bluuebunny/crossref_metadata_2025 huggingface dataset:
Cyclooxygenase-1 and -2–Dependent Prostacyclin Formation in Patients With Atherosclerosis
Authors: Belton Orina , Byrne Dara , Kearney Dermot , Leahy Austin , Fitzgerald Desmond J.
Published: June 2012

Background
—The formation of prostacyclin (PGI
2
), thromboxane (TX) A
2
, and isoprostanes is markedly enhanced in atherosclerosis. We examined the relative contribution of cyclooxygenase (COX)-1 and -2 to the generation of these eicosanoids in patients with atherosclerosis.

Methods and Results
—The study population consisted of 42 patients with atherosclerosis who were undergoing surgical revascularization. COX-2 mRNA was detected in areas of atherosclerosis but not in normal blood vessel walls, and there was evidence of COX-1 induction. The use of immunohistochemical studies localized the COX-2 to proliferating vascular smooth muscle cells and macrophages. Twenty-four patients who did not previously receive aspirin were randomized to receive either no treatment or nimesulide at 24 hours before surgery and then for 3 days. Eighteen patients who were receiving aspirin were continued on a protocol of either aspirin alone or a combination of aspirin and nimesulide. Urinary levels of 11-dehydro-TXB
2
and 2,3-dinor-6-keto-PGF
1α
, metabolites of TXA
2
and PGI
2
, respectively, were elevated in patients with atherosclerosis compared with normal subjects (3211±533 versus 679±63 pg/mg creatinine,
P
<0.001; 594±156 versus 130±22 pg/mg creatinine,
P
<0.05, respectively), as was the level of the isoprostane 8-iso-PGF
2α
. Nimesulide reduced 2,3-dinor-6-keto-PGF
1α
excretion by 46±5% (378.3±103 to 167±37 pg/mg creatinine,
P
<0.01) preoperatively and blunted the increase after surgery. Nimesulide had no significant effect on 11-dehydro-TXB
2
before (2678±694 to 2110±282 pg/mg creatinine) or after surgery. The levels of both products were lower in patients who were taking aspirin, and no further reduction was seen with the addition of nimesulide. None of the treatments influenced urinary 8-iso-PGF
2α
excretion.

Conclusions
—Both COX-1 and -2 are expressed and contribute to the increase in PGI
2
in patients with atherosclerosis, whereas TXA
2
is generated by COX-1.
Perinatal Changes in Myocardial Metabolism in Lambs
Authors: Bartelds Beatrijs , Knoester Hennie , Smid Gioia B. , Takens Janny , Visser G. Henk , Penninga Luit , van der Leij Feike R. , Beaufort-Krol Gertie C. M. , Zijlstra Willem G. , Heymans Hugo S. A. , Kuipers Jaap R. G.
Published: June 2012

Background
—Lactate accounts for a third of myocardial oxygen consumption before and in the first 2 weeks after birth. It is unknown how the remainder of myocardial oxygen is consumed. Glucose is thought to be important before birth, whereas long-chain fatty acids (LC-FA) are the prime substrate for the adult. However, the ability of the myocardium of the newborn to use LC-FA has been doubted.

Methods and Results
—We measured the myocardial metabolism of glucose and LC-FA with [U-
13
C]glucose and [1-
13
C]palmitate in chronically instrumented fetal and newborn lambs. In fetal lambs, myocardial oxidation of glucose was high and that of LC-FA was low. Glucose and LC-FA accounted for 48±4% and 2±2% of myocardial oxygen consumption, respectively. In newborn lambs, oxidation of glucose decreased, whereas oxidation of LC-FA increased. Glucose and LC-FA accounted for 12±3% and 83±19% of myocardial oxygen consumption. To test whether near-term fetal lambs could use LC-FA, we increased the supply of LC-FA with a fat infusion. In fetal lambs during fat infusion, the oxidation of LC-FA increased 15-fold. Although the oxidation of LC-FA was still lower than in newborn lambs, the contribution to myocardial oxygen consumption (70±13%) was the same as in newborn lambs.

Conclusions
—These data show that glucose and lactate account for the majority of myocardial oxygen consumption in fetal lambs, whereas in newborn lambs, LC-FA and lactate account for the majority of myocardial oxygen consumption. Moreover, we showed that the fetal myocardium can use LC-FA as an energy substrate.
Red Wine Intake Prevents Nuclear Factor-κB Activation in Peripheral Blood Mononuclear Cells of Healthy Volunteers During Postprandial Lipemia
Authors: Blanco-Colio Luis Miguel , Valderrama Mónica , Alvarez-Sala Luis Antonio , Bustos Carmen , Ortego Mónica , Hernández-Presa Miguel Angel , Cancelas Pilar , Gómez-Gerique Juan , Millán Jesús , Egido Jesús
Published: June 2012

Background
—Several epidemiological studies have demonstrated the beneficial effect of red wine intake in reducing total and cardiovascular mortality. This effect has been attributed in part to its antioxidant properties. Because the monocytes/macrophages and the nuclear transcription factor κB (NF-κB) are implicated in the pathogenesis of atherosclerotic lesions, we examined the effect of red wine intake on the activation of NF-κB in peripheral blood mononuclear cells.

Methods and Results
—Sixteen healthy volunteers were studied 3 times each: after a moderate dose, a low dose, and no wine with a fat-enriched breakfast. Lipid profile and NF-κB activation (electrophoretic mobility shift assay) were examined in blood samples taken before and 3, 6, and 9 hours after wine intake. In addition, mononuclear cells were incubated with VLDL in the presence of some antioxidants (quercetin and α-tocopherol succinate) contained in red wine to study their effects on NF-κB activation. Subjects receiving a fat-enriched breakfast had increased NF-κB activation in peripheral blood mononuclear cells coinciding with the augmentation in total triglycerides and chylomicrons. Red wine intake prevented NF-κB activity even though it induced a certain increase in serum lipids, particularly VLDL, that did not increase after the fat ingestion alone. However, another form of alcohol intake (vodka) did not modify the NF-κB activation provided by postprandial lipemia. In cultured mononuclear cells, isolated human VLDL caused NF-κB activation in a time-dependent manner that did not occur in the presence of the red wine antioxidants quercetin and α-tocopherol.

