Analysis of bioactive lipids in adipose tissue could lead to better understanding of the pathogenesis of obesity and its complications. However, current MS methods are limited by a high content of triacylglycerols (TAGs), which markedly surpasses the amount of other lipids and suppresses their ionization. The aim of our study was thus to optimize the preanalytical phase of lipid analysis in adipose tissue, focusing in particular on less-abundant lipids. Next, the optimized method was used to describe the differences between epicardial and subcutaneous adipose tissues obtained from patients undergoing cardiac surgery. Lipids were extracted using a modified Folch method with subsequent detachment of TAGs by thin layer chromatography (TLC). The extracts with/without TAGs were analyzed by tandem LC/MS. The repeatability of the presented method expressed by the median of the coefficients of variation was 12/5% for analysis with/without TAGs separation, respectively. The difference in the relative abundance of TAGs gained with/without TLC was, on average, 19% and did not reach significance (p value > 0.05) for any identified TAG. The novel preanalytical step allowed us to detect 37 lipids, which could not have been detected without TAG separation, because their signal to noise ratio is <5 in current methods of untargeted lipidomics. These lipids belong predominately to ceramides, glycerophosphatidylserines, glycerophosphatidylinsitols, sphingomyelins, glycerophosphatidylcholines, glycerophosphatidylethanolamines, diacylglycerols. The two adipose tissue depots differed mainly in the following lipid classes: glycerophosphatidylcholines, glycerophosphatidylinositols, glycerophosphatidylethanolamine, and sphingomyelins. Moreover, other major lipids showed distinctly different distributions between the two adipose tissues. Among these, the changes in TAGs were the most striking, which correspond to previously published data describing the differences between omental and subcutaneous adipose tissue. Implementation of the TLC step for the elimination of TAGs was crucial for enhancing the MS detection limit of minor lipids in adipose tissue. The differences between the overall lipid profiles of subcutaneous and epicardial tissue reflect their different functions arising from their location.
- MeSH
- chromatografie kapalinová metody MeSH
- lidé středního věku MeSH
- lidé MeSH
- lipidy analýza MeSH
- nitrobřišní tuk chemie MeSH
- perikard fyziologie MeSH
- podkožní tuk chemie MeSH
- reprodukovatelnost výsledků MeSH
- senioři MeSH
- tandemová hmotnostní spektrometrie metody MeSH
- Check Tag
- lidé středního věku MeSH
- lidé MeSH
- senioři MeSH
- Publikační typ
- časopisecké články MeSH
In this study, we present a method for the detection of n-3 fatty acid (n-3 FA) signals using MRS in adipose tissue in vivo. This method (called oMEGA-PRESS) is based on the selective detection of the CH3 signal of n-3 FA using the MEGA-PRESS (MEshcher-GArwood Point-RESolved Spectroscopy) J-difference editing technique. We optimized the envelope shape and frequency of spectral editing pulses to minimize the spurious co-editing and incomplete subtraction of the CH3 signal of other FAs, which normally obscure the n-3 FA CH3 signal in MR spectra acquired using standard PRESS techniques. The post-processing of the individual data scans with the phase and frequency correction before data subtraction and averaging was implemented to further improve the quality of in vivo spectra. The technique was optimized in vitro on lipid phantoms using various concentrations of n-3 FA and examined in vivo at 3 T on 15 healthy volunteers. The proportion of n-3 FA estimated by the oMEGA-PRESS method in phantoms showed a highly significant linear correlation with the n-3 FA content determined by gas chromatography. The signal attributed to n-3 FA was observed in all subjects. Comparisons with the standard PRESS technique revealed an enhanced identification of the n-3 FA signal using oMEGA-PRESS. The presented method may be useful for the non-invasive quantification of n-3 FA in adipose tissue, and could aid in obtaining a better understanding of various aspects of n-3 FA metabolism.
