided into four equal groups. Group 1: control group. Group 2 (IR): aorta was occluded. The clamp was removed after 1 hour of ischemia. Blood samples and muscle tissue specimens were collected following a 2-hour reperfusion period. Group 3 (IR + iloprost): during a 1-hour ischemia period, iloprost infusion was initiated from the jugular catheter. During a 2-hour reperfusion period, the iloprost infusion continued. Group 4 (IR + NAC): similar to the iloprost group. Findings. The mean total oxidant status, CK, and LDH levels were highest in Group 2 and lowest in Group 1. The levels of these parameters in Group 3 and Group 4 were lower compared to Group 2 and higher compared to Group 1 (). The histopathological examination showed that Group 3 and Group 4, compared to Group 2, had preserved appearance with respect to hemorrhage, necrosis, loss of nuclei, infiltration, and similar parameters. Conclusion. Iloprost and NAC are effective against ischemia-reperfusion injury and decrease ischemia-related tissue injury. Osman Tiryakioglu, Kamuran Erkoc, Bulent Tunerir, Onur Uysal, H. Firat Altin, Tevfik Gunes, and Selim Aydin Copyright © 2015 Osman Tiryakioglu et al. All rights reserved. Substance P Receptor Antagonism: A Potential Novel Treatment Option for Viral-Myocarditis Mon, 02 Mar 2015 12:27:00 +0000 Viral-myocarditis is an important cause of heart failure for which no specific treatment is available. We previously showed the neuropeptide substance P (SP) is associated with the pathogenesis of murine myocarditis caused by encephalomyocarditis virus (EMCV). The current studies determined if pharmacological inhibition of SP-signaling via its high affinity receptor, NK1R and downstream G-protein, Ras homolog gene family, member-A (RhoA), will be beneficial in viral-myocarditis. Aprepitant (1.2 mg/kg), a SP-receptor antagonist, or fasudil (10 mg/kg), a RhoA inhibitor, or saline control was administered daily to mice orally for 3 days, prior to, or 5 days following, intraperitoneal infection with and without 50 PFU of EMCV, following which disease assessment studies, including echocardiogram and cardiac Doppler were performed in day 14 after infection. Pretreatment and posttreatment with aprepitant significantly reduced mortality, heart and cardiomyocyte size, and cardiac viral RNA levels ( all, ANOVA). Only aprepitant pretreatment improved heart functions; it significantly decreased end systolic diameter, improved fractional shortening, and increased peak aortic flow velocity ( all, ANOVA). Pre- or posttreatment with fasudil did not significantly impact disease manifestations. These findings indicate that SP contributes to cardiac-remodeling and dysfunction following ECMV infection via its high affinity receptor, but not through the Rho-A pathway. These studies suggest that SP-receptor antagonism may be a novel therapeutic-option for patients with viral-myocarditis. Prema Robinson, George E. Taffet, Nikita Engineer, Mitra Khumbatta, Bahrom Firozgary, Corey Reynolds, Thuy Pham, Tushar Bulsara, and Gohar Firozgary Copyright © 2015 Prema Robinson et al. All rights reserved. Vitamin A-Deficient Diet Accelerated Atherogenesis in Apolipoprotein E−/− Mice and Dietary β-Carotene Prevents This Consequence Mon, 23 Feb 2015 11:21:27 +0000 Vitamin A is involved in regulation of glucose concentrations, lipid metabolism, and inflammation, which are major risk factors for atherogenesis. However, the effect of vitamin A deficiency on atherogenesis has not been investigated. Therefore, the objective of the current study was to examine whether vitamin A deficiency accelerates atherogenesis in apolipoprotein E-deficient mice (apoE−/−). ApoE−/− mice were allocated into the following groups: control, fed vitamin A-containing chow diet; BC, fed chow diet fortified with Dunaliella powder containing βc isomers; VAD, fed vitamin A-deficient diet; and VAD-BC group, fed vitamin A-deficient diet fortified with a Dunaliella powder. Following 15 weeks of treatment, liver retinol concentration had decreased significantly in the VAD group to about 30% that of control group. Vitamin A-deficient diet significantly increased both plasma cholesterol concentrations and the atherosclerotic lesion area at the aortic sinus (+61%) compared to the control group. Dietary βc fortification inhibited the elevation in plasma cholesterol and retarded atherogenesis in mice fed the vitamin A-deficient diet. The results imply that dietary vitamin A deficiency should be examined as a risk factor for atherosclerosis and that dietary βc, as a sole source of retinoids, can compensate for vitamin A deficiency. Noa Zolberg Relevy, Dror Harats, Ayelet Harari, Ami Ben-Amotz, Rafael Bitzur, Ralph Rühl, and Aviv Shaish Copyright © 2015 Noa Zolberg Relevy et al. All rights reserved. 18F-Fluorodeoxyglucose Uptake in Abdominal Aortic Aneurysms: A Useful Biomarker of AAA Rupture Risk Thu, 18 Sep 2014 08:17:01 +0000 Kosmas I. Paraskevas, Dimitri P. Mikhailidis, and Frank J. Veith Copyright © 2014 Kosmas I. Paraskevas et al. All rights reserved. Serial Changes of Neointimal Tissue after Everolimus-Eluting Stent Implantation in Porcine Coronary Artery: An Optical Coherence Tomography Analysis Thu, 18 Sep 2014 00:00:00 +0000 Purposes. The serial changes in neointimal tissues were compared between everolimus-eluting stent (EES) and bare-metal stent (BMS) in the porcine coronary artery using optical coherence tomography (OCT). Methods. Serial (1, 3, and 6 month follow-up after stent implantation) OCT examinations were performed in 15 swine with 15 BMS- and 15 EES-treated lesions in porcine coronary arteries. Results. In BMS-implanted lesions, neointimal volume decreased from 7.3 mm3 to 6.9 mm3 and 6.4 mm3 at 1, 3, and 6 months follow-up without statistical significance (). At the time points of 1, 3, and 6 months, neointimal tissue appearance was mainly a homogeneous pattern (80.0%, 93.3%, and 100%, resp.), while the other pattern was layered. In contrast, in EES-implanted lesions, neointimal volume significantly increased from 4.8 mm3 to 9.8 mm3 between 1 and 3 months but significantly decreased to 8.6 mm3 between 3 and 6 months (). Between 1 and 3 months, the layered pattern of neointimal tissue increased from 26.7% to 66.7% but decreased to 20.0% between 3 and 6 months. Conclusions. EES had a biphasic pattern of neointimal amounts that correlated with changes in neointimal morphology. Hoyoun Won, Jung-Sun Kim, Dong-Ho Shin, Byeong-Keuk Kim, Young-Guk Ko, Donghoon Choi, Yangsoo Jang, and Myeong-Ki Hong Copyright © 2014 Hoyoun Won et al. All rights reserved. Anticancer Activities of Citrus Peel Polymethoxyflavones Related to Angiogenesis and Others Thu, 28 Aug 2014 00:00:00 +0000 Citrus is a kind of common fruit and contains multiple beneficial nutrients for human beings. Flavonoids, as a class of plant secondary metabolites, exist in citrus fruits abundantly. Due to their broad range of pharmacological properties, citrus flavonoids have gained increased attention. Accumulative in vitro and in vivo studies indicate protective effects of polymethoxyflavones (PMFs) against the occurrence of cancer. PMFs inhibit carcinogenesis by mechanisms like blocking the metastasis cascade, inhibition of cancer cell mobility in circulatory systems, proapoptosis, and antiangiogenesis. This review systematically summarized anticarcinogenic effect of citrus flavonoids in cancer therapy, together with the underlying important molecular mechanisms, in purpose of further exploring more effective use of citrus peel flavonoids. Liwen Wang, Jinhan Wang, Lianying Fang, Zuliang Zheng, Dexian Zhi, Suying Wang, Shiming Li, Chi-Tang Ho, and Hui Zhao Copyright © 2014 Liwen Wang et al. All rights reserved. SIRT1 Inhibition Affects Angiogenic Properties of Human MSCs Wed, 27 Aug 2014 06:13:21 +0000 Human mesenchymal stem cells (hMSCs) are attractive for clinical and experimental purposes due to their capability of self-renewal and of differentiating into several cell types. Autologous hMSCs transplantation has been proven to induce therapeutic angiogenesis in ischemic disorders. However, the molecular mechanisms underlying these effects remain unclear. A recent report has connected MSCs multipotency to sirtuin families, showing that SIRT1 can regulate MSCs function. Furthermore, SIRT1 is a critical modulator of endothelial angiogenic functions. Here, we described the generation of an immortalized human mesenchymal bone marrow-derived cell line and we investigated the angiogenic phenotype of our cellular model by inhibiting SIRT1 by both the genetic and pharmacological level. We first assessed the expression of SIRT1 in hMSCs under basal and hypoxic conditions at both RNA and protein level. Inhibition of SIRT1 by sirtinol, a cell-permeable inhibitor, or by specific sh-RNA resulted in an increase of premature-senescence phenotype, a reduction of proliferation rate with increased apoptosis. Furthermore, we observed a consistent reduction of tubule-like formation and migration and we found that SIRT1 inhibition reduced the hypoxia induced accumulation of HIF-1α protein and its transcriptional activity in hMSCs. Our findings identify SIRT1 as regulator of hypoxia-induced response in hMSCs and may contribute to the development of new therapeutic strategies to improve regenerative properties of mesenchymal stem cells in ischemic disorders through SIRT1 modulation. Botti Chiara, Caiafa Ilaria, Coppola Antonietta, Cuomo Francesca, Miceli Marco, Altucci Lucia, and Cobellis Gilda Copyright © 2014 Botti Chiara et al. All rights reserved. Retrospective Comparative Study of the Effects of Dendritic Cell Vaccine and Cytokine-Induced Killer Cell Immunotherapy with that of Chemotherapy Alone and in Combination for Colorectal Cancer Mon, 18 Aug 2014 07:41:02 +0000 Purpose. This retrospective study determined the delayed-type hypersensitivity (DTH) skin test and safety of dendritic cell (DC) vaccine and cytokine-induced killer (CIK) cell immunotherapy and the survival compared to chemotherapy in 239 colorectal cancer (CRC) patients. Methods. DTH and safety of the immunotherapy were recorded. The overall survival (OS) and disease free survival curves were compared according to the immunotherapy and/or chemotherapy received with Kaplan-Meier estimates. Results. Of the 70 patients who received immunotherapy, 62.86% had a positive DTH skin test, 38.57% developed fever, 47.14% developed insomnia, 38.57% developed anorexia, 4.29% developed joint soreness, and 11.43% developed skin rash. For 204 resectable CRC patients, median survival time (MST) (198.00 days) was significantly longer in patients with immunotherapy plus chemotherapy than with chemotherapy alone (106.00 days) . For 35 patients with unresectable or postsurgery relapsed CRC and who were confirmed to be dead, no statistical difference was observed in the MST between the patients treated with immunotherapy and with chemotherapy . MST in the patients treated with chemotherapy plus immunotherapy was 154 days longer than that of patients treated with chemotherapy alone . Conclusions. DC vaccination and CIK immunotherapy did not cause severe adverse effects, induce immune response against CRC, and prolong OS. Jingxiu Niu, Yanjie Ren, Tianyu Zhang, Xuejing Yang, Wei Zhu, Hui Zhu, Jing Li, Jiali Li, and Yan Pang Copyright © 2014 Jingxiu Niu et al. All rights reserved. From Sprouting Angiogenesis to Erythrocytes Generation by Cancer Stem Cells: Evolving Concepts in Tumor Microcirculation Mon, 04 Aug 2014 10:56:15 +0000 Angiogenesis is essential for tumor growth and metastasis. Over the last decades, a substantial progress has been achieved in defining different patterns of tumor microcirculation. Sprouting angiogenesis, the oldest model of microcirculation, is the de novo vessel formation from preexisting blood vessels. Vessel splitting and hijacking, also known, respectively, as intussusception and cooption, are alternative models that account for tumor resistance to antiangiogenic therapy. In addition to remodeling the microenvironment, the tumor cell can undergo intrinsic changes and survive hypoxic conditions by acquiring stem cell properties. In line with the concept of pluripotency, tumor