During the past decade, lifestyle health coaching (LHC) has gained increased popularity as a strategy to facilitate behavior change. Few data are available on the effect of LHC on multiple cardiovascular disease (CVD) risk factors. PURPOSE: In this multi-center study, we evaluated the effect of LHC on multiple CVD risk factors in 9,134 consecutive adults and compared the effect of a phase 2 cardiac rehabilitation (CR) program in 11,875 consecutive patients. METHODS: Outcome measures were assessed at baseline and after approximately 12 weeks of LHC and CR. LHC included individualized coaching, predominantly via the telephone and Internet, on exercise training, nutrition, weight management, stress management and tobacco cessation interventions. CR was conducted at multiple centers in the United States. RESULTS: At baseline, CR patients were significantly (p<0.05) older (69 ± 11 years vs. 51 ± 12 years), and had a higher prevalence of atherosclerotic CVD (95.3% vs. 10.6%) and diabetes (23.7% vs. 9.1%) than those participating in LHC. For participants with abnormal baseline risk factors, statistically significant improvements (p <0.05) were observed for multiple variables, as follows: blood pressure (LHC, -9/7 mmHg; CR, -10/10 mmHg; p <0.05 for CR versus LHC); LDL cholesterol (LHC, -22 mg/dl; CR, -50 mg/dl; p <0.05 for CR versus LHC); HDL cholesterol (LHC, 3 mg/dl; CR, 5 mg/dl; p <0.05 for CR versus LHC); triglycerides (LHC, -46 mg/dl; CR, -52 mg/dl; p=NS for CR versus LHC); fasting glucose (LHC, -13 mg/dl; CR, -15 mg/dl; p=NS for CR versus LHC); and body weight (LHC, -5.8 lbs; CR, -2.5 lbs; p <0.05 for LHC versus CR). In participants in the LHC program with a baseline Framingham 10-year coronary heart disease risk score >10%, the score decreased by 20.5% (relative risk reduction, p <0.05). CONCLUSIONS: These data serve to document the magnitude of improvement in multiple CVD risk factors in response to participation in approximately 12 weeks of LHC. The data demonstrate that LHC results in clinically relevant improvements in multiple CVD risk factors and that the precise magnitude of improvement may be similar to, less than or greater than that observed with CR depending on the specific risk factor in question.

Metabolic syndrome (MS) is a constellation of interrelated coronary heart disease and type 2 diabetes risk factors of metabolic origin (MS risk factors) that are associated with increased cardiovascular event rates. Although MS risk factors are strongly influenced by lifestyle, few data are available on the effect of lifestyle health coaching (LHC) on MS risk factors. PURPOSE: In this study, we determined the prevalence of MS and its component MS risk factors on entry into and after ∼ 1 year of participation in LHC. METHODS: Subjects were 7,929 consecutive adults (mean age = 52 ± 12 years) who completed both a baseline evaluation on entry into a LHC program and a follow-up evaluation after ∼1 year of participation in the LHC program. LHC included individualized coaching, predominantly via the telephone and Internet, on exercise training, nutrition counseling, weight management, stress management and tobacco cessation. MS and its 5 individual component MS risk factors were defined in accordance with the National Cholesterol Education Program Adult Treatment Panel III Guidelines; however, a fasting glucose >100 mg/dl (rather than > 110 mg/dl) was used. In individuals with MS at baseline and data on all 5 individual MS risk factors both at baseline and follow-up (n=1,887), results were analyzed to determine the number of individuals with >3, 3, 4, or 5 MS risk factors and the number of individuals with each of the individual MS risk factors. RESULTS: Results were as follows:TABLECONCLUSIONS: LHC was associated with a 30.9% reduction in the prevalence of MS and had a favorable impact on all 5 MS risk factors.

Objective: Methylation of the promoter region of the estrogen receptor gene alpha (ER alpha) occurs as a function of age in human colon, and results in inactivation of gene transcription. In this study, we sought to determine whether such age-related methylation occurs in the cardiovascular system, and whether it is associated with atherosclerotic disease. Methods: We used Southern blot analysis to determine the methylation state of the ER alpha gene in human light atrium, aorta, internal mammary artery, saphenous vein, coronary atherectomy samples, as well as cultured aortic endothelial cells and smooth muscle cells. Results: An age related increase in ER alpha gene methylation occurs in the right atrium (range 6 to 19%, R = 0.36, P<0.05). Significant levels of ER alpha methylation were detected in both veins and arteries. In addition, ER alpha gene methylation appears to be increased in coronary atherosclerotic plaques when compared to normal proximal aorta (10+/-2% versus 4+/-1%, P<0.01). In endothelial cells er;planted from human aorta and grown in vitro, ER alpha gene methylation remains low. In contrast, cultured aortic smooth muscle cells contain a high level of ER alpha gene methylation (19-99%). Conclusions: Methylation associated inactivation of the ER alpha gene in vascular tissue may play a role in atherogenesis and aging of the vascular system. This potentially reversible defect may provide a new target for intervention in heart disease. (C) 1999 Elsevier Science B.V. All rights reserved.

