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1
Shiels, Katie, Alexandros Tsoupras, Ronan Lordan, et al. "Bioactive Lipids of Marine Microalga Chlorococcum sp. SABC 012504 with Anti-Inflammatory and Anti-Thrombotic Activities." Marine Drugs 19, no. 1 (2021): 28. http://dx.doi.org/10.3390/md19010028.
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Microalgae are at the start of the food chain, and many are known producers of a significant amount of lipids with essential fatty acids. However, the bioactivity of microalgal lipids for anti-inflammatory and antithrombotic activities have rarely been investigated. Therefore, for a sustainable source of the above bioactive lipids, the present study was undertaken. The total lipids of microalga Chlorococcum sp., isolated from the Irish coast, were fractionated into neutral-, glyco-, and phospho-lipids, and were tested in vitro for their anti-inflammatory and antithrombotic activities. All tested lipid fractions showed strong anti-platelet-activating factor (PAF) and antithrombin activities in human platelets (half maximal inhibitory concentration (IC50) values ranging ~25–200 μg of lipid) with the highest activities in glyco- and phospho-lipid fractions. The structural analysis of the bioactive lipid fraction-2 revealed the presence of specific sulfoquinovosyl diacylglycerols (SQDG) bioactive molecules and the HexCer-t36:2 (t18:1/18:1 and 18:2/18:0) cerebrosides with a phytosphingosine (4-hydrosphinganine) base, while fraction-3 contained bioactive phosphatidylcholine (PC) and phosphatidylethanolamine (PE) molecules. These novel bioactive lipids of Chlorococcum sp. with putative health benefits may indicate that marine microalgae can be a sustainable alternative source for bioactive lipids production for food supplements and nutraceutical applications. However, further studies are required towards the commercial technology pathways development and biosafety analysis for the use of the microalga.
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Shiels, Katie, Alexandros Tsoupras, Ronan Lordan, et al. "Bioactive Lipids of Marine Microalga Chlorococcum sp. SABC 012504 with Anti-Inflammatory and Anti-thrombotic Activities." Marine Drugs 19, no. 1 (2021): 28. http://dx.doi.org/10.3390/md19010028.
Full textAbstract:
Microalgae are at the start of the food chain, and many are known producers of a significant amount of lipids with essential fatty acids. However, the bioactivity of microalgal lipids for anti-inflammatory and antithrombotic activities have rarely been investigated. Therefore, for a sustainable source of the above bioactive lipids, the present study was undertaken. The total lipids of microalga Chlorococcum sp., isolated from the Irish coast, were fractionated into neutral-, glyco-, and phospho-lipids, and were tested in vitro for their anti-inflammatory and antithrombotic activities. All tested lipid fractions showed strong anti-platelet-activating factor (PAF) and antithrombin activities in human platelets (half maximal inhibitory concentration (IC50) values ranging ~25–200 μg of lipid) with the highest activities in glyco- and phospho-lipid fractions. The structural analysis of the bioactive lipid fraction-2 revealed the presence of specific sulfoquinovosyl diacylglycerols (SQDG) bioactive molecules and the HexCer-t36:2 (t18:1/18:1 and 18:2/18:0) cerebrosides with a phytosphingosine (4-hydrosphinganine) base, while fraction-3 contained bioactive phosphatidylcholine (PC) and phosphatidylethanolamine (PE) molecules. These novel bioactive lipids of Chlorococcum sp. with putative health benefits may indicate that marine microalgae can be a sustainable alternative source for bioactive lipids production for food supplements and nutraceutical applications. However, further studies are required towards the commercial technology pathways development and biosafety analysis for the use of the microalga.
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Tillman-Sutela, Eila, Anu Johansson, Pälvi Laakso, Heikki Kallio, and Anneli Kauppi. "Lipids and their seasonal variation in the nucellar layers of Scots pine (Pinus sylvestris) seeds." Canadian Journal of Botany 74, no. 9 (1996): 1416–24. http://dx.doi.org/10.1139/b96-171.
Full textAbstract:
The content, composition, and seasonal variation of lipids in the nucellar layers of two mature Scots pine seed lots from northern and central Finland were studied. These lipophilic layers, which are located inside the seed coat, are composed of collapsed cells and restrict the imbibition of seeds. In this study, epicuticular wax was found on their surface; in particular, the surface of the nucellar cap was composed of intermeshing wax rods. The content of total lipids of the nucellar layers varied seasonally from 10 to 24%, being highest in April. In addition to simple lipids, which formed the most abundant lipid fraction in nucellar layers, small amounts of phospho- and glyco-lipids were also present. The proportion of simple lipids and their fatty acid composition in the seeds of both provenances remained constant throughout the year, whereas seasonal variation was found in the proportions of phospho- and giyco-lipids and in their fatty acids. In general, the variation was more marked in the northern seed lot. The unsaturated fatty acids dominated in the fractions of simple lipids and phospholipids, with linoleic acid being the most abundant single fatty acid. The saturated fatty acids with more than 22 carbon atoms were most abundant in the glycolipid fractions. The role of lipids for the function of the collapsed nucellar layers in the regulation of imbibition and germination of pine seeds is discussed. Keywords: Pinus sylvestris L., northern seeds, fatty acids, imbibition, germination.
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Hao, Zong Di, Ping Huai Liu, Xun Yang, Jie Shi, and Sen Zhang. "Screening Method for Lipid-Content Microalgae Based on Sulfo-Phospho-Vanillin Reaction." Advanced Materials Research 610-613 (December 2012): 3532–35. http://dx.doi.org/10.4028/www.scientific.net/amr.610-613.3532.
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Studies that address the use of microalgae as biofuels often require the frequent measurement of total lipid content. Traditional methods for the quantification of lipid are time-consuming or involve the use of expensive analytical equipment that is not available in many labs. Here we investigated microalgal culture as the starting material and simple, colorimetric method for quantitative measurement of neutral lipids in microalgae with a relatively high correlation coefficient (R2=0.9038) between gravimetric and spectrophotometric quantification. Linear responses for triolein, vegetable oil and microalgal oil in a concentration range between 0.1 and 1 mg/l were observed. Using this method, Monoraphidium pusillum were screened out of several microalgal strains with the highest lipid content (25.52% dry weight). The color reaction for quantitation of microalgal lipids has significant advantages over traditional methods for screening of high lipid-content strains. Our data implied that the sensitivity and versatility enable this method a useful tool in screening of lipid-content microalgae.
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BheemRao T., Anilkumar V., and Dinesh T.D. "Variations of Lipids in Different Tissues (Liver, Intestine and Brain) of Fresh Water Cat Fish [Heteropneustes fossils (Bloch)]." UTTAR PRADESH JOURNAL OF ZOOLOGY 45, no. 7 (2024): 169–77. http://dx.doi.org/10.56557/upjoz/2024/v45i73989.
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In the present study on characterization and identification of lipids by thin layer chromatography in different tissues of Heteropneustes fossilis showed a considerable variation. The number of lipid spots was highest in brain compared to other tissues. The Rf values of spots indicate the regions of similarity between tissues. Comparison of Rf values with those of standards indicated that the presence of phospholipids, choline lipids, gyolopids, ninhydrine, amino group containing lipids were present. Intestine showed more amount of glycolipids and low amount of ninhydrine containing lipids, While liver possessed Iodine containing lipids.in addition these, whereas brain showed glycolipids, ninhydrine containing lipids, Iodine containing lipids and choline lipids too. The present study revealed that the Brain contain highest amount of different classes of lipids. The Liver tissue has shown less number of lipids. Glycolipids and general class of lipids are present in all tissues with minor variations, choline lipids present in brain but not in remaining tissues, while Phospho lipids are present in intestine and brain only, whereas amino group containing lipids were more in all tissues when compared with other class of lipids.
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Xu, Baomei, Jianhui Li, Shuai Zhang, et al. "The Transport of Charged Molecules across Three Lipid Membranes Investigated with Second Harmonic Generation." Molecules 28, no. 11 (2023): 4330. http://dx.doi.org/10.3390/molecules28114330.
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Subtle variations in the structure and composition of lipid membranes can have a profound impact on their transport of functional molecules and relevant cell functions. Here, we present a comparison of the permeability of bilayers composed of three lipids: cardiolipin, DOPG (1,2-dioleoyl-sn-glycero-3-phospho-(1′-rac-glycerol), and POPG (1-palmitoyl-2-oleoyl-sn-glycero-3-phospho-(1′-rac-glycerol)). The adsorption and cross-membrane transport of a charged molecule, D289 (4-(4-diethylaminostyry)-1-methyl-pyridinium iodide), on vesicles composed of the three lipids were monitored by second harmonic generation (SHG) scattering from the vesicle surface. It is revealed that structural mismatching between the saturated and unsaturated alkane chains in POPG leads to relatively loose packing structure in the lipid bilayers, thus providing better permeability compared to unsaturated lipid bilayers (DOPG). This mismatching also weakens the efficiency of cholesterol in rigidifying the lipid bilayers. It is also revealed that the bilayer structure is somewhat disturbed by the surface curvature in small unilamellar vesicles (SUVs) composed of POPG and the conical structured cardiolipin. Such subtle information on the relationship between the lipid structure and the molecular transport capability of the bilayers may provide clues for drug development and other medical and biological studies.
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Akyol, Sumeyya, Zafer Ugur, Ali Yilmaz, et al. "Lipid Profiling of Alzheimer’s Disease Brain Highlights Enrichment in Glycerol(phospho)lipid, and Sphingolipid Metabolism." Cells 10, no. 10 (2021): 2591. http://dx.doi.org/10.3390/cells10102591.