Conclusions
—Our results provide a new potential mechanism to explain the beneficial effects of red wine intake in the reduction of cardiovascular mortality.
Uptake of Radiolabeled 2′-Fluoro-2′-Deoxy-5-Iodo-1-β-
d
-Arabinofuranosyluracil in Cardiac Cells After Adenoviral Transfer of the Herpesvirus Thymidine Kinase Gene
Authors: Bengel Frank M. , Anton Martina , Avril Norbert , Brill Thomas , Nguyen Ngoc , Haubner Roland , Gleiter Elisabeth , Gansbacher Bernd , Schwaiger Markus
Published: June 2012

Background
—Gene therapy is a promising approach for the treatment of cardiac diseases. Coexpression of therapeutic genes with a suitable marker gene would allow for the noninvasive imaging of successful gene transfer and expression via radiolabeled marker substrates. In the present study, such an approach was first applied to cardiac tissue.

Methods and Results
—The combination of the herpesvirus thymidine kinase reporter gene (HSV1-tk) and radiolabeled 2′-fluoro-2′-deoxy-5-iodo-1-β-
d
-arabinofuranosyluracil (FIAU) was evaluated. H9c2 rat cardiomyoblasts were infected in vitro with a replication-defective HSV1-tk–containing adenovirus and a negative control virus. The intracellular uptake of [
14
C]FIAU increased with increasing multiplicity of infection and with time after infection. Uptake in negative controls remained <15% of positive controls. Additionally, vectors were applied intramyocardially in Wistar rats. The marker substrate [
125
I]FIAU was injected intravenously 3 days later, and animals were killed after 24 hours. Autoradiographically, regional transgene expression was clearly identified in animals receiving the adenovirus containing HSV1-tk (3.4±2.2-fold increase of radioactivity at vector administration site compared with remote myocardium), whereas nonspecific uptake in negative controls was low (<10% of positive controls).

Conclusions
—Using an adenoviral vector, HSV1-tk can be successfully expressed in cardiac cells in vitro and in vivo, yielding high uptake of radiolabeled FIAU. The results suggest that imaging transgene expression in the heart is feasible and may be used to monitor gene therapy noninvasively.
Physical Activity and Coronary Heart Disease in Men
Authors: Sesso Howard D. , Paffenbarger Ralph S. , Lee I-Min
Published: June 2012

Background
—The quantity and intensity of physical activity required for the primary prevention of coronary heart disease (CHD) remain unclear. Therefore, we examined the association of the quantity and intensity of physical activity with CHD risk and the impact of other coronary risk factors.

Methods and Results
—We followed 12 516 middle-aged and older men (mean age 57.7 years, range 39 to 88 years) from 1977 through 1993. Physical activity was assessed at baseline in kilojoules per week (4.2 kJ=1 kcal) from blocks walked, flights climbed, and participation in sports or recreational activities. During follow-up, 2135 cases of incident CHD, including myocardial infarction, angina pectoris, revascularization, and coronary death, occurred. Compared with men expending <2100 kJ/wk, men expending 2100 to 4199, 4200 to 8399, 8400 to 12 599, and ≥12 600 kJ/wk had multivariate relative risks of 0.90, 0.81, 0.80, and 0.81, respectively (
P
for trend=0.003). When we considered the independent effects of specific physical activity components, only total sports or recreational activities (
P
for trend=0.042) and vigorous activities (
P
for trend=0.02) were inversely associated with the risk of CHD. These associations did not differ within subgroups of men defined by coronary risk factors. Finally, among men with multiple coronary risk factors, those expending ≥4200 kJ/wk had reduced CHD risk compared with men expending <4200 kJ/wk.

Conclusions
—Total physical activity and vigorous activities showed the strongest reductions in CHD risk. Moderate and light activities, which may be less precisely measured, showed nonsignificant inverse associations. The association between physical activity and a reduced risk of CHD also extends to men with multiple coronary risk factors.
Effect of Type 2 Diabetes Mellitus on Left Ventricular Geometry and Systolic Function in Hypertensive Subjects
Authors: Palmieri Vittorio , Bella Jonathan N. , Arnett Donna K. , Liu Jennifer E. , Oberman Albert , Schuck Min-Yan , Kitzman Dalane W. , Hopkins Paul N. , Morgan Derek , Rao D. C. , Devereux Richard B.
Published: June 2012

Background
—Type 2 diabetes is a cardiovascular risk factor. It remains to be elucidated in a large, population-based sample whether diabetes is associated with changes in left ventricular (LV) structure and systolic function independent of obesity and systolic blood pressure (BP).

Methods and Results
—Among 1950 hypertensive participants in the HyperGEN Study without overt coronary heart disease or significant valve disease, 20% (n=386) had diabetes. Diabetics were more likely to be women, black, older, and have higher BMI and waist/hip ratio than were nondiabetics. After adjustment for age and sex, diabetics had higher systolic BP, pulse pressure, and heart rate; lower diastolic BP; and longer duration of hypertension than nondiabetics. LV mass and relative wall thickness were higher in diabetic than nondiabetic subjects independent of covariates. Compared with nondiabetic hypertensives, diabetics had lower stress-corrected midwall shortening, independent of covariates, without difference in LV EF. Insulin levels and insulin resistance were higher in non–insulin-treated diabetics (n=195) than nondiabetic (n=1439) subjects (both
P
<0.01). Insulin resistance positively but weakly related to LV mass and relative wall thickness.