The human integrin beta3 participates in a wide range of adhesive biologic functions and is expressed in a selected subset of tissues, but little is known about the cis-acting DNA elements or trans-acting factors responsible for this regulation. Using cell lines characterized for beta3 expression, a number of upstream regulatory regions in the beta3 gene were identified. (1) The three regions from -1159 to -584, -290 to -146, and -126 to -115 demonstrated positive, negative, and negative activity, respectively. (2) The region from -115 to +29 of the beta3 gene was sufficient for cell-specific activity. Deletion of the sequence from -115 to -89 produced a 6- to 40-fold reduction in reporter gene activity in beta3-expressing megakaryocytic cell lines (K562, Dami, and HEL), but only a 1.7- and 2.7-fold reduction, respectively, in beta3-expressing endothelial and melanoma cell lines, and 1.3- and 2. 8-fold reduction, respectively, in non-beta3-expressing Chinese hamster ovary and 293 cell lines. This sequence also bound nuclear proteins in a cell-specific manner in electrophoretic mobility shift assays. Mutational analysis indicated that the sequence GAGGGG (positions -113 to -108) is a megakaryocytic cell line-specific cis-acting element. (3) The region from -89 to +29 promoted lower activity in all cell lines. We also provide evidence that a CCCACCC sequence at position -70 has transcriptional activity, most likely through the Sp1 transcription factor. These data supply the first detailed map of the transcriptional regulatory elements of the 5' region of the beta3 gene, define positive regulatory sequences with potent megakaryocyte preferential activity, and indicate that the ubiquitous transcription factor, Sp1, may augment beta3 gene expression.

Reactive oxygen species play an important role at the site of vascular injuries and arterial thromboses. We studied the mechanism mediating platelet aggregation induced by H2O2, a major cellular oxidant, Exposure to H2O2 triggered platelet aggregation, but only when the platelets were stirred. Strong platelet aggregation induced by H2O2 required the presence of the tyrosine phosphatase inhibitor sodium orthovanadate (NaVO4) and was dependent on the participation of integrin alpha(IIb)beta(3)(glycoprotein IIb-IIIa). A specific inhibitor of alpha(IIb)beta(3) blocked platelet aggregation induced by H2O2 and NaVO4, thus confirming that aggregation requires this receptor. In the presence of H2O2 and NaVO4, multiple platelet substrates were phosphorylated on tyrosine, Such tyrosine kinase response was necessary but not sufficient to activate alpha(IIb)beta(3), as detected by binding of soluble fibrinogen to platelets, Stirring of the platelets exposed to H2O2 and NaVO4 was also needed to allow for binding of fibrinogen to alpha(IIb)beta(3). The tyrosine kinase inhibitor genistein was able to block platelet aggregation induced by H2O2 and NaVO4, thus confirming that tyrosine kinase activity was needed to trigger alpha(IIb)beta(3) activation on stirring. N-Acetyl-L-cysteine, a cell-permeant antioxidant, blocked the tyrosine phosphorylation of platelet substrates and also the platelet aggregation induced by H2O2 and NaVO4. We found that beta(3) was phosphorylated on tyrosine in platelets exposed to H2O2 and NaVO4, even in the absence of aggregation, Hence, tyrosine phosphorylation of beta(3) might contribute to the "priming" of alpha(IIb)beta(3) induced by H2O2 and NaVO4, whereby the receptor can become activated on stirring of the platelets.

