The mechanisms underlying leptin resistance are still being defined. We report here the presence in human blood of several serum leptin-interacting proteins (SLIPs), isolated by leptin-affinity chromatography and identified by mass spectrometry and immunochemical analysis. We confirmed that one of the major SLIPs is C-reactive protein (CRP). In vitro, human CRP directly inhibits the binding of leptin to its receptors and blocks its ability to signal in cultured cells. In vivo, infusion of human CRP into ob/ob mice blocked the effects of leptin upon satiety and weight reduction. In mice that express a transgene encoding human CRP, the actions of human leptin were completely blunted. We also found that physiological concentrations of leptin can stimulate expression of CRP in human primary hepatocytes. Recently, human CRP has been correlated with increased adiposity and plasma leptin. Thus, our results suggest a potential mechanism contributing to leptin resistance, by which circulating CRP binds to leptin and attenuates its physiological functions.

Glutathione (GSH) content and antioxidant enzyme activities were investigated in skeletal muscle of young, adult, and old male Fischer 344 rats. Furthermore, the effect of 10 wk of exercise training on these antioxidant systems was evaluated at all ages. In the soleus muscle, GSH concentration increased markedly with age, with no significant change in glutathione disulfide (GSSG) content. Training caused a 30% decrease of GSH (P < 0.05) in the soleus of young rats and a reduction of the GSH-to-GSSG ratio at all ages. Activity of gamma-glutamyl transpeptidase (GGT), a key enzyme for GSH uptake by muscle, was also significantly decreased with training. GSH, GSSG, and the GSH-to-GSSG ratio were not altered with aging or training in the deep portion of vastus lateralis muscle (DVL). Activities of GSH peroxidase (GPX), GSSG reductase (GR), superoxide dismutase (SOD), catalase (CAT), and GSH sulfur-transferase were increased significantly with aging in both soleus and DVL. In DVL, training increased GPX and SOD activities in the young rats, whereas in soleus, training decreased GR and CAT activities in the adult rats and GGT and CAT activities in the old rats. Muscle lipid peroxidation was significantly increased with aging in both DVL and soleus but was not affected by training. These data indicate that aging may cause not only an overall elevation of antioxidant enzyme activities but also a fiber-specific adaptation of GSH system in skeletal muscle. Exercise training, although increasing selective antioxidant enzymes in the young rats, does not offer additional protection against oxidative stress in the senescent muscle.

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.

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rence H. Young. Activation of AMPK ␣-and ␥-isoform complexes in the intact ischemic rat heart. Am J Physiol Heart Circ Physiol 291: H1927-H1934, 2006. First published April 28, 2006 doi:10.1152/ajpheart.00251.2006.-AMP-activated protein kinase (AMPK) plays a key role in modulating cellular metabolic processes. AMPK, a serine-threonine kinase, is a heterotrimeric complex of catalytic ␣-subunits and regulatory ␤-and ␥-subunits with multiple isoforms. Mutations in the cardiac ␥ 2-isoform have been associated with hypertrophic cardiomyopathy and pre-excitation syndromes. However, physiological regulation of AMPK complexes containing different subunit isoforms is not well defined and is important for an understanding of the function of this signaling pathway in the intact heart. We evaluated the kinase activity associated with heart AMPK complexes containing specific ␣-and ␥-subunit isoforms of AMPK in an in vivo rat model of regional ischemia. Left coronary artery occlusion activated the immunoprecipitated ␣ 1-isoform (6-fold, P Ͻ 0.01) and ␣2-isoform (9-fold, P Ͻ 0.01) in the ischemic left ventricle compared with sham controls. The degree of ␣-subunit activation depended on the extent of ischemia and paralleled echocardiographic contractile dysfunction. The regulatory ␥ 1-and ␥2-isoforms were expressed in the heart. The ␥ 1-and ␥2-isoforms coimmunoprecipitated with ␣1-and ␣ 2-isoforms in proportion to ␣-subunit content. ␥1-Isoform immunocomplexes accounted for 70% of AMPK activity and AMPK phosphorylation (Thr 172 ) in hearts. Ischemia similarly increased AMPK activity associated with the ␥ 1-and ␥2-isoform complexes threefold (P Ͻ 0.01 for each). Thus AMPK catalytic ␣ 1-and ␣ 2-isoforms are activated by regional ischemia in vivo in the heart, irrespective of the regulatory ␥ 1-or ␥2-isoforms to which they are complexed. Despite the pathophysiological importance of ␥ 2-isoform mutations, ␥1-isoform complexes account for most of the AMPK activity in the ischemic heart.

