Increased 90-kDa ribosomal S6 kinase (Rsk) activity is protective against mutant huntingtin toxicity

Uncovering the underlying genetic component of any disease is key to the understanding of its pathophysiology and may open new avenues for development of therapeutic strategies and biomarkers. In the past several years, there has been an explosion of genome-wide association studies (GWAS) resulting in the discovery of novel candidate genes conferring risk for complex diseases, including neurodegenerative diseases. Despite this success, there still remains a substantial genetic component for many complex traits and conditions that is unexplained by the GWAS findings. Additionally, in many cases, the mechanism of action of the newly discovered disease risk variants is not inherently obvious. Furthermore, a genetic region with multiple genes may be identified via GWAS, making it difficult to discern the true disease risk gene. Several alternative approaches are proposed to overcome these potential shortcomings of GWAS, including the use of quantitative, biologically relevant phenotypes. Gene expression levels represent an important class of endophenotypes. Genetic linkage and association studies that utilize gene expression levels as endophenotypes determined that the expression levels of many genes are under genetic influence. This led to the postulate that there may exist many genetic variants that confer disease risk via modifying gene expression levels. Results from the handful of genetic studies which assess gene expression level endophenotypes in conjunction with disease risk suggest that this combined phenotype approach may both increase the power for gene discovery and lead to an enhanced understanding of their mode of action. This review summarizes the evidence in support of gene expression levels as promising endophenotypes in the discovery and characterization of novel candidate genes for complex diseases, which may also represent a novel approach in the genetic studies of Alzheimer's and other neurodegenerative diseases.     

Secretion of a major phosphorylated glycoprotein by hepatocytes. Characterization of specific antibodies and investigations of the processing, excretion kinetics, and phosphorylation

Isolated rat hepatocytes secreted a major phosphorylated glycoprotein (PP63) with apparent Mr = 63,000 and isoelectric point ranging from 4.8 to 5.3. Specific antibodies were raised in a rabbit using material obtained from plasma as an antigen. The biosynthesis of PP63 was studied in vitro in a cell-free system and in intact hepatocytes incubated with or without tunicamycin. The mRNA translation product had a Mr = 43,000 and was of the same size as the major unglycosylated precursor found in intact cells. This precursor was rapidly processed into two major intracellular forms of Mr = 53,000 and 56,000. These species were insensitive to neuraminidase but susceptible to endoglycosidase H, indicating that they contained oligosaccharide side chains of the high mannose-type. Terminal glycosylation gave rise to the mature Mr = 63,000 protein that contained sialic acid and fucose. This species represented the exportable form of the protein and was the only one to be phosphorylated. The charge heterogeneity observed for the mature protein already existed in all the precursors, indicating that it could not be ascribed to sialylation or to phosphorylation. However, these covalent modifications were mainly responsible for the acidic character of PP63. PP63 secretion was altered by tunicamycin. Pulse-chase experiments showed that the phosphorylated glycoprotein was secreted according to kinetics similar to that described for other liver glycoprotein, with slower kinetics than albumin. Permanent phosphorylation did not appear mandatory for excretion since the dephosphorylated PP63 was excreted with an efficacy comparable to that of the phosphorylated protein. Phosphorylation of PP63 was shown to occur on a single tryptic peptide, at a serine residue.     

Forest characteristics and population structure of Glyptostrobus pensilis,a globally endangered relict species of southeastern China

The Chinese water pine Glyptostrobus pensilis is the sole surviving species of the genus Glyptostrobus. It is endemic to southern China, central Vietnam, and eastern Laos, and today it is nearly extinct in the wild. Forest community characteristics and population structure of G. pensilis in China have remained un-known up to now. We investigated six swamp forest stands and analyzed their forest community characteristics (i.e. vertical stratification, species composition, and diversity) and population structure, including the frequency distribution of DBH (diameter at breast height) and age-classes as found in Fujian Province, southeastern China. The vertical stratifications of all the forest stands were rather simple. The remaining wild specimens ranged from roughly 15 to some 357 years for an average of ca. 85 years, with only a few individuals less than 20 years old. Compared with the stands and populations of G. pensilis in Vietnam, the taxonomic compositions of the stands in the two regions were different, except for the dominant species-G. pensilis. The Shannon-Wiener index showed the overstory of each stand had much lower diversity (0.26 on average) in Fujian Province than that (1.97 on average) in Vietnam, whereas the diversity indices were about the same (around 2.41) for the understories in the two regions. Furthermore, we discovered 18 G. pensilis seedlings at the study sites in Fujian Province. This discovery demonstrates that G. pensilis regeneration is extremely poor and its populations are declining, although these populations are rela-tively healthier than those in Vietnam.     