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Alzheimer’s disease (AD) is reported to be closely linked with abnormal lipid metabolism. To gain a more comprehensive understanding of what causes AD and its subsequent development, we profiled the lipidome of postmortem (PM) human brains (neocortex) of people with a range of AD pathology (Braak 0–6). Using high-resolution mass spectrometry, we employed a semi-targeted, fully quantitative lipidomics profiling method (Lipidyzer) to compare the biochemical profiles of brain tissues from persons with mild AD (n = 15) and severe AD (AD; n = 16), and compared them with age-matched, cognitively normal controls (n = 16). Univariate analysis revealed that the concentrations of 420 lipid metabolites significantly (p < 0.05; q < 0.05) differed between AD and controls. A total of 49 lipid metabolites differed between mild AD and controls, and 439 differed between severe AD and mild AD. Interestingly, 13 different subclasses of lipids were significantly perturbed, including neutral lipids, glycerolipids, glycerophospholipids, and sphingolipids. Diacylglycerol (DAG) (14:0/14:0), triacylglycerol (TAG) (58:10/FA20:5), and TAG (48:4/FA18:3) were the most notably altered lipids when AD and control brains were compared (p < 0.05). When we compare mild AD and control brains, phosphatidylethanolamine (PE) (p-18:0/18:1), phosphatidylserine (PS) (18:1/18:2), and PS (14:0/22:6) differed the most (p < 0.05). PE (p-18:0/18:1), DAG (14:0/14:0), and PS (18:1/20:4) were identified as the most significantly perturbed lipids when AD and mild AD brains were compared (p < 0.05). Our analysis provides the most extensive lipid profiling yet undertaken in AD brain tissue and reveals the cumulative perturbation of several lipid pathways with progressive disease pathology. Lipidomics has considerable potential for studying AD etiology and identifying early diagnostic biomarkers.
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Rivas Serna, Irma Magaly, Michal Sitina, Gorazd B. Stokin, et al. "Lipidomic Profiling Identifies Signatures of Poor Cardiovascular Health." Metabolites 11, no. 11 (2021): 747. http://dx.doi.org/10.3390/metabo11110747.
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Ideal cardiovascular health (CVH) is defined for the presence of ideal behavioral and health metrics known to prevent cardiovascular disease (CVD). The association of circulatory phospho- and sphingo-lipids to primary reduction in cardiovascular risk is unclear. Our aim was to determine the association of CVH metrics with the circulating lipid profile of a population-based cohort. Serum sphingolipid and phospholipid species were extracted from 461 patients of the randomly selected prospective Kardiovize study based on Brno, Czech Republic. Lipids species were measured by a hyphenated mass spectrometry technique, and were associated with poor CVH scores, as defined by the American Heart Association. Phosphatidylcholine (PC), phosphatidylethanolamine (PE), lysophosphatidylcholine (LPC), lysophosphatidylethanolamine (LPE) species were significantly lower in ideal and intermediate scores of health dietary metric, blood pressure, total cholesterol and blood fasting glucose compared to poor scores. Current smokers presented higher levels of PC, PE and LPE individual species compared to non-smokers. Ceramide (Cer) d18:1/14:0 was altered in poor blood pressure, total cholesterol and fasting blood glucose metrics. Poor cardiovascular health metric is associated with a specific phospho- and sphingolipid pattern. Circulatory lipid profiling is a potential biomarker to refine cardiovascular health status in primary prevention strategies.
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DEKKERS, David W. C., Paul COMFURIUS, Edouard M. BEVERS, and Robert F. A. ZWAAL. "Comparison between Ca2+-induced scrambling of various fluorescently labelled lipid analogues in red blood cells." Biochemical Journal 362, no. 3 (2002): 741–47. http://dx.doi.org/10.1042/bj3620741.
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Treatment of red blood cells with calcium and ionomycin causes activation of the lipid scramblase, a putative membrane protein catalysing flip-flop of (phospho)lipids. Various fluorescent 1-oleoyl-2-[6(7-nitrobenz-2-oxa-1,3-diazol-4-yl)amino] caproyl (C6-NBD) analogues were tested for transbilayer movement across the plasma membrane of red blood cells. Among these phospholipid analogues were phosphatidylgalactose, phosphatidylmaltose and phosphatidylmaltotriose, which were obtained from C6-NBD-phosphatidylcholine by phospholipase D-catalysed transphosphatidylation. The inward movement after the onset of scrambling was monitored by extraction of the non-internalized probe with BSA. We demonstrate that both the amino group and the size of the headgroup determine the kinetics of lipid scrambling, and that lipids with a ceramide backbone migrate much more slowly than glycerophospholipids with the same headgroup.
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Orr, Amy, William Wickner, Scott F. Rusin, Arminja N. Kettenbach, and Michael Zick. "Yeast vacuolar HOPS, regulated by its kinase, exploits affinities for acidic lipids and Rab:GTP for membrane binding and to catalyze tethering and fusion." Molecular Biology of the Cell 26, no. 2 (2015): 305–15. http://dx.doi.org/10.1091/mbc.e14-08-1298.
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Fusion of yeast vacuoles requires the Rab GTPase Ypt7p, four SNAREs (soluble N-ethylmaleimide–sensitive factor attachment protein receptors), the SNARE disassembly chaperones Sec17p/Sec18p, vacuolar lipids, and the Rab-effector complex HOPS (homotypic fusion and vacuole protein sorting). Two HOPS subunits have direct affinity for Ypt7p. Although vacuolar fusion has been reconstituted with purified components, the functional relationships between individual lipids and Ypt7p:GTP have remained unclear. We now report that acidic lipids function with Ypt7p as coreceptors for HOPS, supporting membrane tethering and fusion. After phosphorylation by the vacuolar kinase Yck3p, phospho-HOPS needs both Ypt7p:GTP and acidic lipids to support fusion.
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Krivoshapko, O. N., A. M. Popov, A. A. Artyukov, and E. Y. Kostetsky. "Particularities of corrective action of polar lipids and bioantioxidants from sea hydrobionts at imbalances of lipid and carbohydrate metabolism." Biomeditsinskaya Khimiya 58, no. 2 (2012): 189–98. http://dx.doi.org/10.18097/pbmc20125802189.
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A total mixture of phospho- and glycolipids from sea macrophytes Sargassum pallidum, Ulva fenestrata, Zostera marina was separated and the fatty acid composition was determined. Biological activity of the mixtures of polar lipids and natural antioxidants echinochrome A from flat sea urchin Scaphechinus mirabilis and polyphenolic complex from sea grass Zostera marina was studied in rats with experimental model of atherosclerosis and diabetes. These experiments revealed optimal compositions for mixtures of polar lipids and antioxidants, which possess high medical-corrective activity. Proposed mechanisms of action of the polar lipids (containing different polyunsaturated fatty acids) and antioxidants studied are presented. These compositions may be used for creation of new biologically-active additives and drugs.
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Engel, Kathrin M., Ulrike Jakop, Karin Müller, Sonja Grunewald, Uwe Paasch, and Jürgen Schiller. "MALDI MS Analysis to Investigate the Lipid Composition of Sperm." Current Analytical Chemistry 16, no. 1 (2020): 79–91. http://dx.doi.org/10.2174/1573411014666181030123256.
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Background: The sperm plasma membrane meets the requirements of sperm transit through the female genital tract and subsequent fertilization. Commonly, the (phospho)lipid composition of sperm is characterized by tremendous amounts of highly unsaturated fatty acyl residues such as docosahexaenoic and docosapentaenoic acid. While human sperm contain almost exclusively diacyl lipids, many animal sperm additionally contain significant amounts of ether lipids such as alkylacyl- and alkenyl-acyl lipids (plasmalogens). Hypothesis/Objective: It is suggested that deviations from the typical lipid composition are indicative of pathological changes. Therefore, simple methods to elucidate the sperm lipid composition are essential. Method: Matrix-assisted laser desorption and ionization (MALDI) mass spectrometry (MS) is a fast and simple method. Since the selection of the most suitable matrix is a crucial step in MALDI MS, this topic will be highlighted. It will also be shown that MALDI MS can be easily combined with thin-layer chromatography to overcome ion suppression effects. Results: The lipid composition of sperm from different species can be elucidated by MALDI MS. However, different matrix compounds have to be used to record positive and negative ion mass spectra. Since some sperm (glyco)lipids are characterized by the presence of sulfate residues which suppress the detection of less acidic lipids in the negative ion mode, previous separation is often necessary. It will be also emphasized that plasmalogens can be easily identified by either enzymatic digestion or treatment with acids. Conclusion: MALDI MS is a reliable method to obtain sperm lipid fingerprints in a simple and convenient way.
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Naoe, Satoko, Hiroshi Tsugawa, Mikiko Takahashi, Kazutaka Ikeda, and Makoto Arita. "Characterization of Lipid Profiles after Dietary Intake of Polyunsaturated Fatty Acids Using Integrated Untargeted and Targeted Lipidomics." Metabolites 9, no. 10 (2019): 241. http://dx.doi.org/10.3390/metabo9100241.
