Phospholipid Composition Research Articles

Fat of the Gut: Epithelial Phospholipids in Inflammatory Bowel Diseases.

Inflammatory bowel diseases (IBD) comprise a distinct set of clinical symptoms resulting from chronic inflammation within the gastrointestinal (GI) tract. Despite the significant progress in understanding the etiology and development of treatment strategies, IBD remain incurable for thousands of patients. Metabolic deregulation is indicative of IBD, including substantial shifts in lipid metabolism. Recent data showed that changes in some phospholipids are very common in IBD patients. For instance, phosphatidylcholine (PC)/phosphatidylethanolamine (PE) and lysophosphatidylcholine (LPC)/PC ratios are associated with the severity of the inflammatory process. Composition of phospholipids also changes upon IBD towards an increase in arachidonic acid and a decrease in linoleic and a-linolenic acid levels. Moreover, an increase in certain phospholipid metabolites, such as lysophosphatidylcholine, sphingosine-1-phosphate and ceramide, can result in enhanced intestinal inflammation, malignancy, apoptosis or necroptosis. Because some phospholipids are associated with pathogenesis of IBD, they may provide a basis for new strategies to treat IBD. Current attempts are aimed at controlling phospholipid and fatty acid levels through the diet or via pharmacological manipulation of lipid metabolism.

Composition and structural characterization of peanut crude oil bodies extracted by aqueous enzymatic method

Peanut is rich in oils and proteins, which are found in oil bodies (OBs) and protein bodies (PBs), respectively. In this study, composition of proteins, neutral lipids, and phospholipids in crude OBs extracted by aqueous enzymatic extraction were analyzed. Distribution of oil, protein, and phospholipid in OBs was observed by confocal laser scanning microscopy (CLSM). Major proteins identified in OBs included oleosin, caleosin, and steroleosin. Lipoxygenase, arachin, conarachin, allergen, and ferritin, were also loosely associated with the surface of crude OBs. There were weak interactions between adsorbed proteins and OBs, such as electrostatic interactions, hydrophobic interactions, and hydrogen bonding. Results showed that fatty acids of crude OBs were rich in oleic acid (40.70 %) and linoleic acid (35.01 %). Phospholipids were mainly composed of phosphatidyl choline, phosphatidyl inositol, phosphatidyl ethanolamine, and phosphatidic acid, with relative contents of 54.87, 23.04, 16.76, and 5.33 %, respectively. CLSM of OBs revealed oil droplets evenly distributed in aqueous phase and uniform distribution of proteins and phospholipids at oil droplets surface. This study provides new insights into composition and structure of peanut OBs and theoretical basis for demulsification.

Dietary intake of n-3 polyunsaturated fatty acids alters the lipid mediator profile of the kidney but does not attenuate renal insufficiency

The efficacy of n-3 polyunsaturated fatty acids (PUFAs) in improving outcomes in a renal ischemia-reperfusion injury (IRI) model has previously been reported. However, the underlying mechanisms remain poorly understood and few reports demonstrate how dietary n-3 PUFAs influence the composition of membrane phospholipids in the kidney. Additionally, it has not been elucidated whether perilla oil (PO), which is mainly composed of the n-3 alpha-linolenic acid, mitigates renal IRI. In this study, we investigated the effect of dietary n-3 PUFAs (PO), compared with an n-6 PUFA-rich soybean oil (SO) diet, on IRI-induced renal insufficiency in a rat model. Levels of membrane phospholipids containing n-3 PUFAs were higher in the kidney of PO-rich diet-fed rats than the SO-rich diet-fed rats. Levels of blood urea nitrogen and serum creatinine were significantly higher in the ischemia-reperfusion group than the sham group under both dietary conditions. However, no significant differences were observed in blood urea nitrogen, serum creatinine, or histological damage between PO-rich diet-fed rats and SO-rich diet-fed rats. In the kidney of PO-rich diet-fed rats, levels of arachidonic acid and arachidonic acid-derived pro-inflammatory lipid mediators were lower than SO-rich diet-fed rats. Eicosapentaenoic acid and eicosapentaenoic acid-derived lipid mediators were significantly higher in the kidney of PO-rich than SO-rich diet-fed rats. These results suggest that dietary n-3 PUFAs alter the fatty acid composition of membrane phospholipids and lipid mediators in the kidney; however, this does not attenuate renal insufficiency or histological damage in a renal IRI model.