cells can form vascular mimicry structures creating their own microcirculation despite a latent vessel growth. The recent identification of the polyploid giant cancer cells and tumor-derived erythrocytes is the most innovative survival mechanism in hypoxia and provides a potential target for more effective therapies. Raafat S. Alameddine, Lana Hamieh, and Ali Shamseddine Copyright © 2014 Raafat S. Alameddine et al. All rights reserved. Immune Response, Safety, and Survival and Quality of Life Outcomes for Advanced Colorectal Cancer Patients Treated with Dendritic Cell Vaccine and Cytokine-Induced Killer Cell Therapy Thu, 17 Jul 2014 09:58:02 +0000 Purpose. To determine the immune response after dendritic cell (DC) vaccine and cytokine-induced killer cells (CIK) therapy and assess its associated toxicity, survival benefit, and changes in the quality of life (QOL) of advanced colorectal cancer (CRC) patients. Methods. We recruited 100 patients with unresectable CRC orrelapsed CRC after surgery who received DC vaccine and CIK cells (group immunotherapy, group I), and, as a control, 251 patients who had similar characteristics and underwent similar treatments, except for this immunotherapy (group nonimmunotherapy, group NI). After a follow-up period of 489.2 ± 160.4 days, overall survival (OS) of the two groups was compared using the Kaplan-Meier method. Results. In group I, 62% of patients developed a positive delayed type hypersensitivity response, and most patients showed an improvement in physical strength (75.2%), appetite (74.2%), sleeping (72.1%), and body weight (70.1%). Adverse events were fever (29.5%), insomnia (19.2%), anorexia (9.1%), sore joints (5.4%), and skin rash (1.0%). No toxicity was observed in patients treated with DC vaccine and CIK therapy. OS was significantly longer in group I than in group NI ( = 0.043). Conclusion. DC vaccine and CIK therapy were safe and could induce an immune response against CRC, thereby improving QOL and prolonging OS. Hui Zhu, Xuejing Yang, Jiali Li, Yanjie Ren, Tianyu Zhang, Chunze Zhang, Jintai Zhang, Jing Li, and Yan Pang Copyright © 2014 Hui Zhu et al. All rights reserved. GABAB Receptors Expressed in Human Aortic Endothelial Cells Mediate Intracellular Calcium Concentration Regulation and Endothelial Nitric Oxide Synthase Translocation Wed, 09 Jul 2014 13:27:41 +0000 GABAB receptors regulate the intracellular Ca2+ concentration ([Ca2+]i) in a number of cells (e.g., retina, airway epithelium and smooth muscle), but whether they are expressed in vascular endothelial cells and similarly regulate the [Ca2+]i is not known. The purpose of this study was to investigate the expression of GABAB receptors, a subclass of receptors to the inhibitory neurotransmitter γ-aminobutyric acid (GABA), in cultured human aortic endothelial cells (HAECs), and to explore if altering receptor activation modified [Ca2+]i and endothelial nitric oxide synthase (eNOS) translocation. Real-time PCR, western blots and immunofluorescence were used to determine the expression of GABAB1 and GABAB2 in cultured HAECs. The effects of GABAB receptors on [Ca2+]i in cultured HAECs were demonstrated using fluo-3. The influence of GABAB receptors on eNOS translocation was assessed by immunocytochemistry. Both GABAB1 and GABAB2 mRNA and protein were expressed in cultured HAECs, and the GABAB1 and GABAB2 proteins were colocated in the cell membrane and cytoplasm. One hundred μM baclofen caused a transient increase of [Ca2+]i and eNOS translocation in cultured HAECs, and the effects were attenuated by pretreatment with the selective GABAB receptor antagonists CGP46381 and CGP55845. GABAB receptors are expressed in HAECs and regulate the [Ca2+]i and eNOS translocation. Cultures of HAECs may be a useful in vitro model for the study of GABAB receptors and vascular biology. Xu-Ping Wang, Zhen-Ying Cheng, and Katrina L. Schmid Copyright © 2014 Xu-Ping Wang et al. All rights reserved. A Vessel Active Contour Model for Vascular Segmentation Tue, 01 Jul 2014 00:00:00 +0000 This paper proposes a vessel active contour model based on local intensity weighting and a vessel vector field. Firstly, the energy function we define is evaluated along the evolving curve instead of all image points, and the function value at each point on the curve is based on the interior and exterior weighted means in a local neighborhood of the point, which is good for dealing with the intensity inhomogeneity. Secondly, a vascular vector field derived from a vesselness measure is employed to guide the contour to evolve along the vessel central skeleton into thin and weak vessels. Thirdly, an automatic initialization method that makes the model converge rapidly is developed, and it avoids repeated trails in conventional local region active contour models. Finally, a speed-up strategy is implemented by labeling the steadily evolved points, and it avoids the repeated computation of these points in the subsequent iterations. Experiments using synthetic and real vessel images validate the proposed model. Comparisons with the localized active contour model, local binary fitting model, and vascular active contour model show that the proposed model is more accurate, efficient, and suitable for extraction of the vessel tree from different medical images. Yun Tian, Qingli Chen, Wei Wang, Yu Peng, Qingjun Wang, Fuqing Duan, Zhongke Wu, and Mingquan Zhou Copyright © 2014 Yun Tian et al. All rights reserved. Administration of the Resveratrol Analogues Isorhapontigenin and Heyneanol-A Protects Mice Hematopoietic Cells against Irradiation Injuries Tue, 24 Jun 2014 12:16:38 +0000 Ionizing radiation (IR) is known not only to cause acute bone marrow (BM) suppression but also to lead to long-term residual hematopoietic injury. These effects have been attributed to IR inducing the generation of reactive oxygen species (ROS) in hematopoietic cells. In this study, we examined if isorhapontigenin and heyneanol-A, two analogues of resveratrol, could mitigate IR-induced BM suppression. The results of cell viability assays, clonogenic assays, and competitive repopulation assays revealed that treatment with these compounds could protect mice BM mononuclear cells (BMMNC), hematopoietic progenitor cells, and hematopoietic stem cells from IR-induced BM suppression. Moreover, the expression of genes related to the endogenous cellular antioxidant system in hematopoietic cells was analyzed. The expression and activity of SOD2 and GPX1 were found to be decreased in irradiated BMMNC, and the application of the resveratrol analogues could ameliorate this damage. Our results suggest that in comparison with resveratrol and isorhapontigenin, treatment with heyneanol-A can protect hematopoietic cells from IR-induced damage to a greater degree; the protective effects of these compounds are probably the result of their antioxidant properties. Hui Wang, Yi-ling Yang, Heng Zhang, Hao Yan, Xiao-jing Wu, and Chun-ze Zhang Copyright © 2014 Hui Wang et al. All rights reserved. The Diagnostic Value of Cervical Lymph Node Metastasis in Head and Neck Squamous Carcinoma by Using Diffusion-Weighted Magnetic Resonance Imaging and Computed Tomography Perfusion Tue, 24 Jun 2014 12:02:00 +0000 Purpose. The aim of this study was to compare diffusion-weighted magnetic resonance imaging (DWI) with computed tomography perfusion (CTP) for preoperative detection of metastases to lymph nodes (LNs) in head and neck squamous cell carcinoma (SCC). Methods. Between May 2010 and April 2012, 30 patients with head and neck SCC underwent preoperative DWI and CTP. Two radiologists measured apparent diffusion coefficient (ADC) values and CTP parameters independently. Surgery and histopathologic examinations were performed on all patients. Results. On DWI, 65 LNs were detected in 30 patients. The mean ADC value of metastatic nodes was lower than benign nodes and the difference was statistically significant (). On CTP images, the mean value in metastatic nodes of blood flow (BF) and blood volume (BV) was higher than that in benign nodes, and mean transit time (MTT) in metastatic nodes was lower than that in benign nodes. There were significant differences in BF and MTT values between metastatic and benign LNs (). There were significant differences between the AUCs of DWI and CTP (, ). Conclusion. DWI with ADC value measurements may be more accurate than CTP for the preoperative diagnosis of cervical LN metastases. Jin Zhong, Zonghong Lu, Liang Xu, Longchun Dong, Hui Qiao, Rui Hua, Yi Gong, Zhenxing Liu, Caixian Hao, Xuehuan Liu, Changqing Zong, Li He, and Jun Liu Copyright © 2014 Jin Zhong et al. All rights reserved. Asymmetric Cell Division in Polyploid Giant Cancer Cells and Low Eukaryotic Cells Sun, 22 Jun 2014 00:00:00 +0000 Asymmetric cell division is critical for generating cell diversity in low eukaryotic organisms. We previously have reported that polyploid giant cancer cells (PGCCs) induced by cobalt chloride demonstrate the ability to use an evolutionarily conserved process for renewal and fast reproduction, which is normally confined to simpler organisms. The budding yeast, Saccharomyces cerevisiae, which reproduces by asymmetric cell division, has long been a model for asymmetric cell division studies. PGCCs produce daughter cells asymmetrically in a manner similar to yeast, in that both use budding for cell polarization and cytokinesis. Here, we review the results of recent studies and discuss the similarities in the budding process between yeast and PGCCs. Dan Zhang, Yijia Wang, and Shiwu Zhang Copyright © 2014 Dan Zhang et al. All rights reserved. Effect of High-Fat Diet upon Inflammatory Markers and Aortic Stiffening in Mice Wed, 11 Jun 2014 13:31:32 +0000 Changes in lifestyle such as increase in high-fat food consumption are an important cause for vascular diseases. The present study aimed to investigate the involvement of ACE and TGF-β in the aorta stiffness induced by high-fat diet. C57BL/6 male mice were divided in two groups according to their diet for 8 weeks: standard diet (ST) and high-fat diet (HF). At the end of the protocol, body weight gain, adipose tissue content, serum lipids and glucose levels, and aorta morphometric and biochemical measurements were performed. Analysis of collagen fibers by picrosirius staining of aorta slices showed that HF diet promoted increase of thin (55%) and thick (100%) collagen fibers deposition and concomitant disorganization of these fibers orientations in the aorta vascular wall (50%). To unravel the mechanism involved, myeloperoxidase (MPO) and angiotensin I converting enzyme (ACE) were evaluated by protein expression and enzyme activity. HF diet increased MPO (90%) and ACE (28%) activities, as well as protein expression of ACE. TGF-β was also increased in aorta tissue of HF diet mice after 8 weeks. Altogether, we have observed that the HF diet-induced aortic stiffening may be associated with increased oxidative stress damage and activation of the RAS in vascular tissue. Andre Bento Chaves Santana, Thais Cristina de Souza Oliveira, Barbara Lobo Bianconi, Valerio Garrone Barauna, Ed Wilson Cavalcante Oliveira Santos, Tatiana P. Alves, Juliane Cristina S. Silva, Patricia Fiorino, Primavera Borelli, Maria Claudia Costa Irigoyen, José Eduardo Krieger, and Silvia Lacchini Copyright © 2014 Andre Bento Chaves Santana et al. All rights reserved. Overexpression of Wnt5a Promotes Angiogenesis in NSCLC Thu, 05 Jun 2014 08:54:27 +0000 To evaluate Wnt5a expression and its role in angiogenesis of non-small-cell lung cancer (NSCLC), immunohistochemistry and CD31/PAS double staining were performed to examine the Wnt5a expression and we analyze the relationships between Wnt5a and microvessel density (MVD), vasculogenic mimicry (VM), and some related proteins. About 61.95% of cases of 205 NSCLC specimens exhibited high expression of Wnt5a. Wnt5a expression level was upregulated in the majority of NSCLC tissues, especially in squamous cell carcinoma, while its expression level in adenocarcinoma was the lowest. Wnt5a was also found more frequently expressed in male patients than in female patients. Except for histological classification and gender, little association