14 research outputs found
Quale modello per la Vita Nova?
No full textThe author reflects on the way in which the various literary models highlighted by critics are compatible with the structure of the Vita Nova. Further, he suggests that it is worth considering the influence that the layout of the manuscripts of those works intended for study may have exerted on Dante’s prosimetru
Full-length transcript amplification and sequencing as universal method to test mRNA integrity and biallelic expression in mismatch repair genes
No full textIsoenergetic feeding of low carbohydrate-high fat diets does not increase brown adipose tissue thermogenic capacity in rats
No full textLow-carbohydrate, high-fat (LC-HF) diets are popular for inducing weight loss in overweighed adults. Adaptive thermogenesis increased by specific effects of macronutrients on energy expenditure has been postulated to induce this weight loss. We studied brown adipose tissue (BAT) morphology and function following exposure to different LC-HF diets.
Methods:
Male Wistar rats were fed a standard control diet ad libitum or pair-fed isoenergetic amounts of three experimental diets for 4 weeks. The diets had the following macronutrient composition (% metabolizable energy: carbohydrates, fat, protein): control (64.3/16.7/19), LC-HF-low protein (LC-HF-LP, 1.7/92.8/5.5), LC-HF-normal-protein (LC-HF-NP, 2.2/78.7/19.1), and a high fat diet with carbohydrates (“high fat”, 19.4/61.9/18.7).
Results:
Body weight gain was reduced in all pair-fed experimental groups as compared to rats fed the control diet, with more pronounced effect in rats on LC-HF diets than on the high fat diet with carbohydrates. High fat diets increased expression of PGC1α and ADRB3 in BAT indicating higher SNS outflow. However, UCP1 mRNA expression and expression of UCP1 assessed by immunohistochemistry was not different between diet groups. In accordance, analysis of mitochondrial function in-vitro by extracellular flux analyser (Seahorse Bioscience) and measurement of inducible thermogenesis in vivo (primary endpoint), explored by indirect calorimetry following norepinephrine injection, did not show significant differences between groups. Histology of BAT revealed increased lipid droplet size in rats fed the high-fat diet and both LC-HF diets.
Conclusion:
All experimental diets upregulated expression of genes which are indicative for increased BAT activity. However, the functional measurements in vivo revealed no increase of inducible BAT thermogenesis. This indicates that lower body weight gain with LC-HF diets and a high fat diet in a pair-feeding setting is not caused by increased adaptive thermogenesis in BAT
Respirometry.
No full text<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
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.
No full text<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
Body core temperature in rats fed control or experimental diets (LC-HF-LP, LC-HF-NP, high fat).
No full text<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
Weight of interscapular brown adipose tissue (iBAT) in rats fed control or experimental diets (LC-HF-LP, LC-HF-NP, high fat).
No full text<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).
No full text<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<sup>0.75</sup>. 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
PRKACA Somatic Mutations Are Rare Findings in Aldosterone-Producing Adenomas.
No full textSomatic mutations have been found causative for endocrine autonomy in aldosterone-producing adenomas (APAs). Whereas mutations of PRKACA (catalytic subunit of protein kinase A) have been identified in cortisol-producing adenomas, the presence of PRKACA variants in APAs is unknown, especially in those that display cosecretion of cortisol.The objective of the study was to investigate PRKACA somatic variants identified in APA cases.Identification of PRKACA somatic variants in APAs by whole-exome sequencing followed by in vitro analysis of the enzymatic activity of PRKACA variants and functional characterization by double immunofluorescence of CYP11B2 and CYP11B1 expression in the corresponding tumor tissues.APA tissues were collected from 122 patients who underwent unilateral adrenalectomy for primary aldosteronism between 2005 and 2015 at a single institution.PRKACA somatic mutations were identified in two APA cases (1.6%). One APA carried a newly identified p.His88Asp variant, whereas in a second case, a p.Leu206Arg mutation was found, previously described only in cortisol-producing adenomas with overt Cushing's syndrome. Functional analysis showed that the p.His88Asp variant was not associated with gain of function. Although CYP11B2 was strongly expressed in the p.His88Asp-mutated APA, the p.Leu206Arg carrying APA predominantly expressed CYP11B1. Accordingly, biochemical Cushing's syndrome was present only in the patient with the p.Leu206Arg mutation. After adrenalectomy, both patients improved with a reduced number of antihypertensive medications and normalized serum potassium levels.We describe for the first time PRKACA mutations as rare findings associated with unilateral primary aldosteronism. As cortisol cosecretion occurs in a subgroup of APAs, other molecular mechanisms are likely to exist
Quantitative real time PCR in iBAT following control or experimental diets (LC-HF-LP, LC-HF-NP, high fat).
No full text<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
