No Kiss1ng by leptin during puberty?

Leptin exerts a permissive action on puberty by stimulating release of gonadotropin-releasing hormone (GnRH) in the hypothalamus. However, GnRH neurons lack leptin receptor (LepR), indicating that leptin must indirectly regulate these neurons. The Kiss1 gene produces kisspeptins that stimulate GnRH secretion. Because Kiss1 neurons express LepR and inactivation of Kiss1 causes hypogonadotropic hypogonadism, Donato et al., in this issue of the JCI, assessed whether deletion of LepR from Kiss1 neurons would prevent sexual maturation. Unexpectedly, mice lacking LepR in Kiss1 neurons had normal pubertal development and fertility. In contrast, deletion of LepR from the ventral premammillary nucleus, a region of the brain involved in sexual behavior, prevented puberty and fertility. These findings highlight the complex biology of leptin in reproduction.     

Cytokine responses at high altitude: effects of exercise and antioxidants at 4300 m

PURPOSE: This study tested the hypothesis that antioxidant supplementation would attenuate plasma cytokine (IL-6, tumor necrosis factor (TNF)-alpha), and C-reactive protein (CRP) concentrations at rest and in response to exercise at 4300-m elevation. METHODS: A total of 17 recreationally trained men were matched and assigned to an antioxidant (N = 9) or placebo (N = 8) group in a double-blinded fashion. At sea level (SL), energy expenditure was controlled and subjects were weight stable. Then, 3 wk before and throughout high altitude (HA), an antioxidant supplement (10,000 IU beta-carotene, 200 IU alpha-tocopherol acetate, 250 mg ascorbic acid, 50 microg selenium, 15 mg zinc) or placebo was given twice daily. At HA, energy expenditure increased approximately 750 kcal.d(-1) and energy intake decreased approximately 550 kcal.d, resulting in a caloric deficit of approximately 1200-1500 kcal.d(-1). At SL and HA day 1 (HA1) and day HA13, subjects exercised at 55% of VO2peak until they expended approximately 1500 kcal. Blood samples were taken at rest, end of exercise, and 2, 4, and 20 h after exercise. RESULTS: No differences were seen between groups in plasma IL-6, CRP, or TNF-alpha at rest or in response to exercise. For both groups, plasma IL-6 concentration was significantly higher at the end of exercise, 2, 4, and 20 h after exercise at HA1 compared with SL and HA13. Plasma CRP concentration was significantly elevated 20 h postexercise for both groups on HA1 compared to SL and HA13. TNF-alpha did not differ at rest or in response to exercise. CONCLUSION: Plasma IL-6 and CRP concentrations were elevated following exercise at high altitude on day 1, and antioxidant supplementation did not attenuate the rise in plasma IL-6 and CRP concentrations associated with hypoxia, exercise, and caloric deficit.     

Foundations of advanced magnetic resonance imaging

During the past decade, major breakthroughs in magnetic resonance imaging (MRI) quality were made by means of quantum leaps in scanner hardware and pulse sequences. Some advanced MRI techniques have truly revolutionized the detection of disease states and MRI can now— within a few minutes—acquire important quantitative information noninvasively from an individual in any plane or volume at comparatively high resolution. This article provides an overview of the most common advanced MRI methods including diffusion MRI, perfusion MRI, functional MRI, and the strengths and weaknesses of MRI at high magnetic field strengths.     

Dynamically-coated capillaries allow for capillary electrophoretic resolution of transferrin sialoforms via direct analysis of human serum

Transferrin sialoforms with fewer than three sialic acid residues (carbohydrate deficient transferrin; CDT) have been implicated as a marker of certain liver pathologies. Transferrin sialoforms in human sera from alcoholic and non-alcoholic patients was analyzed by capillary electrophoresis (CE) using diaminobutane (DAB) to dynamically-coat the capillary wall to minimize protein-wall interactions. Using a DAB concentration of 3 mM, transferrin sialoforms were adequately resolved to allow for direct detection of CDT without extensive treatment of the sera. Serum immunoglobulins, which migrated close to the CDT region, were removed via subtraction with protein A, enhancing the detection of CDT. The reproducibility of sialoform separation in dynamically-coated capillaries was found to be acceptable with run-to-run relative standard deviation values of 0.15% for a sample on a given day and 0.29+/-0.06% for four samples day-to-day. These results suggest that dynamic-coating approaches may provide a simple alternative to the use of covalently-coated capillaries for the CE separation of complex samples.     

Loss of IL-15 receptor �� alters the endurance, fatigability, and metabolic characteristics of mouse fast skeletal muscles

IL-15 receptor α (IL-15Rα) is a component of the heterotrimeric plasma membrane receptor for the pleiotropic cytokine IL-15. However, IL-15Rα is not merely an IL-15 receptor subunit, as mice lacking either IL-15 or IL-15Rα have unique phenotypes. IL-15 and IL-15Rα have been implicated in muscle phenotypes, but a role in muscle physiology has not been defined. Here, we have shown that loss of IL-15Rα induces a functional oxidative shift in fast muscles, substantially increasing fatigue resistance and exercise capacity. IL-15Rα–knockout (IL-15Rα–KO) mice ran greater distances and had greater ambulatory activity than controls. Fast muscles displayed fatigue resistance and a slower contractile phenotype. The molecular signature of these muscles included altered markers of mitochondrial biogenesis and calcium homeostasis. Morphologically, fast muscles had a greater number of muscle fibers, smaller fiber areas, and a greater ratio of nuclei to fiber area. The alterations of physiological properties and increased resistance to fatigue in fast muscles are consistent with a shift toward a slower, more oxidative phenotype. Consistent with a conserved functional role in humans, a genetic association was found between a SNP in the IL15RA gene and endurance in athletes stratified by sport. Therefore, we propose that IL-15Rα has a role in defining the phenotype of fast skeletal muscles in vivo.     

