In vivo characterization of the liver fat 1H MR spectrum

Fourier transform near‐infrared (FT‐NIR) technology is matrix dependent and thus highly dependent on factors that influence the absorption spectra. Ignoring these factors during the development of FT‐NIR models will affect the accuracy and reliability of the classification of fats and oils and the determination of their fatty acid (FA) composition. Four factors were studied: the temperature at which samples are scanned, differences in FA chain length and number of double bonds, and the presence of non‐triacylglycerol components. The results showed that an increase in the recording temperature decreased the absorption peak intensity, but not the position. FT‐NIR spectral differences were linked to variations in molecular vibrations resulting from the number of carbon atoms or double bonds in the FA. The FT‐NIR method could clearly differentiate between chain lengths from 10:0 to 18:0 and numbers of double bonds from zero (18:0) to three (18:3). Contaminants in triacylglycerols altered the FT‐NIR spectra, resulting in increased errors in the FA content. An increased concentration of β‐sitosterol in triolein decreased or increased the observed contents of cis9‐18:1 and cis11‐18:1, respectively. An FT‐NIR model adjusted for the phytosterol content corrected this discrepancy. The revised FT‐NIR model was successfully used to provide the accurate FA compositions of commercial sunflower oils.

Thematic Review Series: Sphingolipids. Biodiversity of sphingoid bases (“sphingosines”) and related amino alcohols

Fatty acids are a major source of structural diversity within the lipidome due to variations in their acyl chain lengths, branching, and cyclization, as well as the number, position, and stereochemistry of double bonds within their mono- and poly-unsaturated species. Here, the utility of 193nm UltraViolet PhotoDissociation tandem mass spectrometry (UVPD-MS/MS) has been evaluated for the detailed structural characterization of a series of unsaturated fatty acid lipid species. UVPD-MS/MS of unsaturated fatty acids is shown to yield pairs of unique diagnostic product ions resulting from cleavages adjacent to their C=C double bonds, enabling unambiguous localization of the site(s) of unsaturation within these lipids. The effect of several experimental variables on the observed fragmentation behaviour and UVPD-MS/MS efficiency, including the position and number of double bonds, the effect of conjugated versus non-conjugated double bonds, the number of laser pulses, and the influence of alkali metal cations (Li, Na, K) as the ionizing adducts, has been evaluated. Importantly, the abundance of the diagnostic ions is shown to enable relative quantitation of mixtures of fatty acid isomers across a range of molar ratios. Finally, the practical application of 193nm UVPD-MS/MS is demonstrated via characterization of changes in the ratios of fatty acid double bond positional isomers in isogenic colorectal cancer cell lines. This study therefore demonstrates the practicality of UVPD-MS/MS for the structural characterization of fatty acid isomers in lipidome analysis workflows.

Influence of steroid structure on the pyrolytic formation of polycyclic aromatic hydrocarbons

To investigate the regional variability of adipose tissue triglyceride composition in vivo using 1 H MRS, examining potential confounders and corrections for artifacts, to allow for adipose tissue spectrum estimation. 1 H magnetic resonance (MR) stimulated echo acquisition mode (STEAM) spectra were acquired in vivo at 3T from 340 adult patients (mean age 48.9 years, range 21-79 years; 172 males, 168 females; mean body mass index [BMI] 34.0, range 22-49 kg/m2 ) with known or suspected nonalcoholic fatty liver disease (NAFLD) in deep (dSCAT), surface (sSCAT) subcutaneous adipose tissue, and visceral adipose tissue (VAT). Triglyceride composition was characterized by the number of double bonds (ndb) and number of methylene-interrupted double bonds (nmidb). A subset of patients (dSCAT n = 80, sSCAT n = 55, VAT n = 194) had the acquisition repeated three times to examine the repeatability of ndb and nmidb estimation. Mean ndb and nmidb showed significant (P < 0.0001) differences between depots except for dSCAT and sSCAT nmidb (dSCAT ndb 2.797, nmidb 0.745; sSCAT ndb 2.826, nmidb 0.737; VAT ndb 2.723, nmidb 0.687). All ndb and nmidb estimates were highly repeatable (VAT ndb ICC = 0.888, nmidb ICC = 0.853; sSCAT: ndb ICC = 0.974, nmidb ICC = 0.964; dSCAT: ndb ICC = 0.959, nmidb ICC = 0.948). Adipose tissue composition can be estimated repeatably using 1 H MRS and different fat depots have different triglyceride compositions. 2 J. MAGN. RESON. IMAGING 2017;45:1455-1463.