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Illuminating the comprehensive lipid profiles after dietary supplementation of polyunsaturated fatty acids (PUFAs) is crucial to revealing the tissue distribution of PUFAs in living organisms, as well as to providing novel insights into lipid metabolism. Here, we performed lipidomic analyses on mouse plasma and nine tissues, including the liver, kidney, brain, white adipose, heart, lung, small intestine, skeletal muscle, and spleen, with the dietary intake conditions of arachidonic acid (ARA), eicosapentaenoic acid (EPA), and docosahexaenoic acid (DHA) as the ethyl ester form. We incorporated targeted and untargeted approaches for profiling oxylipins and complex lipids such as glycerol (phospho) lipids, sphingolipids, and sterols, respectively, which led to the characterization of 1026 lipid molecules from the mouse tissues. The lipidomic analysis indicated that the intake of PUFAs strongly impacted the lipid profiles of metabolic organs such as the liver and kidney, while causing less impact on the brain. Moreover, we revealed a unique lipid modulation in most tissues, where phospholipids containing linoleic acid were significantly decreased in mice on the ARA-supplemented diet, and bis(monoacylglycero)phosphate (BMP) selectively incorporated DHA over ARA and EPA. We comprehensively studied the lipid profiles after dietary intake of PUFAs, which gives insight into lipid metabolism and nutrition research on PUFA supplementation.
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Reshetnikov, V. N., and O. V. Chizhik. "LIPID COMPOSITION OF CALLUSES CELL NUCLEI OF PLANTS OF DIFFERENT SYSTEMATIC GROUPS." Proceedings of the National Academy of Sciences of Belarus, Biological Series 63, no. 3 (2018): 335–38. http://dx.doi.org/10.29235/1029-8940-2018-63-3-335-338.
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The study revealed the results of the comparative analysis of the deoxyribonucleoproteid complex (chromatin) lipid composition in the callus and explant nuclei of plants, belonging to different systematic groups (winter rye, potatoes, peas). It provided the data on the phospho- and neutral lipid composition of the investigated plants and showed that the phospholipid content in the explants from dormant winter rye seed embryos and potato stems, having low metabolic activity, is significantly higher than in the calluses in the stage of active proliferation. Both winter rye and potato calluses tend to have a lower share of neutral lipids, including sterols, in their composition. The revealed factors can be used as markers of metabolic activity of the plant cell biology objects (callus and cell cultures).
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Dargel, Carina, Friederike Gräbitz-Bräuer, Ramsia Geisler, et al. "Stable DOPG/Glycyrrhizin Vesicles with a Wide Range of Mixing Ratios: Structure and Stability as Seen by Scattering Experiments and Cryo-TEM." Molecules 26, no. 16 (2021): 4959. http://dx.doi.org/10.3390/molecules26164959.
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Phosphatidylglycerols represent a large share of the lipids in the plasmamembrane of procaryotes. Therefore, this study investigates the role of charged lipids in the plasma membrane with respect to the interaction of the antiviral saponin glycyrrhizin with such membranes. Glycyrrhizin is a natural triterpenic-based surfactant found in licorice. Vesicles made of 1,2-dioleoyl-sn-glycero-3-phospho-rac-(1’-glycerol) (DOPG)/glycyrrhizin are characterized by small-angle scattering with neutrons and X-rays (SANS and SAXS). Small-angle scattering data are first evaluated by the model-independent modified Kratky–Porod method and afterwards fitted by a model describing the shape of small unilamellar vesicles (SUV) with an internal head-tail contrast. Complete miscibility of DOPG and glycyrrhizin was revealed even at a ratio of lipid:saponin of 1:1. Additional information about the chain-chain correlation distance of the lipid/saponin mixtures in the SUV structures is obtained from wide-angle X-ray scattering (WAXS).
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Leopold, Jenny, Patricia Prabutzki, Kathrin M. Engel, and Jürgen Schiller. "A Five-Year Update on Matrix Compounds for MALDI-MS Analysis of Lipids." Biomolecules 13, no. 3 (2023): 546. http://dx.doi.org/10.3390/biom13030546.
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Matrix-assisted laser desorption and ionization (MALDI) is a widely used soft-ionization technique of modern mass spectrometry (MS). MALDI enables the analysis of nearly all chemical compounds—including polar and apolar (phospho)lipids—with a minimum extent of fragmentation. MALDI has some particular advantages (such as the possibility to acquire spatially-resolved spectra) and is competitive with the simultaneously developed ESI (electrospray ionization) MS. Although there are still some methodological aspects that need to be elucidated in more detail, it is obvious that the careful selection of an appropriate matrix plays the most important role in (lipid) analysis. Some lipid classes can be detected exclusively if the optimum matrix is used, and the matrix determines the sensitivity by which a particular lipid is detected within a mixture. Since the matrix is, thus, crucial for optimum results, we provide here an update on the progress in the field since our original review in this journal in 2018. Thus, only the development during the last five years is considered, and lipids are sorted according to increasing complexity, starting with free fatty acids and ending with cardiolipins and phosphoinositides.
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Zhang, Xinxing, Kevin M. Barraza, and J. L. Beauchamp. "Cholesterol provides nonsacrificial protection of membrane lipids from chemical damage at air–water interface." Proceedings of the National Academy of Sciences 115, no. 13 (2018): 3255–60. http://dx.doi.org/10.1073/pnas.1722323115.
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The role of cholesterol in bilayer and monolayer lipid membranes has been of great interest. On the biophysical front, cholesterol significantly increases the order of the lipid packing, lowers the membrane permeability, and maintains membrane fluidity by forming liquid-ordered–phase lipid rafts. However, direct observation of any influence on membrane chemistry related to these cholesterol-induced physical properties has been absent. Here we report that the addition of 30 mol % cholesterol to 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC) or 1-palmitoyl-2-oleoyl-sn-glycero-3-phospho-(1′-rac-glycerol) (POPG) monolayers at the air–water interface greatly reduces the oxidation and ester linkage cleavage chemistries initiated by potent chemicals such as OH radicals and HCl vapor, respectively. These results shed light on the indispensable chemoprotective function of cholesterol in lipid membranes. Another significant finding is that OH oxidation of unsaturated lipids generates Criegee intermediate, which is an important radical involved in many atmospheric processes.
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Kristoffersen, Kamilla, Kasper Holst Hansen та Maria Andreasen. "Differential Effects of Lipid Bilayers on αPSM Peptide Functional Amyloid Formation". International Journal of Molecular Sciences 25, № 1 (2023): 102. http://dx.doi.org/10.3390/ijms25010102.
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Phenol-soluble modulins (PSMs) are key virulence factors of S. aureus, and they comprise the structural scaffold of biofilm as they self-assemble into functional amyloids. They have been shown to interact with cell membranes as they display toxicity towards human cells through cell lysis, with αPSM3 being the most cytotoxic. In addition to causing cell lysis in mammalian cells, PSMs have also been shown to interact with bacterial cell membranes through antimicrobial effects. Here, we present a study on the effects of lipid bilayers on the aggregation mechanism of αPSM using chemical kinetics to study the effects of lipid vesicles on the aggregation kinetics and using circular dichroism (CD) spectroscopy, Fourier-transform infrared (FTIR) spectroscopy and transmission electron microscopy (TEM) to investigate the corresponding secondary structure of the aggregates. We found that the effects of lipid bilayers on αPSM aggregation were not homogeneous between lipid type and αPSM peptides, although none of the lipids caused changes in the dominating aggregation mechanism. In the case of αPSM3, all types of lipids slowed down aggregation to a varying degree, with 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC) having the most pronounced effect. For αPSM1, lipids had opposite effects, where DOPC decelerated aggregation and lipopolysaccharide (LPS) accelerated the aggregation, while 1,2-dioleoyl-sn-glycero-3-phospho-rac-(1-glycerol) (DOPG) had no effect. For αPSM4, both DOPG and LPS accelerated the aggregation, but only at high concentration, while DOPC showed no effect. None of the lipids was capable of inducing aggregation of αPSM2. Our data reveal a complex interaction pattern between PSMs peptides and lipid bilayers that causes changes in the aggregation kinetics by affecting different kinetic parameters along with only subtle changes in morphology.
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Khaleque, M. A., MA Islam, A. Akhter, and MA Hye. "Studies on the Lipid Classes and Fatty Acid Compositions of Petuli (Trewia nudiflora Linn.) Seed oil." Bangladesh Journal of Scientific and Industrial Research 47, no. 1 (2012): 105–8. http://dx.doi.org/10.3329/bjsir.v47i1.10716.
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Studies were carried out on the lipid classes and fatty acid compositions of petuli seed oil. It was observed that petuli seed grown under the soil and climatic condition of Bangladesh contains about 22% pale yellow coloured oil. The total lipids were fractionated into three major lipid classes, neutral lipid glyco lipid and phospho lipid by silicic acid column chromatography. The neutral lipid was accounted to 92.5% of the total weight of the lipid applied. The oil was also fractionated into mono, di and triglyceride by silicic acid column chromatography. The triglyceride was counted for over 90% of the total weight of the oil. The fatty acid compositions of the oil were analyzed by Gas liquid Chromatography and found the major fatty acid ?-elaeostearic acid 38.50%, oleic acid 34.35%, linoleic acid 26.15% and small amount of arachidic acid 1%. DOI: http://dx.doi.org/10.3329/bjsir.v47i1.10716 Bangladesh J. Sci. Ind. Res. 47(1), 105-108, 2012
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Xu, Li, Xuejie Xu, Ganjun Yuan, Yimin Wang, Yunqiu Qu, and Erxiao Liu. "Mechanism of Azalomycin F5a against Methicillin-Resistant Staphylococcus aureus." BioMed Research International 2018 (2018): 1–5. http://dx.doi.org/10.1155/2018/6942452.
