Background-Chronic alcoholism leads to the onset and progression of alcoholic cardiomyopathy through toxic mechanisms of ethanol and its metabolite, acetaldehyde. This study examined the impact of altered acetaldehyde metabolism through systemic transgenic overexpression of aldehyde dehydrogenase-2 (ALDH2) on chronic alcohol ingestion-induced myocardial damage. Methods and Results-ALDH2 transgenic mice were produced with the chicken ␤-actin promoter. Wild-type FVB and ALDH2 mice were placed on a 4% alcohol diet or a control diet for 14 weeks. Myocardial and cardiomyocyte contraction, intracellular Ca 2ϩ handling, histology (hematoxylin and eosin, Masson trichrome), protein damage, and apoptosis were determined. Western blot was used to monitor the expression of NADPH oxidase, calcineurin, apoptosisstimulated kinase (ASK-1), glycogen synthase kinase-3␤ (GSK-3␤), GATA4, and cAMP-response element binding (CREB) protein. ALDH2 reduced the chronic alcohol ingestion-induced elevation in plasma and tissue acetaldehyde levels. Chronic alcohol consumption led to cardiac hypertrophy, reduced fractional shortening, cell shortening, and impaired intracellular Ca

Background-Elderly patients are more sensitive than younger patients to myocardial ischemia, which results in higher mortality. We investigated how aging affects the cardioprotective AMP-activated protein kinase (AMPK) signaling pathway. Methods and Results-Ischemic AMPK activation was impaired in aged compared with young murine hearts. The expression and secretion of the AMPK upstream regulator, macrophage migration inhibitory factor (MIF), were lower in aged compared with young adult hearts. Additionally, the levels of hypoxia-inducible factor 1␣, a known transcriptional activator of MIF, were reduced in aged compared with young hearts. Ischemia-induced AMPK activation in MIF knockout mice was blunted, leading to greater contractile dysfunction in MIF-deficient than in wild-type hearts. Furthermore, intramyocardial injection of adenovirus encoding MIF in aged mice increased MIF expression and ischemic AMPK activation and reduced infarct size. Conclusions-An impaired MIF-AMPK activation response in senescence thus may be attributed to an agingassociated defect in hypoxia-inducible factor 1␣, the transcription factor for MIF. In the clinical setting, impaired cardiac hypoxia-inducible factor 1␣ activation and consequent reduced MIF expression may play an important role in the increased susceptibility to myocardial ischemia observed in older cardiac patients. (Circulation. 2010;122: 282-292.)

A "longevity " gene, sirtuin 1 (SIRT1), can attenuate age-dependent induction of left ventricular dysfunction. This study aimed to characterize the role of SIRT1 in the tolerance of aged heart to ischemic insults. Male C57BL/6 young (4-6 mo) and aged (24-26 mo) mice were used to determine the role of SIRT1 in myocardial ischemia/reperfusion (I/R) tolerance. SIRT1 localization was assessed by confocal microscopy. Immunoblotting was used to evaluate SIRT1 expression and translocation. The results demonstrated that SIRT1 is expressed predominantly as a sumoylated form in cardiomyocyte nuclei. Moreover, cardiac overexpression of desumoylase, sentrin-specific protease 2 (SENP2), significantly reduces nuclear sumoylated SIRT1 levels (P<0.05). Interestingly, I/R stress leads to desumoylation and translocation of nuclear SIRT1 into the cytoplasm in aged but not in young hearts. SIRT1 activity in ischemic young hearts was 3.2-fold higher than that seen in ischemic aged hearts, which suggests that aging causes impaired nucleocytoplasmic shuttling and activation of SIRT1 during ischemic stress. The infarct size in aged and Sirt1(+/-) knockout hearts was higher than that observed in young and Sirt1(+/+) WT littermate hearts, respectively (all P<0.05). SIRT1 agonist, SRT1720, reduced myocardial infarction in both aged and Sirt1(+/-) hearts. Therefore, impaired cardiac SIRT1 activity plays a critical role in the observed increase in susceptibility of the aged heart to I/R injury. SIRT1 agonist can restore this aging-related loss of cardioprotection.

