Adipose tissue ATGL modifies the cardiac lipidome in pressure-overload-induced left ventricular failure

Adipose tissue lipolysis occurs during the development of heart failure as a consequence of chronic adrenergic stimulation. However, the impact of enhanced adipose triacylglycerol hydrolysis mediated by adipose triglyceride lipase (ATGL) on cardiac function is unclear. To investigate the role of adipose tissue lipolysis during heart failure, we generated mice with tissue-specific deletion of ATGL (atATGL-KO). atATGL-KO mice were subjected to transverse aortic constriction (TAC) to induce pressure-mediated cardiac failure. The cardiac mouse lipidome and the human plasma lipidome from healthy controls (n = 10) and patients with systolic heart failure (HFrEF, n = 13) were analyzed by MS-based shotgun lipidomics. TAC-induced increases in left ventricular mass (LVM) and diastolic LV inner diameter were significantly attenuated in atATGL-KO mice compared to wild type (wt) -mice. More importantly, atATGL-KO mice were protected against TAC-induced systolic LV failure. Perturbation of lipolysis in the adipose tissue of atATGL-KO mice resulted in the prevention of the major cardiac lipidome changes observed after TAC in wt-mice. Profound changes occurred in the lipid class of phosphatidylethanolamines (PE) in which multiple PE-species were markedly induced in failing wt-hearts, which was attenuated in atATGL-KO hearts. Moreover, selected heart failure-induced PE species in mouse hearts were also induced in plasma samples from patients with chronic heart failure. TAC-induced cardiac PE induction resulted in decreased PC/PE-species ratios associated with increased apoptotic marker expression in failing wt-hearts, a process absent in atATGL-KO hearts. Perturbation of adipose tissue lipolysis by ATGL-deficiency ameliorated pressure-induced heart failure and the potentially deleterious cardiac lipidome changes that accompany this pathological process, namely the induction of specific PE species. Non-cardiac ATGL-mediated modulation of the cardiac lipidome may play an important role in the pathogenesis of chronic heart failure

Influence of organic versus inorganic dietary selenium supplementation on the concentration of selenium in colostrum, milk and blood of beef cows

<p>Abstract</p> <p>Background</p> <p>Selenium (Se) is important for the postnatal development of the calf. In the first weeks of life, milk is the only source of Se for the calf and insufficient level of Se in the milk may lead to Se deficiency. Maternal Se supplementation is used to prevent this.</p> <p>We investigated the effect of dietary Se-enriched yeast (SY) or sodium selenite (SS) supplements on selected blood parameters and on Se concentrations in the blood, colostrum, and milk of Se-deficient Charolais cows.</p> <p>Methods</p> <p>Cows in late pregnancy received a mineral premix with Se (SS or SY, 50 mg Se per kg premix) or without Se (control – C). Supplementation was initiated 6 weeks before expected calving. Blood and colostrum samples were taken from the cows that had just calved (Colostral period). Additional samples were taken around 2 weeks (milk) and 5 weeks (milk and blood) after calving corresponding to Se supplementation for 6 and 12 weeks, respectively (Lactation period) for Se, biochemical and haematological analyses.</p> <p>Results</p> <p>Colostral period. Se concentrations in whole blood and colostrum on day 1 <it>post partum </it>and in colostrum on day 3 <it>post partum </it>were 93.0, 72.9, and 47.5 μg/L in the SY group; 68.0, 56.0 and 18.8 μg/L in the SS group; and 35.1, 27.3 and 10.5 μg/L in the C group, respectively. Differences among all the groups were significant (<it>P </it>< 0.01) at each sampling, just as the colostrum Se content decreases were from day 1 to day 3 in each group. The relatively smallest decrease in colostrum Se concentration was found in the SY group (<it>P </it>< 0.01).</p> <p>Lactation period. The mean Se concentrations in milk in weeks 6 and 12 of supplementation were 20.4 and 19.6 μg/L in the SY group, 8.3 and 11.9 μg/L in the SS group, and 6.9 and 6.6 μg/L in the C group, respectively. The values only differed significantly in the SS group (<it>P </it>< 0.05). The Se concentrations in the blood were similar to those of cows examined on the day of calving. The levels of glutathione peroxidase (GSH-Px) activity were 364.70, 283.82 and 187.46 μkat/L in the SY, SS, and C groups, respectively. This was the only significantly variable biochemical and haematological parameter.</p> <p>Conclusion</p> <p>Se-enriched yeast was much more effective than sodium selenite in increasing the concentration of Se in the blood, colostrum and milk, as well as the GSH-Px activity.</p