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To investigate the mechanism of azalomycin F5a against methicillin-resistant Staphylococcus aureus (MRSA), the conductivity of MRSA suspension and the adenylate kinase activity of MRSA culture were determined with the intervention of azalomycin F5a, which were significantly increased compared to those of blank controls. This inferred that azalomycin F5a could lead to the leakage of cellular substances possibly by increasing permeability to kill MRSA. As phospholipid bilayer was mainly responsible for cell-membrane permeability, the interaction between azalomycin F5a and cell-membrane lipids was further researched by determining the anti-MRSA activities of azalomycin F5a combined with cell-membrane lipids extracted from test MRSA or with 1,2-dipalmitoyl-sn-glycero-3-phospho-glycerol (DPPG) for possible molecular targets lying in MRSA cell-membrane. The results indicated that the anti-MRSA activity of azalomycin F5a remarkably decreased when it combined with membrane lipids or DPPG. This indicated that cell-membrane lipids especially DPPG might be important targets of azalomycin F5a against MRSA.
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Pavlek, Ana-Marija, Barbara Pem, and Danijela Bakarić. "The Signature of Fluctuations of the Hydrogen Bond Network Formed by Water Molecules in the Interfacial Layer of Anionic Lipids." Biophysica 4, no. 1 (2024): 92–106. http://dx.doi.org/10.3390/biophysica4010007.
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As the water molecules found at the interface of lipid bilayers exhibit distinct structural and reorientation dynamics compared to water molecules found in bulk, the fluctuations in their hydrogen bond (HB) network are expected to be different from those generated by the bulk water molecules. The research presented here aims to gain an insight into temperature-dependent fluctuations of a HB network of water molecules found in an interfacial layer of multilamellar liposomes (MLVs) composed of anionic 1,2-dimyristoyl-sn-glycero-3-phospho-L-serine (DMPS) lipids. Besides suspending DMPS lipids in phosphate buffer saline (PBS) of different pH values (6.0, 7.4, and 8.0), the changes in HB network fluctuations were altered by the incorporation of a non-polar flavonoid molecule myricetin (MCE) within the hydrocarbon chain region. By performing a multivariate analysis on the water combination band observed in temperature-dependent FTIR spectra, the results of which were further mathematically analyzed, the temperature-dependent fluctuations of interfacial water molecules were captured; the latter were the greatest for DMPS in PBS with a pH value of 7.4 and in general were greater for DMPS multibilayers in the absence of MCE. The presence of MCE made DMPS lipids more separated, allowing deeper penetration of water molecules towards the non-polar region and their restricted motion that resulted in decreased fluctuations. The experimentally observed results were supported by MD simulations of DMPS (+MCE) lipid bilayers.
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Dorsch, Anna Daniela, Walison Augusto da Silva Brito, Mihaela Delcea, Kristian Wende, and Sander Bekeschus. "Lipid Corona Formation on Micro- and Nanoplastic Particles Modulates Uptake and Toxicity in A549 Cells." Materials 16, no. 14 (2023): 5082. http://dx.doi.org/10.3390/ma16145082.
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Plastic waste is a global issue leaving no continents unaffected. In the environment, ultraviolet radiation and shear forces in water and land contribute to generating micro- and nanoplastic particles (MNPP), which organisms can easily take up. Plastic particles enter the human food chain, and the accumulation of particles within the human body is expected. Crossing epithelial barriers and cellular uptake of MNPP involves the interaction of plastic particles with lipids. To this end, we generated unilamellar vesicles from POPC (1-palmitoyl-2-oleoyl-glycero-3-phosphocholine) and POPS (1-palmitoyl-2-oleoyl-sn-glycero-3-phospho-L-serine) and incubated them with pristine, carboxylated, or aminated polystyrene spheres (about 1 µm in diameter) to generate lipid coronas around the particles. Lipid coronas enhanced the average particle sizes and partially changed the MNPP zeta potential and polydispersity. In addition, lipid coronas led to significantly enhanced uptake of MNPP particles but not their cytotoxicity, as determined by flow cytometry. Finally, adding proteins to lipid corona nanoparticles further modified MNPP uptake by reducing the uptake kinetics, especially in pristine and carboxylated plastic samples. In conclusion, our study demonstrates for the first time the impact of different types of lipids on differently charged MNPP particles and the biological consequences of such modifications to better understand the potential hazards of plastic exposure.
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Sanina, N. M., E. Y. Kostetsky, and S. N. Goncharova. "Thermotropic behaviour of membrane lipids from brown marine alga Laminaria japonica." Biochemical Society Transactions 28, no. 6 (2000): 894–97. http://dx.doi.org/10.1042/bst0280894.
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Microcalorimetry was used to show that transition-temperature ranges of phospho- and glycolipids of the brown marine alga Laminaria japonica were similar (from –62 to –47°C up to 62– 65°C), except for monogalactosyldiacylglycerol, the low-temperature limit of which was shifted to –78°C. As was shown by polarizing microscopy, the low-enthalpy peaks at temperatures of approx. 30–45°C corresponded to isotropic melting of galactolipids and coincided with the hightemperature limit for photosynthetic and mitochondrial activity of the algae. As a whole, a classical interrelation was observed between thermotropic behaviour and the fatty acid unsaturation of lipids.
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SAGARMAY, SAHA, BANDYOPADHYAYA ISHANI, and DUTTA JYOTIRMOY. "Lipid of Hilsa Fish (Hilsa ilisa)." Journal of Indian Chemical Society Vol. 67, May 1990 (1990): 414–16. https://doi.org/10.5281/zenodo.6202798.
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Bose Institute, 93/1 Acharya Prafulla Chandra Road, Caloutta-700 009 <em>Manuscript received 10 May 1989, revised 15 December 1989, accepted 1 January 1990</em> The major fatty acids of the flesh-lipid of hilsa fish <em>(Hilsa ilisa) </em>are 16 : 0 (32.0±5.0%), 18 : 0 (8.2±0.5%), 16 : 1 n7 (17.7±1.0%), 18 : 1 n9 (29.6±1.9%), 20 : 5 n3 (4.9±0.5%), 20 : 4 n6 (1.7±0.3%) and 22 : 6 n3 (1.5±0.6%). In this respect this lipid resembles the dietary fat of the Greenland Eskimos. In fish at minimum egg formation, the flesh contains 15.0±0.7% lipid, composed of neutral (88.4±1.7%), phospho-(5.1 ±1 6%) and glyco-(6.5 ± 1.2%) lipids. Lipid contents and lipid-class compositions of the flesh-lipid of hilsa fish at different degrees of egg maturation apparently indicate that the energy required for upstream migration by this fish, for spawning, is supplied by the oxidation of flesh-lipid without any preference for a particular lipid class.
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Chen, Zhipeng, Lingfeng Wang, Shuang Qiu, and Shijian Ge. "Determination of Microalgal Lipid Content and Fatty Acid for Biofuel Production." BioMed Research International 2018 (2018): 1–17. http://dx.doi.org/10.1155/2018/1503126.
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Biofuels produced from microalgal biomass have received growing worldwide recognition as promising alternatives to conventional petroleum-derived fuels. Among the processes involved, the downstream refinement process for the extraction of lipids from biomass greatly influences the sustainability and efficiency of the entire biofuel system. This review summarizes and compares the current techniques for the extraction and measurement of microalgal lipids, including the gravimetric methods using organic solvents, CO2-based solvents, ionic liquids and switchable solvents, Nile red lipid visualization method, sulfo-phospho-vanillin method, and the thin-layer chromatography method. Each method has its own competitive advantages and disadvantages. For example, the organic solvents-based gravimetric method is mostly used and frequently employed as a reference standard to validate other methods, but it requires large amounts of samples and is time-consuming and expensive to recover solvents also with low selectivity towards desired products. The pretreatment approaches which aimed to disrupt cells and support subsequent lipid extraction through bead beating, microwave, ultrasonication, chemical methods, and enzymatic disruption are also introduced. Moreover, the principles and procedures for the production and quantification of fatty acids are finally described in detail, involving the preparation of fatty acid methyl esters and their quantification and composition analysis by gas chromatography.
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Mironov, O. A., and I. P. Muraviova. "Oil hydrocarbons and total lipids in the coastal casts of Cystoseira barbata (Stackh.) C. Agardh in the littoral zone of Sevastopol (Black Sea)." Marine Biological Journal 2, no. 2 (2017): 49–54. http://dx.doi.org/10.21072/mbj.2017.02.2.05.
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The coastal zone of Sevastopol, especially the recreational part, feels great anthropogenic impact due to hosting a large number of tourists, but also because of the urban and stormwater discharge, containing oil hydrocarbons in its composition. Brown algae Cystoseira barbata forms vast underwater thickets here. Cut off from the substrate algae are at the edge of the water, mixed with coastal casts, resulting in secondary pollution of both coastline and coastal waters. At the same time freshly detached plants are used by representatives of different taxonomic groups and are actively involved in the food chain. In this context, the aim of this work is to study the lipid-hydrocarbon composition of Cystoseira from the coastal storm casts. The amount of oil hydrocarbons was determined by gravimetric method and the amount of lipids by color reaction with phospho-vanillin reagent. The results obtained indicate higher levels of chloroform-extracted substances, total lipids and oil hydrocarbons in macrophytes of Kruglaya Bay in comparison with the levels in water area of Park Pobedy. Onshore casts of Cystoseira may be the source of secondary pollution of the coastal waters by oil hydrocarbons.
