Plasma α-MSH levels in women with low body weight.

<p>(A) Daily mean values of plasma α-MSH levels in young women with anorexia nervosa (AN), constitutional thinness (CT) and in controls (Controls). Data represent means ± SEM of 4 independent experiments. ns, non significant; **, p<0.01. (B) Twelve-point circadian plasma levels of α-MSH in AN, CT and controls. The arrows indicate meal schedule. Each point represents mean ± SEM of 4 independent experiments. Significant point-by-point intergroup differences (p< 0.05): *, AN <i>vs</i> C; #, CT <i>vs</i> C; &, AN <i>vs</i> CT.</p

Plasma NPY levels in women with low body weight.

<p>(A) Daily mean values of plasma NPY levels in young women with anorexia nervosa (AN), constitutional thinness (CT) and in controls (Controls). Data represent means ± SEM of 4 independent experiments. ns, non significant. (B) Twelve-point circadian plasma levels of NPY in AN, CT and controls. The arrows indicate meal schedule. Each point represents mean ± SEM of 4 independent experiments. Significant point-by-point intergroup differences (p< 0.05): *, AN <i>vs</i> C; #, CT <i>vs</i> C; &, AN <i>vs</i> CT.</p

Antropometric and hormonal parameters in studied groups.

<p>a) AN <i>vs</i> C; b) AN <i>vs</i> CT; c) C <i>vs</i> CT</p><p>Antropometric and hormonal parameters in studied groups.</p

WE-14 plasma concentrations in healthy volunteers and patients with pheochromocytoma.

<p>Comparison of the diagnostic sensitivity of WE-14, CgA and EM66 assays.</p><p>*, median value of controls <i>vs</i> median value of patients, pheochromocytoma, paraganglioma, benign, malignant, sporadic or hereditary tumors. * <i>p</i><0.05, *** <i>p</i><0.001; <i>n</i>, number of individuals.</p

CgA levels in the plasma of patients with pheochromocytoma.

<p>(<b>A</b>) Scattergram of CgA concentrations in plasma samples of patients with pheochromocytoma before surgical removal of the tumor (preop, <b>+</b>, n = 37). (<b>B</b>) Distribution of CgA preoperative concentrations of patients reported in (A), depending on the adrenal (PHEO, ▴, n = 26) or extra-adrenal (PGL, ▾, n =  11) location of the tumor, the benign (•, n = 32) or malignant (▪, n = 5), and the sporadic (♦, n = 24) or hereditary (•, n = 13) nature of the neoplasms. The <i>grey zone</i> represents the cut-off level for the CgA test assay as indicated by the manufacturer. See legends of <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0088698#pone-0088698-g002" target="_blank">Figure 2</a> for other designations.</p

Cytochrome oxidase activity in cerebral areas of vehicle and everolimus-treated mice.

<p>Percentage of optical densities of cytochrome oxidase activity staining after everolimus treatment in the telencephalon, the diencephalon, the mesencephalon, the metencephalon, the myelecephalon and the cerebellum.</p><p>Data are means +SEM. (Student <i>t</i> test, *<i>p</i><.05). (a, area; ant, anterior; Diag, diagonal; horiz, horizontal; genic, geniculate; mesenc, mesencephalic; n, nucleus; ret, reticular; vert, vertical).</p><p>Cytochrome oxidase activity in cerebral areas of vehicle and everolimus-treated mice.</p

Characterization of WE-14 in plasma.

<p>(<b>A</b>) Semilogarithmic plots comparing competitive inhibition of antibody-bound ¹²⁵I-labeled Tyr₀-WE-14 by synthetic human WE-14 (•) and serial dilutions of plasma samples from healthy volunteer (▴), post-operative (▪) and preoperative (♦) patients with benign pheochromocytoma. (<b>B, C</b>) Reversed-phase HPLC analysis of WE-14 immunoreactivity in plasma samples from a healthy volunteer (B) and a patient with benign pheochromocytoma (C). The <i>bars</i> above the peaks indicate the elution position of synthetic human WE-14 (peak I) and its oxidized form (peak II) chromatographed the same day as the extracts. The <i>dashed line</i> shows the concentration of acetonitrile in the eluting solvent.</p

Evaluation of the Impact of the Cancer Therapy Everolimus on the Central Nervous System in Mice

