Benzyl Amino Purine and Gibberellic Acid Coupled to Nitrogen-Limited Stress Induce Fatty Acids,Biomass Accumulation,and Gene Expression in <i>Scenedesmus Obliquus</i>

The need for renewable energy sources makes microalgae an essential feedstock for biofuels production. The molecular aspects and the response to nitrogen (N)-limited conditions with a phytohormone stimulus in microalgae have been slightly explored. In this work, Scenedesmus obliquus was used as a study model to analyze the effect of benzyl amino purine (BAP) and gibberellic acid (GA) coupled to nitrogen limitation on cell growth, biomass and fatty acids. The selected 10^-5 M BAP increased the biomass by 1.44-fold, and 10^-6 M GA by 1.35-fold. The total lipids also increased by 2.8 and 1.11-fold, respectively. The 10^-5 M BAP and
10^-6 M GA addition to S. obliquus cultures at different initial nitrogen percentages (N-0, N-25, and N-50) showed a significant increase in cell growth and biomass productivity compared to the unstimulated cultures. BAP N-0 and GA N-0 produced the highest lipid yields with 55% and 50%, respectively. The lipid profile analysis revealed an increase, particularly in C18:1 and C16:0 fatty acids. Gene expression analysis showed an over-expression of acyl carrying protein (ACP), stearoyl-ACP desaturase (SAD), fatty acid acyl-ACP thioesterase (FATA), and diacylglycerol acyltransferase (DGAT) genes, which were mainly induced by nitrogen limitation. Furthermore, BAP and GA produced a significant over-expression on these genes in the N-replete cultures. This study shows that BAP and GA, coupled to N limitation stress, can be used to increase the biomass and lipid production in S. obliquus for sustainable biofuels.     

Biomass production,total protein,chlorophylls, lipids and fatty acids of freshwater green and blue-green algae under different nitrogen regimes

Two green algae (Chlorella vulgaris and Scenedesmus obliquus) and four blue-green algae (Anacystis nidulans, Microcystis aeruginosa, Oscillatoria rubescens and Spirulina platensis) were grown in 81 batch cultures at different nitrogen levels. In all the algae increasing N levels led to an increase in the biomass (from 8 to 450 mg/l), in protein content (from 8 to 54 %) and in chlorophyll. At low N levels, the green algae contained a high percentage of total lipids (45 % of the biomass). More than 70 % of these were neutral lipids such as triacylglycerols (containing mainly 16:0 and 18:1 fatty acids) and trace amounts of hydrocarbons. At high N levels, the percentage of total lipids dropped to about 20 % of the dry weight. In the latter case the predominant lipids were polar lipids containing polyunsaturated C₁₆ and C₁₈ fatty acids. The blue-green algae, however, did not show any significant changes in their fatty acid and lipid compositions, when the nitrogen concentrations in the nutrient medium were varied. Thus the green but not the blue-green algae can be manipulated in mass cultures to yield a biomass with desired fatty acid and lipid compositions. The data may indicate a hitherto unrecognized distinction between prokaryotic and eukaryotic organisms.     

Lipid class composition of bleached and recovering Porites compressa Dana, 1846 and Montipora capitata Dana, 1846 corals from Hawaii

Corals rely on stored reserves, especially lipids, to survive bleaching events. Lipid class composition reveals the lipid source, and provides evidence of metabolic changes (i.e., photoautotrophic or heterotrophic) during bleaching and recovery. Porites compressa Dana, 1846 and Montipora capitata Dana, 1846 corals were experimentally bleached in outdoor tanks with seawater temperature elevated to 30 °C (treatment corals). Additional control fragments were maintained in separate tanks at ambient temperatures (27 °C). After one month, all fragments were returned to the reef for 0, 1.5, 4, or 8 months. Lipid class composition was analyzed by Iatroscan (thin layer chromatography-flame ionization detection). In treatment P. compressa, triacylglycerol (TG) decreased at 0 and 1.5 months, phospholipid (PL) also decreased at 1.5 months, and both remained lower relative to controls along with wax esters (WE) after 8 months. Neither treatment nor control P. compressa had any detectable monoacylglycerol (MG) or diacylglycerol (DG). Overall, P. compressa first consumed available storage, then structural lipids, and all lipid classes remained low at the end of the study. In treatment M. capitata, TG and PL decreased, while MG increased relative to controls at 0 months. At 4 months, free fatty acid (FFA), sterol (ST), and PL in treatment M. capitata were two to ten times higher than controls. Treatment and control lipid class composition were not different from each other at 8 months. In contrast to P. compressa, M. capitata consumed some lipid classes and augmented others, probably due to sequential metabolism of storage lipids and increased heterotrophy. Overall, lipid class assimilation was more rapid in treatment M. capitata corals that switch between heterotrophy and photoautotrophy, than in treatment P. compressa corals that rely mostly on photoautotrophy.     