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Sprott, G. Dennis, Giulio Ferrante, and Irena Ekiel. "Tetraether lipids of Methanospirillum hungatei with head groups consisting of phospho-N,N-dimethylaminopentanetetrol, phospho-N,N,N-trimethylaminopentanetetrol, and carbohydrates." Biochimica et Biophysica Acta (BBA) - Lipids and Lipid Metabolism 1214, no. 3 (1994): 234–42. http://dx.doi.org/10.1016/0005-2760(94)90069-8.
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Tian, Huidan, Qiaoling Wang, Xingying Yan, et al. "The Disruptions of Sphingolipid and Sterol Metabolism in the Short Fiber of Ligon-Lintless-1 Mutant Revealed Obesity Impeded Cotton Fiber Elongation and Secondary Cell Wall Deposition." International Journal of Molecular Sciences 26, no. 3 (2025): 1375. https://doi.org/10.3390/ijms26031375.
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Boosting evidence indicated lipids play important roles in plants. To explore lipid function in cotton fiber development, the lipid composition and content were detected by untargeted and targeted lipidomics. Compared with rapid elongation fibers, the lipid intensity of 16 sub-classes and 56 molecular species decreased, while only 7 sub-classes and 26 molecular species increased in the fibers at the stage of secondary cell wall deposition. Unexpectedly, at the rapid elongation stage, 20 sub-classes and 60 molecular species increased significantly, while only 5 sub-classes and 8 molecular species decreased in the ligon lintless-1 (li-1) mutant compared with its wild-type Texas Maker-1 (TM-1). Particularly, campesteryl, sitosteryl, and total steryl ester increased by 21.8-, 48.7-, and 45.5-fold in the li-1 fibers, respectively. All the molecular species of sphingosine-1-P, phytoceramide-OHFA, and glucosylceramide increased while all sphingosine, phytosphingosine, and glycosyl inositol phospho ceramides decreased in the li-1 fibers. Similarly, the different expression genes between the mutant and wild type were enriched in many pathways involved in the lipid metabolism. Furthermore, the number of lipid droplets also increased in the li-1 leaf and fiber cells when compared with the wild type. These results illuminated that fiber cell elongation being blocked in the li-1 mutant was not due to a lack of lipids, but rather lipid over-accumulation (obesity), which may result from the disruption of sphingolipid and sterol metabolism. This study provides a new perspective for further studying the regulatory mechanisms of fiber development.
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Swana, Kathleen W., Terri A. Camesano, and Ramanathan Nagarajan. "Formation of a Fully Anionic Supported Lipid Bilayer to Model Bacterial Inner Membrane for QCM-D Studies." Membranes 12, no. 6 (2022): 558. http://dx.doi.org/10.3390/membranes12060558.
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Supported lipid bilayers (SLBs) on quartz crystals are employed as versatile model systems for studying cell membrane behavior with the use of the highly sensitive technique of quartz crystal microbalance with dissipation monitoring (QCM-D). Since the lipids constituting cell membranes vary from predominantly zwitterionic lipids in mammalian cells to predominantly anionic lipids in the inner membrane of Gram-positive bacteria, the ability to create SLBs of different lipid compositions is essential for representing different cell membranes. While methods to generate stable zwitterionic SLBs and zwitterionic-dominant mixed zwitterionic–anionic SLBs on quartz crystals have been well established, there are no reports of being able to form predominantly or fully anionic SLBs. We describe here a method for forming entirely anionic SLBs by treating the quartz crystal with cationic (3-aminopropyl) trimethoxysilane (APTMS). The formation of the anionic SLB was tracked using QCM-D by monitoring the adsorption of anionic lipid vesicles to a quartz surface and subsequent bilayer formation. Anionic egg L-α-phosphatidylglycerol (PG) vesicles adsorbed on the surface-treated quartz crystal, but did not undergo the vesicle-to-bilayer transition to create an SLB. However, when PG was mixed with 10–40 mole% 1-palmitoyl-2-hydroxy-sn-glycero-3-phospho-(1′-rac-glycerol) (LPG), the mixed vesicles led to the formation of stable SLBs. The dynamics of SLB formation monitored by QCM-D showed that while SLB formation by zwitterionic lipids followed a two-step process of vesicle adsorption followed by the breakdown of the adsorbed vesicles (which in turn is a result of multiple events) to create the SLB, the PG/LPG mixed vesicles ruptured immediately on contacting the quartz surface resulting in a one-step process of SLB formation. The QCM-D data also enabled the quantitative characterization of the SLB by allowing estimation of the lipid surface density as well as the thickness of the hydrophobic region of the SLB. These fully anionic SLBs are valuable model systems to conduct QCM-D studies of the interactions of extraneous substances such as antimicrobial peptides and nanoparticles with Gram-positive bacterial membranes.
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Xu, Yanping, Yajamana Ramu, and Zhe Lu. "Removal of phospho-head groups of membrane lipids immobilizes voltage sensors of K+ channels." Nature 451, no. 7180 (2008): 826–29. http://dx.doi.org/10.1038/nature06618.
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McMahon, Anne, Hua Lu, and Igor A. Butovich. "The Spectrophotometric Sulfo-Phospho-Vanillin Assessment of Total Lipids in Human Meibomian Gland Secretions." Lipids 48, no. 5 (2013): 513–25. http://dx.doi.org/10.1007/s11745-013-3755-9.
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Mardešić, Ivan, Zvonimir Boban, and Marija Raguz. "Electroformation of Giant Unilamellar Vesicles from Damp Films in Conditions Involving High Cholesterol Contents, Charged Lipids, and Saline Solutions." Membranes 14, no. 10 (2024): 215. http://dx.doi.org/10.3390/membranes14100215.
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Giant unilamellar vesicles (GUVs) are frequently used as membrane models in studies of membrane properties. They are most often produced using the electroformation method. However, there are a number of parameters that can influence the success of the procedure. Some of the most common conditions that have been shown to have a negative effect on GUV electroformation are the presence of high cholesterol (Chol) concentrations, the use of mixtures containing charged lipids, and the solutions with an elevated ionic strength. High Chol concentrations are problematic for the traditional electroformation protocol as it involves the formation of a dry lipid film by complete evaporation of the organic solvent from the lipid mixture. During drying, anhydrous Chol crystals form. They are not involved in the formation of the lipid bilayer, resulting in a lower Chol concentration in the vesicle bilayer compared to the original lipid mixture. Motivated primarily by the issue of artifactual Chol demixing, we have modified the electroformation protocol by incorporating the techniques of rapid solvent exchange (RSE), ultrasonication, plasma cleaning, and spin-coating for reproducible production of GUVs from damp lipid films. Aside from decreasing Chol demixing, we have shown that the method can also be used to produce GUVs from lipid mixtures with charged lipids and in ionic solutions used as internal solutions. A high yield of GUVs was obtained for Chol/1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC) samples with mixing ratios ranging from 0 to 2.5. We also succeeded in preparing GUVs from mixtures containing up to 60 mol% of the charged lipid 1-palmitoyl-2-oleoyl-sn-glycero-3-phospho-L-serine (POPS) and in NaCl solutions with low ionic strength (<25 mM).
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Tukendorf, Anna, Zbigniew Krupa та Tadeusz Baszyński. "β-Carotene as a factor in the reconstitution of cyclic phospho rylation in damaged chloroplast membranes". Acta Societatis Botanicorum Poloniae 49, № 4 (2014): 435–43. http://dx.doi.org/10.5586/asbp.1980.039.
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Phenazine methosulphate mediated cyclic phosphorylation suppressed by heptane extraction or galactolipase treatment of spinach chloroplasts is restored by β -carotene, in 100% and 50%, respectively. Xanthophylls are not able to reconstitute this reaction. β-Carotene replaces galactolipids in reactivation of galactolipase treated chloroplasts, indicating a nonspecific effect of lipids in photosystem I dependent reactions.
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Xiao, Qing, Shujun Zhang, Cheng Yang та ін. "Ginsenoside Rg1 Ameliorates Palmitic Acid-Induced Hepatic Steatosis and Inflammation in HepG2 Cells via the AMPK/NF-κB Pathway". International Journal of Endocrinology 2019 (28 липня 2019): 1–11. http://dx.doi.org/10.1155/2019/7514802.
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Nonalcoholic fatty liver disease (NAFLD) is one of the common diseases in the world, and it can progress from simple lipid accumulation to sustained inflammation. The present study was designed to investigate the effects and underlying mechanisms of ginsenoside Rg1 (G-Rg1) treatment on NAFLD in vitro. HepG2 cells were treated with palmitic acid (PA) to induce steatosis and inflammation and then successively incubated with G-Rg1. Lipids accumulation was analyzed by Oil Red O staining and intracellular triglyceride (TG) quantification. Inflammatory conditions were examined by quantifying the levels of cell supernatant alanine transaminase/aspartate aminotransferase (ALT/AST) and secretory proinflammatory cytokines, including IL-1β, IL-6, and TNF-α in the cell supernatants. Quantitative RT-PCR and western blotting were used to measure the expressions of genes and proteins associated with lipogenic synthesis and inflammation, including AMP-activated protein kinase (AMPK) and nuclear factor-kappa B (NF-κB) pathways. HepG2 cells were pretreated with an AMPK inhibitor; then, Oil Red O staining and TG quantification were performed to study the lipid deposition. Phospho-AMPK (Thr172) (p-AMPK) and phospho-acetyl-CoA carboxylase (Ser79) (p-ACCα) were quantified by immunoblotting. Immunofluorescence was performed to demonstrate the nuclear translocation of NF-κB P65. The present study showed that PA markedly increased the intracellular lipid droplets accumulation and TG levels, but decreased AMPK phosphorylation and the expressions of its downstream lipogenic genes. However, G-Rg1 alleviated hepatic steatosis and reduced the intracellular TG content; these changes were accompanied by the activation of the AMPK pathway. In addition, blocking AMPK by using the AMPK inhibitor markedly abolished the G-Rg1-mediated protection against PA-induced lipid deposition in HepG2 cells. Furthermore, G-Rg1 reduced the ALT/AST levels and proinflammatory cytokines release, which were all enhanced by PA. These effects were correlated with the inactivation of the NF-κB pathway and translocation of P65 from the cytoplasm to the nucleus. Overall, these results suggest that G-Rg1 effectively ameliorates hepatic steatosis and inflammation, which might be associated with the AMPK/NF-κB pathway.