Tennis Play Intensity Distribution and Relation with Aerobic Fitness in Competitive Players

The aims of this study were (i) to describe the relative intensity of simulated tennis play based on the cumulative time spent in three metabolic intensity zones, and (ii) to determine the relationships between this play intensity distribution and the aerobic fitness of a group of competitive players. 20 male players of advanced to elite level (ITN) performed an incremental on-court specific endurance tennis test to exhaustion to determine maximal oxygen uptake (VO_2max) and the first and second ventilatory thresholds (VT₁, VT₂). Ventilatory and gas exchange parameters were monitored using a telemetric portable gas analyser (K4 b², Cosmed, Rome, Italy). Two weeks later the participants played a simulated tennis set against an opponent of similar level. Intensity zones (1: low, 2: moderate, and 3: high) were delimited by the individual VO₂ values corresponding to VT₁ and VT₂, and expressed as percentage of maximum VO₂ and heart rate. When expressed relative to VO_2max, percentage of playing time in zone 1 (77 ± 25%) was significantly higher (p < 0.001) than in zone 2 (20 ± 21%) and zone 3 (3 ± 5%). Moderate to high positive correlations were found between VT₁, VT₂ and VO_2max, and the percentage of playing time spent in zone 1 (r = 0.68–0.75), as well as low to high inverse correlations between the metabolic variables and the percentage of time spent in zone 2 and 3 (r = -0.49–0.75). Players with better aerobic fitness play at relatively lower intensities. We conclude that players spent more than 75% of the time in their low-intensity zone, with less than 25% of the time spent at moderate to high intensities. Aerobic fitness appears to determine the metabolic intensity that players can sustain throughout the game.     

Modulation of adenosine A1 and A2A receptors in C6 glioma cells during hypoxia: involvement of endogenous adenosine

During hypoxia, extracellular adenosine levels are increased to prevent cell damage, playing a neuroprotective role mainly through adenosine A₁ receptors. The aim of the present study was to analyze the effect of hypoxia in both adenosine A₁ and A_2A receptors endogenously expressed in C6 glioma cells. Two hours of hypoxia (5% O₂) caused a significant decrease in adenosine A₁ receptors. The same effect was observed at 6 h and 24 h of hypoxia. However, adenosine A_2A receptors were significantly increased at the same times. These effects were not due to hypoxia-induced alterations in cells number or viability. Changes in receptor density were not associated with variations in the rate of gene expression. Furthermore, hypoxia did not alter HIF-1α expression in C6 cells. However, HIF-3α, CREB and CREM were decreased. Adenosine A₁ and A_2A receptor density in normoxic C6 cells treated with adenosine for 2, 6 and 24 h was similar to that observed in cells after oxygen deprivation. When C6 cells were subjected to hypoxia in the presence of adenosine deaminase, the density of receptors was not significantly modulated. Moreover, DPCPX, an A₁ receptor antagonist, blocked the effects of hypoxia on these receptors, while ZM241385, an A_2A receptor antagonist, was unable to prevent these changes. These results suggest that moderate hypoxia modulates adenosine receptors and cAMP response elements in glial cells, through a mechanism in which endogenous adenosine and tonic A₁ receptor activation is involved.     

Intestinal gluconeogenesis is a key factor for early metabolic changes after gastric bypass but not after gastric lap-band in mice

Unlike the adjustable gastric banding procedure (AGB), Roux-en-Y gastric bypass surgery (RYGBP) in humans has an intriguing effect: a rapid and substantial control of type 2 diabetes mellitus (T2DM). We performed gastric lap-band (GLB) and entero-gastro anastomosis (EGA) procedures in C57Bl6 mice that were fed a high-fat diet. The EGA procedure specifically reduced food intake and increased insulin sensitivity as measured by endogenous glucose production. Intestinal gluconeogenesis increased after the EGA procedure, but not after gastric banding. All EGA effects were abolished in GLUT-2 knockout mice and in mice with portal vein denervation. We thus provide mechanistic evidence that the beneficial effects of the EGA procedure on food intake and glucose homeostasis involve intestinal gluconeogenesis and its detection via a GLUT-2 and hepatoportal sensor pathway.     