The purpose of this study was to evaluate the effect of endurance exercise training on both locomotor skeletal muscle collagen characteristics and passive stiffness properties in the young adult and old rat. Young (3-mo-old) and senescent (23-mo-old) male Fischer 344 rats were randomly assigned to either a control or exercise training group [young control (YC), old control (OC), young trained (YT), old trained (OT)]. Exercise training consisted of treadmill running at approximately 70% of maximal oxygen consumption (45 min/day, 5 days/wk, for 10 wk). Passive stiffness (stress/strain) of the soleus (Sol) muscle from all four groups was subsequently measured in vitro at 26 degreesC. Stiffness was significantly greater for Sol muscles in OC rats compared with YC rats, but in OT rats exercise training resulted in muscles with stiffness characteristics not different from those in YC rats. Sol muscle collagen concentration and the level of the nonreducible collagen cross-link hydroxylysylpyridinoline (HP) significantly increased from young adulthood to senescence. Although training had no effect on Sol muscle collagen concentration in either age group, it resulted in a significant reduction in the level of Sol muscle HP in OT rats. In contrast, exercise had no effect on HP in the YT animals. These findings indicate that 10 wk of endurance exercise significantly alter the passive viscoelastic properties of Sol muscle in old but not in young adult rats. The coincidental reduction in the principal collagen cross-link HP also observed in response to training in OT muscle highlights the potential role of collagen in influencing passive muscle viscoelastic properties.

Objective-Vascular smooth muscle cell (VSMC) migration, proliferation, and collagen synthesis are key events involved in the pathogenesis of cardiovascular disease. Growth factors, such as platelet-derived growth factor (PDGF) and fibroblast growth factor, released during vascular injury plays a pivotal role in regulating these events. Curcumin (diferuloyl methane), a major component of the spice turmeric (Curcuma longa), has been shown recently to have beneficial effects in chronic conditions, such as inflammation, cancer, cystic fibrosis, and Alzheimer's disease. The objective of this study was to investigate the ability of curcumin to inhibit PDGF-stimulated migration, proliferation, and collagen synthesis in cultured VSMCs and neointima formation after carotid artery injury in rats. Methods and Results-Curcumin (1 to 25 M) produced a concentration-dependent inhibition of PDGF-elicited VSMC migration, proliferation, and collagen synthesis assessed by chemotaxis, [ 3 H]thymidine incorporation, and [ 3 H]-L-proline incorporation, respectively. Curcumin blocked PDGF-induced VSMC actin-cytoskeleton reorganization, attenuated PDGF signal transduction, and inhibited the binding of PDGF to its receptors. Carotid artery neointima formation was significantly attenuated by perivascular curcumin compared with vehicle controls 14 days after injury, characterized by reduced DNA synthesis, collagen synthesis, and PDGF receptor phosphorylation. Conclusions-These data suggest that curcumin is a potent inhibitor of key PDGF-stimulated VSMC functions and may play a critical role in regulating these events after vascular injury. T he primary event in the development of atherosclerosis and restenosis after percutaneous transluminal coronary angioplasty is thought to involve injury to the endothelium, leading to a response that may be similar to wound healing requiring migration of vascular smooth muscle cells (VSMCs) from the media to the intima and subsequent proliferation. [1][2][3] Intimal smooth muscle cells (SMCs) in the intima assume a synthetic phenotype vis-à-vis the normal contractile phenotype, resulting in the deposition of extracellular matrix within the neointimal tissue. 1,2 Although the mechanisms responsible for migration and proliferation of VSMCs are not fully understood, several factors produced in response to vascular injury have been implicated in this process. 3 Platelet-derived growth factor (PDGF) is a potent growth factor produced by platelets, VSMCs, and endothelial cells in the injured vascular wall. 4,5 PDGF initiates a multitude of biological effects through the activation of intracellular signal transduction pathways that contribute to VSMC proliferation, migration, and collagen synthesis. 6 The importance of PDGF in the development of neointima has been established in arterial injury models. 6 PDGF is also a potent stimulant of extracellular matrix synthesis by VSMCs. Accordingly, inhibition of PDGF-stimulated VSMC migration, proliferation, and extracellular matrix synthesis represents an important...

