Serum Metabolic Profiles Based on Nuclear Magnetic Resonance Spectroscopy among Patients with Deep Vein Thrombosis and Healthy Controls

Deep venous thrombosis (DVT) is associated with significant morbidity and mortality. Studies on changes in the level of metabolites could have the potential to reveal biomarkers that can assist in the early detection, diagnosis, monitoring of DVT progression, response to treatment, or recurrence of DVT. In this scenario, the metabolomic analysis can provide a better understanding of the biochemical dysregulations of thrombosis. Using an untargeted metabolomic approach through magnetic resonance spectroscopy and multi- and univariate statistical analysis, we compared 40 patients with previous venous thrombosis and 40 healthy individuals, and we showed important serum differences between patients and controls, especially in the spectral regions that correspond to glucose, lipids, unsaturated lipids, and glycoprotein A. Considering the groups depending on risk factors and the local of the previous episode (lower limbs or cerebral system), we also noticed differences in metabolites linked to lipids and lactate. Comparative analyses pointed to altered ratios of glucose/lactate and branched-chain amino acids (BCAAs)/alanine, which might be associated with the fingerprints of thrombosis. Although samples for metabolomic analysis were collected months after the acute episode, these results highlighted that, alterations can still remain and may contribute to a better understanding of the complications of the disease.

Visualizing Cancer Cell Metabolic Dynamics Regulated With Aromatic Amino Acids Using DO-SRS and 2PEF Microscopy

Oxidative imbalance plays an essential role in the progression of many diseases that include cancer and neurodegenerative diseases. Aromatic amino acids (AAA) such as phenylalanine and tryptophan have the capability of escalating oxidative stress because of their involvement in the production of Reactive Oxygen Species (ROS). Here, we use D2O (heavy water) probed stimulated Raman scattering microscopy (DO-SRS) and two Photon Excitation Fluorescence (2PEF) microscopy as a multimodal imaging approach to visualize metabolic changes in HeLa cells under excess AAA such as phenylalanine or trytophan in culture media. The cellular spatial distribution of de novo lipogenesis, new protein synthesis, NADH, Flavin, unsaturated lipids, and saturated lipids were all imaged and quantified in this experiment. Our studies reveal ∼10% increase in de novo lipogenesis and the ratio of NADH to flavin, and ∼50% increase of the ratio of unsaturated lipids to saturated lipid in cells treated with excess phenylalanine or trytophan. In contrast, these cells exhibited a decrease in the protein synthesis rate by ∼10% under these AAA treatments. The cellular metabolic activities of these biomolecules are indicators of elevated oxidative stress and mitochondrial dysfunction. Furthermore, 3D reconstruction images of lipid droplets were acquired and quantified to observe their spatial distribution around cells’ nuceli under different AAA culture media. We observed a higher number of lipid droplets in excess AAA conditions. Our study showcases that DO-SRS imaging can be used to quantitatively study how excess AAA regulates metabolic activities of cells with subcellular resolution in situ.

Selective arrangement of vesicles on artificial lipid membrane by biotin-avidin interaction

Lipid bilayers suspended over microwells on Si substrates are promising platforms for nanobiodevices that mimic cell membranes. Using the biotin-avidin interaction, we have succeeded in selectively arranging vesicles on the freestanding region of a lipid bilayer. When ternary lipid mixtures of saturated lipid, unsaturated lipid, and cholesterol are used, they separate into liquid-order (Lo) and liquid-crystalline (Lα) domains. A freestanding lipid bilayer prefers the Lα-phase over the Lo-phase because of the difference in their flexibility. In addition, the type of biotinylated lipid determines whether it is localized in the Lα-phase domain or the Lo-phase domain. As a result, the biotinylated unsaturated lipids localized in the Lα-phase domain aggregate in the freestanding lipid bilayer, and vesicles labeled with biotin selectively bind to the freestanding lipid bilayer by the biotin-avidin interaction. This technique helps to introduce biomolecules into the freestanding lipid bilayer of nanobiodevices via vesicles.