Replacement of Extrusion by Temperature-Controlled Ultrasonication in Emulsome Production

Emulsomes are lipid-based drug delivery systems comprising of a solid fat core surrounded by phospholipid multi-layers. Stability, prolonged release profile, biocompatibility and high encapsulation capacity for lipophilic compounds are their most prominent features. This study investigates the suitability of a temperature-controlled ultrasonication procedure to replace extrusion step that is widely applied for production and homogenization of emulsomes. In this study, emulsomes homogenized through ultrasonication were evaluated based on physicochemical properties including size, morphology and zeta potential. Temperature-controlled ultrasonication yielded production of tripalmitin-based emulsomes with size 285.6 ± 68.7 nm and zeta potential 31.6 ± 9.3 mV in average. Hexadecylamine in the phospholipid composition conferred the formulation a net positive surface charge. The morphology of emulsomes were investigated with scanning electron microscope. To further investigate whether ultrasonication alters the encapsulation capacity of emulsomes for lipophilic compounds, curcumin was added to the formulation as a model lipophilic substance. The physicochemical features as well as loading capacity of emulsomes produced with the introduced approach were compared with the values in the literature, where emulsomes of the same composition were homogenized through extrusion during their production. The results suggested that the control on temperature at ultrasonication benefited on emulsome production with high reproducibility in size and zeta potential and regarded as the solid feature of the proposed approach. In conclusion, the study provides evidence that the temperature-controlled ultrasonication methodology can safely replace the extrusion step as a robust, reliable approach for production of emulsomes.

Physicochemical and rheological properties of peanut oil body following alkaline pH treatment

Effects of alkaline pH treatment on composition, physicochemical and rheological properties, and microstructures of peanut oil body (OB) were investigated. Alkaline pH treatment could remove parts of proteins and phospholipids from OBs surface, and adsorbed proteins on OBs surface were almost completely removed at pH 11, yielding pure OBs. Essential amino acids and hydrophobic amino acids of OBs increased with the increase of alkaline pH. Although alkaline pH had no effect on fatty acid composition, it did have a significant effect on phospholipid composition. With the decrease of adsorbed proteins content, isoelectric point increased, and stability of OBs decreased, which is benefit for demulsification. pH and NaCl are important factors affecting stability of OBs by changing electrostatic repulsion, while effect of temperature on stability of OBs is small and irregular. OBs were stable system dominated by elasticity, and apparent viscosities of pH 8.0-OB, pH 9.5-OB, and pH 11.0-OB were obviously lower than crude-OB. It was also found that alkaline pH treatment had little effect on microstructures of peanut OBs. Results from this study suggest that improvement of OBs extraction technology to remove adsorbed proteins is important for demulsification.

Commensal microbiota and host metabolic divergence are associated with the adaptation of Diploderma vela to spatially heterogeneous environments.

Heterogeneous environment adaptation is critical to understand the species evolution and response to climate change. However, how narrow-range species adapt to micro-geographic heterogeneity has been overlooked, and there is a lack of insights from metabolism and commensal microbiota. Here, we studied the environmental adaptation for 3 geographic populations (>40 km apart) of Diploderma vela, a lizard endemic to dry-hot valleys of the Hengduan Mountain Region. The climatic boundary caused a cooler, droughtier, and barren environment for northernmost population (RM) than the middle (QZK) and southernmost populations (FS). Correspondingly, significant divergences in liver and muscle metabolism and commensal microbiota were detected between RM and QZK or FS individuals, but not between QZK and FS individuals. Phospholipid composition, coenzyme level (i.e. pyridoxal and NAD+ ), and cholesterol metabolism (e.g. androgen and estriol synthesis) constituted the major metabolic difference between RM and QZK/FS groups. FS and QZK individuals kept abundant Proteobacteria and antifungal strains, while RM individuals maintained more Firmicutes and Bacteroidota. Strong associations existed between varied host metabolite and gut microbes. How were these interpopulation variations associated to the environment adaptation were discussed. These results provided some novel insights into the environmental adaptation and implicated the consequence of climate change on narrow-range species.

Mitochondrial Dynamics in SARS-COV2 Spike Protein Treated Human Microglia: Implications for Neuro-COVID.