Conclusions
—In a relatively healthy, population-based sample of hypertensive adults, type 2 diabetes was associated with higher LV mass, more concentric LV geometry, and lower myocardial function, independent of age, sex, body size, and arterial BP. structural and functional abnormalities in addition to, and independent of, atherosclerosis.

13

14

In the Framingham cohort, diabetes was associated with higher LV mass in women but not men.

15

High blood pressure (BP), obesity, and abnormal lipid profile, which often coexist with diabetes, tend to be associated with preclinical cardiovascular abnormalities

16

and may contribute to the association of diabetes with cardiovascular events. Cardiac features of diabetic and nondiabetic hypertensive subjects remain incompletely described in population-based samples. Therefore, we compared clinical and metabolic characteristics, LV geometry, and systolic function between diabetic and nondiabetic hypertensive participants in the Hypertension Genetic Epidemiology Network (HyperGEN) Study.
Nerve Sprouting and Sympathetic Hyperinnervation in a Canine Model of Atrial Fibrillation Produced by Prolonged Right Atrial Pacing
Authors: Chang Che-Ming , Wu Tsu-Juey , Zhou Shengmei , Doshi Rahul N. , Lee Moon-Hyoung , Ohara Toshihiko , Fishbein Michael C. , Karagueuzian Hrayr S. , Chen Peng-Sheng , Chen Lan S.
Published: June 2012

Background
—Long-term rapid atrial pacing may result in atrial fibrillation (AF) in dogs. Whether there is histological evidence for neural remodeling is unclear.

Method and Results
—We performed rapid right atrial pacing in 6 dogs for 111±76 days to induce sustained AF. Tissues from 6 healthy dogs were used as controls. Immunocytochemical staining of cardiac nerves was performed using anti–growth-associated protein 43 (GAP43) and anti–tyrosine hydroxylase (TH) antibodies. In dogs with AF, the density of GAP43-positive and TH-positive nerves in the right atrium was 470±406 and 231±126 per mm
2
, respectively, which was significantly (
P
<0.001) higher than the nerve density in control tissues (25±32 and 88±40 per mm
2
, respectively). The density of GAP43-positive and TH-positive nerves in the atrial septum was 317±36 and 155±85 per mm
2
, respectively, and was significantly (
P
<0.001) higher than the nerve density in control tissues (9±13 and 30±7 per mm
2
, respectively). Similarly, the density of GAP43-positive and TH-positive nerves in the left atrium of dogs with AF was 119±61 and 91±40 per mm
2
, respectively, which was significantly (
P
<0.001) higher than the nerve density in control tissues (10±15 and 38±39 per mm
2
, respectively). Furthermore, in dogs with AF, the right atrium had a significantly higher nerve density than the left atrium. Microscopic examinations revealed an inhomogeneous distribution of cardiac nerves within each sampling site.

Conclusions
—Significant nerve sprouting and sympathetic hyperinnervation are present in a canine model of sustained AF produced by prolonged right atrial pacing. The magnitude of nerve sprouting and hyperinnervation was higher in the right atrium than in the left atrium.
Women’s Healthy Lifestyle Project: A Randomized Clinical Trial
Authors: Kuller Lewis H. , Simkin-Silverman Laurey R. , Wing Rena R. , Meilahn Elaine N. , Ives Diane G.
Published: June 2012

Background
—The Women’s Healthy Lifestyle Project Clinical Trial tested the hypothesis that reducing saturated fat and cholesterol consumption and preventing weight gain by decreased caloric and fat intake and increased physical activity would prevent the rise in LDL cholesterol and weight gain in women during perimenopause to postmenopause.

Methods and Results
—There were 275 premenopausal women randomized into the assessment only group and 260 women into the intervention group. The mean age of participants at baseline was 47 years, and 92% of the women were white. The mean LDL cholesterol was 115 mg/dL at baseline, and mean body mass index was 25 kg/m
2
. The follow-up through 54 months was excellent. By 54 months, 35% of the women had become postmenopausal. At the 54-month examination, there was a 3.5-mg/dL increase in LDL cholesterol in the intervention group and an 8.9-mg/dL increase in the assessment-only group (
P
=0.009). Weight decreased 0.2 lb in the intervention and increased 5.2 lb in the assessment-only group (
P
=0.000). Triglycerides and glucose also increased significantly more in the assessment-only group than in the intervention group. Waist circumference decreased 2.9 cm in the intervention compared with 0.5 cm in the assessment-only group (
P
=0.000).

Conclusions
—The trial was successful in reducing the rise in LDL cholesterol during perimenopause to postmenopause but could not completely eliminate the rise in LDL cholesterol. The trial was also successful in preventing the increase in weight from premenopause to perimenopause to postmenopause. The difference in LDL cholesterol between the assessment and intervention groups was most pronounced among postmenopausal women and occurred among hormone users and nonusers.
Use of Lipid-Lowering Medications at Discharge in Patients With Acute Myocardial Infarction
Authors: Fonarow Gregg C. , French William J. , Parsons Lori S. , Sun Haili , Malmgren Judith A.
Published: June 2012

Background
—The present study aimed to assess use of lipid-lowering medication at discharge in a current national sample of patients hospitalized with acute myocardial infarction and to evaluate factors associated with prescribing patterns.

Methods and Results
—Demographic, procedural, and discharge medication data were collected from 138 001 patients with acute myocardial infarction discharged from 1470 US hospitals participating in the National Registry of Myocardial Infarction 3 from July 1998 to June 1999. Lipid-lowering medications were part of the discharge regimen in 31.7%. Among patients with prior history of CAD, revascularization, or diabetes, less than one half of the patients were discharged on treatment. In multivariate analysis, factors independently related to lipid-lowering use included history of hypercholesterolemia (odds ratio [OR] 4.93; 95% CI 4.79 to 5.07), cardiac catheterization during hospitalization (OR 1.29; 95% CI 1.24 to 1.34), care provided at a teaching hospital, (OR 1.26; 95% CI 1.22 to 1.32), use of β-blocker (OR 1.43; 95% CI 1.39 to 1.48), and smoking cessation counseling (OR 1.51; 95% CI 1.44 to 1.59). Lipid-lowering medications were given less often to patients who were older (65 to 74 versus <55 years of age; OR 0.82; 95% CI 0.78 to 0.86), those with a history of hypertension (OR 0.92; 95% CI 0.89 to 0.95), and those undergoing coronary artery bypass graft surgery (OR 0.58; 95% CI 0.55 to 0.60).