The aim of this work was to validate a sequential method for quantifying the triglyceride fatty acid composition with 3.0 T MRI. The image acquisition was performed with a 3D spoiled gradient multiple echo sequence. A specific phase correction algorithm was implemented to correct the native phase images for wrap, zero- and first-order phase and rebuild the real part images. Then, using a model of a fat (1)H MR spectrum integrating nine components, the number of double bonds (ndb) and the number of methylene-interrupted double bonds (nmidb) were derived. The chain length (CL) was obtained from these parameters using heuristic approximation. Validations were performed on different vegetable oils whose theoretical fatty acid composition was used as reference and in five human subjects. In vivo measurements were made in the liver and in the subcutaneous and visceral adipose tissues. Linear regressions showed strong correlations between ndb and nmidb quantified with MRI and the theoretical values calculated using oil composition. Mean ndb/nmidb/CL were 1.80 ± 0.25/0.51 ± 0.21/17.43 ± 0.07, 2.72 ± 0.31/0.94 ± 0.16/17.47 ± 0.08 and 2.53 ± 0.21/0.84 ± 0.14/17.43 ± 0.07 in the liver, subcutaneous and visceral adipose tissues respectively. The results suggest that the triglyceride fatty acid composition can be assessed in human fatty liver and adipose tissues with a clinically relevant MRI method at 3.0 T.

Chemical shift magnetic resonance imaging (MRI) is commonly used to estimate the amount of fat in tissues, namely the proton density fat fraction (PDFF). In addition to PDFF, the type of fat can be inferred and characterized in terms of the number of double bonds (NDB), number of methylene-interrupted double bonds (NMIDB) and the chain length (CL) of the fatty acid chains. The saturation index is potentially a marker for metabolic disorders. This study assesses the feasibility of estimating these parameters independently or in a constrained manner. Correlations with spectroscopy were measured in 109 subjects’ subcutaneous and visceral fat depots (p = 2 × 10−28), and with the NAFLD Activity Score (NAS) from histological evaluation of biopsies. The findings indicate that imaging estimates are comparable to spectroscopy (p = 0.0002), but there is no significant association of NDB with NAS (p = 0.1).

Identification of fatty acids in gas chromatography by application of different temperature and pressure programs on a single capillary column

Rat liver microsomes contain a delta 6 desaturase that is active not only with the unsaturated acids of 18 carbons: oleic, linoleic and alpha-linolenic but also with similar acids of 20 carbons with the double bonds in delta 9, 12 and delta 12, 15. The rate of the reaction increases with the number of double bonds of the substrate but the acids of 20 carbons are less desaturated than the corresponding homologs of 18 carbons proving that the enzyme recognizes the number and position of double bonds as well as the chain length. The investigation of the inhibitory effect of different poly-unsaturated acids suggests that the acids would not only bind to the enzyme by means of the -CoA but also by other parts of the molecule different of the active center. The geometry of the molecule would be a substantial factor.

Dietary mono- or di-trans fatty acids with chain lengths of 18-22 increase the risk of cardiovascular diseases because they increase LDL cholesterol and decrease HDL cholesterol in the plasma. However, the effects of trans isomers of PUFAs on lipid metabolism remain unknown. Dietary PUFAs, especially eicosapentaenoic acid (EPA) in marine oils, improve serum lipid profiles by suppressing liver X receptor alpha (LXRalpha) activity in the liver. In this study, we compared the effects of trans geometric isomers of eicosapentaenoic acid (TEPA) on triacylglycerol synthesis induced by a synthetic LXRalpha agonist (T0901317) with the effects of EPA in HepG2 cells. TEPA significantly decreased the amount of cellular triacylglycerol and the expression of mRNAs encoding fatty acid synthase, stearoyl-CoA desaturase-1, and glycerol-3-phosphate acyltransferase induced by T0901317 compared with EPA. However, there was no significant difference between the suppressive effect of TEPA or EPA on the expression of sterol-regulatory element binding protein-1c (SREBP-1c) induced by T0901317. We found that TEPA, but not EPA, decreased the mRNA expression of peroxisome proliferator-activated receptor gamma coactivator 1beta (PGC-1beta), which is a coactivator of both LXRalpha and SREBP-1. These results suggest that the hypolipidemic effect of TEPA can be attributed to a decrease not only in SREBP-1 but also in PGC-1beta expression.