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Lai, Ying, and Yeon-Kyun Shin. "The importance of an asymmetric distribution of acidic lipids for synaptotagmin 1 function as a Ca2+ sensor." Biochemical Journal 443, no. 1 (2012): 223–29. http://dx.doi.org/10.1042/bj20112044.
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Syt1 (synaptotagmin 1) is a major Ca2+ sensor for synaptic vesicle fusion. Although Syt1 is known to bind to SNARE (soluble N-ethylmaleimide-sensitive fusion protein-attachment protein receptor) complexes and to the membrane, the mechanism by which Syt1 regulates vesicle fusion is controversial. In the present study we used in vitro lipid-mixing assays to investigate the Ca2+-dependent Syt1 function in proteoliposome fusion. To study the role of acidic lipids, the concentration of negatively charged DOPS (1,2-dioleoyl-sn-glycero-3-phospho-L-serine) in the vesicle was varied. Syt1 stimulated lipid mixing by 3–10-fold without Ca2+. However, with Ca2+ there was an additional 2–5-fold enhancement. This Ca2+-dependent stimulation was observed only when there was excess PS (phosphatidylserine) on the t-SNARE (target SNARE) side. If there was equal or more PS on the v-SNARE (vesicule SNARE) side the Ca2+-dependent stimulation was not observed. We found that Ca2+ at a concentration between 10 and 50 μM was sufficient to give rise to the maximal enhancement. The single-vesicle-fusion assay indicates that the Ca2+-dependent enhancement was mainly on docking, whereas its effect on lipid mixing was small. Thus for Syt1 to function as a Ca2+ sensor, a charge asymmetry appears to be important and this may play a role in steering Syt1 to productively trans bind to the plasma membrane.
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Merdzhanova, A., V. Panayotova, D. A. Dobreva, and K. Peycheva. "Proximate composition, lipid quality and heavy metals content in the muscle of two carp species." Agricultural Science and Technology 10, no. 4 (2018): 358–69. http://dx.doi.org/10.15547/10.15547/ast.2018.04.066.
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Abstract. The aim of the presented study was to characterize the quality of edible tissue of freshwater common carp (Cyprinus carpio) and bighead carp (Aristichthys nobilis), based on their proximate and lipid composition (lipid classes, fatty acid profile, fat soluble vitamins, carotenoids and cholesterol). Health risk assessment was evaluated based on the analysis of some toxic elements (As, Cd, Ni, Pb and total Hg). Proximate composition (moisture, crude protein and total lipid) was determined using standard procedures. Lipids were subsequently separated into neutral (NL) and polar lipids: Phospho- (PL) and Glycolipids (GL) by means of column and thin-layer chromatography. Lipid classes were derivatized into fatty acid methyl esters (FAMEs) which were analysed by gas chromatography–mass spectrometry (GC-MS). Vitamins A, D3 and E, beta-carotene, astaxanthin and cholesterol were analysed simultaneously using high performance liquid chromatography (HPLC). Heavy metals (As, Pb, Cd, Hg and Ni) were determined by optical emission spectrometry with inductively coupled plasma (ICP-OES) following a microwave digestion procedure. Protein content was higher in bighead carp (18.5%) and lower for common carp (15.5%), whereas lipid content showed opposite trend. Similarities in lipid classes distribution were observed for both species: NL>GL>PL. Neutral lipids constituted approximately 70% of TL in both species, as FAs profile was dominated by monounsaturated fatty acids (MUFA), whereas polyunsaturated FAs (PUFA) prevailed in polar fractions. Omega-3 PUFAs were higher in all lipid classes compared to omega-6 PUFAs. Cholesterol content was low (17-24 mg.100-1g ww). Astaxanthin was detected only in bighead carp, whereas beta-carotene, vitamin D3 and vitamin A showed similar concentrations in both samples. Vitamin E content was higher in bighead carp (10.4 mg.100 g-1 w.w.). Trace elements content was higher in bighead carp showing a maximum value of As (0.312 mg.kg-1 w.w). All determined toxic elements were found below the recommended value in carp muscle. The results of the present study confirmed the high quality and safety of common carp and bighead carp meat. These freshwater species are valuable sources of essential nutrients such as proteins, vitamin D3 and long chain omega-3 PUFAs. Together with the nutrients, the information for low concentrations of toxic elements makes them valuable components of a healthy human diet.
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Klemm, Robin W. "Getting in Touch Is an Important Step: Control of Metabolism at Organelle Contact Sites." Contact 4 (January 2021): 251525642199370. http://dx.doi.org/10.1177/2515256421993708.
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Metabolic pathways are often spread over several organelles and need to be functionally integrated by controlled organelle communication. Physical organelle contact-sites have emerged as critical hubs in the regulation of cellular metabolism, but the molecular understanding of mechanisms that mediate formation or regulation of organelle interfaces was until recently relatively limited. Mitochondria are central organelles in anabolic and catabolic pathways and therefore interact with a number of other cellular compartments including the endoplasmic reticulum (ER) and lipid droplets (LDs). An interesting set of recent work has shed new light on the molecular basis forming these contact sites. This brief overview describes the discovery of unanticipated functions of contact sites between the ER, mitochondria and LDs in de novo synthesis of storage lipids of brown and white adipocytes. Interestingly, the factors involved in mediating the interaction between these organelles are subject to unexpected modes of regulation through newly uncovered Phospho-FFAT motifs. These results suggest dynamic regulation of contact sites between organelles and indicate that spatial organization of organelles within the cell contributes to the control of metabolism.
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Schäbler, Stefan, Kelechi M. Amatobi, Melanie Horn, et al. "Loss of function in the Drosophila clock gene period results in altered intermediary lipid metabolism and increased susceptibility to starvation." Cellular and Molecular Life Sciences 77, no. 23 (2020): 4939–56. http://dx.doi.org/10.1007/s00018-019-03441-6.
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Abstract The fruit fly Drosophila is a prime model in circadian research, but still little is known about its circadian regulation of metabolism. Daily rhythmicity in levels of several metabolites has been found, but knowledge about hydrophobic metabolites is limited. We here compared metabolite levels including lipids between period01 (per01) clock mutants and Canton-S wildtype (WTCS) flies in an isogenic and non-isogenic background using LC–MS. In the non-isogenic background, metabolites with differing levels comprised essential amino acids, kynurenines, pterinates, glycero(phospho)lipids, and fatty acid esters. Notably, detectable diacylglycerols (DAG) and acylcarnitines (AC), involved in lipid metabolism, showed lower levels in per01 mutants. Most of these differences disappeared in the isogenic background, yet the level differences for AC as well as DAG were consistent for fly bodies. AC levels were dependent on the time of day in WTCS in phase with food consumption under LD conditions, while DAGs showed weak daily oscillations. Two short-chain ACs continued to cycle even in constant darkness. per01 mutants in LD showed no or very weak diel AC oscillations out of phase with feeding activity. The low levels of DAGs and ACs in per01 did not correlate with lower total food consumption, body mass or weight. Clock mutant flies showed higher sensitivity to starvation independent of their background-dependent activity level. Our results suggest that neither feeding, energy storage nor mobilisation is significantly affected in per01 mutants, but point towards impaired mitochondrial activity, supported by upregulation of the mitochondrial stress marker 4EBP in the clock mutants.
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Pakiet, Alicja, Agnieszka Jakubiak, Paulina Mierzejewska, et al. "The Effect of a High-Fat Diet on the Fatty Acid Composition in the Hearts of Mice." Nutrients 12, no. 3 (2020): 824. http://dx.doi.org/10.3390/nu12030824.
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The Western diet can lead to alterations in cardiac function and increase cardiovascular risk, which can be reproduced in animal models by implementing a high-fat diet (HFD). However, the mechanism of these alterations is not fully understood and may be dependent on alterations in heart lipid composition. The aim of this study was to evaluate the effect of an HFD on the fatty acid (FA) composition of total lipids, as well as of various lipid fractions in the heart, and on heart function. C57BL/6 mice were fed an HFD or standard laboratory diet. The FA composition of chow, serum, heart and skeletal muscle tissues was measured by gas chromatography–mass spectrometry. Cardiac function was evaluated by ultrasonography. Our results showed an unexpected increase in polyunsaturated FAs (PUFAs) and a significant decrease in monounsaturated FAs (MUFAs) in the heart tissue of mice fed the HFD. For comparison, no such effects were observed in skeletal muscle or serum samples. Furthermore, we found that the largest increase in PUFAs was in the sphingolipid fraction, whereas the largest decrease in MUFAs was in the phospholipid and sphingomyelin fractions. The hearts of mice fed an HFD had an increased content of triacylglycerols. Moreover, the HFD treatment altered aortic flow pattern. We did not find significant changes in heart mass or oxidative stress markers between mice fed the HFD and standard diet. The above results suggest that alterations in FA composition in the heart may contribute to deterioration of heart function. A possible mechanism of this phenomenon is the alteration of sphingolipids and phospholipids in the fatty acid profile, which may change the physical properties of these lipids. Since phospho- and sphingolipids are the major components of cell membranes, alterations in their structures in heart cells can result in changes in cell membrane properties.