Adaptability of the digestive function according to age at weaning in the rabbit: I. Effect on feed intake and digestive functionality

The functional adaptability of the digestive system to the level of feed intake was investigated in the young rabbits by comparing two groups of 12 litters each, weaned at 21 (W21) or 35 (W35) days of age. From 14 days onwards, rabbits were fed a pelleted feed (NDF: 332 g/kg, CP: 177 g/kg, starch: 98 g/kg, as-fed basis). Until 49 days of age, the profile of digestive enzymes was weekly determined in the small intestinal content and mucosa, as well as caecal fermentation traits and fibrolytic activities. In the W21 group, the solid feed intake was increased by 57% between 21 and 35 days (P < 0.01), while the daily body growth was lower from 21 till 42 days (-17%, P < 0.05) when compared with the W35 group. Activities of enzymes of pancreatic origin were only scarcely influenced by the weaning age. In the W21 group, amylase activity tended to be lower at 28 days of age (-36%, P = 0.064), and trypsin activity was decreased by 31% at 49 days of age (P < 0.01). Lipase activity was similar in both weaning groups. Duodenal and jejunal activities of maltase and aminopeptidase N (APN) were higher on day 28 in the W21 group as compared with the W35 group (×1.4 to ×2.4, respectively, P < 0.05). On day 35, duodenal APN activity was twice as higher in the W21 group than in the W35 group (P < 0.01). In caecum, major differences between both weaning groups were observed at 28 days of age with a decrease in ammonia concentration (-43%, P < 0.01) in W21 compared with W35 rabbits. Conversely, the acetate proportion was 5% higher in the W21 group (P < 0.01) on day 28. In conclusion, the digestive tract of early-weaned rabbits showed some adaptative properties in response to nutritional environment changes, but they were insufficient to maintain their growth rate.     

Effect of organic and inorganic selenium sources in sow diets on colostrum production and piglet response to a poor sanitary environment after weaning

The objectives of this study were to determine the effect of the chemical form of selenium (Se) fed to sows (1) on production and immune quality of colostrum and (2) on piglet response to a deterioration of sanitary conditions after weaning. Twenty-two pregnant sows were assigned to receive a diet supplemented with 0.3 ppm Se from either sodium selenite (inorganic Se) or Se-enriched yeast (organic Se as Sel-Plex®; Alltech Inc., Nicholasville, KY, USA). Dietary treatments were applied during the last month of pregnancy and lactation. Blood samples were collected on sows before dietary treatment, on the day of weaning and 6 weeks later, and on three to five piglets within litters at birth, at weaning and 6 weeks post weaning. Whole blood was analysed for Se concentration. Colostrum samples were collected at 0, 3, 6 and 24 h post partum and milk samples on days 14 and 27 of lactation. Colostrum and milk were analysed for Se and immunoglobulin G (IgG) concentrations. At weaning, 40 pairs of littermate piglets were moved to rooms where sanitary conditions were good or purposely deteriorated. Piglets were reared individually and fed ad libitum. After 15 days, piglets and feed refusals were weighed and a blood sample was collected to measure plasma haptoglobin concentration. When sows were fed organic Se, Se concentrations were increased by 33% in colostrum (P < 0.05), 89% in milk (P < 0.001) and by 28% in whole blood of piglets at weaning (P < 0.001). Colostrum production during the 24 h after the onset of farrowing and IgG concentrations in colostrum and milk did not significantly differ between the two groups of sows. Weaned piglets reared in good sanitary conditions grew faster (P < 0.001) than piglets housed in poor conditions. Sanitary conditions did not influence mean plasma haptoglobin concentrations of piglets (P > 0.1). The source of Se fed to the dams did not influence piglet performance or haptoglobin concentrations after weaning. These findings confirm that, compared with inorganic Se, organic Se fed to the dam is better transferred to colostrum and milk, and consequently to piglets. They indicate that the Se source influences neither colostrum production nor IgG concentrations in colostrum, and that the higher Se contents of piglets does not limit the reduction of growth performance when weaning occurs in experimentally deteriorated sanitary conditions.