Comparison of Multivendor Single-Voxel MR Spectroscopy Data Acquired in Healthy Brain at 26 Sites

Background: The hardware and software differences between MR vendors and individual sites influence the quantification of MR spectroscopy data. An analysis of a large data set may help to better understand sources of the total variance in quantified metabolite levels. Purpose: To compare multisite quantitative brain MR spectroscopy data acquired in healthy participants at 26 sites by using the vendor-supplied single-voxel point-resolved spectroscopy (PRESS) sequence. Materials and Methods: An MR spectroscopy protocol to acquire short-echo-time PRESS data from the midparietal region of the brainwas disseminated to 26 research sites operating 3.0-T MR scanners from three different vendors. In this prospective study, healthy participants were scanned between July 2016 and December 2017. Data were analyzed by using software with simulated basis sets customized for each vendor implementation. The proportion of total variance attributed to vendor-, site-, and participant-related effects was estimated by using a linear mixed-effects model. P values were derived through parametric bootstrapping of the linearmixed-effects models (denoted P-boot). Results: In total, 296 participants (mean age, 26 years +/- 4.6; 155 women and 141 men) were scanned. Good-quality data were recorded from all sites, as evidenced by a consistent linewidth of N-acetylaspartate (range, 4.4-5.0 Hz), signal-to-noise ratio (range,174-289), and low Cramer-Rao lower bounds ( .90), N-acetylaspartate and N-acetylaspartylglutamate (P-boot =.13), or glutamate and glutamine (P-boot =.11). Among the smaller resonances, no vendor effects were found for ascorbate (P-boot =.08), aspartate (P-boot >.90), glutathione (P-boot > .90), or lactate (P-boot =.28). Conclusion: Multisite multivendor single-voxel MR spectroscopy studies performed at 3.0 T can yield results that are coherent across vendors, provided that vendor differences in pulse sequence implementation are accounted for in data analysis. However, the site related effects on variability were more profound and suggest the need for further standardization of spectroscopic protocols. (C) RSNA, 202

Deletion of ATGL in adipose tissue attenuates pressure overload-induced LV failure.

<p><b>A:</b> Representative images of the hearts. <b>B</b>: Heart weight (HW)/ body weight (BW) ratio (mean and SEM, n = 5–6). <b>C</b>: Representative microscopic cross-sections of the hearts stained with hematoxylin/ eosin (H/E). <b>D</b>: Myocardial area, calculated based on microscopic sections of heart tissue, stained with H/E, analogue to the images presented in C (mean and SEM, n = 5–6). <b>E</b>: Representative M-Mode images of the echocardiographic analysis. <b>F-J</b>: cardiac echocardiographic analysis of mice (mean and SEM, n = 7): <b>F</b>: Left-ventricular mass (LVM). <b>G</b>: LVM relative to tibia length (LVM/TL). <b>H</b>: Left-ventricular internal diameter in diastole (LVID-d). <b>I</b>: Ejection fraction [%] (EF). <b>J</b>: Fractional shortening [%] (FS). <b>K</b>: Analysis of mRNA expression of beta-cardiac myosin heavy chain isogene (βMHCH), qRT-PCR studies were carried out using total RNA isolated from LV tissue. Data are presented as x-fold over wt-sham mice (mean and SEM, n = 5–6). <b>L</b>: Representative microscopic cross-sections of the hearts stained with picrosirius red. <b>M</b>: Representative high magnification images from picrosirius red-stained sections. <b>N:</b> Cardiac fibrosis calculated based on microscopic sections of the heart tissue, stained with picrosirius red, analogue to the images presented in L: 0 = no fibrosis, 1 = mild fibrosis, 2 = moderate fibrosis, 3 = severe fibrosis (mean and SEM, n = 5–6). <b>O and P:</b> Analysis of mRNA expression of collagen (Col) 1a1 (O) and Col3 (P), qRT-PCR studies were carried out using total RNA isolated from LV tissue. Data are presented as x-fold over wt-sham mice (mean and SEM, n = 5–6).*p<0.05 vs. wt sham, **p<0.01 vs. wt sham, ***p<0.001 vs. wt sham, ****p<0.0001 vs. wt sham, $p<0.05 vs. wt TAC,$ $p<0.01 vs. wt TAC,$ $$ p<0.001 vs. wt TAC, $$ $$ p<0.0001 vs. wt TAC, ##p<0.01 vs. atATGL-KO sham; 2-way ANOVA (Bonferroni post-test).</p