Conclusions
—Analysis of current practice patterns for the use of lipid-lowering medications in patients hospitalized with acute myocardial infarction reveals that a significant proportion of high-risk patients did not receive treatment at time of discharge.
Prospective Study of Pathogen Burden and Risk of Myocardial Infarction or Death
Authors: Zhu Jianhui , Nieto F. Javier , Horne Benjamin D. , Anderson Jeffrey L. , Muhlestein Joseph B. , Epstein Stephen E.
Published: June 2012

Background
—We previously demonstrated that the risk of coronary artery disease (CAD) increased in relation to the number of pathogens (the “pathogen burden”) in a cross-sectional study. In the present prospective study with a different patient cohort, we evaluated the effect of pathogen burden on the risk of myocardial infarction (MI) or death among CAD patients.

Methods and Results
—IgG antibodies to cytomegalovirus (CMV), hepatitis A virus (HAV), herpes simplex virus type 1 (HSV1), HSV type 2 (HSV2),
Chlamydia pneumoniae
and
Helicobacter pylori
, and C-reactive protein (CRP) levels were tested in baseline blood samples from 890 patients who had significant CAD on angiography. The mean follow-up period was 3 years. The baseline prevalence of antibodies directed against CMV, HAV, HSV1, or HSV2, but not
C pneumoniae
and
H pylori,
was significantly higher among patients who subsequently developed MI or death than among control subjects. After adjustment for traditional risk factors, number of diseased vessels, and clinical presentation, relative hazards (95% confidence limits) for MI or death were 2.0 (1.4 to 3.2) for CMV, 1.6 (1.1 to 2.3) for HAV, and 1.5 (1.0 to 2.2) for HSV2. Increasing pathogen burden was significantly associated with increasing risk of MI or death in a dose-response fashion. Adjusted relative hazards of MI or death associated with pathogen burden were significant among individuals with low or high CRP levels.

Conclusions
—The results suggest that infection plays an important role in incident MI or death and that the risk posed by infection is independently related to the pathogen burden.
Cardiac Troponin I Is Modified in the Myocardium of Bypass Patients
Authors: McDonough J. L. , Labugger R. , Pickett W. , Tse M. Y. , MacKenzie S. , Pang S. C. , Atar D. , Ropchan G. , Van Eyk J. E.
Published: June 2012

Background
—Selective proteolysis of cardiac troponin I (cTnI) is a proposed mechanism of contractile dysfunction in stunned myocardium, and the presence of cTnI degradation products in serum may reflect the functional state of the remaining viable myocardium. However, recent swine and canine studies have not demonstrated stunning-dependent cTnI degradation.

Methods and Results
—To address the universality of cTnI modification, myocardial biopsy samples were obtained from coronary artery bypass patients (n=37) before and 10 minutes after removal of cross-clamp. Analysis of biopsy samples for cTnI by Western blotting revealed a spectrum of modified cTnI products in myocardium both before and after cross-clamp, including degradation products (7 products resulting from differential N- and C-terminal processing) and covalent complexes (3 products). In particular, a 22-kDa cTnI degradation product with C-terminal proteolysis was identified, which may represent an initial ischemia-dependent cTnI modification, similar to cTnI
1–193
observed in stunned rat myocardium. Although no systematic change in amount of modified cTnI was observed, subgroups of patients displayed an increase (n=10, 85±5% of cTnI remaining intact before cross-clamp versus 75±5% after) or a decrease (n=12, 67±5% before versus 78±5% after). Electron microscopy demonstrated normal ultrastructure in biopsy samples, which suggests no necrosis was present. In addition, cTnI modification products were observed in serum through a modified SDS-PAGE methodology.

Conclusions
—cTnI modification, in particular proteolysis, occurs in myocardium of bypass patients and may play a key role in stunning in some bypass patients.
Evidence for Joint Genetic Control of Insulin Sensitivity and Systolic Blood Pressure in Hispanic Families With a Hypertensive Proband
Authors: Xiang Anny H. , Azen Stanley P. , Raffel Leslie J. , Tan Sylvia , Cheng Li Shu-Chuan , Diaz Justo , Toscano Edgar , Henderson Paula C. , Hodis Howard N. , Hsueh Willa A. , Rotter Jerome I. , Buchanan Thomas A.
Published: June 2012

Background
—The clustering of hypertension, insulin resistance, and obesity remains unexplained. We tested for genetic and nongenetic influences on the association among these traits in Hispanic families with hypertension.

Methods and Results
—Blood pressure and body mass index (BMI) were measured in 331 members of 73 Hispanic families in which an index case (proband) had hypertension. Insulin sensitivity (S
I
) was measured by euglycemic clamp in 287 probands and their spouses (parents’ generation) or their adult offspring. Correlation analysis examined relationships among traits within and between generations. Path analysis estimated genetic and nongenetic contributions to variability in systolic blood pressure (SBP), S
I
, and the correlation between them. In the offspring, there was a significant correlation between individuals for each trait, as well as significant correlations within and between individuals for all possible pairs of traits. Between generations, SBP, S
I
, and BMI in parents correlated with the same traits in their offspring; BMI in parents correlated with S
I
and SBP in offspring; and S
I
in parents correlated with SBP in offspring. Path analysis estimated that among offspring, genetic effects unrelated to BMI accounted for 60.8% of the variation in SBP, 36.8% of the variation in S
I
, and 31.5% of the correlation between SBP and S
I
after adjustment for age and sex. Heritable effects related to BMI accounted for an additional 14.0% of variation in SBP, 26.8% of variation in S
I
, and 56.3% of variation in their correlation.