- MeSH
- chromatografie plynová MeSH
- fantomy radiodiagnostické MeSH
- index tělesné hmotnosti MeSH
- lidé MeSH
- lněný olej chemie MeSH
- magnetická rezonanční spektroskopie metody MeSH
- oleje rostlin chemie MeSH
- omega-3 mastné kyseliny analýza MeSH
- podkožní tuk chemie MeSH
- tuková tkáň chemie MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
The objective of the study was to investigate the effects of sex and slaughter age on the fatty acid (FA) composition and stearoyl-CoA desaturase gene expression in muscle and adipose tissue. Twenty-four Charolais × Simmental crossbred bulls and heifers were raised under similar conditions and slaughtered at 14 or 18 months of age. The total amount of FA in muscle increased markedly in older animals with higher contents of monounsaturated FA (MUFA) in heifers than bulls. The proportions of MUFA and desaturation indices were higher in heifers, whereas polyunsaturated FA were higher in bulls in both muscle and subcutaneous adipose tissue. The results of this study demonstrated sex-dependent differences in the FA composition of muscle and subcutaneous adipose tissue from cattle slaughtered at different ages. The expression of the stearoyl-CoA desaturase gene was higher in the adipose tissue of heifers compared to bulls, and its variation partly contributed to sex- and age-differences in the FA composition of bovine adipose tissue.
- MeSH
- kosterní svaly chemie enzymologie MeSH
- kyseliny mastné mononenasycené analýza MeSH
- mastné kyseliny analýza MeSH
- messenger RNA genetika metabolismus MeSH
- nenasycené mastné kyseliny analýza MeSH
- podkožní tuk chemie enzymologie MeSH
- polymerázová řetězová reakce s reverzní transkripcí MeSH
- regulace genové exprese MeSH
- sexuální faktory MeSH
- skot MeSH
- stearyl-CoA-desaturasa genetika metabolismus MeSH
- věkové faktory MeSH
- zvířata MeSH
- Check Tag
- mužské pohlaví MeSH
- skot MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- srovnávací studie MeSH
The previously reported genetic polymorphisms of the stearoyl-CoA desaturase (SCD1) and sterol regulatory element binding protein-1 (SREBP-1) genes were investigated in Fleckvieh bulls using the PCR-RFLP and AS-PCR methods, respectively. The genomic DNA was obtained from a total of 370 bulls. The frequencies of alleles A and V of the single nucleotide polymorphism in exon 5 of the SCD1 gene (SNP 878C>T) were 0.555 and 0.445, respectively. In the 84-bp Ins/Del polymorphism in intron 5 of the SREBP-1 gene, the frequency of the L allele (insertion) was markedly higher (0.920) than that of the S allele (deletion; 0.080). Fatty acid profile was determined in a total of 367 samples of muscle fat (MSF) and 150 samples of subcutaneous fat (SCF). The AA genotype of SCD1 polymorphism showed a lower content of C18:0 (P<0.01) and higher contents of C14:1 cis-9 (P<0.001) and C18:1 cis-9 (P<0.05) in MSF compared to the VV genotype. As a result, the bulls with genotypes AA or AV had lower SFA (P<0.01), higher MUFA (P<0.05) and higher MUFA/SFA (P<0.01) than VV animals. The results obtained for SCF were similar. The SREBP-1 polymorphism was associated with a higher content of C14:1 cis-9 (P<0.01) in the LS compared to LL genotype in SCF. The results of this study demonstrated the existence of the polymorphisms in the SCD1 and SREBP-1 genes in the population of Fleckvieh cattle and their associations with the concentrations of several MSF and SCF fatty acids.
- MeSH
- DNA analýza MeSH
- exony MeSH
- frekvence genu MeSH
- genom MeSH
- genotyp MeSH
- introny MeSH
- jednonukleotidový polymorfismus MeSH
- kosterní svaly chemie MeSH
- kyseliny mastné mononenasycené analýza MeSH
- mastné kyseliny analýza genetika MeSH
- podkožní tuk chemie MeSH
- polymerázová řetězová reakce metody MeSH
- protein SREBP1 genetika MeSH
- skot MeSH
- stearyl-CoA-desaturasa genetika MeSH
- uhlík MeSH
- zvířata MeSH
- Check Tag
- mužské pohlaví MeSH
- skot MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Cíl studie: Cílem výzkumu je posoudit vhodnost miniinvazivních technologií, umožňujících kontrolu metabolismu glukózy kontinuálním monitorováním hladiny intersticiální glukózy, jako alternativ ke stanovení hladiny glukózy v krvi (intersticiální glukóza je hodnocena dvěma metodami kontinuálního monitorování – mikrodialýzou a systémem Guardian Real Time) u kriticky nemocných, ventilovaných pacientů. Typ studie: Lokální, prospektivní. Typ pracoviště: Klinika anesteziologie, resuscitace a intenzivní medicíny, Fakultní nemocnice. Materiál a metodika: K monitorování hladiny intersticiální glukózy byly použity mikrodialýza a Guardian RT glucose monitoring system. Mikrodialyzační katétr, zavedený do podkožní tukové tkáně, byl promýván izotonickým roztokem rychlostí 0,3 µl . min-1 a získaný mikrodialyzát byl sbírán v hodinových intervalech. Z hodnot glykémie naměřených systémem Guardian RT byly do studie zahrnuty pouze hodnoty naměřené v čase výměny mikrodialyzačních vzorků. Vzorky arteriální a venózní krve ke stanovení plasmatické glykémie byly odebírány v hodinových intervalech, v čase výměny mikrodialyzačních vzorků. Sběr dat probíhal 24 hodin. Získaná data byla analyzována pomocí stanovení Pearsonova korelačního koeficientu a Bland Altmanova grafu. Výsledky: Korelační koeficient mezi arteriální glykémii a hladinou glukózy v mikrodialyzátu byl r = 0,7044 (p < 0,0001). Korelační koeficient mezi arteriální glykémii a hladinou glukózy ze systému Guardian RT byl r = 0,6938 (p < 0,0001). Korelační koeficient mezi glukózou systému Guardian RT a mikrodialýzou byl r = 0,5736 (p < 0,0001). Závěr: Uvedené výsledky nepodporují použití systému Guardian RT a mikrodialýzy ke sledování hladin intersticiální glukózy jako alternativ k monitorování systémové glykémie u kriticky nemocných ventilovaných pacientů s těsnou kontrolou glykémie.
Objective: The aim of study is to investigate whether the glucose values obtained by miniinvasive techniques of interstitial glucose monitoring (Guardian Real Time (RT) continuous glucose monitoring system and microdialysis) differ substantially from blood glucose readings and whether these methods could be used instead of blood glucose sampling in mechanically ventilated patients with tight glycemic control. Design: Local, prospective study. Setting: Department of Anesthesiology and Intensive Care, University Hospital. Materials and methods. Interstitial glucose concentrations were measured by the Guardian RT system and by the microdialysis of subcutaneous adipose tissue. The microdialysis catheter was constantly perfused with isotonic solution at a flow rate of 0.3 µL/min. Dialysate was collected in hourly fractions. Arterial and venous blood samples were collected in hourly intervals during 24 hours and paired with Guardian RT and microdialysis readings. Data were analyzed using the Bland-Altman method and the correlation coefficient was calculated. Results: Correlation coefficient between arterial glycemia and microdialysis interstitial glucose level was r = 0.7044 (P < 0.0001). Correlation coefficient between arterial glycemia and Guardian RT readings was r = 0.6938 (P < 0.0001). Correlation between interstitial glucose level obtained by microdialysis and Guardian RT glucose readings was r = 0.5736 (P < 0.0001). Conclusion: Due to obtained results, microdialysis and Guardian Real Time continuous glucose monitoring system could not be considered equivalent to blood glucose measurement and findings do not support their use for tight glycemic control management in mechanically ventilated, critically ill patients.
- MeSH
- ambulantní monitorování přístrojové vybavení MeSH
- financování organizované MeSH
- krevní glukóza analýza MeSH
- krevní plazma chemie MeSH
- lidé MeSH
- mikrodialýza metody MeSH
- péče o pacienty v kritickém stavu metody MeSH
- podkožní tuk chemie MeSH
- prediktivní hodnota testů MeSH
- prospektivní studie MeSH
- referenční hodnoty MeSH
- Check Tag
- lidé MeSH
Retinol binding protein 4 (RBP4) is a novel adipokine which might be involved in the development of insulin resistance. The aim of the study was to investigate the expression of RBP4 mRNA in subcutaneous and visceral fat depots and the relationship between RBP4 plasma and mRNA levels relative to indices of adiposity and insulin resistance. In 59 Caucasian women (BMI 20 to 49 kg/m2) paired samples of subcutaneous and visceral fat were obtained for RBP4, leptin and GLUT 4 mRNA analysis using reverse transcription-quantitative PCR. Euglycemic hyperinsulinemic clamp and computed tomography scans were performed. RBP4 mRNA levels as well as GLUT 4 mRNA and leptin mRNA levels were lower (P<0.001, P<0.01 and P<0.001, respectively) in visceral compared to subcutaneous fat. No differences were found in RBP4 mRNA expression in the two fat depots or in RBP4 plasma levels between subgroups of non-obese subjects (n=26), obese subjects without metabolic syndrome (n=17) and with metabolic syndrome (n=16). No correlations between RBP4 mRNA or plasma levels relative to adiposity, glucose disposal rate and GLUT 4 mRNA expression in adipose tissue were found. There was a weak positive correlation between plasma RBP4 and plasma triglycerides (r = 0.30, p<0.05) and between plasma RBP4 and blood glucose (r = 0.26, p<0.05). Regardless of the state of adiposity or insulin resistance, RBP4 expression in humans was lower in visceral than in subcutaneous fat. We found no direct relationship between either RBP4 mRNA or its plasma levels and the adiposity or insulin resistance.
