Additional file 1: Table S1. of Design of the Growth hormone deficiency and Efficacy of Treatment (GET) score and non-interventional proof of concept study

GET score point allocation for HRQoL parameters (which comprise 50 of the total of 100 points of the GET score). Table S2 Example of a calculation of SF-36 GET score component points. Table S3 GET score point allocation for somatic parameters (which comprise 50 of the total of 100 points of the GET score).Table S4 Example of calculation of GET score including SF-36 subtotal and addition to other components of the score. GET score calculation. Table S5 Example of calculation of an adjusted GET score due to a missing value (DOCX 39 kb

Weight curves of rats on the control diet (circles, fed ad libitum), compared to the three experimental diets (iso-energetic pair feeding): low-carbohydrate high-fat, low-protein diet (LC-HF-LP; squares), low-carbohydrate high-fat, normal-protein diet (LC-HF-NP; upward triangles) or high fat diet (downward triangles); p<0.0005, repeated measurements ANOVA.

<p>The cumulative average weight gain for each group during the four week feeding period is given in the insert, analyzed by global one-way ANOVA and Dunnett tests for pairwise comparison vs. control: * p<0.05, *** p<0.001. Data are shown as means±SEM; n = 7 animals/group.</p

Respirometry.

<p>Measurement of mitochondrial oxygen consumption using microplate based extracellular flux analyzer in tissues from rats fed the control or experimental diets (LC-HF-LP, LC-HF-NP). A: brown adipocyte mitochondria. B: muscle mitochondria. Stage II, basal; stage III, ADP; stage IVo, oligomycin. N = 3/group.</p

Primer sequences (5′-3′).

<p>Primer sequences (5′-3′).</p

Weight of interscapular brown adipose tissue (iBAT) in rats fed control or experimental diets (LC-HF-LP, LC-HF-NP, high fat).

<p>A: total weight (absolute); B: percentage of body weight (relative). Data are shown as means±SEM, n = 7/group; analyzed by global one-way ANOVA and Dunnett tests for pairwise comparison vs. control. * p<0.05, ** p<0.01 vs. control.</p

In-vivo measurement of maximal adaptive thermogenic capacity in rats fed the control or experimental diets (LC-HF-LP, LC-HF-NP, high fat).

<p>A: respiratory quotient after feeding control or experimental diets for 4 weeks: basal and following norepinephrine injection. B: Basal and norepinephrine stimulated energy expenditure (EE), expressed as kcal/h/kg BW^0.75. The time points 0 min and 36 min are not given, as cages had to be opened for animal handling and measures of RQ and EE are not reliable at these time points. The insert shows the mean EE of timepoints 77, 84 and 91 min. Data are shown as means±SEM; n = 7 (control), 6 (LC-HF-LP), 5 (LC-HF-NP), 7 (high fat). Statistical analysis was performed on the average of three time points at baseline and during the plateau phase after NE injection, respectively, using global one-way ANOVA and Dunnett tests for pairwise comparison vs. control.</p

Body core temperature in rats fed control or experimental diets (LC-HF-LP, LC-HF-NP, high fat).

<p>A: morning, B: evening. Data are shown as means±SEM; n = 7/group; analyzed by global one-way ANOVA and by Dunnett tests for pairwise comparison vs. control.</p

DataSheet_1_Steroid profiling using liquid chromatography mass spectrometry during adrenal vein sampling in patients with primary bilateral macronodular adrenocortical hyperplasia.pdf

IntroductionAdrenal vein sampling (AVS) is not a routine procedure in patients with primary bilateral macronodular adrenocortical hyperplasia (PBMAH), but has been used to determine lateralization of cortisol secretion in order to guide decision of unilateral adrenalectomy. Our aim was to characterize the steroid fingerprints in AVS samples of patients with PBMAH and hypercortisolism and to identify a reference hormone for AVS interpretation.MethodRetrospectively, we included 17 patients with PBMAH from the German Cushing’s registry who underwent AVS. 15 steroids were quantified in AVS and peripheral blood samples using LC-MS/MS. We calculated lateralization indices and conversion ratios indicative of steroidogenic enzyme activity to elucidate differences between individual adrenal steroidomes and in steroidogenic pathways.ResultsAdrenal volume was negatively correlated with peripheral cortisone (r=0.62, pConclusionSteroid profiling by LC-MS/MS led us to select DHEA as a candidate reference hormone for cortisol secretion. Lateralization and different steroid ratios showed that each steroid and all three steroidogenic pathways may be affected in PBMAH patients. In patients with germline ARMC5 mutations, the androgen pathway was particularly dysregulated.</p

Quantitative real time PCR in iBAT following control or experimental diets (LC-HF-LP, LC-HF-NP, high fat).

<p>ADRB3: β3 adrenergic receptor. Gene expression was normalized to 18 S rRNA. Data are shown as means±SEM; n = 7/group; analyzed by global one-way ANOVA and by Dunnett tests for pairwise comparison vs. control. * p<0.05, ** p<0.01 vs. control.</p

Analysis of body weight, contents of DNA, RNA and protein in <i>M. quadriceps femoris</i> of Fzt:DU (29 weeks) and DU6P (11 and 29 weeks) female mice (n = 5).

<p>a,b - different superscripts indicate significant differences (p<0.05);</p>*<p>- significantly different if compared to 11-week DU6P or Fzt:DU, respectively as evaluated using the Wilcoxon-signed rank test.</p><p>Furthermore the non-polysomal and polysomal RNA fraction in <i>M. quadriceps femoris</i> of Fzt:DU (29 weeks) and DU6P (11 and 29 weeks) female mice (n = 4) was analysed.</p