Conclusions
—Clustering of hypertension and insulin resistance in Hispanic Americans is accounted for in part by heritable factors both associated with and independent of BMI.
Gene Expression Profiling of Human Stent-Induced Neointima by cDNA Array Analysis of Microscopic Specimens Retrieved by Helix Cutter Atherectomy
Authors: Zohlnhöfer Dietlind , Klein Christoph A. , Richter Thomas , Brandl Richard , Murr Alexander , Nührenberg Thomas , Schömig Albert , Baeuerle Patrick A. , Neumann Franz-Josef
Published: June 2012

Background
—Restenosis due to neointima formation is the major limitation of stent-supported balloon angioplasty. Despite abundant animal data, molecular mechanisms of neointima formation have been investigated on only a limited basis in patients. This study sought to establish a method for profiling gene expression in human in-stent neointima and to identify differentially expressed genes that may serve as novel therapeutic targets.

Methods and Results
—We retrieved tissue specimens from patients with symptomatic in-stent restenosis using a novel helix cutter atherectomy device. cDNA samples prepared from neointima (n=10) and, as a control, from the media of normal arteries (n=14) were amplified using a novel polymerase chain reaction protocol and hybridized to cDNA arrays. Immunohistochemistry characterized the atherectomy material as neointima. cDNA arrays readily identified differentially expressed genes. Some of the differentially expressed genes complied with expected gene expression patterns of neointima, including downregulation of desmin and upregulation of thrombospondin-1, cyclooxygenase-1, and the 70-kDa heat shock protein B. Additionally, we discovered previously unknown gene expression patterns, such as downregulation of mammary-derived growth inhibitor and upregulation of FK506-binding protein 12 (FKBP12). Upregulation of FKBP12 was confirmed at the protein level in neointimal smooth muscle cells.

Conclusions
—Gene expression patterns of human neointima retrieved by helix-cutter atherectomy can be reliably analyzed by cDNA array technology. This technique can identify therapeutic targets in patients, as exemplified by the findings regarding FKBP12. FKBP12 is the receptor for Rapamycin (sirolimus), which in animal models reduced neointima formation. Our study thus yields a rationale for the use of Rapamycin to prevent restenosis in patients.
Polycystic Ovary Syndrome Is Associated With Endothelial Dysfunction
Authors: Paradisi Giancarlo , Steinberg Helmut O. , Hempfling Annette , Cronin Jessica , Hook Ginger , Shepard Marguerite K. , Baron Alain D.
Published: June 2012

Background
—We recently reported endothelial dysfunction as a novel cardiovascular risk factor associated with insulin resistance/obesity. Here, we tested whether hyperandrogenic insulin-resistant women with polycystic ovary syndrome (PCOS) who are at increased risk of macrovascular disease display impaired endothelium-dependent vasodilation and whether endothelial function in PCOS is associated with particular metabolic and/or hormonal characteristics.

Methods and Results
—We studied leg blood flow (LBF) responses to graded intrafemoral artery infusions of the endothelium-dependent vasodilator methacholine chloride (MCh) and to euglycemic hyperinsulinemia in 12 obese women with PCOS and in 13 healthy age- and weight-matched control subjects (OBW). LBF increments in response to MCh were 50% lower in the PCOS group than in the OBW group (
P
<0.01). Euglycemic hyperinsulinemia increased LBF above baseline by 30% in the PCOS and 60% in OBW group (
P
<0.05 between groups). Across all subjects, the maximal LBF response to MCh exhibited a strong inverse correlation with free testosterone levels (
r
=−0.52,
P
<0.007). This relationship was stronger than with any other parameter, including insulin sensitivity.

Conclusions
—PCOS is characterized by (1) endothelial dysfunction and (2) resistance to the vasodilating action of insulin. This endothelial dysfunction appears to be associated with both elevated androgen levels and insulin resistance. Given the central vasoprotective role of endothelium, these findings could explain, at least in part, the increased risk for macrovascular disease in women with PCOS.
Testing Platelet Activation With a Shear-Dependent Platelet Function Test Versus Aggregation-Based Tests
Authors: Osende Julio I. , Fuster Valentin , Lev Eli I. , Shimbo Daichi , Rauch Ursula , Marmur Jonathan D. , Richard Merwin , Varon David , Badimon Juan Jose
Published: June 2012

Background
—Tests developed to monitor glycoprotein (GP) IIb/IIIa blockade do not properly reflect platelet function in vivo and need a baseline (pretreatment) value. Because GP IIb/IIIa is essential in platelet aggregation and thrombosis under shear conditions, a flow-dependent approach to monitor its inhibition can be used.

Methods and Results
—We compared a test based on flow-dependent platelet deposition, the Cone and Platelet Analyzer (CPA), with in vitro platelet aggregometry and the Rapid Platelet Function Assay (RPFA) on platelet function after GP IIb/IIIa inhibition. In vitro, increasing concentrations of abciximab (0% to 100% receptor occupancy) were tested. Ex vivo, platelet function was monitored with the CPA and with aggregometry for up to 1 week after abciximab administration. The CPA was better correlated with the percentage of free GP IIb/IIIa receptors than was aggregometry or the RPFA. Only the RPFA, when expressed as a ratio over baseline (pretreatment), was comparable to the CPA. Ex vivo, the CPA, but not aggregometry, showed prolonged platelet inhibition with gradual recovery from GP IIb/IIIa receptor blockade in the first week after abciximab administration.