was found between Wnt5a and clinicopathological features. Moreover, Wnt5a was significantly correlated with prognosis. Overall, Wnt5a-positive expression in patients with NSCLC indicated shorter survival time. As for vascularization in NSCLC, Wnt5a showed close association with VM and MVD. In addition, Wnt5a was positively related with β-catenin-nu, VE-cadherin, MMP2, and MMP9. The results demonstrated that overexpression of Wnt5a may play an important role in NSCLC angiogenesis and it may function via canonical Wnt signal pathway. This study will provide evidence for further research on NSCLC and also will provide new possible target for NSCLC diagnosis and therapeutic strategies. Lingli Yao, Baocun Sun, Xiulan Zhao, Xueming Zhao, Qiang Gu, Xueyi Dong, Yanjun Zheng, Junying Sun, Runfen Cheng, Hong Qi, and Jindan An Copyright © 2014 Lingli Yao et al. All rights reserved. Gold Nanoparticles Inhibit VEGF165-Induced Migration and Tube Formation of Endothelial Cells via the Akt Pathway Sun, 01 Jun 2014 11:29:43 +0000 The early stages of angiogenesis can be divided into three steps: endothelial cell proliferation, migration, and tube formation. Vascular endothelial growth factor (VEGF) is considered the most important proangiogenic factor; in particular, VEGF165 plays a critical role in angiogenesis. Here, we evaluated whether gold nanoparticles (AuNPs) could inhibit the VEGF165-induced human umbilical vein endothelial cell (HUVEC) migration and tube formation. AuNPs and VEGF165 were coincubated overnight at 4°C, after which the effects on cell migration and tube formation were assessed. Cell migration was assessed using a modified wound-healing assay and a transwell chamber assay; tube formation was assessed using a capillary-like tube formation assay and a chick chorioallantoic membrane (CAM) assay. We additionally detected the cell surface morphology and ultrastructure using atomic force microscopy (AFM). Furthermore, Akt phosphorylation downstream of VEGFR-2/PI3K in HUVECs was determined in a Western blot analysis. Our study demonstrated that AuNPs significantly inhibited VEGF165-induced HUVEC migration and tube formation by affecting the cell surface ultrastructure, cytoskeleton and might have inhibited angiogenesis via the Akt pathway. Yunlong Pan, Qing Wu, Li Qin, Jiye Cai, and Bin Du Copyright © 2014 Yunlong Pan et al. All rights reserved. A New Measure of Decompression Sickness in the Rat Sun, 25 May 2014 06:10:22 +0000 In this study we assessed the reliability of a tilting-board grip score as a measure of decompression sickness in rats. In experiments using a hyperbaric compression/decompression protocol, rats were observed for signs of decompression sickness and their grip strength measured on a tilting particle board hinged to a metal frame. Angles at which rats lost grip were converted to gravitational vectors. Decreased mean grip scores following decompression were fitted to a logistic regression model with strain, age, and weight. Decrease in grip score was significantly associated with observed decompression sickness (). The log odds ratio for decompression sickness = 1.40 (decrease in grip score). In rats with no decrease in mean grip score there was a 50% probability of decompression sickness (pDCS). This increased steadily with decreases in mean grip score. A decrease of 0.3 had a 60% pDCS, a decrease of 0.6 had a 70% pDCS, and a decrease of 2.1 had a 95% pDCS. The tilting board grip score is a reliable measure of the probability of decompression sickness. Peter Buzzacott, Aleksandra Mazur, Qiong Wang, Kate Lambrechts, Michael Theron, Jacques Mansourati, and François Guerrero Copyright © 2014 Peter Buzzacott et al. All rights reserved. Stem Cell-Like Circulating Tumor Cells Indicate Poor Prognosis in Gastric Cancer Thu, 22 May 2014 15:48:16 +0000 Circulating tumor cells (CTCs), which have stem cell-like characteristics, might play a crucial role in cancer metastasis. CD44 has been identified as gastric cancer (GC) stem cell (CSC) marker. Here, the prognostic significance of CD44-positive CTCs in GC patients was investigated. CTCs were detected in 27 of 45 GC patients. The presence of CTCs was significantly associated with lymph node metastasis, distant metastasis, and recurrence (, , and , resp.). Nineteen of the 27 CTC-positive patients had CD44-positive CTCs. These patients were more likely to develop metastasis and recurrence than patients with CD44-negative CTCs. CD44-positive CTC counts were higher in recurrent patients than in the nonrecurrent ones (means 4.8 and 1.9, resp.; ). Furthermore, 13 of 19 patients with CD44-positive CTCs developed recurrent disease, and the mean time to recurrence was shorter than that in patients with CD44-negative CTCs ( and months, resp.; ). COX proportional hazards model indicated that the presence of CD44-positive CTCs and TNM stage were independent predictors of recurrence for GC ( and 0.008). So identifying the stem cell-like CTC subset may provide more clinically useful prognostic information than only detecting CTCs. Man Li, Baogang Zhang, Zhiguang Zhang, Xia Liu, Xiangjuan Qi, Jianqiu Zhao, Yong Jiang, Haoyu Zhai, Yinglan Ji, and Dan Luo Copyright © 2014 Man Li et al. All rights reserved. Influence of Egr-1 in Cardiac Tissue-Derived Mesenchymal Stem Cells in Response to Glucose Variations Thu, 22 May 2014 11:47:34 +0000 Mesenchymal stem cells (MSCs) represent a promising cell population for cell therapy and regenerative medicine applications. However, how variations in glucose are perceived by MSC pool is still unclear. Since, glucose metabolism is cell type and tissue dependent, this must be considered when MSCs are derived from alternative sources such as the heart. The zinc finger transcription factor Egr-1 is an important early response gene, likely to play a key role in the glucose-induced response. Our aim was to investigate how short-term changes in in vitro glucose concentrations affect multipotent cardiac tissue-derived MSCs (cMSCs) in a mouse model of Egr-1 KO (Egr-1−/−). Results showed that loss of Egr-1 does not significantly influence cMSC proliferation. In contrast, responses to glucose variations were observed in wt but not in Egr-1−/− cMSCs by clonogenic assay. Phenotype analysis by RT-PCR showed that cMSCs Egr-1−/− lost the ability to regulate the glucose transporters GLUT-1 and GLUT-4 and, as expected, the Egr-1 target genes VEGF, TGFβ-1, and p300. Acetylated protein levels of H3 histone were impaired in Egr-1−/− compared to wt cMSCs. We propose that Egr-1 acts as immediate glucose biological sensor in cMSCs after a short period of stimuli, likely inducing epigenetic modifications. Daniela Bastianelli, Camilla Siciliano, Rosa Puca, Andrea Coccia, Colin Murdoch, Antonella Bordin, Giorgio Mangino, Giulio Pompilio, Antonella Calogero, and Elena De Falco Copyright © 2014 Daniela Bastianelli et al. All rights reserved. Novel Biomarkers of Abdominal Aortic Aneurysm Disease: Identifying Gaps and Dispelling Misperceptions Tue, 20 May 2014 00:00:00 +0000 Abdominal aortic aneurysm (AAA) is a prevalent and potentially life-threatening disease. Early detection by screening programs and subsequent surveillance has been shown to be effective at reducing the risk of mortality due to aneurysm rupture. The aim of this review is to summarize the developments in the literature concerning the latest biomarkers (from 2008 to date) and their potential screening and therapeutic values. Our search included human studies in English and found numerous novel biomarkers under research, which were categorized in 6 groups. Most of these studies are either experimental or hampered by their low numbers of patients. We concluded that currently no specific laboratory markers allow screeing for the disease and monitoring its progression or the results of treatment. Further studies and studies in larger patient groups are required in order to validate biomarkers as cost-effective tools in the AAA disease. Demetrios Moris, Eleftherios Mantonakis, Efthymios Avgerinos, Marinos Makris, Chris Bakoyiannis, Emmanuel Pikoulis, and Sotirios Georgopoulos Copyright © 2014 Demetrios Moris et al. All rights reserved. May Renal Resistive Index Be an Early Predictive Tool of Postoperative Complications in Major Surgery? Preliminary Results Tue, 20 May 2014 00:00:00 +0000 Background. Patients who undergo high-risk surgery represent a large amount of post-operative ICU-admissions. These patients are at high risk of experiencing postoperative complications. Renal Resistive Index was found to be related with renal dysfunction, hypertension, and posttraumatic hemorrhagic shock, probably due to vasoconstriction. We explored whether Renal Resistive Index (RRI), measured after awakening from general anesthesia, could have any relationship with postoperative complications. Methods. In our observational, stratified dual-center trial, we enrolled patients who underwent general anesthesia for high-risk major surgery. After awakening in recovery room (or during awakening period in subjects submitted to cardiac surgery) we measured RRI by echo-color-Doppler method. Primary endpoint was the association of altered RRI (>0.70) and outcome during the first postoperative week. Results. 205 patients were enrolled: 60 (29.3%) showed RRI > 0.70. The total rate of adverse event was 27 (18.6%) in RRI ≤ 0.7 group and 19 (31.7%) in RRI > 0.7 group (). Significant correlation between RRI > 0.70 and complications resulted in pneumonia (), septic shock (), and acute renal failure () subgroups. Patients with RRI > 0.7 showed longer ICU stay () and lasting of mechanical ventilation (). These results were confirmed in cardiothoracic surgery subgroup. RRI > 0.7 duplicates triplicates the risk of complications, both in general (OR 2.03 93 95% CI 1.02–4.02, ) and in cardiothoracic (OR 2.62 95% CI 1.11–6.16, ) population. Furthermore, we found RRI > 0.70 was associated with a triplicate risk of postoperative septic shock (OR 3.04, CI 95% 1.5–7.01; ). Enrico Giustiniano, Massimo Meco, Emanuela Morenghi, Nadia Ruggieri, Daniele Cosseta, Silvia Cirri, Orazio Difrancesco, Paola Cosma Zito, Yari Gollo, and Ferdinando Raimondi Copyright © 2014 Enrico Giustiniano et al. All rights reserved. Human Cytomegalovirus-Encoded miR-US25-1 Aggravates the Oxidised Low Density Lipoprotein-Induced Apoptosis of Endothelial Cells Thu, 08 May 2014 00:00:00 +0000 Human cytomegalovirus (HCMV) infection is linked to the development and severity of the cardiovascular disease atherosclerosis; however, there is little known about the promotion of atherosclerosis. miR-US25-1 is one of HCMV-encoded miRNAs and targets cellular genes that are essential for virus growth to control the life cycle of the virus and host cells. The prominent regulation on cell cycle genes of the miR-US25-1 attracts us to explore its role in the atherosclerosis promotion. It was indicated that miR-US25-1 level was upregulated in subjects or in endothelial cells with HCMV infection; and the miR-US25-1 downregulated the expression of BRCC 3 by targeting the 5′ UTR of BRCC 3. And a miR-US25-1 mimics transfection could reduce the EAhy926 cell viability but did not induce apoptosis in EAhy926 cells. And what is more, miR-US25-1 mimicis transfection deteriorated the ox-LDL-induced apoptosis and aggravated the upregulation of apoptosis-associated molecules by oxidised low density lipoprotein (ox-LDL) in EAhy926 cells. And we have also confirmed the deregulation of BRCC 3 expression by miR-US25-1 by targeting the 5′ UTR of it. Given the vital role of BRCC 3 in DNA damage repairing, we speculated that the targeting inhibition of BRCC 3 by miR-US25-1 may contribute to the aggravation of ox-LDL-promoted apoptosis of endothelial EAhy926 cells. Jianmin Fan, Wen Zhang, and Qiming Liu Copyright © 2014 Jianmin Fan et al. All rights reserved. A Survey of Italian Physicians' Opinion about Stem Cells Research: What Doctors Prefer and What the Law Requires Wed, 30 Apr 2014 07:26:55 +0000 To evaluate the Italian physicians' knowledge/information level about the therapeutic potential of stem cells, the research choice between embryonic and cordonal stem cells, and the preference between autologous