Simulation of mass spectral intensities by regression analysis of calculated structural characteristics

To compare MR-based fatty acid composition (FAC) quantification methods against the gold standard technique, gas chromatography (GC), with comparison of a free and a constrained signal model. The FAC was measured in the healthy and edematous legs of lymphedema patients. In vivo MRS and MRI data were acquired from 19 patients at 3 T. Biopsies were collected from subcutaneous adipose tissue of both thighs during liposuction. The saturated, monounsaturated, and polyunsaturated fatty acid fractions (fSFA , fMUFA and fPUFA , respectively) were estimated with the MR-based methods using two signal models: free and constrained (number of methylene-interrupted double bonds expressed in number of double bonds, based on GC data). Linear regression, Bland-Altman plots, and correlation coefficients were used to evaluate the MR methods against the GC of the biopsies. Paired t-test was used to compare the FAC difference between edematous and healthy legs. The estimated parameters correlated well with the GC data (rSFA , rMUFA , and rPUFA = 0.82, 0.81 and 0.89, respectively) using the free model MRI-based approach. In comparison, the MRS-based method resulted in weaker correlations and larger biases compared with MRI. In both cases, correct estimation of fMUFA and fPUFA fractions were not possible using the constrained model. The difference in FAC of healthy and edematous legs were estimated to 0.008 (P = .01), -0.009 (P = .005), and 0.002 (P = .03) for fSFA , fMUFA , and fPUFA . In this study, MRI-based FAC quantification was highly correlated, although slightly biased, compared with GC, whereas the MRS-based approach resulted in weaker correlations. Small but significant differences could be found between the healthy and edematous legs of lymphedema patients using GC analysis.

Hydrogen isotope ratios (2H/H or D/H) of long-chain unsaturated ketones (alkenones) preserved in lake and marine sediments hold great promise for paleoclimate studies. However, compound-specific hydrogen isotope analysis of individual alkenones has not been possible due to chromatographic coelution of alkenones with the same carbon chain length but different numbers of double bonds. Published studies have only reported the deltaD values of the mixture of coeluting alkenones. We developed an efficient procedure to isolate individual alkenones based on double-bond numbers using silica gel impregnated with silver nitrate. The chromatographic procedure is simple, inexpensive, and highly reproducible, offers 87-100% sample recovery, and allows for the first time hydrogen isotopic measurement on individual alkenones. deltaD values of specific di-, tri- and tetraunsaturated C37 alkenones produced by an Emiliania huxleyi culture, as well as those isolated from Greenland lake sediments, differ consecutively by 43-65 per thousand. These findings suggest that alkenones with different numbers of carbon-carbon double bonds express significantly different deltaD values and that coelution of different alkenones may lead to erroneous source water deltaD reconstructions. Our alkenone isolation approach opens a new avenue for paleoclimate reconstructions using hydrogen isotope ratios of individual alkenones.

Mechanism of Long-Chain Free Fatty Acid Protonation at the Membrane-Water Interface

Streptomyces sp. strain No. 560 produces four kinds of DNA methyltransferase inhibitors in the culture filtrate. One of them, DMI-4 was distinguished from DMI-1, -2 and -3 previously reported with respect to certain properties, DMI-4 is considered to be a triglyceride consisting of the fatty acids anteisopentadecanoic acid (C15:0), isopalmitic acid (C16:0) and isostearic acid (C18:0) from the results of gas chromatography analysis. Since DMI-4 contains three molecules of fatty acid, and the previously reported DMI-1, 8-methylpentadecanoic acid, is analogous to a fatty acid, the inhibitory activity has been examined of various fatty acids and their methyl esters against Eco RI DNA methyltransferase (M. Eco RI). Oleic acid (C18:1) was found to be a potent inhibiton of M. Eco RI. The inhibitory activity of oleic acid was shown to be pH- and temperature-dependent and inhibited M. Eco RI in a noncompetitive manner with respect to DNA or S-adenosylmethionine (SAM). The number of carbon atoms and double bonds in the fatty acid molecule affected the inhibitory activity, but their methyl esters were not inhibitors. Our results suggest that the length of the carbon chain, the number of double bonds and the presence of a carboxyl group and branched methyl group in the fatty acid molecule may play an important role in the inhibition of DNA methyltransferase.