Emerging clinical data from the current COVID-19 pandemic suggests that ~ 40% of COVID-19 patients develop neurological symptoms attributed to viral encephalitis while in COVID long haulers chronic neuro-inflammation and neuronal damage result in a syndrome described as Neuro-COVID. We hypothesize that SAR-COV2 induces mitochondrial dysfunction and activation of the mitochondrial-dependent intrinsic apoptotic pathway, resulting in microglial and neuronal apoptosis. The goal of our study was to determine the effect of SARS-COV2 on mitochondrial biogenesis and to monitor cell apoptosis in human microglia non-invasively in real time using Raman spectroscopy, providing a unique spatio-temporal information on mitochondrial function in live cells. We treated human microglia with SARS-COV2 spike protein and examined the levels of cytokines and reactive oxygen species (ROS) production, determined the effect of SARS-COV2 on mitochondrial biogenesis and examined the changes in molecular composition of phospholipids. Our results show that SARS- COV2 spike protein increases the levels of pro-inflammatory cytokines and ROS production, increases apoptosis and increases the oxygen consumption rate (OCR) in microglial cells. Increases in OCR are indicative of increased ROS production and oxidative stress suggesting that SARS-COV2 induced cell death. Raman spectroscopy yielded significant differences in phospholipids such as Phosphatidylinositol (PI), phosphatidylserine (PS), phosphatidylethanolamine (PE) and phosphatidylcholine (PC), which account for ~ 80% of mitochondrial membrane lipids between SARS-COV2 treated and untreated microglial cells. These data provide important mechanistic insights into SARS-COV2 induced mitochondrial dysfunction which underlies neuropathology associated with Neuro-COVID.Graphical

31-Phosphorus Magnetic Resonance Spectroscopy in Evaluation of Glioma and Metastases in 3T MRI.

Background: 31-Phosphorus magnetic resonance spectroscopy (31-P MRS) has excellent potential for clinical neurological practice because of its noninvasive in-vivo assessment of cellular energy metabolism and the indirect evaluation of the phospholipid composition of the cell membrane, intracellular pH, and intracellular Mg2+ concentration. Purpose: The aim of this study was to evaluate the metabolic characteristics of glioma and metastases using 31-P MRS and assess utility to differentiate both. Study Type: Prospective study. Population: Fifteen consecutive patients with brain tumor. Field Strength/Sequence: Three-tesla magnetic resonance imaging/three-dimensional MRS imaging sequence. Statistical Tests: Unpaired sample t -test, and one-way analysis of variance with Tukey's post-hoc test. Results: Significantly decreased values of phosphomonoesters/inorganic phosphate (PME/Pi) in the tumor group (1.22 ± 0.72) compared with controls (2.28 ± 1.44) with a p -value of 0.041 were observed. There is a significant decrease in phosphocreatine (PCr)/Pi values (energy demand) in the tumor group (2.76 ± 0.73) compared with controls (4.13 ± 1.75) with a p -value of 0.050. Significant increase in Pi/adenosine triphosphate (ATP) was noted in tumor group (0.28 ± 0.09) compared with controls (0.22 ± 0.08) with p -value 0.049. Among tumor group, PME/PCr values were significantly decreased in gliomas (0.35 ± 0.17) than metastasis (0.58 ± 0.23) compared with controls with a p -value of 0.047. A significant decrease in PME/ATP was noted in gliomas (0.25 ± 0.12) than metastasis (0.41 ± 0.14) compared with controls with a p -value of 0.034. The tumor group exhibits alkaline pH (7.12 ± 0.10) compared with the normal parenchyma (7.04 ± 0.06) with a significant p -value of 0.025. Glioma and metastasis could not be differentiated with pH. However, the perilesional edema of glioma shows alkaline pH (7.09 ± 0.06) and metastasis shows acidic pH (7.02 ± 0.05) with a significant p -value of 0.030. Conclusion: Our study provides new insight into the cellular constituents and pH of gliomas and metastases and results were significant in differentiation between these two. However, due to the additional high expense, it is available as a research tool in very few institutions in India.

Plasmalogens regulate the AKT-ULK1 signaling pathway to control the position of the axon initial segment.