and heterologous storage of cordonal stem cells, we performed a national survey. The questionnaire—distributed to 3361 physicians—involved physicians of different religious orientations and of different medical specialities. Most of the physicians involved (67%) were Catholics, and the majority were gynaecologists and paediatricians (43%) who are mainly in charge to inform future mothers about the possibility of cordonal stem cells conservation. The majority of the physicians interviewed do not have specific knowledge about stem cells (59%), most of them having only generic information (92%). The largest part of physicians prefer to use umbilical cord blood cells rather than embryonic stem cells. Nevertheless, a large percentage of physicians were in favour of embryo research, especially when embryos are supernumerary (44% versus 34%). Eighty-seven % of the physicians interviewed proved to have a general knowledge about stem cells and believe in their therapeutic potential. They prefer research on cordonal stem cells rather than on embryo stem cells. Although they are in favour of heterologous stem cells donation, they still prefer cryopreservation for personal use. Paola Frati, Matteo Gulino, Arianna Pacchiarotti, Stefano D'Errico, Lorella Sicuro, and Vittorio Fineschi Copyright © 2014 Paola Frati et al. All rights reserved. The Multifaceted Functions of CXCL10 in Cardiovascular Disease Wed, 23 Apr 2014 09:28:28 +0000 C-X-C motif ligand 10 (CXCL10), or interferon-inducible protein-10, is a small chemokine belonging to the CXC chemokine family. Its members are responsible for leukocyte trafficking and act on tissue cells, like endothelial and vascular smooth muscle cells. CXCL10 is secreted by leukocytes and tissue cells and functions as a chemoattractant, mainly for lymphocytes. After binding to its receptor CXCR3, CXCL10 evokes a range of inflammatory responses: key features in cardiovascular disease (CVD). The role of CXCL10 in CVD has been extensively described, for example for atherosclerosis, aneurysm formation, and myocardial infarction. However, there seems to be a discrepancy between experimental and clinical settings. This discrepancy occurs from differences in biological actions between species (e.g. mice and human), which is dependent on CXCL10 signaling via different CXCR3 isoforms or CXCR3-independent signaling. This makes translation from experimental to clinical settings challenging. Furthermore, the overall consensus on the actions of CXCL10 in specific CVD models is not yet reached. The purpose of this review is to describe the functions of CXCL10 in different CVDs in both experimental and clinical settings and to highlight and discuss the possible discrepancies and translational difficulties. Furthermore, CXCL10 as a possible biomarker in CVD will be discussed. Pleunie van den Borne, Paul H. A. Quax, Imo E. Hoefer, and Gerard Pasterkamp Copyright © 2014 Pleunie van den Borne et al. All rights reserved. Epithelial-Mesenchymal Transition Regulated by EphA2 Contributes to Vasculogenic Mimicry Formation of Head and Neck Squamous Cell Carcinoma Thu, 17 Apr 2014 13:34:19 +0000 Purpose. Vasculogenic mimicry (VM) was related to invasion and metastasis of head and neck squamous cell carcinoma (HNSCC) patients. This study was designed to investigate the role of EphA2 in VM formation of HNSCC. Methods. The SiRNA technique was used to knock down the expression of EphA2 in vitro. The ability of cell migration and invasion were measured by transwell and wound healing assays; three-dimensional culture was used to detect the ability of channel-like structure formation; Western blot was used to detect the expression of epithelial-mesenchymal transition- (EMT-) related molecules in vitro. Further semiquantitative real-time RT-PCR assays and immunohistochemistry were used to demonstrate expression of EphA2 and EMT-related molecules according to VM presence or not in human tissue. Results. Knocking down EphA2 in vitro leads to disabled channel-like structure formation, reduction of invasion and migration ability, and reverse of EMT-related markers. Both semiquantitative real-time RT-PCR and immunohistochemistry showed that expressions of EphA2, Twist, and Vimentin were higher in the VM-positive group than in the VM-negative group significantly, while expressions of E-cadherin, claudin4, and DSG-3 were reverse. Conclusions. EphA2 played a key role in VM formation of HNSCC through regulation of EMT. Wei Wang, Peng Lin, Baocun Sun, Shiwu Zhang, Wenjuan Cai, Chunrong Han, Li Li, Honghua Lu, and Xiulan Zhao Copyright © 2014 Wei Wang et al. All rights reserved. Scattering Coefficients of Mice Organs Categorized Pathologically by Spectral Domain Optical Coherence Tomography Sun, 13 Apr 2014 15:21:51 +0000 Differences in tissue density cause a variety of scattering coefficients. To quantify optical coherence tomography (OCT) images for diagnosis, the tissue's scattering coefficient is estimated by curve fitting the OCT signals to a confocal single backscattering mode. The results from a group of 30 mice show that the scattering coefficients of bone, skin, liver, brain, testis, and spleen can be categorized into three groups: a scattering coefficient between 1.947 and 2.134 mm−1: bone and skin; a scattering coefficient between 1.303 and 1.461 mm−1: liver and brain; a scattering coefficient between 0.523 and 0.634 mm−1: testis and spleen. The results indicate that the scattering coefficient is tissue specific and could be used in tissue diagnosis. Q. Q. Zhang, X. J. Wu, C. Wang, S. W. Zhu, Y. L. Wang, Bruce Z. Gao, and X.-C. Yuan Copyright © 2014 Q. Q. Zhang et al. All rights reserved. The Evolution of Cardiovascular Surgery in Elderly Patient: A Review of Current Options and Outcomes Thu, 10 Apr 2014 00:00:00 +0000 Due to the increase in average life expectancy and the higher incidence of cardiovascular disease with advancing age, more elderly patients present for cardiac surgery nowadays. Advances in pre- and postoperative care have led to the possibility that an increasing number of elderly patients can be operated on safely and with a satisfactory outcome. Currently, coronary artery bypass surgery, aortic and mitral valve surgery, and major surgery of the aorta are performed in elderly patients. The data available show that most cardiac surgical procedures can be performed in elderly patients with a satisfactory outcome. Nevertheless, the risk for these patients is only acceptable in the absence of comorbidities. In particular, renal dysfunction, cerebrovascular disease, and poor clinical state are associated with a worse outcome in elderly patients. Careful patient selection, flawless surgery, meticulous hemostasis, perfect anesthesia, and adequate myocardial protection are basic requirements for the success of cardiac surgery in elderly patients. The care of elderly cardiac surgical patients can be improved only through the strict collaboration of geriatricians, anesthesiologists, cardiologists, and cardiac surgeons, in order to obtain a tailored treatment for each individual patient. Francesco Nicolini, Andrea Agostinelli, Antonella Vezzani, Tullio Manca, Filippo Benassi, Alberto Molardi, and Tiziano Gherli Copyright © 2014 Francesco Nicolini et al. All rights reserved. Engineered Tumor Cell Apoptosis Monitoring Method Based on Dynamic Laser Tweezers Tue, 01 Apr 2014 12:30:53 +0000 Monitoring the cells’ apoptosis progression could provide a valuable insight into the temporal events that initiate cell death as well as the potential for rescue of apoptotic cells. In this paper, we engineered a novel and robust method for monitoring apoptosis of tumor cells based on dynamic laser tweezers, using A549 and HeLa cell line as typical samples. The entire experiment can be completed in a few hours with small amount of fluid sample, presenting great advantages of celerity, microscaled measurement, and label-free explorations without perturbing experimental conditions in combination with other probes. Validity and stability of this method are verified experimentally in terms of physical parameters of the system. The proposed technique has great potential in improving cancer treatment by monitoring the objective efficacy of tumor cell killing. Yuquan Zhang, Xiaojing Wu, Changjun Min, Siwei Zhu, H. Paul Urbach, and Xiaocong Yuan Copyright © 2014 Yuquan Zhang et al. All rights reserved. An Experimental Study to Replace the Thoracic Descending Aorta for Pigs with a Self-Made Sutureless Blood Vessel Tue, 18 Feb 2014 11:38:49 +0000 To simplify the procedure of blood vessel replacement operation and shorten the vascular anastomosis time, we developed a special artificial blood vessel which can be connected to native blood vessels without suture. The self-made sutureless blood vessel (SMSBV) was made from two titanium connectors and a Gore-Tex graft. To investigate blood compatibility and histocompatibility of the SMSBV, we carried thoracic descending aorta replacement using either SMSBV or Gore-Tex, respectively, in pigs. The aortic clamp time and the operative blood loss in the experimental group (using SMSBV) were less than those in the control group (using Gore-Tex). The whole blood hematocrit, platelet count, plasma soluble P-selectin, plasma free hemoglobin, and interleukins 2, 6 at each time point were not different significantly between the two groups. Light microscopy and transmission electron microscopy examination showed there were layers of vascular smooth muscle cells and endothelial cells adhered in the inner wall of artificial blood vessel without any signs of thrombosis. Based on the result, we have drawn the conclusion that the application of SMSBV can significantly shorten the vascular anastomosis time, reduce operative blood loss, and show good blood and tissue compatibility. Fenglin Song, Wenwu Zhou, Tao Tang, Xiaobing Li, Xiaoming Wu, and Jinfu Yang Copyright © 2014 Fenglin Song et al. All rights reserved. Comparison of Semi-Automated and Manual Measurements of Carotid Intima-Media Thickening Tue, 21 Jan 2014 12:09:10 +0000 Carotid intima-media thickening (CIMT) is a marker of both arteriosclerotic and atherosclerotic risks. Technological advances have semiautomated CIMT image acquisition and quantification. Studies comparing manual and automated methods have yielded conflicting results possibly due to plaque inclusion in measurements. Low atherosclerotic risk subjects were recruited to minimise the effect of focal atherosclerotic lesions on CIMT variability. CIMT was assessed by high-resolution B-mode ultrasound (Philips HDX7E, Phillips, UK) images of the common carotid artery using both manual and semiautomated methods (QLAB, Phillips, UK). Intraclass correlation coefficient (ICC) and the mean differences of paired measurements (Bland-Altman method) were used to compare both methodologies. The ICC of manual ( mm) and automated (0.524 ± 0.068 mm) methods was and an absolute mean bias ± SD of  mm was observed. Interobserver and intraobserver ICC were greater for automated ( and 0.99) compared to manual ( and 0.88) methods. Although not considered to be clinically significant, manual measurements yielded higher values compared to automated measurements. Automated measurements were more reproducible and showed lower interobserver variation compared to manual measurements. These results offer important considerations for large epidemiological studies. Oscar Mac Ananey, Greg Mellotte, and Vincent Maher Copyright © 2014 Oscar Mac Ananey et al. All rights reserved. Roles of Bone-Marrow-Derived Cells and Inflammatory Cytokines in Neointimal Hyperplasia after Vascular Injury Tue, 14 Jan 2014 09:26:00 +0000 Bone-marrow-derived cells can generate vascular progenitor cells that contribute to pathological remodeling in models of restenosis after percutaneous coronary intervention (PCI). We created models of vascular injury in mice with bone marrow transplants (BMT) to determine relationships between bone-marrow-derived cells and subsequent biological factors. Mesenchymal stromal cells (MSCs) seemed to inhibit the inflammatory reaction and help stabilize injured vascular regions through mobilizing more endogenous bone-marrow-derived (EBMD) cells to the peripheral circulation. Granulocyte-colony stimulating factor (G-CSF) mobilized more EBMD cells to the peripheral circulation, and they accumulated on the injured side of the vascular lumen. The inflammatory cytokines, tumor necrosis factor (TNF)-alpha, and interleukin (IL)-6 mobilized EBMD cells that play an important role in the process of neointimal hyperplasia after vascular injury. These factors might comprise a mechanism that alters the transdifferentiation or paracrine capabilities of EBMD cells and are potential targets of treatment for patients with cardiovascular diseases. Makoto Shoji, Shinji Koba, and Youichi Kobayashi Copyright © 2014 Makoto Shoji et al. All rights reserved. Impact of Mean Platelet Volume on Combined Safety Endpoint and Vascular and Bleeding Complications following Percutaneous Transfemoral Transcatheter Aortic Valve Implantation Mon, 23 Dec 2013 18:13:15 +0000 Background. Vascular and bleeding complications remain important complications in patients undergoing percutaneous transfemoral transcatheter aortic valve implantation (TF-TAVI). Platelets play an important role in bleeding events. Mean platelet volume (MPV) is an indicator of platelet activation. The objective of this study was to assess whether low MPV is an indicator of major vascular and bleeding complications following TF-TAVI. Methods. A retrospective cohort study of 330 subjects undergoing TF-TAVI implantation was performed. The primary study endpoint was the occurrence of combined safety endpoint (CSEP); secondary endpoints included major vascular complications and life-threatening bleeding. Endpoints were defined according to Valve Academic Research Consortium 2. Results. The CSEP at 30 days was reached in 30.9%; major vascular complications were observed in 14.9% while life-threatening bleeding occurred in 20.6%. Logistic Euroscore and MPV were independent predictors of CSEP. Predictors of vascular complications were female sex, previous myocardial infarction, red blood cell distribution width (RDW), and MPV while predictors of life-threatening bleeding were peripheral arterial disease, RDW, and MPV. Conclusion. A low baseline MPV was shown for the first time to be a significant predictor of CSEP, major vascular complications, and life-threatening bleeding following TF-TAVI. Caroline J. Magri, Alaide Chieffo, Alessandro Durante, Azeem Latib, Matteo Montorfano, Francesco Maisano, Michela Cioni, Eustachio Agricola, Remo Daniel Covello, Chiara Gerli, Annalisa Franco, Pietro Spagnolo, Ottavio Alfieri, and Antonio Colombo Copyright © 2013 Caroline J. Magri et al. All rights reserved. Alleviation of Hyperglycemia Induced Vascular Endothelial Injury by Exenatide Might Be Related to the Reduction of Nitrooxidative Stress Tue, 26 Nov 2013 10:19:58 +0000 We will investigate the effects of exenatide on vascular endothelial injury and nitrooxidative stress in hyperglycemia both in vivo and in vitro and explore the role of nitrooxidative stress in endothelium-protective action of exenatide. Healthy male Wistar rats were randomly divided into 4 groups: control, diabetes mellitus (DM) model, low dose of exenatide treatment, and high dose of exenatide treatment. In vitro study showed that, compared with control group, the DM rats exhibited a lowered endothelium-dependent relaxation and damaged structural integrity of thoracic aortas, and there was a significant increase in plasma nitrotyrosine concentration. These parameters were improved after treatment with either low dose or high dose of exenatide for 45 days. In vitro study showed that exendin-4 (the active ingredient of exenatide) attenuated HUVECs injury induced by high glucose, with improving cell viability and attenuating cell apoptosis. Exendin-4 also significantly alleviated the increased malondialdehyde (MDA), nitrotyrosine content, and inducible nitric oxide synthase (iNOS) expression induced by high glucose in HUVECs. In conclusion, this study demonstrates that exenatide treatment can alleviate the vascular endothelial injury, as well as attenuating the nitrooxidative stress in hyperglycemia, implying that the endothelium-protective effect of exenatide might be related to the reduction of nitrooxidative stress. Qian Zhao, Chun-ling Xu, Hai-yan Xiong, Wen Huang, Mei Zhang, Yun Wang, Si-yu Wang, and Wen Wang Copyright © 2013 Qian Zhao et al. All rights reserved. Clinical Application of Vascular Regenerative Therapy for Peripheral Artery Disease Sun, 24 Nov 2013 09:16:06 +0000 Prognosis of peripheral artery disease (PAD), especially critical limb ischemia, is very poor despite the development of endovascular therapy and bypass surgery. Many patients result in leg amputation and, therefore, vascular regenerative therapy is expected in this field. Gene therapy using vascular endothelial growth factor is the first step of vascular regenerative therapy, but did not confirm effectiveness in a large-scale randomized comparative study. Based on animal experiments, bone marrow mononuclear cells (MNCs), peripheral blood MNCs were used as the cell source for regenerative therapy. Those cells were confirmed to be effective to decrease rest pain and ulcer size, but its effect was not fully satisfied. Mesenchymal stem cells (MSCs) are expected as an effective cell source for vascular regeneration and clinical studies are ongoing, because the cells are able to differentiate into various cell types and produce a significant amount of vascular growth factors. Of vascular regeneration therapy, peripheral MNCs and bone marrow MNCs were recognized as advanced medical technology but do not attain to the standard therapy. However, clinical use of MSCs have already started, and induced pluripotent stem cells are surely promising tool for vascular regeneration therapy although further basic studies are required for clinical application. Hiroshi Suzuki and Yoshitaka Iso Copyright © 2013 Hiroshi Suzuki and Yoshitaka Iso. All rights reserved. CX3CR1 Receptor Polymorphisms, Th1 Cell Recruitment, and Acute Myocardial Infarction Outcome: Looking for a Link Thu, 07 Nov 2013 08:50:18 +0000 Fractalkine is a proinflammatory chemokine that participates in atherosclerotic process mediating the interactions of vascular cells and leukocytes and selective recruitment of Th1 lymphocytes, through interaction with CX3CR1 receptor. The polymorphism of the fractalkine receptor 280M-containing haplotype, which codifies for a receptor with minor expression and with a reduced binding capability, represents a novel protective factor of atherosclerotic disease. We investigated the association among CX3CR1 genotype, the inflammatory infiltrate subpopulations recruited in the plaque, and the in situ expression of fractalkine and its receptor, in patients who died of myocardial infarction (AMI) compared with subjects who died of noncardiac causes. Patients with nonlethal AMI (AMI survivors) were also investigated to correlate the CX3CR1 polymorphisms and the incidence of lethal AMI. A strong T cells infiltrate was found in infarct related artery (IRA) plaques of AMI patients presenting the V249 T280 haplotype (84%). Conversely, a decreased T cell recruitment was associated with I249T280 haplotype in the controls (64%). The significant higher presence of the variant allele I249 in homo- and heterozygosis, found in controls (91%) and in AMI survivors (94%), with respect to the patients who died of AMI (48%), showed the relevance of this polymorphism both in the onset and outcome of acute myocardial infarction. The presence of CX3CR1 polymorphisms could influence the incidence and the outcome of acute myocardial infarction, altering the inflammation of the whole coronary tree by the impaired recruitment of Th1 polarized subpopulation in the coronary plaque. S. Pucci, P. Mazzarelli, M. J. Zonetti, T. Fisco, E. Bonanno, L. G. Spagnoli, and A. Mauriello Copyright © 2013 S. Pucci et al. All rights reserved. Isolation, Characterization, and Transplantation of Cardiac Endothelial Cells Sun, 27 Oct 2013 15:04:51 +0000 Isolation and ex vivo expansion of cardiac endothelial cells have been a recurrent challenge due to difficulties in isolation, cell heterogeneity, lack of specific markers to identify myocardial endothelial cells, and inadequate conditions to maintain long-term cultures. Herein, we developed a method for isolation, characterization, and expansion of cardiac endothelial cells applicable to study endothelial cell biology and clinical applications such as neoangiogenesis. First, we dissociated the cells from murine heart by mechanical disaggregation and enzymatic digestion. Then, we used flow cytometry coupled with specific markers to isolate endothelial cells from murine hearts. CD45+ cells were gated out to eliminate the hematopoietic cells. CD31+/Sca-1+ cells were isolated as endothelial cells. Cells isolated from atrium grew faster than those from ventricle. Cardiac endothelial cells maintain endothelial cell function such as vascular tube formation and acetylated-LDL uptake in vitro. Finally, cardiac endothelial cells formed microvessels in dorsal matrigel plug and engrafted in cardiac microvessels following intravenous and intra-arterial injections. In conclusion, our multicolor flow cytometry method is an effective method to analyze and purify endothelial cells from murine heart, which in turn can be ex vivo expanded to study the biology of endothelial cells or for clinical applications such as therapeutic angiogenesis. Busadee Pratumvinit, Kanit Reesukumal, Kajohnkiart Janebodin, Nicholas Ieronimakis, and Morayma Reyes Copyright © 2013 Busadee Pratumvinit et al. All rights reserved. Paired Measurements of Paraoxonase 1 and Serum Amyloid A as Useful Disease Markers Tue, 22 Oct 2013 14:31:09 +0000 Paraoxonase 1 (PON1) and serum amyloid A (SAA) are proteins carried by high-density lipoprotein (HDL) particles. Among the HDL-associated protein molecules, SAA, an inflammation-related marker, and PON1, an antioxidant marker, tend to change in relatively clear opposite directions in physiological situations. In clinical chemistry, paired measurements of both markers may provide useful information to understand dysfunctional HDL in diseases with inflammation and oxidative stress conditions. Actually, limited clinical studies have suggested that the combined use of PON1 and SAA may be a tool for observing the pathophysiology of some disease entities. From the findings of experimental studies, PON1 appears to be cooperatively regulated by inflammation- and oxidative stress-related molecules linked with SAA regulation in humans. More studies remain to be performed to ascertain the value of paired measurements of both promising markers in clinical practice. Kazuhiko Kotani, Toshiyuki Yamada, and Alejandro Gugliucci Copyright © 2013 Kazuhiko Kotani et al. All rights reserved. Prevention of Atherosclerosis Progression by 9-cis-β-Carotene Rich Alga Dunaliella in apoE-Deficient Mice Mon, 23 Sep 2013 08:23:13 +0000 Introduction. β-Carotene-rich diet has been shown to be inversely associated with the risk of coronary heart disease. However, clinical trials using synthetic all-trans-β-carotene failed to demonstrate a beneficial effect. We therefore sought to study the effect of natural source of β-carotene, the alga Dunaliella, containing both all-trans and 9-cis-β-carotene on atherosclerosis. In a previous study we showed that 9-cis-β-carotene-rich powder of the alga Dunaliella inhibits early atherogenesis in low-density lipoprotein receptor knockout mice. Aims. The aims of the current work were to study whether diet enriched with Dunaliella powder would inhibit the progression of established atherosclerosis in old male apoE-deficient mice and to compare the effect of Dunaliella on lipid profile and atherosclerosis in a low-versus high-fat diet fed mice. Methods. In the first experiment, young mice (12 weeks old) were allocated into 3 groups: (1) low-fat diet; (2) low-fat diet + Dunaliella powder (8%); (3) low-fat diet + β-carotene-deficient Dunaliella. In the second experiment, old mice (7 months old) with established atherosclerotic lesions were allocated into 4 groups: (1) low-fat diet; (2) low-fat