Reductive Stress of Sulfur-Containing Amino Acids within Proteins and Implication of Tandem Protein–Lipid Damage

Reductive radical stress represents the other side of the redox spectrum, less studied but equally important compared to oxidative stress. The reactivity of hydrogen atoms (H•) and hydrated electrons (e–aq) connected with peptides/proteins is summarized, focusing on the chemical transformations of methionine (Met) and cystine (CysS–SCys) residues into α-aminobutyric acid and alanine, respectively. Chemical and mechanistic aspects of desulfurization processes with formation of diffusible sulfur-centered radicals, such as methanethiyl (CH3S•) and sulfhydryl (HS•) radicals, are discussed. These findings are further applied to biomimetic radical chemistry, modeling the occurrence of tandem protein–lipid damages in proteo-liposomes and demonstrating that generation of sulfur-centered radicals from a variety of proteins is coupled with the cis–trans isomerization of unsaturated lipids in membranes. Recent applications to pharmaceutical and pharmacological contexts are described, evidencing novel perspectives in the stability of formulations and mode of action of drugs, respectively.

HR-MS Based Untargeted Lipidomics Reveals Characteristic Lipid Signatures of Wilson’s Disease

Background and Aims: The diagnosis of Wilson’s disease (WD) is challenging by clinical or genetic criteria. A typical early pathological change of WD is the increased liver lipid deposition and lowered serum triglyceride (TG). Therefore, the contents of serum lipids may provide evidence for screening of biomarkers for WD.Methods: 34 WD patients, 31 WD relatives, and 65 normal controls were enrolled in this study. Serum lipidomics data was acquired by an ultra-high-performance liquid chromatography high-resolution mass spectrometry system, and the data were analyzed by multivariate statistical methods.Results: Of all 510 identified lipids, there are 297 differential lipids between the WD and controls, 378 differential lipids between the relatives and controls, and 119 differential lipids between the patients and relatives. In WD, the abundances of most saturated TG were increased, whereas other unsaturated lipids decreased, including phosphatidylcholine (PC), sphingomyelin (SM), lysophosphatidylcholine (LPC), ceramide (Cer), and phosphatidylserine (PS). We also found many serum lipid species may be used as biomarkers for WD. The areas under the receiver operating characteristic curve (AUC) of PS (35:0), PS (38:5), and PS (34:0) were 0.919, 0.843, and 0.907. The AUCs of TG (38:0) and CerG1 (d42:2) were 0.948 and 0.915 and the AUCs of LPC (17:0) and LPC (15:0) were 0.980 and 0.960, respectively. The lipid biomarker panel exhibits good diagnostic performance for WD. The correlation networks were built among the different groups and the potential mechanisms of differential lipids were discussed. Interestingly, similar lipid profile of WD is also found in their relatives, which indicated the changes may also related to the mutation of the ATP7B gene.Conclusions: Lipid deregulation is another important hallmark of WD besides the deposition of copper. Our lipidomic results provide new insights into the diagnostic and therapeutic targets of WD.