The axon initial segment (AIS) is a specialized region in neurons that encompasses two essential functions, the generation of action potentials and the regulation of the axodendritic polarity. The mechanism controlling the position of the axon initial segment to allow plasticity and regulation of neuron excitability is unclear. Here we demonstrate that plasmalogens, the most abundant ether-phospholipid, are essential for the homeostatic positioning of the AIS. Plasmalogen deficiency is a hallmark of Rhizomelic Chondrodysplasia Punctata (RCDP) and Zellweger spectrum disorders, but Alzheimer's and Parkinson's disease, are also characterized by plasmalogen defects. Neurons lacking plasmalogens displaced the AIS to more distal positions and were characterized by reduced excitability. Treatment with a short-chain alkyl glycerol was able to rescue AIS positioning. Plasmalogen deficiency impaired AKT activation, and we show that inhibition of AKT phosphorylation at Ser473 and Thr308 is sufficient to induce a distal relocation of the AIS. Pathway analysis revealed that downstream of AKT, overtly active ULK1 mediates AIS repositioning. Rescuing the impaired AKT signaling pathway was able to normalize AIS position independently of the biochemical defect. These results unveil a previously unknown mechanism that couples the phospholipid composition of the neuronal membrane to the positional assembly of the AIS.

Branched-chain amino acid metabolism controls membrane phospholipid structure in Staphylococcus aureus

Branched-chain amino acids (primarily isoleucine) are important regulators of virulence and are converted to precursor molecules used to initiate fatty acid synthesis in Staphylococcus aureus. Defining how bacteria control their membrane phospholipid composition is key to understanding their adaptation to different environments. Here, we used mass tracing experiments to show that extracellular isoleucine is preferentially metabolized by the branched-chain ketoacid dehydrogenase complex, in contrast to valine, which is not efficiently converted to isobutyryl-CoA. This selectivity creates a ratio of anteiso:iso C5-CoAs that matches the anteiso:iso ratio in membrane phospholipids, indicating indiscriminate utilization of these precursors by the initiation condensing enzyme FabH. Lipidomics analysis showed that removal of isoleucine and leucine from the medium led to the replacement of phospholipid molecular species containing anteiso/iso 17- and 19-carbon fatty acids with 18- and 20-carbon straight-chain fatty acids. This compositional change is driven by an increase in the acetyl-CoA:C5-CoA ratio, enhancing the utilization of acetyl-CoA by FabH. The acyl carrier protein (ACP) pool normally consists of odd carbon acyl-ACP intermediates, but when branched-chain amino acids are absent from the environment, there was a large increase in even carbon acyl-ACP pathway intermediates. The high substrate selectivity of PlsC ensures that, in the presence or the absence of extracellular Ile/Leu, the 2-position is occupied by a branched-chain 15-carbon fatty acid. These metabolomic measurements show how the metabolism of isoleucine and leucine, rather than the selectivity of FabH, control the structure of membrane phospholipids.

Effects of quercetin and artemetin prenylation on bioavailability and bioactivity

Flavonoids are a huge class of polyphenolic compounds ubiquitous in higher plants, in most food and beverages of natural origin. They could be considered as dietary phenols, which exert many health-promoting effects on human and animal physiology with a wide range of biomedical and nutritional functions such as activation or inhibition of enzymes like lipoxygenase and cyclooxygenase, the detoxification of carcinogens and chemoprevention. From a chemical point of view, these aromatic compounds can be divided in six subgroups depending on the position of aromatic B ring on C ring, the degree of unsaturation and oxidation, the position of hydroxyl groups and their functionalization. Between flavonoids, the prenylated ones represent a unique class occurring in nature where the C-prenylation is the most common, whereas O-prenylation is rarely present. The presence of this lipophilic functional group in different positions on the scaffold of flavonoids can sometimes lead to relevant changes in their biological activity due to an increased bioavailability. Capitalizing on the restricted incidence in nature of prenylated flavonoids, we have assessed the synthesis of C- and O-prenylated derivatives starting from two flavonoids, quercetin and artemetin, aimed at the exploration of structure-activity relationships. Results showed that prenylation significantly increased the cytotoxic effect of flavonoids in cancer HeLa cells, also improving their capacity to affect cell phospholipid and fatty acid composition. A marked cell bioavailability increase was demonstrated for the artemetin C-prenylated derivative.

Protein arginine methyltransferase 8 modulates mitochondrial bioenergetics and neuroinflammation after hypoxic stress.