diet + Dunaliella; (3) high fat-diet; (4) high-fat diet + Dunaliella. Results. In young mice fed a low-fat diet, a trend toward lower atherosclerotic lesion area in the aortic sinus was found in the Dunaliella group compared with the control group. In old mice with established atherosclerotic lesion, Dunaliella inhibited significantly plasma cholesterol elevation and atherosclerosis progression in mice fed a high-fat diet. Conclusion. The results of this study suggest that a diet containing natural carotenoids, rich in 9-cis-β-carotene, has the potential to inhibit atherosclerosis progression, particularly in high-fat diet regime. Ayelet Harari, Revital Abecassis, Noa Relevi, Zohar Levi, Ami Ben-Amotz, Yehuda Kamari, Dror Harats, and Aviv Shaish Copyright © 2013 Ayelet Harari et al. All rights reserved. Preexisting High Expression of Matrix Metalloproteinase-2 in Tunica Media of Saphenous Vein Conduits Is Associated with Unfavorable Long-Term Outcomes after Coronary Artery Bypass Grafting Mon, 16 Sep 2013 15:45:35 +0000 Introduction. Migration of the smooth muscle cells (SMCs) to the tunica media in the saphenous vein (SV) transplants is facilitated by matrix metalloproteinases (MMPs). The aim of this study was to identify any associations between expression of MMP-2 or endogenous tissue inhibitors (TIMP-2 and TIMP-3) in the SV segments and late failure of the SV grafts. Methods. Two hundred consecutive patients with a mean age of 63.1 ± 8.9 years who underwent primary isolated venous CABG were examined. Patients were retrospectively split into two subgroups, with the SV graft disease (SVGD (+); ) or without it (SVGD (−); ). In the SV segments, immunohistochemical analysis of the expression of the MMP-2, TIMP-2, and -3 was performed. Results. In the SVGD (+) patients, tissue expression of MMP-2 was stronger, whereas that of both TIMPs was weaker than in the SVGD (−) patients. In majority of the SV segments obtained from the SVGD (−) individuals, a balance in MMP and TIMP expressions was found, whereas an upregulation of MMP-2 expression was usually noted in the SVGD (+) subjects. Conclusion. The strong expression of MMP-2 accompanied by reduced immunostaining of both TIMPs is associated with the development of the SV graft disease and unfavorable CABG outcomes. Bartlomiej Perek, Agnieszka Malinska, Marcin Misterski, Danuta Ostalska-Nowicka, Maciej Zabel, Anna Perek, and Michal Nowicki Copyright © 2013 Bartlomiej Perek et al. All rights reserved. Incidence and Risk Factors of Early Delirium after Cardiac Surgery Thu, 12 Sep 2013 14:55:30 +0000 Introduction. The aim of our study was to identify the incidence and risk factors of delirium after cardiac surgery implementing Intensive Care Delirium Screening Checklist (ICDSC). Material and Methods. 87 patients, undergoing cardiac surgery at Vilnius University hospital, were prospectively monitored for postoperative delirium development, during intensive care unit stay. Results. The incidence of postoperative delirium was 13.30%. No statistically relevant preoperative predictors of delirium were found. The duration of surgery was significantly longer in delirium group ( versus hours, ). Patients in delirium group more often had blood product transfusions (1.50 (± 1.57) versus 0.49 (± 0.91) ) and had a higher incidence of low cardiac output syndrome (33.30% versus 3.00%, ); they were significantly longer mechanically ventilated ( versus 8.78 ± 4.77 ()) hours (OR = 1.15 ()) and had twice longer ICU stay ( versus 2.60 ± 1.10 ()) days (OR = 1.91 ()). Conclusions. The incidence of delirium after cardiac surgery was 13.3%. Independent predictors of delirium were duration of postoperative mechanical ventilation and intensive care unit stay. Ieva Norkienė, Donata Ringaitienė, Vilma Kuzminskaitė, and Jūratė Šipylaitė Copyright © 2013 Ieva Norkienė et al. All rights reserved. Doxycycline Prevents Intimal Hyperplasia In Vitro and May Improve Patency of the Internal Thoracic Artery Thu, 22 Aug 2013 08:18:57 +0000 Objectives. The development of intimal hyperplasia and graft failure is an important problem in cardiac surgery. A fundamental process in intimal hyperplasia is the degradation of extracellular matrix by metalloproteases which induces the vascular smooth-muscle cells migration and sets the scene for graft atherosclerosis. This study investigated whether doxycycline, a metalloproteases inhibitor, can prevent the intimal hyperplasia occurrence in cultured human internal mammary artery, thus extending graft patency. Methods. Segments of internal mammary artery from 20 consecutive patients were prepared and cultured for 2 weeks in serum-supplemented medium (control) or in medium supplemented with 10−5 M and 10−6 M doxycycline concentrations. Tissues were fixed, sectioned, and stained, and neointimal thickness was measured by computer-aided image analysis. Further sections were cultured and prepared for gel enzymography to measure the matrix metalloproteinase-2 and -9 levels. Results. At the end of the culture period, neointimal thickness was significantly () dose-dependently reduced in samples treated with doxycycline when compared with controls. Gelatin enzymography demonstrated a reduction in values for both latent and active forms of metalloproteases. Conclusions. Doxycycline, in a model of internal mammary artery intimal hyperplasia, has a specific role in inhibiting metalloproteases activity and may prevent graft stenosis. Vito Mannacio, Luigi Di Tommaso, Anita Antignano, Ettorino Di Tommaso, Paolo Stassano, and Carlo Vosa Copyright © 2013 Vito Mannacio et al. All rights reserved. Knockout of Density-Enhanced Phosphatase-1 Impairs Cerebrovascular Reserve Capacity in an Arteriogenesis Model in Mice Tue, 06 Aug 2013 09:18:49 +0000 Collateral growth, arteriogenesis, represents a proliferative mechanism involving endothelial cells, smooth muscle cells, and monocytes/macrophages. Here we investigated the role of Density-Enhanced Phosphatase-1 (DEP-1) in arteriogenesis in vivo, a protein-tyrosine-phosphatase that has controversially been discussed with regard to vascular cell biology. Wild-type C57BL/6 mice subjected to permanent left common carotid artery occlusion (CCAO) developed a significant diameter increase in distinct arteries of the circle of Willis, especially in the anterior cerebral artery. Analyzing the impact of loss of DEP-1 function, induction of collateralization was quantified after CCAO and hindlimb femoral artery ligation comparing wild-type and DEP-1−/− mice. Both cerebral collateralization assessed by latex perfusion and peripheral vessel growth in the femoral artery determined by microsphere perfusion and micro-CT analysis were not altered in DEP-1−/− compared to wild-type mice. Cerebrovascular reserve capacity, however, was significantly impaired in DEP-1−/− mice. Cerebrovascular transcriptional analysis of proarteriogenic growth factors and receptors showed specifically reduced transcripts of PDGF-B. SiRNA knockdown of DEP-1 in endothelial cells in vitro also resulted in significant PDGF-B downregulation, providing further evidence for DEP-1 in PDGF-B gene regulation. In summary, our data support the notion of DEP-1 as positive functional regulator in vascular cerebral arteriogenesis, involving differential PDGF-B gene expression. Daniel Hackbusch, André Dülsner, Nora Gatzke, Janine Krüger, Philipp Hillmeister, Stephanie Nagorka, Florian Blaschke, Zully Ritter, Christa Thöne-Reineke, Frank-D. Böhmer, Ivo Buschmann, and Kai Kappert Copyright © 2013 Daniel Hackbusch et al. All rights reserved. Aorta Structural Alterations in Term Neonates: The Role of Birth and Maternal Characteristics Thu, 25 Jul 2013 14:12:12 +0000 Aim. To evaluate the influence of selected maternal and neonatal characteristics on aorta walls in term, appropriately grown-for-gestational age newborns. Methods. Age, parity, previous abortions, weight, height, body mass index before and after delivery, smoking, and history of hypertension, of diabetes, of cardiovascular diseases, and of dyslipidemia were all assessed in seventy mothers. They delivered 34 males and 36 females healthy term newborns who underwent ultrasound evaluation of the anteroposterior infrarenal abdominal aorta diameter (APAO), biochemical profile (glucose, insulin, total cholesterol, HDL and LDL cholesterol, triglycerides, fibrinogen, and D-dimers homeostasis model assessment [HOMAIR]index), and biometric parameters. Results. APAO was related to newborn length (; ), head circumference (; ), gestational age (, ), HOMA index (; ), and D-dimers (, ). Smoke influenced APAO values (odds ratio: 1.80; confidence interval 95%: 1.05–3.30), as well as diabetes during pregnancy (, ). Maternal height influenced neonatal APAO (, ). Multiple regression analysis outlined neonatal D-dimers as still significantly related to neonatal APAO values. Conclusions. Many maternal and neonatal characteristics could influence aorta structures. Neonatal D-dimers are independently related to APAO. Marco Matteo Ciccone, Pietro Scicchitano, Christian Salerno, Michele Gesualdo, Fara Fornarelli, Annapaola Zito, Lucia Filippucci, Roberta Riccardi, Francesca Cortese, Francesca Pini, Lucia Angrisani, Antonio Di Mauro, Federico Schettini, and Nicola Laforgia Copyright © 2013 Marco Matteo Ciccone et al. All rights reserved. Aortic Dissection Type A in Alpine Skiers Sun, 21 Jul 2013 11:29:07 +0000 Patients and Methods. 140 patients with aortic dissection type A were admitted for cardiac surgery. Seventy-seven patients experienced their dissection in the winter season (from November to April). We analyzed cases of ascending aortic dissection associated with alpine skiing. Results. In 17 patients we found skiing-related aortic dissections. Skiers were taller (180 (172–200) cm versus 175 (157–191) cm, ) and heavier (90 (68–125) kg versus 80 (45–110) kg, ) than nonskiers. An extension of aortic dissection into the aortic arch, the descending thoracic aorta, and the abdominal aorta was found in 91%, 74%, and 69%, respectively, with no significant difference between skiers and nonskiers. Skiers experienced RCA ostium dissection requiring CABG in 17.6% while this was true for 5% of nonskiers (). Hospital mortality of skiers was 6% versus 13% in nonskiers (). The skiers live at an altitude of 170 (0–853) m.a.s.l. and experience their dissection at 1602 (1185–3105; ) m.a.s.l. In 82% symptom start was during recreational skiing without any trauma. Conclusion. Skiing associated aortic dissection type A is usually nontraumatic. The persons affected live at low altitudes and practice an outdoor sport at unusual high altitude at cold temperatures. Postoperative outcome is good. Thomas Schachner, Nikolaus Fischler, Julia Dumfarth, Nikolaos Bonaros, Christoph Krapf, Wolfgang Schobersberger, and Michael Grimm Copyright © 2013 Thomas Schachner et al. All rights reserved. Association of Atherosclerotic Peripheral Arterial Disease with Adiponectin Genes SNP+45 and SNP+276: A Case-Control Study Mon, 03 Jun 2013 15:43:19 +0000 Objectives. We hypothesized that adiponectin gene SNP+45 (rs2241766) and SNP+276 (rs1501299) would be associated with atherosclerotic peripheral arterial disease (PAD). Furthermore, the association between circulating adiponectin levels, fetuin-A, and tumoral necrosis factor-alpha (TNF-) in patients with atherosclerotic peripheral arterial disease was investigated. Method. Several blood parameters (such as adiponectin, fetuin-A, and TNF-) were measured in 346 patients, 226 with atherosclerotic peripheral arterial disease (PAD) and 120 without symptomatic PAD (non-PAD). Two common SNPs of the ADIPOQ gene represented by +45T/G 2 and +276G/T were also investigated. Results. Adiponectin concentrations showed lower circulating levels in the PAD patients compared to non-PAD patients (). Decreasing adiponectin concentration was associated with increasing serum levels of fetuin-A in the PAD patients. None of the investigated adiponectin SNPs proved to be associated with the subjects’ susceptibility to PAD (). Conclusion. The results of our study demonstrated that neither adiponectin SNP+45 nor SNP+276 is associated with the risk of PAD. Claudia D. Gherman, Doru