Conclusions
—Platelet function assessment by shear-induced deposition is a reliable test to monitor a wide range of GP IIb/IIIa inhibition. Its accuracy does not require a baseline reference. The effects of GP IIb/IIIa blockade on platelet function should be examined under high shear conditions.
Independent and Joint Effects of Antibodies to Human Heat-Shock Protein 60 and
Chlamydia pneumoniae
Infection in the Development of Coronary Atherosclerosis
Authors: Burian Katalin , Kis Zoltan , Virok Dezso , Endresz Valeria , Prohaszka Zoltan , Duba Jeno , Berencsi Klara , Boda Krisztina , Horvath Laura , Romics Laszlo , Fust George , Gonczol Eva
Published: June 2012

Background
—Studies have suggested that the prevalence of antibodies against heat-shock proteins (HSPs),
Chlamydia pneumoniae
(
Cpn
), and cytomegalovirus (CMV) is associated with coronary artery disease (CAD), but the independent or joint effects of human (h) HSP60 antibodies and these pathogens in patients have not been fully elucidated.

Methods and Results
—A total of 405 subjects (276 patients with CAD and 129 control individuals) were tested for serum antibodies to hHSP60,
Cpn
, and CMV immediate-early-1 (IE1) antigens. Patients were also assessed for serum cholesterol, triglyceride levels, and smoking habit. Significantly elevated levels of antibodies to hHSP60 and
Cpn
but not to CMV-IE1 antigens were documented in CAD patients. Multiple logistic regression analysis and subanalyses of selected subjects showed that these associations were independent of age, sex, smoking, and serum lipid levels. Antibodies to hHSP60 and
Cpn
did not correlate quantitatively; however, the relative risk of disease development was substantially increased in subjects with high antibody levels to both hHSP60 and
Cpn
, reaching an odds ratio of 82.0 (95% CI 10.6 to 625.0).

Conclusions
—High levels of antibodies to hHSP60 and
Cpn
are independent risk factors for coronary atherosclerosis, but their simultaneous presence substantially increases the risk for disease development.
Abdominal Aortic Calcific Deposits Are an Important Predictor of Vascular Morbidity and Mortality
Authors: Wilson Peter W. F. , Kauppila Leena I. , O’Donnell Christopher J. , Kiel Douglas P. , Hannan Marian , Polak Joseph M. , Cupples L. Adrienne
Published: June 2012

Background
—The impact of abdominal arterial calcific deposits on the prediction of cardiovascular disease (CVD) over a long follow-up interval deserves greater scrutiny.

Methods and Results
—Lateral lumbar radiographs were studied as a predictor of incident coronary heart disease (CHD), CVD, and CVD mortality in 1049 men and 1466 women (mean age, 61 years) who were followed from 1967 to 1989. Anterior and posterior wall calcific deposits in the aorta at the level of the first through fourth lumbar vertebrae were graded according to increasing severity using a previously validated rating scale for abdominal aortic calcium (AAC) that ranges from 0 to 24 points. There were 454 cases of CHD, 709 cases of CVD, and 365 CVD deaths. Proportional hazards logistic regression was used to test for associations between AAC and later events after adjustment for age, cigarette use, diabetes mellitus, systolic blood pressure, left ventricular hypertrophy, body mass index, cholesterol, and HDL cholesterol. In comparisons with the lowest AAC tertile, the multivariate age-adjusted relative risks (RR) for CVD were increased in tertile 2 (men: RR, 1.33; 95% confidence interval [CI], 1.02 to 1.74; women: RR, 1.25; 95% CI, 0.95 to 1.65) and tertile 3 (men: RR, 1.68; 95% CI, 1.25 to 2.27; women: RR, 1.78; 95% CI, 1.33 to 2.38). Similar results were obtained with CHD and CVD mortality.

Conclusions
—AAC deposits, detected by lateral lumbar radiograms, are a marker of subclinical atherosclerotic disease and an independent predictor of subsequent vascular morbidity and mortality.
Isolated Mitral Valve Replacement With St Jude Medical Prosthesis
Authors: Remadi J. P. , Baron O. , Roussel C. , Bizouarn P. , Habasch Al , Despins P. , Michaud J. L. , Duveau D.
Published: June 2012

Background
—In this retrospective study, ≈440 patients received mitral valve replacements with the St Jude Medical prothesis. The last patient was operated on 10 years before the beginning of the follow-up. The extended follow-up was 19 years.

Methods and Results
—Four hundred forty patients (sex ratio, 1.32 [men to women]; age, 60±11.4 years; age range, 7 to 75 years) were operated on from 1979 to 1987. All patients underwent isolated mitral valve replacement. Tricuspid plasty was the only associated procedure. The follow-up at 19 years was 98% complete. The overall actuarial survival rate was 63±3.3% at 19 years, and the actuarial survival rate (only valve related) was 83±2.7%. The operative mortality rate (0 to 30 days) was 4.09%. We found that 89.4% of the patients alive at 19 years were in NYHA class I/II. Multivariate analysis showed that age and sex were significantly correlated with valve-related mortality and that age, sex, NYHA class, and atrial fibrillation were significantly correlated with overall mortality. The linearized rates (percent patient-years) of thromboembolism, thrombosis, and hemorrhage were 0.69, 0.2, and 1, respectively. At 19 years, freedom from endocarditis and reoperation was 98.6±1% and 90±3%, respectively.

Conclusions
—In this study, the very-long-term results confirm the excellent durability of the St Jude Medical prosthesis in the mitral position and show the difficulty of adjusting the anticoagulation protocol, even after long-term treatment.
Effects of Nonlipid Risk Factors on Atherosclerosis in Youth With a Favorable Lipoprotein Profile
Authors: McGill Henry C. , McMahan C. Alex , Zieske Arthur W. , Malcom Gray T. , Tracy Richard E. , Strong Jack P.
Published: June 2012

Background
—The strong association between coronary heart disease and dyslipoproteinemia has often overshadowed the effects of the nonlipid risk factors–smoking, hypertension, obesity, and diabetes and impaired glucose tolerance–and even led to questioning the importance of these risk factors in the presence of a favorable lipoprotein profile.

