Consumption of a high-fat diet (HFD) in experimental animal models initiates a series of molecular events and outcomes, including insulin resistance and obesity, that mimic the metabolic syndrome in humans. The relationship among, and order of, the molecular events linking a diet high in fat to pathologies is often unclear. In the present study, we provide several novel insights into the relationship between a HFD and AMP-activated protein kinase (AMPK), a key regulator of cellular metabolism and whole-body energy balance. HFD substantially decreased the activities of both iso-forms of AMPK in white adipose tissue, heart, and liver. These decreases in AMPK activity occurred in the absence of decreased AMPK transcription, systemic inflammation, hyperglycemia, or elevated levels of free fatty acids. The HFD-induced decrease in AMPK activity was associated with systemic insulin resistance and hyperleptinemia. In blood, >98 % of AMPK activity was localized in agranulocytes as the α1 isoform. In contrast to the solid tissues studied, AMPK activities were not altered by HFD in granulocytes or agranulocytes. We conclude that HFD-induced obesity causes a broad, non-tissue, or isoform-specific lowering of AMPK activity. Given the central position AMPK plays in whole-body energy balance, this decreased AMPK activity may play a previously unrecognized role in obesity and its associated pathologies.

Purpose Demonstrate a novel manufacturing method to generate extracellular matrix scaffolds from cardiac fibroblasts (CF-ECM) as a therapeutic mesenchymal stem cell-transfer device. Materials and Methods Rat CF were cultured at high-density (~1.6×105/cm2) for 10–14 days. Cell sheets were removed from the culture dish by incubation with EDTA and decellularized with water and peracetic acid. CF-ECM was characterized by mass spectrometry, immunofluorescence and scanning electron microscopy. CF-ECM seeded with human embryonic stem cell derived mesenchymal stromal cells (hEMSCs) were transferred into a mouse myocardial infarction model. 48 hours later, mouse hearts were excised and examined for CF-ECM scaffold retention and cell transfer. Results CF-ECM scaffolds are composed of fibronectin (82%), collagens type I (13%), type III (3.4%), type V (0.2%), type II (0.1%) elastin (1.3%) and 18 non-structural bioactive molecules. Scaffolds remained intact on the mouse heart for 48 hours without the use of sutures or glue. Identified hEMSCs were distributed from the epicardium to the endocardium. Conclusions High density cardiac fibroblast culture can be used to generate CF-ECM scaffolds. CF-ECM scaffolds seeded with hEMSCs can be maintained on the heart without suture or glue. hEMSC are successfully delivered throughout the myocardium.

Bauwens JD, Schmuck EG, Lindholm CR, Ertel RL, Mulligan JD, Hovis I, Viollet B, Saupe KW. Cold tolerance, cold-induced hyperphagia, and nonshivering thermogenesis are normal in ␣ 1-AMPK Ϫ/Ϫ mice.

The aged heart displays a loss of cardiomyocyte number and function, possibly due to the senescence and decreased regenerative potential that has been observed in some cardiac progenitor cells. An important cardiac progenitor that has not been studied in the context of aging is the cardiac side population (CSP) cell. To address this, flow cytometry-assisted cell sorting was used to isolate CSP cells from adult (6-10 months old) and aged (24-32 months old) C57Bl/6 mice that were fed either a control diet or an anti-aging diet (caloric restriction, CR). Aging caused a 2.3-fold increase in the total number of CSP cells and a 3.2-fold increase in the cardiomyogenic sca1(+)/CD31(-) subpopulation. Aging did not affect markers of proliferation or senescence, including telomerase activity and expression of cell cycle genes, in sca1(+)/CD31(-) CSP cells. In contrast, the aged cells had reduced expression of genes associated with differentiation, including smooth muscle actin and cardiac muscle actin (5.1- and 3.2-fold, respectively). None of these age effects were altered by CR diet. Therefore, it appears that the manner in which CSP cells age is distinct from the aging of post-mitotic tissue (and perhaps other progenitor cells) that can often be attenuated by CR.

The aged heart displays a loss of cardiomyocyte number and function, possibly due to decreased regenerative potential of cardiac progenitor cells. An important cardiac progenitor population that has not been studied in the context of aging is cardiac side population cells (CSPC). To address this, flow cytometry‐assisted cell sorting was used to isolate CSPC from adult (~5‐months old) and aged (~25‐months old) C57Bl/6 mice that were fed either a control diet or an anti‐aging (caloric restriction, CR) diet. Aging caused a 2.0‐fold increase in both the total number of CSPC and in the highly cardiomyogenic sca1+/CD31‐ subpopulation (n=8–10 mice/group). Telomerase activity (a marker of proliferation) in CSPC was not altered by aging (n=4 mice/group). In contrast, aged CSPC showed reduced expression of several genes associated with differentiation, including smooth muscle actin, cardiac muscle actin, and bone collagen type I (5.1‐, 3.2‐, and 2.2‐fold respectively; n=4/group). These data suggest that any loss of regenerative potential in aged CSPC is likely due to inefficiency of differentiation into cardiac lineages rather than poor proliferation rate. None of these age effects were altered by CR diet. Therefore it appears that the manner in which CSPC (and perhaps other types of progenitor cells) age is distinct from the post‐mitotic tissue aging that can often be attenuated by CR.