Selected lipid species are altered in plasma samples from patients with HFrEF MS-based shotgun lipidomics analysis of human plasma samples from HFrEF-patients (n = 13) and non-HFrEF controls (n = 10).

<p>A: Box plots show distribution of total mole percent values for each lipid class corrected for age and BMI (see supplementary methods). To test for differential changes a Mann-Whitney U test between HFrEF-patients and controls was performed. Adjusted p-values are indicated: *p<0.05, **p<0.01, **p<0.001. B: Estimated mean log2 fold change (HFrEF vs. control) vs. estimated mean mole percent of lipid species in the control group (see supplementary methods). Triangles represent significantly changed lipid species (FDR adjusted p-value < 0.1 and absolute value of log2-fold change ≥ 0.5), bubbles show those which are not significantly changed, size indicates log-transformed adjusted p-values. C: Bar graph shows the estimated log2-fold change (HFrEF vs. control) ± regression standard error of differentially changed lipid species (see B.). Colors represent log10 adjusted p-values as indicated. Lipid classes: Cer: ceramide, DAG: diacylglycerol, LPC: lyso-phosphatidylcholine, LPE: lyso-phosphatidylethanolamine, PC: phosphatidylcholine, PC O-: phosphatidylcholine-ether, PE: phosphatidylethanolamine, PE O-: phosphatidylethanolamine-ether, PI: phosphatidylinositol, SE: sterol ester, SM: sphingomyelin, ST: sterols, TAG: triacylglycerol.</p

Induction of pressure-mediated cardiac PE species is attenuated in atATGL-KO mice.

<p>MS-based shotgun lipidomics analysis of heart tissue samples (LV) isolated 11 weeks after intervention (sham or TAC) from wild-type (wt) or atATGL-KO mice. Lipid class denotations see <a href="http://www.plosgenetics.org/article/info:doi/10.1371/journal.pgen.1007171#pgen.1007171.g003" target="_blank">Fig 3</a>. <b>A-F:</b> Mean log2-fold change (TAC vs. sham) vs. mean mole percent of lipid species. Triangles represent significantly changed lipid species (FDR adjusted p-value < 0.1 and absolute value of log2-fold change ≥ 0.5); bubbles represent those which are not significantly changed; size indicates log-transformed adjusted p-values. <b>A, C, E:</b> wt-mice. <b>B, D, F:</b> atATGL-KO-mice. <b>G+H:</b> Significantly changed PC-PE ratios of matched FAs in wt-mice (<b>G</b>) or atATGL-KO mice (<b>H</b>). The mean ratio ± SEM is shown on a logarithmic scale, Mann-Whitney U test for TAC vs. sham: *p<0.05, **p<0.01 (adjusted) in wt-mice, no significant changes were found in at ATGL-KO mice. <b>I:</b> upper panels: WB analysis of heart lysates using antibodies against cleaved caspase 3 and Bcl-associated X protein (Bax); lower panel: WB analysis of HL-1 cardiomyocytes lysates from cells stimulated with vehicle (Veh) or fatty acid (FA) mix (C16:0, C18:1,C18:2 in different concentrations) using antibodies against cleaved caspase 3; loading control: β–actin, glyceraldehyde 3-phosphate dehydrogenase (GAPDH).</p

Metabolic phenotype and analysis of blood FA-profile in wt- and atATGL-KO mice.