Methods and Results
—A cooperative multicenter study, the Pathobiological Determinants of Atherosclerosis in Youth (PDAY), examined the relation of the nonlipid risk factors to atherosclerosis in 629 men and 227 women 15 to 34 years of age who died of external causes and who had a favorable lipoprotein profile (non-HDL cholesterol <4.14 mmol/L [<160 mg/dL] and HDL cholesterol ≥0.91 mmol/L [≥35 mg/dL]). In the abdominal aorta, smokers had more extensive fatty streaks and raised lesions than nonsmokers, and hypertensive blacks had more raised lesions than normotensive blacks. In the right coronary artery, hypertensive blacks had more raised lesions than normotensive blacks, obese men (body mass index ≥30 kg/m
2
) had more extensive fatty streaks and raised lesions than nonobese men, and individuals with impaired glucose intolerance had more extensive fatty streaks. Obese men had more severe lesions (American Heart Association grade 2 through 5) of the left anterior descending coronary artery.

Conclusions
—These substantial effects of the nonlipid risk factors on the extent and severity of coronary and aortic atherosclerosis, even in the presence of a favorable lipoprotein profile, support the need to control all cardiovascular risk factors.
Impaired Conduction in the Bundle Branches of Mouse Hearts Lacking the Gap Junction Protein Connexin40
Authors: van Rijen Harold V. M. , van Veen Toon A. B. , van Kempen Marjan J. A. , Wilms-Schopman Francien J. G. , Potse Mark , Krueger Olaf , Willecke Klaus , Opthof Tobias , Jongsma Habo J. , de Bakker Jacques M. T.
Published: June 2012

Background
—Connexin (Cx)40 and Cx45 are the major protein subunits of gap junction channels in the conduction system of mammals. To determine the role of Cx40, we correlated cardiac activation with
C
onnexin distribution in normal and Cx40-deficient mice hearts.

Methods and Results
—Epicardial and septal activation was recorded in Langendorff-perfused adult mice hearts with a 247-point compound electrode (interelectrode distance, 0.3 mm). After electrophysiological measurements, hearts were prepared for immunohistochemistry and histology to determine
C
onnexin distribution and fibrosis. In both wild-type and Cx40-deficient animals, epicardial activation patterns were similar. The right and left ventricular septum was invariably activated from base to apex. Histology revealed a continuity of myocytes from the common bundle to the septal myocardium. Within this continuity, colocalization was found of Cx43 and Cx45 but not of Cx40 and Cx43. Both animals showed similar His-bundle activation. In Cx40-deficient mice, the proximal bundle branches expressed Cx45 only. The absence of Cx40 in the proximal bundles correlated with right bundle-branch block. Conduction in the left bundle branch was impaired as compared with wild-type animals.

Conclusions
—Our data show that (1) in mice, a continuity exists between the common bundle and the septum, and (2) Cx40 deficiency results in right bundle-branch block and impaired left bundle-branch conduction.
Distribution of
Chlamydia pneumoniae
in the Human Arterial System and Its Relation to the Local Amount of Atherosclerosis Within the Individual
Authors: Vink Aryan , Poppen Marieke , Schoneveld Arjan H. , Roholl Paul J. M. , de Kleijn Dominique P. V. , Borst Cornelius , Pasterkamp Gerard
Published: June 2012

Background
—
Chlamydia pneumoniae
has been suggested to play a role in the origin of atherosclerosis. We studied the prevalence of
C pneumoniae
at multiple locations in the arterial system within the same individual. Studying the association between atherosclerosis and
C pneumoniae
within the individual excludes confounding by interindividual variability.

Methods and Results
—Postmortem, the presence in the intima/plaque and media of
C pneumoniae
membrane protein was determined by use of a
C pneumoniae
–specific monoclonal antibody. In 24 individuals, 33 arterial locations were studied (n=738 segments). Area stenosis was determined in adjacent cross sections. In all individuals, immunostaining of
C pneumoniae
was observed in ≥1 artery. The highest prevalences were observed in the abdominal aorta (67%), internal and common iliac arteries (41%), and coronary arteries (33%). The lowest prevalences were observed in the radial (0%) and cerebral (2%) arteries. Within the individual, area stenosis was larger in cross sections with immunoreactivity compared with cross sections without immunoreactivity (31.0±11.9% versus 14.3±6.1%, respectively;
P
<0.001). In the individual, immunoreactivity was observed in 15±10% of the arteries (range, 3% to 45%). Between individuals, the percentage of arteries with immunoreactivity to
C pneumoniae
was associated with the average area stenosis throughout the arterial system (
r
2
=0.56,
P
<0.001).

Conclusions
—
C pneumoniae
was mostly observed at locations that are related to clinically relevant features. Within the individual, the distribution of
C pneumoniae
is associated with the distribution of atherosclerosis. The role of the microorganism in atherosclerotic disease remains to be elucidated.
Pharmacogenetic Interactions Between β-Blocker Therapy and the Angiotensin-Converting Enzyme Deletion Polymorphism in Patients With Congestive Heart Failure
Authors: McNamara Dennis M. , Holubkov Richard , Janosko Karen , Palmer Amy , Wang Jue J. , MacGowan Guy A. , Murali Srinivas , Rosenblum Warren D. , London Barry , Feldman Arthur M.
Published: June 2012

Background
—Activation of the renin-angiotensin and sympathetic nervous systems adversely affect heart failure progression. The
ACE
deletion allele (
ACE D
) is associated with increased renin-angiotensin activation; however, its influence on patient outcomes remains uncertain, and the pharmacogenetic interactions with β-blocker therapy have not been previously evaluated.