227 Awardee Talk: Advances in Lipid Nutrition of Modern Sows

The modern lactating sow is a highly productive female capable of producing large quantities of milk to support large, rapidly growing litters. The associated high energy and nutritional demands are challenging to meet, especially during heat stress, when tissue loss is extensive. Supplemental lipids are efficient energy sources with a high energy density and low heat increment associated with digestion and metabolism. Besides serving as concentrated energy sources, unsaturated lipids are important sources of essential fatty acids (EFA; linoleic acid, C18:2n-6; and α-linolenic acid, C18:3n-3). These EFA are precursors of various prostanoids that are important regulators of reproductive processes. Lactating sows fed diets devoid of supplemental lipids exhibited poor reproduction but was significantly improved by inclusion of only 2% unsaturated lipids. It was shown that lactation diets were deficient in both linoleic and α-linolenic acid and tissue reserves were inadequate in mature sows. A multi-trial meta-analysis showed that more sows were successfully bred and maintained pregnancy when they consumed supplemental linoleic acid. Further, increased linoleic acid intake during lactation progressively reduced the number of sows removed from the herd due to reproductive failure (sows not returning to estrus, returning to estrus after breeding, pregnancy loss). Based on dose-response curves, the near maximum response was achieved when sows consumed a minimum of 125 g/d of linoleic acid and 10 g/d of α-linolenic acid during lactation. Recent studies also demonstrated improved subsequent reproduction with supplemental linoleic and α-linolenic acid during lactation, but not during the post-weaning period. Responses appeared to be related to heavier fetoplacental units, elevated allantoic fluid volume, and higher embryo survival measured on day 30 of gestation in sows supplemented with linoleic acid during lactation and early gestation. Research demonstrated the importance of both EFA in lactating sows and their critical role in improving reproduction, thus resolving seasonal anestrus.

Effects of Surfactant Volume Fraction on the Antioxidant Efficiency and on The Interfacial Concentrations of Octyl and Tetradecyl p-Coumarates in Corn Oil-in-Water Emulsions

Surfactants have been used for decades in the food industry for the preparation of lipid-based emulsified food stuffs. They play two main roles in the emulsification processes: first they decrease the interfacial tension between the oil and water, facilitating droplet deformation and rupture; second, they reduce droplet coalescence by forming steric barriers. However, addition of surfactants to binary oil-water mixtures also brings up the formation of three-dimensional interfacial layers, surrounding each emulsion droplet, that significantly alter chemical reactivity. This is the case, for instance, in the inhibition reaction between antioxidants and the lipid radicals formed in the course of the spontaneous oxidation reaction of unsaturated lipids, which are commonly employed in the preparation of food-grade emulsions. The rate of the inhibition reaction depends on the effective concentrations of antioxidants, which are mostly controlled by the amount of surfactant employed in the preparation of the emulsion. In this work, we analyze the effects of the surfactant Tween 20 on the oxidative stability and on the effective concentrations of two model antioxidants derived from cinnamic acid, determining their interfacial concentrations in the intact emulsions to avoid disrupting the existing equilibria and biasing results. For this purpose, a recently developed methodology was employed, and experimental results were interpreted on the grounds of a pseudophase kinetic model.

The Effect of Climate and Diet on Body Lipid Composition in the Oriental Hornet (Vespa orientalis)

Fatty acids (FA) are the primary metabolic fuel for many organisms and the fundamental component of membranes of all living organisms. FAs can be saturated (SFA), monounsaturated (MUFA), or polyunsaturated (PUFA). PUFA are not synthesized by most animals and are considered as essential nutrients. We examined the effect of climate on the saturation level of polar (mostly membranal) and neutral lipids in the body of the Oriental hornet (Vespa orientalis) from two extreme climatic zones: Mediterranean high elevation; and hot arid desert. In contrast to previous reports, the environmental temperature was shown to affect the hornet colonies’ thermal environments. The hornets nonetheless maintained their colony temperature within a narrow range. Analyses of the hornets’ unsaturation levels of polar and non-polar body lipids revealed caste differences: gynes and males contained less unsaturated lipids than workers. However, there were no differences in the respective castes between the two different climate zones tested. Experimentally manipulating the diet of queenless hornet colonies to a high Omega-3 diet (salmon) or a high Omega-6 diet (crickets) had only a minor effect on the worker-born males’ lipid composition. Although salmon-fed males had a higher Omega-3 content than cricket-fed ones, the proportion of these fatty acids was still low (below 1%). Cricket-fed males had significantly higher levels of Omega-6 than salmon-fed males. Our data show that the specific lipid composition of the hornet body is highly regulated and deficient in essential PUFA, even under different climates or high Omega-3 or Omega-6 PUFA diet. PUFA, especially Omega-3, is considered to have a beneficial effect on physiological processes. Our finding that these FA, when common in the diet, are almost absent in the body raises questions about how they affect animals’ physiology.