<p><b>A:</b> Intraperitoneal Glucose Tolerance Test (ipGTT), (n = 4–6, 2-way ANOVA (Bonferroni post-test) from AUC). <b>B</b>: Area under the curve (AUC) of ipGTT, (mean and SEM, n = 4–6, 2-way ANOVA (Bonferroni posttest)). <b>C</b>: Insulin Tolerance Test (ITT), (n = 7–8, 2-way ANOVA (Bonferroni posttest) from AUC). <b>D</b>: Area under the curve (AUC) of ITT, (mean and SEM, n = 7–8, 2-way ANOVA (Bonferroni posttest)). <b>E</b>: Profile of selected serum FAs in TAC-operated mice analyzed by rapid resolution HPLC/ Tandem MS. <b>F</b>: Serum level of non-esterified fatty acids (NEFAs) in wt-TAC and atATGL-KO-TAC mice. <b>G</b>: Serum level of triacylglyerols (TAGs) in wt-TAC and atATGL-KO-TAC mice. (mean and SEM, n = 5, or as indicated, unpaired t-test). ***p<0.001 vs. wt sham, $p<0.05 vs. wt TAC,$ $$ p<0.001 vs. wt TAC, $$ $$ p<0.001 vs. wt TAC.</p

Comparison of Multivendor Single-Voxel MR Spectroscopy Data Acquired in Healthy Brain at 26 Sites

Background: The hardware and software differences between MR vendors and individual sites influence the quantification of MR spectroscopy data. An analysis of a large data set may help to better understand sources of the total variance in quantified metabolite levels. Purpose: To compare multisite quantitative brain MR spectroscopy data acquired in healthy participants at 26 sites by using the vendor-supplied single-voxel point-resolved spectroscopy (PRESS) sequence. Materials and Methods: An MR spectroscopy protocol to acquire short-echo-time PRESS data from the midparietal region of the brain was disseminated to 26 research sites operating 3.0-T MR scanners from three different vendors. In this prospective study, healthy participants were scanned between July 2016 and December 2017. Data were analyzed by using software with simulated basis sets customized for each vendor implementation. The proportion of total variance attributed to vendor-, site-, and participant-related effects was estimated by using a linear mixed-effects model. P values were derived through parametric bootstrapping of the linear mixed-effects models (denoted Pboot). Results: In total, 296 participants (mean age, 26 years ± 4.6; 155 women and 141 men) were scanned. Good-quality data were recorded from all sites, as evidenced by a consistent linewidth of N-acetylaspartate (range, 4.4–5.0 Hz), signal-to-noise ratio (range, 174–289), and low Cramér-Rao lower bounds (≤5%) for all of the major metabolites. Among the major metabolites, no vendor effects were found for levels of myo-inositol (Pboot > .90), N-acetylaspartate and N-acetylaspartylglutamate (Pboot = .13), or glutamate and glutamine (Pboot = .11). Among the smaller resonances, no vendor effects were found for ascorbate (Pboot = .08), aspartate (Pboot > .90), glutathione (Pboot > .90), or lactate (Pboot = .28). Conclusion: Multisite multivendor single-voxel MR spectroscopy studies performed at 3.0 T can yield results that are coherent across vendors, provided that vendor differences in pulse sequence implementation are accounted for in data analysis. However, the site-related effects on variability were more profound and suggest the need for further standardization of spectroscopic protocols