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Nave, J. F., H. d'Orchymont, J. B. Ducep, F. Piriou, and M. J. Jung. "Mechanism of the inhibition of cholesterol biosynthesis by 6-fluoromevalonate." Biochemical Journal 227, no. 1 (1985): 247–54. http://dx.doi.org/10.1042/bj2270247.
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6-Fluoromevalonate blocks the incorporation of mevalonic acid, but not that of isopentenyl pyrophosphate, into non-saponifiable lipids in a rat liver multienzyme system. With 3H-labelled 6-fluoromevalonate, it was found that 6-fluoromevalonate is converted to its phospho and pyrophospho derivatives in this system. The kinetics of the two kinases were studied. 6-Fluoromevalonate 5-pyrophosphate is a potent competitive inhibitor of pyrophosphomevalonate decarboxylase (Ki 37 nM). In the multienzyme assay for cholesterol biosynthesis, there is accumulation of mevalonate 5-phosphate and mevalonate 5-pyrophosphate in the presence of 5 microM-6-fluoromevalonate, and 6-fluoromevalonate 5-pyrophosphate is more effective than 6-fluoromevalonate in inhibiting cholesterol biosynthesis. We suggest therefore that 6-fluoromevalonate blocks cholesterol biosynthesis at the level of pyrophosphomevalonate decarboxylase after being pyrophosphorylated.
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Vengaiah, V., Naik A. Govardhan, and C. Changamma. "ALTERATIONS IN LIPID METABOLISM INDUCED BY THE BETEL LEAF STALK EXTRACT IN MALE ALBINO RATS." Biolife 2, no. 4 (2022): 1100–1109. https://doi.org/10.5281/zenodo.7229227.
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<strong>ABSTRACT</strong> The aim of this study was to investigate the antifertility efficacy of betel leaf stalk extract on lipid metabolites. The betel leaf stalk extract was administered at the dose of 50 mg/Kg/day through oral gavages method for 15 days. The impaired lipid metabolisms were observed in testis. The higher lipids in seminal vesicle and prostate gland indicate the elevation in lipid metabolism and lipogenesis, suggesting some alterations in the chemical composition of the seminal plasma and prostatic fluid. The accumulation of lipids in prostate causes the inhibition in lipase activity. The elevated testicular Phospholipids due to administration of extract results the disruption in integrity of the spermatozoal membrane and its adequate phospholipid composition leads to infertility. Significant reductions in triglycerides represent no risk of cancer. Increased concentration of cholesterol in testes suggests the impairment of spermatogenesis. The decreased cholesterol content of the epididymis and prostate gland indicated impairment in its synthesis and may hinder steroidogenesis, thus suggesting anti-steroidogenic potential of the plant extract. <strong>KEYWORDS: </strong>Betel leaf stalk, lipogenesis, Phospho lipids, Infertility, Spermatogenisis, Steroidogenisis. <strong>REFERENCES</strong> Thejashwini M.S, Krishna Ram H, Shivabasavaiah. 2012. Reversible antifertility effect of <em>cyamposis psoralioides </em>in male swiss albino mice. International Journal of advanced biological research<em>. </em>2(4):657-665. Chakraborty D, Shah B.2011. Antimicrobial antioxidative and antihemolytic activity of Piper betel leaf extracts. Int J Pharm Pharm Sci. 3:192-199. Mishra R.K, Singh S.K. 2009. Antispermatogenic and antifertility effects of fruits of <em>Piper nigrum </em>L. in mice. Indian journal of Experimental Biology. 47: 706-714. Sarkar M, Gangopadhyay P, Basak B, Chakrabarty K, Banerji J, Adhikary P, Chatterjee A. 2000. 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Ładniak, Agata, Małgorzata Jurak, Marta Palusińska-Szysz, and Agnieszka Ewa Wiącek. "The Influence of Polysaccharides/TiO2 on the Model Membranes of Dipalmitoylphosphatidylglycerol and Bacterial Lipids." Molecules 27, no. 2 (2022): 343. http://dx.doi.org/10.3390/molecules27020343.
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The aim of the study was to determine the bactericidal properties of popular medical, pharmaceutical, and cosmetic ingredients, namely chitosan (Ch), hyaluronic acid (HA), and titanium dioxide (TiO2). The characteristics presented in this paper are based on the Langmuir monolayer studies of the model biological membranes formed on subphases with these compounds or their mixtures. To prepare the Langmuir film, 1,2-dipalmitoyl-sn-glycero-3-phospho-rac-(1-glycerol) (DPPG) phospholipid, which is the component of most bacterial membranes, as well as biological material-lipids isolated from bacteria Escherichia coli and Staphylococcus aureus were used. The analysis of the surface pressure-mean molecular area (π-A) isotherms, compression modulus as a function of surface pressure, CS−1 = f(π), relative surface pressure as a function of time, π/π0 = f(t), hysteresis loops, as well as structure visualized using a Brewster angle microscope (BAM) shows clearly that Ch, HA, and TiO2 have antibacterial properties. Ch and TiO2 mostly affect S. aureus monolayer structure during compression. They can enhance the permeability of biological membranes leading to the bacteria cell death. In turn, HA has a greater impact on the thickness of E. coli film.
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Liao, Sijia, André Gollowitzer, Lisa Börmel та ін. "α-Tocopherol-13′-Carboxychromanol Induces Cell Cycle Arrest and Cell Death by Inhibiting the SREBP1-SCD1 Axis and Causing Imbalance in Lipid Desaturation". International Journal of Molecular Sciences 24, № 11 (2023): 9229. http://dx.doi.org/10.3390/ijms24119229.
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α-Tocopherol-13′-carboxychromanol (α-T-13′-COOH) is an endogenously formed bioactive α-tocopherol metabolite that limits inflammation and has been proposed to exert lipid metabolism-regulatory, pro-apoptotic, and anti-tumoral properties at micromolar concentrations. The mechanisms underlying these cell stress-associated responses are, however, poorly understood. Here, we show that the induction of G0/G1 cell cycle arrest and apoptosis in macrophages triggered by α-T-13′-COOH is associated with the suppressed proteolytic activation of the lipid anabolic transcription factor sterol regulatory element-binding protein (SREBP)1 and with decreased cellular levels of stearoyl-CoA desaturase (SCD)1. In turn, the fatty acid composition of neutral lipids and phospholipids shifts from monounsaturated to saturated fatty acids, and the concentration of the stress-preventive, pro-survival lipokine 1,2-dioleoyl-sn-glycero-3-phospho-(1′-myo-inositol) [PI(18:1/18:1)] decreases. The selective inhibition of SCD1 mimics the pro-apoptotic and anti-proliferative activity of α-T-13′-COOH, and the provision of the SCD1 product oleic acid (C18:1) prevents α-T-13′-COOH-induced apoptosis. We conclude that micromolar concentrations of α-T-13′-COOH trigger cell death and likely also cell cycle arrest by suppressing the SREBP1-SCD1 axis and depleting cells of monounsaturated fatty acids and PI(18:1/18:1).
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Sachinidis, Agapios, Claudia Seul, Stefan Seewald, Ioanna Gouni-Berthold, Yon Ko, and Hans Vetter. "Gangliosides GM1 and GM2 Stimulate Vascular Smooth Muscle Cell Growth and the Mitogen-Activated Protein Kinase Signaling Transduction Pathway." Hypertension 36, suppl_1 (2000): 721. http://dx.doi.org/10.1161/hyp.36.suppl_1.721-c.
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P156 The gangliosides GM1, GM2 and GM3 appear to be involved in the development of cardiovascular diseases. However, little is known about the direct effects of gangliosides on vascular smooth muscle cell growth (VSMC) growth and mitogenic signal transduction pathway. Stimulation of quiescent VSMCs with GM1 (5, 20, 50 and 100 μM) resulted in a dose-dependent increase in [ 3 H]thymidine incorporation from 41±3.5 (basal value) to 64±3.7, 69.6±7.1, 180±11 and 190±10.7 (mean±SE, n=3), respectively. GM3 (1 to 50 μM) had no effects on DNA synthesis whereas 100 μM GM3 caused a reduction in DNA synthesis from 41±3.5 to 12 ±1.4 cpm/μg protein (mean±SE, n=3). Stimulation of quiescent VSMCs with 50 μM GM1 and GM2 caused a 95±7% and 100±4% increase in cell counts (mean±SE, n=3) whereas 50 μM GM3 had no effects. Phosphorylated (activated) MAP kinases were detected by the enhanced chemiluminescence western blotting method using a phospho-specific extracellular signal-response kinases (ERK)1/2 antibody, a phospho-specific p38 MAPK antibody and a phospho-specific c-Jun N-terminal kinase/stress-activated protein kinases (JNK/SAPK) (the p54 and p46 isoforms) antibody. Stimulation of the cells with 50 μM GM1 and GM2 for 1 to 30 min resulted in time-dependent increase of phosphorylated ERK1/2 and p54/p46 with a maximum at 15 min (5-fold increase). The effect of GM1 and GM2 on the ERK1/2 and JNK/SAPK was also dose-dependent with maximal effect at 50 μM. Gangliosides had no effect on p38 MAPK phosphorylation. Treatment of the cells with 100 ng/ml pertussis toxin (PTX) (G i protein inhibitor) and 20 μM PD 98059 (MEK inhibitor) caused a complete inhibition of the phosphorylation of the ERK1/2 and JNK/SAPK. We conclude that the gangliosides may be involved in the development of cardiovascular diseases via proliferation of VSMCs. Furthermore, like bioactive lipids, gangliosides are able to stimulate the MAP kinase pathway via a PTX-sensitive G i coupled receptor.