Background: Chronic migraine (CM) is a common and disabling neurological disorder that affects 1-2% of the global population. The aim of the present study was to identify the functional characteristics of the CM brain using static functional connectivity (s-FC), static functional network connectivity (s-FNC), and dynamic functional network connectivity (d-FNC) analyses. Methods: In the present study, 17 CM patients and 20 sex-and age-matched healthy controls (HCs) underwent resting-state functional magnetic resonance imaging. We utilized independent component (IC) analysis to identify 13 ICs. These 13 ICs were then classified into the following 6 resting-state networks (RSNs): the default mode network (DMN), executive control network (ECN), dorsal attention network, auditory network (AN), visual network (VN), and cerebellum network. Subsequently, s-FC, s-FNC, and d-FNC analyses of 13 ICs were employed for between-group comparisons. Three temporal metrics (fraction of time spent, mean dwell time, and number of transitions), which were derived from the state-transition vector, were calculated for group comparisons. In addition, correlation analyses were performed between these dynamic metrics and clinical characteristics [mean visual analog scale (VAS) scores, days with headache per month, days with migraine pain feature per month, and disease duration]. Results: In the comparison of s-FC of 13 ICs within RSNs between the CM and HC groups, increased connectivity was observed in the left angular gyrus (Angular_L) of the ECN (IC 2) and the right superior parietal gyrus (Parietal_Sup_R) of the AN (IC 5), and reduced connectivity was found in the left superior frontal gyrus (Frontal_Sup_2_L) of the AN (IC 5) and DMN (IC 19), the right calcarine sulcus (Calcarine_ R) of the VN (IC 7), and the left precuneus (Precuneus_L) of the DMN (IC 17) in CM patients. In the comparison of the d-FNC of 13 IC pairs within RSNs between the two groups, the CM group exhibited significantly decreased connections between the DMN (IC 11) and AN (IC 5), and increased connections between the ECN (IC 2, IC 4) and DMN (IC 19), ECN (IC 4) and AN (IC 5), and ECN (IC 4) and VN (IC 13) in state 1. However, no significant differences in s-FNC were observed between the two groups during the s-FNC analysis. Between-group comparisons of three dynamic metrics between the CM and HC groups showed a longer fraction of time spent and mean dwell time in state 2 for CM patients. Furthermore, from the correlation analyses between these metrics and clinical characteristics, we observed a significant positive correlation between the number of transitions and mean VAS scores. Conclusions: Our findings suggest that functional features of the CM brain may fluctuate over time instead of remaining static, and provide further evidence that migraine chronification may be related to abnormal pattern connectivity between sensory and cognitive brain networks.

Modulation of KRAS activity by upstream signals has revealed a promising new approach for pancreatic cancer therapy; however, it is not clear whether microRNA-associated KRAS axis is involved in the carcinogenesis of pancreatic cancer. Here, we identified miR-193b as a tumor-suppressive miRNA in pancreatic ductal adenocarcinoma (PDAC). Expression analyses revealed that miR-193b was downregulated in (10/11) PDAC specimens and cell lines. Moreover, we found that miR-193b functioned as a cell-cycle brake in PDAC cells by inducing G1-phase arrest and reducing the fraction of cells in S phase, thereby leading to dampened cell proliferation. miR-193b also modulated the malignant transformation phenotype of PDAC cells by suppressing anchorage-independent growth. Mechanistically, KRAS was verified as a direct effector of miR-193b, through which the AKT and ERK pathways were modulated and cell growth of PDAC cells was suppressed. Taken together, our findings indicate that miR-193b-mediated deregulation of the KRAS axis is involved in pancreatic carcinogenesis, and suggest that miR-193b could be a potentially effective target for PDAC therapy.