- Klíčová slova
- Obesity, Insulin resistance, Visceral and subcutaneous adipose tissue, Retinol-binding protein 4,
- MeSH
- adipozita MeSH
- dospělí MeSH
- financování organizované MeSH
- inzulin krev MeSH
- inzulinová rezistence MeSH
- krevní glukóza analýza MeSH
- leptin analýza MeSH
- lidé středního věku MeSH
- lidé MeSH
- messenger RNA analýza MeSH
- metabolický syndrom metabolismus patofyziologie radiografie MeSH
- mladý dospělý MeSH
- nitrobřišní tuk chemie patologie radiografie MeSH
- obezita metabolismus patofyziologie radiografie MeSH
- plazmatické proteiny vázající retinol analýza genetika MeSH
- počítačová rentgenová tomografie MeSH
- podkožní tuk chemie patofyziologie radiografie MeSH
- polymerázová řetězová reakce s reverzní transkripcí MeSH
- přenašeč glukosy typ 4 analýza MeSH
- senioři MeSH
- Check Tag
- dospělí MeSH
- lidé středního věku MeSH
- lidé MeSH
- mladý dospělý MeSH
- mužské pohlaví MeSH
- senioři MeSH
- ženské pohlaví MeSH
OBJECTIVE: To study the influence of chronic malnutrition in patients with anorexia nervosa on endocrine function of adipose tissue on both circulating and subcutaneous fat mRNA expression level. PATIENTS AND DESIGN: A total of 12 patients with anorexia nervosa and 18 normal weight age-matched women underwent anthropometric examination, single blood drawing and subcutaneous adipose tissue biopsy. MEASUREMENTS: Serum concentrations of high-sensitive CRP (hsCRP), leptin, soluble leptin receptor, adiponectin, resistin, interleukin-6 and insulin were measured by Luminex, enzyme-linked immunosorbent assay (ELISA) and radioimmunoassay (RIA) kits. Subcutaneous adipose tissue mRNA expression of the same adipokines, adiponectin receptors 1 and 2 and immunocompetent cells marker CD68 was measured by real-time polymerase chain reaction (PCR). RESULTS: Decreased body fat content of patients with anorexia nervosa was accompanied by reduced hsCRP, leptin and increased adiponectin and soluble leptin receptor. Resistin, interleukin-6 and insulin levels did not differ from those of the control group. Fat mRNA adiponectin, leptin, interleukin-6 and CD68 expression was reduced, resistin mRNA expression was increased and adiponectin receptor 1 and 2 expression were unchanged as compared to the control group. CONCLUSIONS: Local perturbations in resistin, adiponectin and interleukin-6 mRNA expression in subcutaneous adipose tissue are not reflected by its circulating levels. These changes could be involved in some local metabolic disturbances in subcutaneous adipose tissue of anorexia nervosa patients.
- MeSH
- adiponektin genetika krev MeSH
- antigeny diferenciační myelomonocytární genetika MeSH
- C-reaktivní protein analýza MeSH
- CD antigeny genetika MeSH
- dospělí MeSH
- exprese genu MeSH
- financování organizované MeSH
- fyziologická adaptace MeSH
- interleukin-6 genetika MeSH
- inzulin krev MeSH
- krevní glukóza analýza MeSH
- leptinové receptory krev MeSH
- lidé MeSH
- mentální anorexie imunologie metabolismus MeSH
- messenger RNA analýza MeSH
- neparametrická statistika MeSH
- parakrinní signalizace MeSH
- podkožní tuk chemie imunologie metabolismus MeSH
- podvýživa imunologie metabolismus MeSH
- resistin genetika krev MeSH
- studie případů a kontrol MeSH
- Check Tag
- dospělí MeSH
- lidé MeSH
- ženské pohlaví MeSH
- Publikační typ
- srovnávací studie MeSH