<div><p>Cancer and treatments may induce cognitive impairments in cancer patients, and the causal link between chemotherapy and cognitive dysfunctions was recently validated in animal models. New cancer targeted therapies have become widely used, and their impact on brain functions and quality of life needs to be explored. We evaluated the impact of everolimus, an anticancer agent targeting the mTOR pathway, on cognitive functions, cerebral metabolism, and hippocampal cell proliferation/vascular density in mice. Adult mice received everolimus daily for 2 weeks, and behavioral tests were performed from 1 week after the last treatment. Everolimus-treated mice displayed a marked reduction in weight gain from the last day of the treatment period. <i>Ex vivo</i> analysis showed altered cytochrome oxidase activity in selective cerebral regions involved in energy balance, food intake, reward, learning and memory modulation, sleep/wake cycle regulation, and arousal. Like chemotherapy, everolimus did not alter emotional reactivity, learning and memory performances, but in contrast to chemotherapy, did not affect behavioral flexibility or reactivity to novelty. <i>In vivo</i> hippocampal neural cell proliferation and vascular density were also unchanged after everolimus treatments. In conclusion, two weeks daily everolimus treatment at the clinical dose did not evoke alteration of cognitive performances evaluated in hippocampal- and prefrontal cortex-dependent tasks that would persist at one to four weeks after the end of the treatment completion. However, acute everolimus treatment caused selective CO modifications without altering the mTOR effector P70S6 kinase in cerebral regions involved in feeding behavior and/or the sleep/wake cycle, at least in part under control of the solitary nucleus and the parasubthalamic region of the hypothalamus. Thus, this area may represent a key target for everolimus-mediating peripheral modifications, which has been previously associated with symptoms such as weight loss and fatigue.</p></div

WE-14 levels in the plasma of patients with pheochromocytoma.

<p>(A) Scattergram of WE-14 concentrations in plasma samples of healthy volunteers (controls, ×, n = 21), and patients with pheochromocytoma (+, n = 37). (<b>B</b>) Distribution of WE-14 preoperative concentrations in patients reported in (A), depending on the adrenal (PHEO, ▴, n = 26) or extra-adrenal (PGL, ▾, n =  11) location of the tumor, the benign (•, n = 32) or malignant (▪, n = 5), and the sporadic (♦, n = 24) or hereditary (•, n = 13) nature of the neoplasms. (C) Distribution of WE-14 levels (n = 12) in preoperative (•, preop) and postoperative patients for which the tumor was resected (○, postop). The <i>bars</i> represent the median value for each group. The <i>grey zone</i> corresponds to the distribution of control values and indicates the cut-off level for WE-14 test assay. The * symbol refers to statistical difference between median values of controls <i>vs</i> each group. <i>PHEO</i>, pheochromocytoma; <i>PGL</i>, paraganglioma; <i>post-op</i>, post-operative patients; <i>preop</i>, preoperative patients. <i>ns</i>, not significant. ***, <i>p</i><0.001; **, <i>p</i><0.01. The numbers refer to the gene mutation as follow : <i>1</i>, SDHD ; <i>2</i>, SDHB ; <i>3</i>, NF1 ; <i>4</i>, RET ; <i>5</i>, VHL.</p

Spatial learning and memory, and behavioral flexibility of mice treated with vehicle or everolimus in the Morris water maze test.

<p>(A) Motivation and visuo-motor abilities after vehicle or everolimus treatment (Student <i>t</i> test, <i>p</i>>.05). (B and C) During the training and retrieval (R) phases, latency (B) and distance crossed (C) after vehicle or everolimus treatment (ANOVA, Day effect ***<i>p</i><.001). (D) During the probe test, time spent by animals of both groups in the quadrant where the platform was located during the training phase (χ², ***<i>p</i><.001 <i>vs</i> chance). During the transfer phase, escape latency (E), distance crossed (F), and swimming speed (G) in vehicle- or everolimus-treated mice (ANOVA, Treatment effect *<i>p</i><.05, Trial effect *** <i>p</i><.001 followed by LSD post hoc ^##<i>p</i><.01). (H) During the 1st day of the transfer phase, time spent in the previously correct northwest quadrant after vehicle or everolimus treatment (Student <i>t</i> test, <i>p</i>>.05). Data are means +SEM.</p