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The human blood platelet fibrinogen receptor, integrin alphaIIbbeta3 (glycoprotein IIb-IIIa) is an archetypal member of the integrin family of adhesive molecules and is the only integrin encoded by genes physically linked in the genome. Because studies on the normal and abnormal expression of any gene require a thorough understanding of its organization, the initial goals of the current study were to determine the size and complete the genomic organization for the beta3 gene. We now report the isolation of the entire beta3 gene in a single P1 plasmid and for the first time have linked the first and second exons on a contiguous fragment of DNA. Using pulsed-field gel analysis, we determined the full size of the beta3 gene to be 63 kb and show a large (16.7 kb) first intron; based on this information, we propose a uniform numbering system for the beta3 exons. We have completed the 5' genomic structure and generated a long-range restriction map. The promoter and the 5' end of the first intron were found to have approximately 50% sequence identity with a region of the avian beta3 gene known to possess functional transcriptional activity. Analysis of three different homologous regions led to the identification of a sequence in the 5'-UTR of the human gene, CCGCGGGAGG, which shares 90% identity with the avian gene and which bound nuclear proteins in DNaseI and electrophoretic mobility shift assay studies. Mutating this sequence caused a 2.6-fold reduction in reporter gene activity. In these studies we have (1) determined the full length and 5' organization of the beta3 gene, (2) identified a large region of homology between the 5' regions of the avian and human genes, and (3) identified a sequence in the 5'-UTR that augments gene expression. Knowing the genomic structure of beta3 has permitted the uncovering of new mechanisms of mutagenesis causing Glanzmann thrombasthenia (Jin et al, J Clin Invest 98:1745, 1996), and our findings will be valuable for such genetic analyses as well as for studies on the transcriptional regulation of beta3 and other integrin genes.

Endothelial cells provide an antithrombotic and anti-inflammatory barrier for the normal vessel wall. Dysfunction of endothelial cells has been shown to promote atherosclerosis, and normalization of previously dysfunctional endothelial cells can inhibit the genesis of atheroma. In normal arteries, endothelial cells are remarkably quiescent. Acceleration of the turnover rate of endothelial cells can lead to their dysfunction. Apoptosis is a physiological process that contributes to vessel homeostasis, by eliminating damaged cells from the vessel wall. However, increased endothelial cell turnover mediated through accelerated apoptosis may alter the function of the endothelium and therefore, promote atherosclerosis. Apoptotic endothelial cells can be detected on the luminal surface of atherosclerotic coronary vessels, but not in normal vessels. This finding links endothelial cell apoptosis and the process of atherosclerosis, although a causative role for apoptosis in this process remains hypothetical. Estrogen metabolites have been shown to be among the most potent anti-atherogenic agents available to date for post-menopausal women. The mechanism of estrogen's protective effect is currently incompletely characterized. Here we show that 17β-estradiol, a key estrogen metabolite, inhibits apoptosis in cultured endothelial cells. Our data support the hypothesis that 17β-estradiol's anti-apoptotic effect may be mediated via improved endothelial cell interaction with the substratum, increased tyrosine phosphorylation of pp125 focal adhesion kinase, and a subsequent reduction in programmed cell death of endothelial cells. Inhibition of apoptosis by estrogens may account for some of the anti-atherogenic properties of these compounds.

The maturation of megakaryocytes in vivo requires polyploidization or repeated duplication of DNA without cytokinesis. As DNA replication and cytokinesis are tightly regulated in somatic cells by cyclins and cyclin-dependent kinases, we sought to determine the pattern of cyclin gene expression in cells that undergo megakaryocytic differentiation and polyploidization. The Dami megakaryocytic cell line differentiates and increases ploidy in response to phorbol 12-myristate 13-acetate (PMA) stimulation in vitro. We used Northern blotting to analyze mRNA levels of cyclins A, B, C, D1, and E in PMA-induced Dami cells and found that cyclin D1 mRNA levels increased dramatically (18-fold). Similar increases in cyclin D1 mRNA were obtained for other cell lines (HEL and K562) with megakaryocytic properties, but not in HeLa cells. The increase in cyclin D1 was confirmed by Western immunoblotting of PMA-treated Dami cells. This finding suggested that cyclin D1 might participate in megakaryocyte differentiation by promoting endomitosis and/or inhibiting cell division. To address these possibilities, we constructed two stable Zn+2-inducible, cyclin D1-overexpressing Dami cell lines. Cyclin D1 expression alone was not sufficient to induce polyploidy, but in conjunction with PMA-induced differentiation, polyploidization was slightly enhanced. However, unlike other cell systems, cyclin D1 overexpression caused cessation of cell growth. Although the mechanism by which cyclin D1 may affect megakaryocyte differentiation is not clear, these data demonstrate that cyclin D1 is upregulated in differentiating megakaryocytic cells and may contribute to differentiation by arresting cell proliferation.