The seasonal and interannual variability of the water masses, nutrients, phytoplankton biomass and primary productivity of the waters off the eastern coast of Tasmania are described. The seasonal and interannual variability in the water masses on the east coast could be explained by the varying influence of tropical and subantarctic waters and the presence of the northern edge of the subtropical convergence north-east from Maria Island. The physical oceanography was dominated by mesoscale events and the influence of the two parent water masses was highly episodic. Subtropical water rarely extended as far south as Tasman Island in summer and subantarctic water never extended as far north as Flinders Island. Data from satellite Advanced Very High Resolution Radiometer (AVHRR) images confirmed the hydrographic data and were used to interpret the seasonal and interannual variability. Interannual variability in maximum summer temperatures at Maria Island was correlated with the southerly extension of subtropical waters and with El Nino/Southern Oscillation events at least until the mid-1970s. There was evidence of a long-term warming at Maria Island and a cyclic fluctuation of 10-15 years in smoothed maximum summer temperatures. The decline in nitrate, and hence the phytoplankton spring bloom, occurred a month earlier inshore than offshore. Long-term data records from the inshore station at Maria Island showed strong interannual changes in the timing of the nitrate decline. The timing of the spring bloom varied from year to year by as much as 4 months. The seasonal cycle of phytoplankton biomass in 1984 and 1985 showed spring (October) and autumnal (April) blooms, with an early summer bloom in December. Low chlorophyll levels in February and March coincided with the influence of subtropical water. The seasonal cycle of phytoplankton biomass was therefore a complex function of seasonal and episodic events. Primary productivity data from the spring of 1984 showed low productivity on the west coast of Tasmania but high productivity on the east coast around Maria Island and the islands in Bass Strait.

Cellular hypertrophy is regulated by coordinated pro- and antigrowth machineries. Foxo transcription factors initiate an atrophy-related gene program to counter hypertrophic growth. This study was designed to evaluate the role of Akt, the forkhead transcription factor Foxo3a, and atrophy genes muscle-specific RING finger (MuRF)-1 and atrogin-1 in cardiac hypertrophy and contractile dysfunction associated with high-fat diet-induced obesity. Mice were fed a low- or high-fat diet for 6 mo along with a food-restricted high-fat weight control group. Echocardiography revealed decreased fractional shortening and increased end-systolic diameter and cardiac hypertrophy in high-fat obese but not in weight control mice. Cardiomyocytes from high-fat obese but not from weight control mice displayed contractile and intracellular Ca2+ defects including depressed maximal velocity of shortening/relengthening, prolonged duration of shortening/relengthening, and reduced intracellular Ca2+ rise and clearance. Caspase activities were greater in high-fat obese but not in weight control mouse hearts. Western blot analysis revealed enhanced basal Akt and Foxo3a phosphorylation and reduced insulin-stimulated phosphorylation of Akt and Foxo3a without changes in total protein expression of Akt and Foxo3a in high-fat obese hearts. RT-PCR and immunoblotting results displayed reduced levels of the atrogens atrogin-1 and MuRF-1, the upregulated hypertrophic markers GATA4 and ciliary neurotrophic factor receptor-alpha, as well as the unchanged calcineurin and proteasome ubiquitin in high-fat obese mouse hearts. Transfection of H9C2 myoblast cells with dominant-negative Foxo3a adenovirus mimicked palmitic acid (0.8 mM for 24 h)-induced GATA4 upregulation without an additive effect. Dominant-negative Foxo3a-induced upregulation of pAkt and repression of phosphatase and tensin homologue were abrogated by palmitic acid. These results suggest a cardiac hypertrophic response in high-fat diet-associated obesity at least in part through inactivation of Foxo3a by the Akt pathway.

This study evaluated the single and interactive effects of age and training status on selected collagen parameters in two rodent locomotor skeletal muscles contrasting in fiber type composition. Gastrocnemius (GAST) and soleus (SOL) muscles from both trained (10 wk of daily treadmill running) and sedentary young adult (5-mo-old), middle-aged (15-mo-old), and old (23-mo-old) female Fischer 344 rats were evaluated for concentrations of collagen (measured by hydroxyproline concentration ([OH-Pro])) and of the predominant nonreducible lysine aldehyde-derived collagen cross-link hydroxylysylpyridinoline ([HP]). Maximal aerobic capacity was significantly elevated in all three trained groups compared with sedentary age-matched control groups. Slow-twitch SOL had a significantly higher [OH-Pro] than fast-twitch GAST (P < 0.05). Although aging had no effect on [OH-Pro] in GAST, in SOL a significant increase with age was seen (P < 0.02). In sedentary rats both GAST and SOL [HP] increased with age, with this increase being more pronounced for SOL. Additionally, although training had no effect on the aging-associated increase in GAST [HP], it prevented the rise seen in SOL. The observed training-induced reduction in SOL [HP] presumably reflects exercise recruitment and subsequent stimulation of collagen synthesis and degradation rates in this muscle. We conclude that both aging and training affect the extracellular matrix in rodent limb skeletal muscle.