Pamfil, and Sorana D. Bolboacă Copyright © 2013 Claudia D. Gherman et al. All rights reserved. PPAR Agonist GW501516 Inhibits PDGF-Stimulated Pulmonary Arterial Smooth Muscle Cell Function Related to Pathological Vascular Remodeling Wed, 27 Mar 2013 10:43:00 +0000 Pulmonary arterial hypertension (PAH) is a severe and progressive disease, a key feature of which is pulmonary vascular remodeling. Growth factors, cytokines, and lipid mediators are involved in this remodeling process. Recent reports suggest that the peroxisome proliferator-activated receptors (PPARs) play important roles in the regulation of cell growth and differentiation as well as tissue wounding and repair. In this study, we examined the role of PPARδ in the regulation of proliferation, migration, collagen synthesis, and chemokine production in human pulmonary arterial smooth muscle cells (HPASMCs). The data showed that PPARδ was the most abundant isoform in HPASMCs. PPARδ was upregulated in HPASMCs treated with PDGF, which is the major mediator in pulmonary vascular remodeling. Activation of PPARδ by GW501516, a specific PPARδ ligand, significantly inhibited PDGF-induced proliferation in HPASMCs. The inhibitory effect of GW501516 on HPASMCs was associated with decreased expression of cyclin D1, cyclin D3, CDK2, and CDK4 as well as increased expression of the cell cycle inhibitory genes G0S2 and . Pretreatment of HPASMCs with GW501516 significantly inhibited PDGF-induced cell migration and collagen synthesis. GW501516 also significantly attenuated TNF-mediated expression of MCP-1. These results suggest that PPARδ may be a potential therapeutic target against the progression of vascular remodeling in PAH. Guangjie Liu, Xuan Li, Yan Li, Xin Tang, Jie Xu, Ran Li, Peng Hao, and Yongchang Sun Copyright © 2013 Guangjie Liu et al. All rights reserved. In Vivo Imaging of Leukocyte Recruitment to the Atheroprone Femoral Artery Reveals Anti-Inflammatory Effects of Rosuvastatin Sun, 30 Dec 2012 16:06:29 +0000 Objective. To monitor the anti-inflammatory effect of rosuvastatin in leukocyte endothelial interactions in the atheroprone femoral artery in vivo. Methods and Results. Male Apolipoprotein E null mice (ApoE−/− mice, 6 weeks old) were fed a high-fat diet (20% fat, 1.25% cholesterol) with or without the HMG CoA reductase inhibitor rosuvastatin (10 mg/kg/day) for 6 weeks. Significant leukocyte adhesion was observed in the femoral artery of ApoE−/− mice, but not of wild type mice, in the absence of rosuvastatin. Interestingly, no obvious plaque formation was observed in the artery at this time point. The number of adherent leukocytes was dramatically diminished in ApoE−/− mice treated with rosuvastatin. DHE-associated oxidative stress and the expression of gp91-phox, a component of NADPH oxidase, were induced in ApoE−/− mice and were abolished by rosuvastatin treatment. Conclusion. Our data documented leukocyte recruitment prior to lipid accumulation and subsequent inhibition by rosuvastatin. The underlying mechanism seemed to involve oxidative stress and an anti-inflammatory effect on the endothelium of atheroprone vessels. Mizuko Osaka, Sumihiko Hagita, and Masayuki Yoshida Copyright © 2013 Mizuko Osaka et al. All rights reserved. Genistein Attenuates Vascular Endothelial Impairment in Ovariectomized Hyperhomocysteinemic Rats Tue, 06 Nov 2012 13:19:33 +0000 Hyperhomocysteinemia (HHcy) is a well-known independent risk factor for vascular diseases in the general population. This study was to explore the effect of genistein (GST), a natural bioactive compound derived from legumes, on HHcy-induced vascular endothelial impairment in ovariectomized rats in vivo. Thirty-two adult female Wistar rats were assigned randomly into four groups (): (a) Con: control; (b) Met: 2.5% methionine diet; (c) OVX + Met: ovariectomy + 2.5% methionine diet; (d) OVX + Met + GST: ovariectomy + 2.5% methionine diet + supplementation with genistein. After 12 wk of different treatment, the rats' blood, toracic aortas and liver samples were collected for analysis. Results showed that high-methionine diet induced both elevation of plasma Hcy and endothelial dysfunction, and ovariectomy deteriorated these injuries. Significant improvement of both functional and morphological changes of vascular endothelium was observed in OVX + Met + GST group; meanwhile the plasma Hcy levels decreased remarkably. There were significant elevations of plasma ET-1 and liver MDA levels in ovariectomized HHcy rats, and supplementation with genistein could attenuate these changes. These results implied that genistein could lower the elevated Hcy levels, and prevent the development of endothelial impairment in ovariectomized HHcy rats. This finding may shed a novel light on the anti-atherogenic activities of genistein in HHcy patients. Panpan Zhen, Qian Zhao, Dandan Hou, Teng Liu, Dongqiao Jiang, Jinhong Duan, Lingqiao Lu, and Wen Wang Copyright © 2012 Panpan Zhen et al. All rights reserved. Validation of a New Animal Model of Vulnerable Plaques by Intravascular Optical Coherence Tomography In Vivo Wed, 03 Oct 2012 10:18:18 +0000 We aimed to establish a rabbit model of vulnerable plaques (VPs) with the morphology and component characteristics of human VPs and to evaluate the microstructural features of VPs in vivo using intravascular optical coherence tomography (OCT). Twelve rabbits underwent endothelial denudation of the carotid artery and consumed a 1% high-cholesterol diet (HCD). They were equally divided into two groups: group A (modified needle injury) and group B (balloon injury). OCT was undertaken thrice before injury as well as 1 h and 12 weeks after injury. The degree of acute artery injury after endothelial denudation was detected by OCT. Twelve weeks after injury, OCT showed that both groups generated VPs which had thin fibrous caps and a large lipid core, whereas plaques in group A had smaller lipid arcs (). Histological findings demonstrated that a larger eccentricity index (EI) () and greater infiltration of macrophages () in group A than in group B. Qualitative and morphometric analyses of plaques showed a significant correlation between histological and OCT measurements. A combination of modified endothelial denudation and an HCD in rabbits produced more eccentric lesions similar to those seen in humans. These data suggest that OCT could be a useful tool for evaluation of the degree of injury and VPs in vivo. Yan Fang, Sining Hu, Jingbo Hou, Lingbo Meng, Shaosong Zhang, and Bo Yu Copyright © 2012 Yan Fang et al. All rights reserved. Role of Exogenous Nitric Oxide Donor in Treatment of Decompensated Hemorrhagic Shock in Normotensive and Hypertensive Rats Tue, 12 Jun 2012 15:24:49 +0000 Introduction. In this study, we investigated the role of exogenous NO donor, sodium nitroprusside (SNP), on hemodynamic responses and survival rate during decompensated hemorrhagic shock in normotensive and hypertensive rat. Methods. Male wistar rats were divided into normotensive and hypertensive groups (𝑛=12 each). Then, the animals were subjected to decompensated hemorrhagic shock by withdrawing blood until the mean arterial pressure (MAP) reached to 40 mmHg. After the shock period, the animals were randomly assigned to SNP-treated (0.5 mg/kg) and control groups (𝑛=6 each). MAP and heart rate (HR) were monitored throughout the experiment and 60 min after the administration of drug. Serum NO concentrations were measured. The survival rate was counted during next 72 h. Results. Infusion of SNP caused no significant changes in MAP and HR in normotensive and hypertensive animals. Hemorrhagic shock increased serum NO concentration and SNP administration reduced serum NO concentration in either normotensive or hypertensive groups. Survival counts during 72 h after experiment did not improve by SNP administration, and there were no significant differences between normotensive and hypertensive groups. Conclusion. SNP administration cannot improve hemodynamic responses and survival count during decompensated hemorrhagic shock in normotensive and hypertensive animals. Majid Khazaei and Babak Barmaki Copyright © 2012 Majid Khazaei and Babak Barmaki. All rights reserved. A Novel Model of Atherosclerosis in Rabbits Using Injury to Arterial Walls Induced by Ferric Chloride as Evaluated by Optical Coherence Tomography as well as Intravascular Ultrasound and Histology Mon, 14 May 2012 14:11:28 +0000 This study aim was to develop a new model of atherosclerosis by FeCl3-induced injury to right common carotid arteries (CCAs) of rabbits. Right CCAs were induced in male New Zealand White rabbits (𝑛=15) by combination of a cholesterol-rich diet and FeCl3-induced injury to arterial walls. The right and left CCAs were evaluated by histology and in vivo intravascular ultrasound (IVUS) and optical coherence tomography (OCT) examinations of 24 hours (𝑛=3), 8 weeks (𝑛=6), and 12 weeks (𝑛=6) after injury. Each right CCA of the rabbits showed extensive white-yellow plaques. At eight and 12 weeks after injury, IVUS, OCT, and histological findings demonstrated that the right CCAs had evident eccentric plaques. Six plaques (50%) with evident positive remodeling were observed. Marked progression was clearly observed in the same plaque at 12 weeks after injury when it underwent repeat OCT and IVUS. We demonstrated, for the first time, a novel model of atherosclerosis induced by FeCl3. The model is simple, fast, inexpensive, and reproducible and has a high success rate. The eccentric plaques and remodeling of plaques were common in this model. We successfully carried out IVUS and OCT examinations twice in the same lesion within a relatively long period of time. Jinwei Tian, Sining Hu, Yanli Sun, Xiang Ban, Huai Yu, Nana Dong, Jian Wu, and Bo Yu Copyright © 2012 Jinwei Tian et al. All rights reserved. Transfer of Bone-Marrow-Derived Mesenchymal Stem Cells Influences Vascular Remodeling and Calcification after Balloon Injury in Hyperlipidemic Rats Mon, 14 May 2012 09:21:52 +0000 Bone-marrow-derived mesenchymal stem cells (BM-MSCs) were found to markedly increase atherosclerotic lesion size. The aim of this paper was to investigate whether BM-MSCs contribute to vascular remodeling and calcification after balloon injury in hyperlipidemic rats. Labeled BM-MSCs were found in the lesion of hyperlipidemic rats after balloon injury. Comparing injury group, transferred BM-MSCs significantly triggered vascular negative remodeling, characterized by the changes of remodeling index (0.628±0.0293 versus 0.544±0.0217), neointimal area (0.078±0.015 mm2 versus 0.098±0.019 mm2), PCNA index (23.91±6.59% versus 43.11±5.31%), and percentage of stenosis (18.20±1.09% versus 30.58±1.21%). Apparent vascular calcification was detected in medial layers at 6 weeks after balloon angioplasty, which may be associated with upregulation of bone morphogenetic protein-2 (BMP-2). Our data indicated that unselected BM-MSCs transfer may induce vascular remodeling and calcification after balloon injury in hyperlipidemic rats. Jianquan Liao, Xiaochun Chen, Yongdong Li, Zhiping Ge, Hongyu Duan, Yunzeng Zou, and Junbo Ge Copyright © 2012 Jianquan Liao et al. All rights reserved. Temporal and Quantitative Analysis of Atherosclerotic Lesions in Diet-Induced Hypercholesterolemic Rabbits Wed, 14 Mar 2012 11:41:23 +0000 The diet-induced atherosclerotic rabbit is an ideal model for atherosclerosis study, but temporal changes in atherosclerotic development in hypercholesterolemic rabbits are poorly understood. Japanese white rabbits were fed a high-cholesterol diet to induce sustained hypercholesterolemia, and each group of 10–12 animals was then sacrificed at 6, 12, 16, or 28 weeks. The rabbit aortas were harvested, and the sizes of the gross and intima atherosclerotic lesions were quantified. The cellular component of macrophages (Mφs) and smooth muscle cells (SMCs) in aortic intimal lesions was also quantified by immunohistochemical staining, and the correlation between plasma cholesterol levels and the progress of atherosclerotic lesions was studied. The ultrastructure of the atherosclerotic lesions was observed by transmission electron microscopy (TEM). Widely variable atherosclerotic