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Panzenboeck, Lisa, Nina Troppmair, Sara Schlachter, Gunda Koellensperger, Jürgen Hartler, and Evelyn Rampler. "Chasing the Major Sphingolipids on Earth: Automated Annotation of Plant Glycosyl Inositol Phospho Ceramides by Glycolipidomics." Metabolites 10, no. 9 (2020): 375. http://dx.doi.org/10.3390/metabo10090375.
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Glycosyl inositol phospho ceramides (GIPCs) are the major sphingolipids on earth, as they account for a considerable fraction of the total lipids in plants and fungi, which in turn represent a large portion of the biomass on earth. Despite their obvious importance, GIPC analysis remains challenging due to the lack of commercial standards and automated annotation software. In this work, we introduce a novel GIPC glycolipidomics workflow based on reversed-phase ultra-high pressure liquid chromatography coupled to high-resolution mass spectrometry. For the first time, automated GIPC assignment was performed using the open-source software Lipid Data Analyzer (LDA), based on platform-independent decision rules. Four different plant samples (salad, spinach, raspberry, and strawberry) were analyzed and the results revealed 64 GIPCs based on accurate mass, characteristic MS2 fragments and matching retention times. Relative quantification using lactosyl ceramide for internal standardization revealed GIPC t18:1/h24:0 as the most abundant species in all plants. Depending on the plant sample, GIPCs contained mainly amine, N-acetylamine or hydroxyl residues. Most GIPCs revealed a Hex-HexA-IPC core and contained a ceramide part with a trihydroxylated t18:0 or a t18:1 long chain base and hydroxylated fatty acid chains ranging from 16 to 26 carbon atoms in length (h16:0–h26:0). Interestingly, four GIPCs containing t18:2 were observed in the raspberry sample, which was not reported so far. The presented workflow supports the characterization of different plant samples by automatic GIPC assignment, potentially leading to the identification of new GIPCs. For the first time, automated high-throughput profiling of these complex glycolipids is possible by liquid chromatography-high-resolution tandem mass spectrometry and subsequent automated glycolipid annotation based on decision rules.
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Sevrain, Charlotte M., Jean-Pierre Haelters, Aurélie Chantôme, et al. "Glyco-Phospho-Glycero Ether Lipids (GPGEL): synthesis and evaluation as small conductance Ca2+-activated K+ channel (SK3) inhibitors." MedChemComm 3, no. 11 (2012): 1471. http://dx.doi.org/10.1039/c2md20207g.
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Balasubramanyam, Ashok, Harry Mersmann, Farook Jahoor, et al. "Effects of transgenic expression of HIV-1 Vpr on lipid and energy metabolism in mice." American Journal of Physiology-Endocrinology and Metabolism 292, no. 1 (2007): E40—E48. http://dx.doi.org/10.1152/ajpendo.00163.2006.
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HIV infection is associated with abnormal lipid metabolism, body fat redistribution, and altered energy expenditure. The pathogenesis of these complex abnormalities is unclear. Viral protein R (Vpr), an HIV-1 accessory protein, can regulate gene transcription mediated by the glucocorticoid receptor and peroxisome proliferator-activated receptor-γ and affect mitochondrial function in vitro. To test the hypothesis that expression of Vpr in liver and adipocytes can alter lipid metabolism in vivo, we engineered mice to express Vpr under control of the phospho enolpyruvate carboxykinase promoter in a tissue-specific and inducible manner and investigated the effects of dietary fat, indinavir, and dexamethasone on energy metabolism and body composition. The transgenic mice expressed Vpr mRNA in white and brown adipose tissues and liver and immunoaffinity capillary electrophoresis revealed that they had free Vpr protein in the plasma. Compared with wild-type (WT) animals, Vpr mice had lower plasma triglyceride levels after 6 wk ( P < 0.05) but not after 10 wk of a high-fat diet and lower plasma cholesterol levels after 10 wk of high-fat diet ( P < 0.05). Treatment with dexamethasone obviated group differences, whereas indinavir had no significant independent effect on lipids. In the fasted state, Vpr mice had a higher respiratory quotient than WT mice ( P < 0.05). These data provide the first in vivo evidence that HIV-1 Vpr expressed at low levels in adipose tissues and liver can 1) circulate in the blood, 2) regulate lipid and fatty acid metabolism, and 3) alter fuel selection for oxidation in the fasted state.
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Pašalić, Lea, Barbara Pem, and Danijela Bakarić. "Lamellarity-Driven Differences in Surface Structural Features of DPPS Lipids: Spectroscopic, Calorimetric and Computational Study." Membranes 13, no. 1 (2023): 83. http://dx.doi.org/10.3390/membranes13010083.
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Although single-lipid bilayers are usually considered models of eukaryotic plasma membranes, their research drops drastically when it comes to exclusively anionic lipid membranes. Being a major anionic phospholipid in the inner leaflet of eukaryote membranes, phosphatidylserine-constituted lipid membranes were occasionally explored in the form of multilamellar liposomes (MLV), but their inherent instability caused a serious lack of efforts undertaken on large unilamellar liposomes (LUVs) as more realistic model membrane systems. In order to compensate the existing shortcomings, we performed a comprehensive calorimetric, spectroscopic and MD simulation study of time-varying structural features of LUV made from 1,2-dipalmitoyl-sn-glycero-3-phospho-L-serine (DPPS), whereas the corresponding MLV were examined as a reference. A substantial uncertainty of UV/Vis data of LUV from which only Tm was unambiguously determined (53.9 ± 0.8 °C), along with rather high uncertainty on the high-temperature range of DPPS melting profile obtained from DSC (≈50–59 °C), presumably reflect distinguished surface structural features in LUV. The FTIR signatures of glycerol moiety and those originated from carboxyl group serve as a strong support that in LUV, unlike in MLV, highly curved surfaces occur continuously, whereas the details on the attenuation of surface features in MLV were unraveled by molecular dynamics.
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Araújo, Vilma Barbosa da Silva, Vânia Maria Barboza Silva, Evandro Bernardo Lira, et al. "Metabolites produced by microalgae from northeastern Brazil with potential food industry uses." Research, Society and Development 11, no. 6 (2022): e7411628724. http://dx.doi.org/10.33448/rsd-v11i6.28724.
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The production potential of metabolites of interest to the food industry was evaluated in 17 microalgae species isolated from natural sources in northeastern Brazil. The species were cultivated to their stationary phase under controlled conditions, when the experiments were interrupted and the dry biomass harvested. We observed differences in their growth parameters, productivity, and the biochemical compositions of their biomasses, with high levels of protein productivity in Monoraphidium litorale D296WC (48.96%), Kirchneriella concorta D498WC (42.49%), Monoraphidium griffithi D499WC (48.37%), Chlamydomonas sp. D530WC (44.80%), and Cosmarium sp cf. depressum D578WC (49.32). The greatest carbohydrate productivities were observed in Xanthonema sp. D464WC (34.15%), K. concorta D498WC (38.95%), and Scenedesmus acuminatus D514WC (36.54%). The three different extraction techniques of microalgae lipids all gave slightly different results, with the method utilizing phospho-vanillin being considered the most rapid and it requires only small quantities of biomass. Unsaturated fatty acids (oleic, linoleic, and linolenic) were encountered at high levels in most of the species, especially α-linolenic acid (ω3), which reached concentrations above 30% in Golenkinia radiata (D325WC). Due to their high productivity, rapid growth, and the large numbers of important dietary metabolites they produce, the species Monoraphidium litorale (D296WC), Xanthonema sp. (D464WC) and Monoraphidium griffithi (D499WC) show significant potential for utilization by the food industry as sources of proteins, lipids, and carbohydrates.
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Feulner, J. Amelia, Mingfang Lu, JohnM Shelton, Mei Zhang, James A. Richardson, and Robert S. Munford. "Identification of Acyloxyacyl Hydrolase, a Lipopolysaccharide- Detoxifying Enzyme, in the Murine Urinary Tract." Infection and Immunity 72, no. 6 (2004): 3171–78. http://dx.doi.org/10.1128/iai.72.6.3171-3178.2004.
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ABSTRACT Acyloxyacyl hydrolase (AOAH) is an unusual but highly conserved lipase, previously described only in myeloid cells, that removes secondary fatty acyl chains from bacterial lipopolysaccharides (LPS) and may also act on various glycero(phospho)lipids. Deacylation by AOAH greatly reduces the ability of LPS to stimulate cells via CD14-MD-2-Toll-like receptor 4. We report here that renal cortical tubule cells produce AOAH and secrete it into urine, where it can deacylate LPS. In vitro studies revealed that proximal tubule cells secrete pro-AOAH, which can be taken up by bladder cells and processed to the heterodimeric, more enzymatically active, mature form of AOAH. AOAH can then be used by the recipient cells to deacylate LPS. The enzyme produced by proximal tubule epithelium may thus be shared with downstream cells. In addition, mature AOAH is found in the urine. We suggest that cortical tubule cells may produce and secrete AOAH to limit inflammatory responses to gram-negative bacteria throughout the urinary tract.