Protein arginine methyltransferases (PRMTs) are a family of enzymes involved in gene regulation and protein/histone modifications. PRMT8 is primarily expressed in the central nervous system, specifically within the cellular membrane and synaptic vesicles. Recently, PRMT8 has been described to play key roles in neuronal signaling such as a regulator of dendritic arborization, synaptic function and maturation, and neuronal differentiation and plasticity. Here, we examined the role of PRMT8 in response to hypoxia-induced stress in brain metabolism. Our results from liquid chromatography mass spectrometry, mitochondrial oxygen consumption rate, and protein analyses indicate that PRMT8(-/-) knockout mice presented with altered membrane phospholipid composition, decreased mitochondrial stress capacity, and increased neuroinflammatory markers, such as tumor necrosis factor alpha and ionized calcium binding adaptor molecule 1 (Iba1, a specific marker for microglia/macrophage activation) after hypoxic stress. Furthermore, adenovirus-based overexpression of PRMT8 reversed the changes in membrane phospholipid composition, mitochondrial stress capacity, and neuroinflammatory markers. Together, our findings establish PRMT8 as an important regulatory component of membrane phospholipid composition, short-term memory function, mitochondrial function, and neuroinflammation in response to hypoxic stress.

Effects of temperature and food availability on liver fatty acid composition and plasma cortisol concentration in age-0 lake sturgeon: Support for homeoviscous adaptation

Overwintering survival in north temperate fishes involves a series of adaptive responses to multiple environmental stressors. Homeoviscous adaptation includes changes in membrane lipid composition in response to reduced environmental temperature, which may be driven by changes in hormones involved in the endocrine stress response. We examined how reduced temperature and food availability may act in concert to influence hepatic fatty acid composition of phospholipids and triglycerides, in addition to plasma concentration of cortisol in age-0 lake sturgeon (A. fulvescens). At 153 days post hatch (dph), temperature was decreased from 16 °C to 1 °C at a rate of 0.5 °C per day, and at 200 dph, fish were either fed every other day or deprived of food for 45 days to simulate an overwintering event. Liver fatty acid composition of phospholipids and triglycerides were assessed before temperature manipulation (16 °C; 153 dph), when fish had been at 1 °C for 16 days (199 dph), 25 days of overwintering (225 dph) and 45 days of overwintering (245 dph). Plasma cortisol concentration was assessed at 153, 225 and 245 dph. When temperature was decreased, both mono- and polyunsaturated fatty acids significantly increased in phospholipids and triglycerides. Total omega-6 fatty acids significantly increased in phospholipids while total omega-3 fatty acids did not. During the simulated overwintering, there was no obvious difference in fatty acids of phospholipids and triglycerides between diet treatments and no difference in circulating cortisol concentration between baseline and post-stressed fish in the fasted group. Our results provide support for homeoviscous adaptation to cold temperatures in lake sturgeon.

Low-salted salmon: Effects of salt reduction on physicochemical, lipidomic, and sensory characteristics

In this study, sodium-reduced salted salmon (SRSS) were prepared from six treatment groups formed using sodium replacers and flavor enhancers. The first three groups were prepared by replacing NaCl with 30% KCl (S2), 20% KCl + 10% CaCl2 (S3), and 50% KCl (S4), while the other three groups were prepared by the addition of the flavor enhancers of 5% yeast extract (F1), 3% lysine (F2), and 0.25% taurine (F3) along with 70% NaCl + 30% KCl. And their effects on the physicochemical properties, fatty acid, and phospholipid composition, and sensory characteristics of SRSS were investigated. The results demonstrated that replacing 30% NaCl with KCl (S2) had lower sensory damage compared with that obtained using 100% NaCl (S1). Moreover, this group exhibited higher physicochemical and lipidomic effects compared with other replacement treatments. The samples of S2 contained 0.8 mg of MDA/kg TBARS and 28.28 μg mg−1 phospholipids. Furthermore, the addition of yeast extract best improved the sensory defects caused by KCl. In conclusion, the results of this study can guide the production of healthier salted salmons with good sensory acceptance and low sodium content.