Fractalkine is a novel human adipochemokine associated with type 2 diabetes

OBJECTIVE

Leukocyte infiltration of adipose is a critical determinant of obesity-related metabolic diseases. Fractalkine (CX3CL1) and its receptor (CX3CR1) comprise a chemokine system involved in leukocyte recruitment and adhesion in atherosclerosis, but its role in adipose inflammation and type 2 diabetes is unknown.

RESEARCH DESIGN AND METHODS

CX3CL1 mRNA and protein were quantified in subcutaneous adipose and blood during experimental human endotoxemia and in lean and obese human adipose. CX3CL1 cellular source was probed in human adipocytes, monocytes, and macrophages, and CX3CL1-blocking antibodies were used to assess its role in monocyte-adipocyte adhesion. The association of genetic variation in CX3CR1 with metabolic traits was determined in a community-based sample. Finally, plasma CX3CL1 levels were measured in a case-control study of type 2 diabetes.

RESULTS

Endotoxemia induced adipose CX3CL1 mRNA (32.7-fold, P < 1 × 10⁻⁵) and protein (43-fold, P = 0.006). Obese subjects had higher CX3CL1 levels in subcutaneous adipose compared with lean (0.420 ± 0.387 vs. 0.228 ± 0.187 ng/mL, P = 0.04). CX3CL1 was expressed and secreted by human adipocytes and stromal vascular cells. Inflammatory cytokine induction of CX3CL1 in human adipocytes (27.5-fold mRNA and threefold protein) was completely attenuated by pretreatment with a peroxisome proliferator–activated receptor-γ agonist. A putative functional nonsynonymous single nucleotide polymorphism (rs3732378) in CX3CR1 was associated with adipose and metabolic traits, and plasma CX3CL1 levels were increased in patients with type 2 diabetes vs. nondiabetics (0.506 ± 0.262 vs. 0.422 ± 0.210 ng/mL, P < 0.0001).

CONCLUSIONS

CX3CL1-CX3CR1 is a novel inflammatory adipose chemokine system that modulates monocyte adhesion to adipocytes and is associated with obesity, insulin resistance, and type 2 diabetes. These data provide support for CX3CL1 as a diagnostic and therapeutic target in cardiometabolic disease.Adipose tissue inflammation plays a central role in obesity-related metabolic and cardiovascular complications, including type 2 diabetes (1). A key event in adipose inflammation is recruitment of leukocytes (2–4), which produces local inflammatory signaling molecules creating a feed-forward cycle of adipose and systemic inflammation and insulin resistance. Upregulation of adipose chemokines has emerged as an important mechanism in leukocyte recruitment, early adipose inflammation, and insulin resistance in obesity. In particular, monocyte chemotactic protein-1 (MCP-1 or CCL2) and its receptor CCR2 are significant contributors to adipose macrophage recruitment and insulin resistance in obesity (3,5). However, adverse metabolic consequences are only partially attenuated in CCL2- or CCR2-deficient mice, suggesting involvement of additional chemokine pathways (6). Several other CC and CXC chemokines implicated in recruitment of inflammatory T-cells and monocytes (4), including CCL5 (also called RANTES [regulated on activation, normal T-cell expressed and secreted]), CXCL8 (interleukin-8) and CXCL 10 (interferon γ-induced protein) are increased in obesity in rodents (7). However, which chemokines play a causal role in human adipose inflammation and its metabolic complications remains uncertain.Fractalkine (CX3CL1), a chemokine that signals through a single known receptor (CX3CR1), is implicated in recruitment and adhesion of both monocytes and T-cells in atherosclerosis, rheumatologic disorders, and HIV-1 (8). CX3CL1, the sole CX3C chemokine, is unique among chemokines in having both soluble and transmembrane forms, the latter of which mediates firm cell adhesion (9). Many leukocyte subtypes, in particular monocytes, T-cells, and NK cells, express CX3CR1 (9,10), a G-protein–coupled receptor that promotes leukocyte activation and survival (11). In fact, CX3CR1 is required for vascular recruitment of inflammatory monocytes and development of macrophage-rich atherosclerotic lesions (12,13). Knowledge of CX3CL1’s role in adipose biology is limited but recent data suggest that CX3CL1 is expressed in adipocytes and that CX3CR1 signaling in macrophages is downregulated by peroxisome proliferator–activated receptor (PPAR)-γ agonists (14). Furthermore, the fact that manipulation of the CX3CL1/CX3CR1 system can modulate chronic inflammatory diseases, including atherosclerosis, independent of CCL2/CCR2 (15) suggests that this may also occur in adipose inflammation and its complications.Recently, using microarray of adipose mRNA during experimental endotoxemia, we found that CX3CL1 is one of several genes markedly upregulated in human adipose by in vivo inflammation (16). Here, we define CX3CL1 as a novel inflammatory adipose chemokine in humans. First, adipose CX3CL1 is increased in obesity as well as in evoked adipose inflammation. Second, CX3CL1 promotes monocyte adhesion to human adipocytes. Third, genetic variation in CX3CR1 is associated with metabolic traits in humans while plasma CX3CL1 levels are higher in type 2 patients with diabetes compared with control subjects.