plaques were found from 6 weeks to 28 weeks, and the lesional progress was closely correlated with cholesterol exposure. Interestingly, a relatively reduced accumulation of Mφ, an increased numbers of SMCs, and a damaged endothelial layer were presented in advanced lesions. Moreover, SMCs were closely correlated with cholesterol exposure and lesional progress for the whole period. Cholesterol exposure directly determines atherosclerotic progress in a rabbit model, and the changes in the cellular component of advanced lesions may affect plaque stability in an atherosclerotic rabbit model. Qi Yu, Yafeng Li, Ahmed Bilal Waqar, Yanli Wang, Bingqiao Huang, Yulong Chen, Sihai Zhao, Peigang Yang, Jianglin Fan, and Enqi Liu Copyright © 2012 Qi Yu et al. All rights reserved. All-trans-Retinoic Acid Ameliorated High Fat Diet-Induced Atherosclerosis in Rabbits by Inhibiting Platelet Activation and Inflammation Thu, 01 Mar 2012 09:28:17 +0000 Background. All-trans-retinoic acid (atRA) is effective for many proliferative diseases. We investigated the protective effects of atRA against atherosclerosis. Methods. Rabbits were randomly allocated to receive basal diet or an HFD for 4 weeks. HFD group then received rosuvastatin (3 mg/day), atRA (5 mg/kg/day), or the same volume of vehicle, respectively, for next 8 weeks. Results. HFD group showed increases in plasma lipids and aortic plaque formation. P-selectin expression and fibrinogen binding on platelets or deposition on the intima of the aorta also increased significantly as did the levels of TNF-α, IL-6, and fibrinogen in plasma. After 8 weeks of treatment with atRA, there was a significant decrease in plasma lipids and improvement in aortic lesions. AtRA also inhibited the expression of P-selectin and fibrinogen binding on platelets and deposition on the intima of the aorta. Conclusion. AtRA can ameliorate HFD-induced AS in rabbits by inhibiting platelet activation and inflammation. Birong Zhou, Ying Pan, Zeping Hu, Xiaobian Wang, Jianxiong Han, Qing Zhou, Zhimin Zhai, and Yuan Wang Copyright © 2012 Birong Zhou et al. All rights reserved. The Role of Costimulatory Receptors of the Tumour Necrosis Factor Receptor Family in Atherosclerosis Thu, 22 Dec 2011 09:08:25 +0000 Atherosclerosis is a chronic inflammatory disease that is mediated by both the innate and adaptive immune responses. T lymphocytes, that together with B cells are the cellular effectors of the adaptive immune system, are currently endowed with crucial roles in the development and progression of atherosclerosis. Costimulatory receptors are a class of molecules expressed by T lymphocytes that regulate the activation of T cells and the generation of effector T-cell responses. In this review we present the roles of costimulatory receptors of the tumour necrosis factor receptor (TNFR) superfamily in atherosclerosis and discuss the implications for future therapies that could be used to specifically modulate the immune response of pathogenic T cells in this disease. Ricardo F. Antunes, Juan Carlos Kaski, and Ingrid E. Dumitriu Copyright © 2012 Ricardo F. Antunes et al. All rights reserved. Novel Approaches to Treat Experimental Pulmonary Arterial Hypertension: A Review Mon, 22 Mar 2010 10:22:51 +0000 Background. Pulmonary arterial hypertension (PAH) is a life-threatening disease characterized by an increase in pulmonary artery pressure leading to right ventricular (RV) hypertrophy, RV failure, and ultimately death. Current treatments can improve symptoms and reduce severity of the hemodynamic disorder but gradual deterioration in their condition often necessitates a lung transplant. Methods and Results. In experimental models of PAH, particularly the model of monocrotaline-induced pulmonary hypertension, efficacious treatment options tested so far include a spectrum of pharmacologic agents with actions such as anti-mitogenic, proendothelial function, proangiogenic, antiinflammatory and antioxidative. Emerging trends in PAH treatment are gene and cell therapy and their combination, like (progenitor) cells enriched with eNOS or VEGF gene. More animal data should be collected to investigate optimal cell type, in vitro cell transduction, route of administration, and number of cells to inject. Several recently discovered and experimentally tested interventions bear potential for therapeutic purposes in humans or have been shown already to be effective in PAH patients leading to improved life expectation and better quality of life. Conclusion. Since many patients remain symptomatic despite therapy, we should encourage research in animal models of PAH and implement promising treatments in homogeneous groups of PAH patients. S. Umar, P. Steendijk, D. L. Ypey, D. E. Atsma, E. E. van der Wall, M. J. Schalij, and A. van der Laarse Copyright © 2010 S. Umar et al. All rights reserved. Glucagon Effects on Ischemic Vasodilatation in the Isolated Rat Heart Thu, 18 Mar 2010 16:08:12 +0000 The myocardial reperfusion following ischemia leads to the ischemic vasodilation by affecting the release of various vasoactive substances, such as free radicals, NO, and histamine. In addition, some evidences suggest that glucagon itself may alter the release of those substances. In this study, we investigated the ischemic vasodilation of the isolated rat heart, as well as the concentrations of NO, TBARS, and histamine in the coronary venous effluent either in the presence or in the absence of glucagon. Our results showed that in the presence of glucagon, there was a faster restoration of coronary perfusion pressure during ischemic vasodilation compared to the absence of glucagon (124±5.6 versus 81±5.2 s) with no apparent changes in TBARS concentration. The glucagon's administration leads to the decreased release of histamine by approximately 35%. Biphasic release of NO in the presence of glucagon initially showed augmentation by 60%, followed by the significant attenuation of 45%. Mirko Rosic, Suzana Pantovic, Gvozden Rosic, Aleksandra Tomic-Lucic, Tatjana Labudovic, Vladimir Zivkovic, and Vladimir Jakovljevic Copyright © 2010 Mirko Rosic et al. All rights reserved. Characterization of Arterial Wave Reflection in Healthy Bonnet Macaques: Feasibility of Applanation Tonometry Tue, 10 Mar 2009 14:47:57 +0000 Nonhuman primates are commonly used in cardiovascular research. Increased arterial stiffness is a marker of subclinical atherosclerosis and higher CV risk. We determined the augmentation index (AI) using applanation tonometry in 61 healthy monkeys (59% female, age 1–25 years). Technically adequate studies were obtained in all subjects and required minutes. The brachial artery provided the highest yield (95%). AI was correlated with heart rate (HR) (), crown rump length (CRL) (r = 0.42, P = .001), and left ventricular (LV) mass determined using echocardiography (). On multivariate analysis, HR () and CRL (P = .005) were independent predictors of AI (). Body Mass Index (BMI) and AI were independent predictors of higher LV mass on multivariate analysis ( and ). In conclusion, applanation tonometry is feasible for determining AI. Reference values are provided for AI in bonnet macaques, in whom higher AI is related to HR and CRL, and in turn contributes to higher LV mass. Jason Lazar, Ghazanfar Qureshi, Haroon Kamran, Leonard A. Rosenblum, John G. Kral, and Louis Salciccioli Copyright © 2009 Jason Lazar et al. All rights reserved. Thermolabile Methylenetetrahydrofolate Reductase C677T Polymorphism and Homocysteine Are Risk Factors for Coronary Artery Disease in Moroccan Population Wed, 07 Mar 2007 00:00:00 +0000 Increased plasma total homocysteine (tHcy) levels have been shown to be a risk factor for coronary artery disease (CAD). The common methylenetetrahydrofolate reductase C677T (MTHFR C677T) polymorphism has been reported to be a strong predictor of mild hyperhomocysteinaemia (HHcy). We assessed whether this mutation was associated with increased risk of CAD and plasma levels of tHcy. We also evaluated interactions between this polymorphism, mild elevated tHcy levels and conventional risk factors of CAD. Method. Using PCR-RFLP analysis, we studied the frequency of the C677T genotypes and its effect on CAD and on tHcy concentrations in 400 subjects without and with CAD angiographically confirmed. There were 210 subjects with CAD and 190 subjects without CAD. Results. The frequencies of the C677T genotypes were 53% (59.5% in controls versus 48.1% in cases), 34.8% (32.1 in controls versus 37.1 in cases), and 11.8% (8.4% in controls versus 14.8% in cases), respectively, for 677CC, 677CT, and 677TT. The genotype frequencies were significantly different between case and control groups (P<.05). The 677T allele enhances the risk of CAD associated to HHcy (P<.01). In multivariate analysis models, MTHFR C677T polymorphism effect on CAD was masked by other risk factors. HHcy was only and independently influenced by MTHFR polymorphism and smoking habits, and it is a strong predictor of CAD independently of conventional risk factors. Conclusion. Our data suggest that HHcy is strongly and independently associated to CAD risk increase; and MTHFR C677T polymorphism and smoking habits were the main predictors of tHcy levels. The CAD risk increase is mainly associated with mild HHcy in 677TT, whereas in 677CT and 677CC it is mainly associated with the conventional risk factors. Nawal Bennouar, Abdellatif Allami, Houssine Azeddoug, Abdenbi Bendris, Abdelilah Laraqui, Amal El Jaffali, Nizar El Kadiri, Rachid Benzidia, Anwar Benomar, Seddik Fellat, and Mohamed Benomar Copyright © 2007 Nawal Bennouar et al. All rights reserved. Reduced Atherosclerotic Lesion Size in P-Selectin Deficient Apolipoprotein E-Knockout Mice Fed a Chow but Not a Fat Diet Tue, 04 Apr 2006 00:00:00 +0000 Endothelial cells lining atherosclerotic, but not healthy sites, on human arteries express P-selectin. We investigated the role of P-selectin on the development of vascular lesions in an ApoE−/− male mice. Double-knockout (ApoE−/−, P-selectin-/-; DKO) were compared to single-knockout (ApoE−/−; SKO) mice. They were fed a chow or fat diet for 3, 6, 15, and 20 weeks, without any differences in cholesterol levels. DKO mice fed a chow diet exhibited a ratio of lesion area over media lower than SKO mice, for 3 (P<.03) , 6 (P<.001), and 15 (P<.02) weeks. DKO mice fed a fat diet showed a lower ratio only at 3 weeks. P-selectin deficiency in ApoE−/− mice has a protective effect in atherosclerotic lesions development. Reduction of lesion size depends on diet type and duration. A fat diet could neutralize the beneficial effects of P-selectin deficiency, inducing atherosclerotic lesions via probably other adhesion molecules. Marie-Claude Bourdillon, Jacques Randon, Lydie Barek, Kazem Zibara, Chantal Covacho, Robin N Poston, Elza Chignier, and John L. McGregor Copyright © 2006 Marie-Claude Bourdillon et al. All rights reserved. Identification, Structural, and Functional Characterization of a New Early Gene (6A3-5, 7 kb): Implication in the Proliferation and Differentiation of Smooth Muscle Cells Mon, 01 Jan 1900 00:00:00 +0000 Arterial smooth muscle cells (SMCs) play a major role in atherosclerosis and restenosis. Differential display was used to compare transcription profiles of synthetic SMCs to proliferating rat cultured SMC line. An isolated cDNA band (6A3-5) was shown by northern (7 kb) to be upregulated in the proliferating cell line. A rat tissue northern showed differential expression of this gene in different tissues. Using 5’ RACE and screening of a rat brain library, part of the cDNA was cloned and sequenced (5.4 kb). Sequence searches showed important similarities with a new family of transcription factors, bearing ARID motifs. A polyclonal antibody was raised and showed a protein band of 175 kd, which is localized intracellularly. We also showed that 6A3-5 is upregulated in dedifferentiated SMC (P9) in comparison to contractile SMC ex vivo (P0). This work describes cloning, structural, and functional characterization of a new early gene involved in SMC phenotype modulation. Kazem Zibara, Gwenaële Garin, and John L. McGregor Copyright © 2005 Hindawi Publishing Corporation. All rights reserved.