Methods and Results
—We prospectively followed 328 patients (age, 56.1±11.9 years) with systolic dysfunction (left ventricular ejection fraction, 0.24±0.08) to assess the impact of the
ACE D
allele on transplant-free survival (median follow-up, 21 months). Transplant-free survival was compared by genotype for the whole cohort and separately in patients with (n=120) and those without β-blocker therapy (n=208) at the time of entry. Transplant-free survival was significantly poorer for patients with the
D
allele (1-year percent survival
II/ID/DD
=94/77/75; 2-year=78/65/60; ordered log-rank test,
P
=0.044). In patients not treated with β-blockers, the adverse impact of
ACE D
allele was dramatically increased (1-year percent survival
II/ID/DD
=95/75/67; 2-year=81/61/48;
P
=0.005). In contrast, in patients receiving β-blocker therapy, no influence of
ACE
genotype on transplant-free survival was evident (1-year percent survival
II/ID/DD
=91/80/86; 2-year=70/71/77;
P
=0.73).

Conclusions
—In a cohort of patients with systolic dysfunction, the
ACE D
allele was associated with a significantly poorer transplant-free survival. This effect was primarily evident in patients not treated with β-blockers and was not seen in patients receiving therapy. These findings suggest a potential pharmacogenetic interaction between the
ACE D/I
polymorphism and therapy with β-blockers in the determination of heart failure survival.
Upregulation of β
3
-Adrenoceptors and Altered Contractile Response to Inotropic Amines in Human Failing Myocardium
Authors: Moniotte Stéphane , Kobzik Lester , Feron Olivier , Trochu Jean-Noël , Gauthier Chantal , Balligand Jean-Luc
Published: June 2012

Background
—Contrary to β
1
- and β
2
-adrenoceptors, β
3
-adrenoceptors mediate a negative inotropic effect in human ventricular muscle. To assess their functional role in heart failure, our purpose was to compare the expression and contractile effect of β
3
-adrenoceptors in nonfailing and failing human hearts.

Methods and Results
—We analyzed left ventricular samples from 29 failing (16 ischemic and 13 dilated cardiomyopathic) hearts (ejection fraction 18.6±2%) and 25 nonfailing (including 12 innervated) explanted hearts (ejection fraction 64.2±3%). β
3
-Adrenoceptor proteins were identified by immunohistochemistry in ventricular cardiomyocytes from nonfailing and failing hearts. Contrary to β
1
-adrenoceptor mRNA, Western blot analysis of β
3
-adrenoceptor proteins showed a 2- to 3-fold increase in failing compared with nonfailing hearts. A similar increase was observed for Gα
i-2
proteins that couple β
3
-adrenoceptors to their negative inotropic effect. Contractile tension was measured in electrically stimulated myocardial samples ex vivo. In failing hearts, the positive inotropic effect of the nonspecific amine isoprenaline was reduced by 75% compared with that observed in nonfailing hearts. By contrast, the negative inotropic effect of β
3
-preferential agonists was only mildly reduced.

Conclusions
—Opposite changes occur in β
1
- and β
3
-adrenoceptor abundance in the failing left ventricle, with an imbalance between their inotropic influences that may underlie the functional degradation of the human failing heart.
Cardiac Output and Central Distribution of Blood Flow in the Human Fetus
Authors: Mielke Gunther , Benda Norbert
Published: June 2012

Background
—The objectives of this study were to establish reference ranges for left and right cardiac output and to investigate blood flow distribution through the foramen ovale, ductus arteriosus, and pulmonary bed in human fetuses.

Methods and Results
—A prospective study was performed in 222 normal fetuses from 13 to 41 weeks of gestation with high-resolution color Doppler ultrasound. Cardiac output and ductal flow were calculated by use of vessel diameter and the time-velocity integral. Pulmonary blood flow was expressed as the difference between right cardiac output and ductal flow. Foramen ovale flow was estimated as the difference between pulmonary flow and left cardiac output. Gestational age–specific reference ranges are given for left, right, and biventricular output and volume of ductal blood flow, showing an exponential increase with gestational age. Median ratio of right to left cardiac output was 1.42 and was not associated with gestational age. Right cardiac output was 59% and left cardiac output was 41% of biventricular cardiac output. Median biventricular cardiac output was estimated to be 425 mL · min
−1
· kg
−1
fetal weight. Ductal blood flow was 46%, estimated pulmonary flow was 11%, and estimated foramen ovale flow was 33% of biventricular output.

Conclusions
—The study establishes reference ranges for fetal cardiac output and offers insights into the central blood flow distribution in human fetuses from 13 weeks to term. There is a clear right heart dominance. The estimated ratio of pulmonary blood flow to cardiac output is higher than in fetal lamb studies.
Prevention of Autoimmune Myocarditis Through the Induction of Antigen-Specific Peripheral Immune Tolerance
Authors: Godsel Lisa M. , Wang Kegiang , Schodin Beth A. , Leon Juan S. , Miller Stephen D. , Engman David M.
Published: June 2012

Background
—Autoimmunity to cardiac antigens, in particular cardiac myosin, has been observed in humans with myocarditis and in animals with experimental inflammatory heart disease. Current treatments for myocarditis are in many cases immunosuppressive and might lead to increased cardiac damage by reducing host defenses against infectious agents. Therefore, we sought to develop an antigen-specific approach to inhibit autoimmunity in mice with myosin-induced experimental autoimmune myocarditis.

Methods and Results
—Syngeneic splenocytes, coupled with cardiac myosin by use of ethylene carbodiimide, were administered intravenously before disease induction, and the effects of this peripheral tolerization on myosin-induced myocarditis were assessed. This antigen-specific immunotherapy significantly reduced both the incidence and severity of myocarditis, with the prevention of myocyte necrosis, mononuclear cell infiltration, and fibrosis. Myosin-specific delayed-type hypersensitivity and antibody production were significantly reduced, demonstrating that peripheral tolerance affected both T- and B-cell responsiveness to the autoantigen.

Conclusions
—These results suggest that the induction of antigen-specific peripheral immune tolerance may be an effective approach for the treatment of myocarditides with autoimmune involvement.