Aerosol, chemical and physical properties of dry powder synthetic lung surfactant for noninvasive treatment of neonatal respiratory distress syndrome

Inhalation of dry powder synthetic lung surfactant may assist spontaneous breathing by providing noninvasive surfactant therapy for premature infants supported with nasal continuous positive airway pressure. Surfactant was formulated using spray-drying with different phospholipid compositions (70 or 80 total weight% and 7:3 or 4:1 DPPC:POPG ratios), a surfactant protein B peptide analog (KL4, Super Mini-B, or B-YL), and Lactose or Trehalose as excipient. KL4 surfactant underperformed on initial adsorption and surface activity at captive bubble surfactometry. Spray-drying had no effect on the chemical composition of Super Mini-B and B-YL peptides and surfactant with these peptides had excellent surface activity with particle sizes and fine particle fractions that were well within the margins for respiratory particles and similar solid-state properties. Prolonged exposure of the dry powder surfactants with lactose as excipient to 40 °C and 75% humidity negatively affected hysteresis during dynamic cycling in the captive bubble surfactometer. Dry powder synthetic lung surfactants with 70% phospholipids (DPPC and POPG at a 7:3 ratio), 25% trehalose and 3% of SMB or B-YL showed excellent surface activity and good short-term stability, thereby qualifying them for potential clinical use in premature infants.

Lipidomic Profiling of Clinical Prostate Cancer Reveals Targetable Alterations in Membrane Lipid Composition.

Dysregulated lipid metabolism is a prominent feature of prostate cancer that is driven by androgen receptor (AR) signaling. Here we used quantitative mass spectrometry to define the "lipidome" in prostate tumors with matched benign tissues (n = 21), independent unmatched tissues (n = 47), and primary prostate explants cultured with the clinical AR antagonist enzalutamide (n = 43). Significant differences in lipid composition were detected and spatially visualized in tumors compared with matched benign samples. Notably, tumors featured higher proportions of monounsaturated lipids overall and elongated fatty acid chains in phosphatidylinositol and phosphatidylserine lipids. Significant associations between lipid profile and malignancy were validated in unmatched samples, and phospholipid composition was characteristically altered in patient tissues that responded to AR inhibition. Importantly, targeting tumor-related lipid features via inhibition of acetyl-CoA carboxylase 1 significantly reduced cellular proliferation and induced apoptosis in tissue explants. This characterization of the prostate cancer lipidome in clinical tissues reveals enhanced fatty acid synthesis, elongation, and desaturation as tumor-defining features, with potential for therapeutic targeting. SIGNIFICANCE: This study identifies malignancy and treatment-associated changes in lipid composition of clinical prostate cancer tissues, suggesting that mediators of these lipidomic changes could be targeted using existing metabolic agents.

Adipocytes Promote Breast Cancer Cell Survival and Migration through Autophagy Activation.

Simple SummaryBreast tumours are in direct contact with the adipose tissue of the mammary gland. Although the interactions between breast cancer cells and adipocytes that secrete tumour-promoting factors are well known, the molecular mechanisms remain under investigation. The aim of our study was to understand whether and how adipocytes regulate a cell-recycling pathway in breast cancer cells—autophagy. We show that adipocytes promote autophagy in breast cancer cells through the acidification of lysosomes, leading to cancer cell survival in nutrient-deprived conditions and to cancer cell migration. In this study, we have identified a new mechanism, which can link adipose tissue with breast cancer progression.White adipose tissue interacts closely with breast cancers through the secretion of soluble factors such as cytokines, growth factors or fatty acids. However, the molecular mechanisms of these interactions and their roles in cancer progression remain poorly understood. In this study, we investigated the role of fatty acids in the cooperation between adipocytes and breast cancer cells using a co-culture model. We report that adipocytes increase autophagy in breast cancer cells through the acidification of lysosomes, leading to cancer cell survival in nutrient-deprived conditions and to cancer cell migration. Mechanistically, the disturbance of membrane phospholipid composition with a decrease in arachidonic acid content is responsible for autophagy activation in breast cancer cells induced by adipocytes. Therefore, autophagy might be a central cellular mechanism of white adipose tissue interactions with cancer cells and thus participate in cancer progression.

Content and Composition of Phospholipids, Fatty Acids and Sterols in Commercial Natural Phospholipid Excipients

Background: Phospholipids are widely used in food and pharmaceutical industry as functional excipients. In spite of the many analytical methods reported, there are very limited reports concerning systematic research and comparison of phospholipid excipients. Objective: To present a comprehensive evaluation of commercial natural phospholipid excipients (CNPEs). Methods: Seventeen batches of CNPEs from five manufacturing enterprises, isolated either from soybean or egg yolk, were investigated. The content and composition of phospholipids, fatty acids and sterols as a whole were considered as the evaluative index of CNPEs. Eight kinds of phospholipids were determined by Supercritical Fluid Chromatography (SFC), twenty-one kinds of fatty acids were determined by Gas Chromatography (GC) after boron trifluoride-methanol derivatization, and nine kinds of sterols were determined by High Performance Liquid Chromatography (HPLC) after separation and derivatization of the unsaponifiable matter. Cluster analysis was employed for the classification and identification of the CNPEs. Results: The results showed that each kind of CNPEs had its characteristic content and composition of phospholipids, fatty acids and sterols. Seventeen batches of samples were divided into eight groups in cluster analysis. CNPEs of the same type from different source (soybean or egg yolk) or enterprises presented different content and composition of phospholipids, fatty acids and sterols. Conclusion: Each type of CNPEs had its characteristic content and composition of phospholipid, fatty acid and sterol. The compositions of phospholipid, fatty acid and sterol as a whole can be applied as an indicator of the quality and characteristics for CNPEs.

LPEATs Tailor Plant Phospholipid Composition through Adjusting Substrate Preferences to Temperature.

Acyl-CoA:lysophosphatidylethanolamine acyltransferases (LPEATs) are known as enzymes utilizing acyl-CoAs and lysophospholipids to produce phosphatidylethanolamine. Recently, it has been discovered that they are also involved in the growth regulation of Arabidopsis thaliana. In our study we investigated expression of each Camelina sativa LPEAT isoform and their behavior in response to temperature changes. In order to conduct a more extensive biochemical evaluation we focused both on LPEAT enzymes present in microsomal fractions from C. sativa plant tissues, and on cloned CsLPEAT isoforms expressed in yeast system. Phylogenetic analyses revealed that CsLPEAT1c and CsLPEAT2c originated from Camelina hispida, whereas other isoforms originated from Camelina neglecta. The expression ratio of all CsLPEAT1 isoforms to all CsLPEAT2 isoforms was higher in seeds than in other tissues. The isoforms also displayed divergent substrate specificities in utilization of LPE; CsLPEAT1 preferred 18:1-LPE, whereas CsLPEAT2 preferred 18:2-LPE. Unlike CsLPEAT1, CsLPEAT2 isoforms were specific towards very-long-chain fatty acids. Above all, we discovered that temperature strongly regulates LPEATs activity and substrate specificity towards different acyl donors, making LPEATs sort of a sensor of external thermal changes. We observed the presented findings not only for LPEAT activity in plant-derived microsomal fractions, but also for yeast-expressed individual CsLPEAT isoforms.

Eicosapentaenoic acid membrane incorporation stimulates ABCA1-mediated cholesterol efflux from human THP-1 macrophages

A high intake in polyunsaturated fatty acids (PUFAs), especially eicosapentaenoic acid (EPA) (C20:5 n-3), is cardioprotective. Dietary PUFAs incorporate into membrane phospholipids, which may modify the function of membrane proteins. We investigated the consequences of the membrane incorporation of several PUFAs on the key antiatherogenic ABCA1-mediated cholesterol efflux pathway. Human THP-1 macrophages were incubated with EPA, arachidonic acid (AA) (C20:4 n-6) or docosahexaenoic acid (DHA) (C22:6 n-3) for a long time to mimic a chronic exposure. EPA 70 μM, but not AA 50 μM or DHA 15 μM, increased ABCA1-mediated cholesterol efflux to apolipoprotein (apo) AI by 28% without altering aqueous diffusion. No variation in ABCA1 expression or localization was observed after EPA treatment. EPA incorporation did not affect the phenotype of THP-1 macrophages. The membrane phospholipids composition of EPA cells displayed higher levels of both EPA and its elongation product docosapentaenoic acid, which was associated with drastic lower levels of AA. Treatment by EPA increased the ATPase activity of the transporter, likely through a PKA-dependent mechanism. Eicosanoids were not involved in the stimulated ABCA1-mediated cholesterol efflux from EPA-enriched macrophages. In addition, EPA supplementation increased the apo AI binding capacity from macrophages by 38%. Moreover, the increased apo AI binding in EPA-enriched macrophages can be competed. In conclusion, EPA membrane incorporation increased ABCA1 functionality in cholesterol-normal human THP-1 macrophages, likely through a combination of different mechanisms. This beneficial in vitro effect may partly contribute to the cardioprotective effect of a diet enriched with EPA highlighted by several recent clinical trials.