The assimilation of glycerol into lipid acyl chains and associated carbon backbones of Nannochloropsis salina varies under nitrogen replete and deplete conditions

Mixotrophic cultivation can increase microalgae productivity, yet the associated lipid metabolism remains mostly unknown. Stable isotope labeling was used to track assimilation of glycerol into the triacylglyceride (TAG) and membrane lipids of Nannochloropsis salina. In N-replete media, glycerol uptake and ¹³C incorporation into acyl chains were, respectively, 6-fold and 12-fold higher than in N-deplete conditions. In N-replete cultures, 42% of the carbon in the consumed glycerol was assimilated into lipid acyl chains, mostly in membrane lipids rather than TAG. In N-deplete cultures, only 11% of the limited amount of consumed glycerol was fixed into lipid acyl chains. Labeled lipid-associated glycerol backbones were predominantly ¹³C₃ labeled, suggesting that intact glycerol molecules were directly esterified with fatty acids/polar head groups. However, the presence of singly and doubly labeled lipid-bound glycerol species suggested that some glycerol also went through the central carbon metabolism before forming glycerol-3-phosphate destined for lipid esterification. ¹³C incorporation was higher in the saturated and monounsaturated than the polyunsaturated acyl chains of TAG, indicating the flux of carbon from glycerol went first to de novo fatty acid synthesis before acyl editing reactions. The results demonstrate that nitrogen availability influences both glycerol consumption and utilization for lipid synthesis in Nannochloropsis, providing novel insights for developing mixotrophic cultivation strategies.     

Changes in Composition of Non-polar Lipids of the Axis and Root of Germinating Soybeans

Soybean seedlings were grown at 28°C under dark or light conditions for 12 days. Non-polar lipids (NPL) were separated by silicic acid column chromatography from total lipids in epicotyl containing young leaves, hypocotyl and root. The glyceride (TG, DG, and MG), free fatty acid (FFA) and sterol lipid (SE) components in NPL were analyzed mainly by thin-layer and gas-liquid chromatographies (TLC and GLC).

During germination, the amounts of polar lipids (PL) markedly increased in the tissues of soybean seedlings, especially in light-grown seedlings, whereas these of NPL increased slightly or maintained constant values. The features of the compositions and changing patterns of NPL in the tissues were more clarified in light-grown seedlings than in dark-grown ones. The pattern of change in fatty acid composition was similar in TG and 1,2-DG, which showed higher proportions of linoleic and linolenic acids, whereas FFA, 1,3-DG or MG had high proportions of saturated fatty acids. These results indicate that the compositions and changing patterns of NPL and their fatty acids in the tissues depend on the differences under two germinating conditions tested.     

Seasonal changes in oleosomic lipids and fatty acids of perennial root nodules of beach pea

Seasonal changes in the fatty acid composition of phospholipids (PL), monoglycerides (MG), diglycerides (DG), free fatty acids (FA) and triglycerides (TG) separated from oleosomes (lipid bodies) of perennial root nodules of beach pea (Lathyrus maritimus) were analysed. Thin layer chromatography (TLC) revealed that PL and MG are the major lipids in nodule oleosomes. The fatty acid profile and overall double bond index (DBI) varied among lipid classes depending upon the season. High DBI in PL and MG found during late winter and early spring indicated that they may play a major role in winter survival and regeneration of perennial nodules. The DBI of DG was high at the end of the fall season and the DBI of FA and TG was high in summer months. The dominant fatty acids are C16:0 followed by C18:0 and C18:1. The levels of many unsaturated fatty acids such as C18:1, C18:2 and C18:3 increased while saturated fatty acid C18:0 decreased during winter. These unsaturated fatty acids possibly play an important role in the protection of nodule cells from cold stress. Nodules seem to retain some fatty acids and selectively utilize specific fatty acids to survive the winter and regenerate in spring.     

Nitrogen deficiency in Arabidopsis affects galactolipid composition and gene expression and results in accumulation of fatty acid phytyl esters

Nitrogen is an essential nutrient for plants because it represents a major constituent of numerous cellular compounds, including proteins, amino acids, nucleic acids and lipids. While N deprivation is known to have severe consequences for primary carbon metabolism, the effect on chloroplast lipid metabolism has not been analysed in higher plants. Nitrogen limitation in Arabidopsis led to a decrease in the chloroplast galactolipid monogalactosyldiacylglycerol (MGDG) and a concomitant increase in digalactosyldiacylglycerol (DGDG), which correlated with an elevated expression of the DGDG synthase genes DGD1 and DGD2. The amounts of triacylglycerol and free fatty acids increased during N deprivation. Furthermore, phytyl esters accumulated containing medium-chain fatty acids (12:0, 14:0) and a large amount of hexadecatrienoic acid (16:3). Fatty acid phytyl esters were localized to chloroplasts, in particular to thylakoids and plastoglobules. Different polyunsaturated acyl groups were found in phytyl esters accumulating in Arabidopsis lipid mutants and in other plants, including 16:3 and 18:3 species. Therefore N deficiency in higher plants results in a co-ordinated breakdown of galactolipids and chlorophyll with deposition of specific fatty acid phytyl esters in thylakoids and plastoglobules of chloroplasts.     

Effects of Growth Temperature on Fatty Acid and Alk-1-Enyl Group Compositions of Veillonella parvula and Megasphaera elsdenii Phospholipids

Veillonella parvula ATCC 10790, an anaerobic gram-negative coccus, contains diacyl and alk-1-enyl acyl (plasmalogen) forms of phosphatidylethanolamine and phosphatidylserine. We studied the effect of growth temperature on the lipid composition of this strain. There was a small increase in the phosphatidylethanolamine content but no change in the content of plasmalogens at the lower growth temperatures tested. The total acyl chains and the plasmalogen acyl chains contained between 73 and 80% mono-unsaturated fatty acids at all growth temperatures. The plasmalogen alk-1-enyl chains were significantly more unsaturated in cells grown at 30 and 25°C than in cells grown at 37°C. Differential scanning calorimetry of the hydrated phospholipids showed lower phase transition temperatures for the lipids from the cells grown at the lower temperatures. In Megasphaera elsdenii lipids, which are similar in composition to the lipids of V. parvula, the proportion of phosphatidylethanolamine also increased slightly at lower growth temperatures, with no significant change in the content of plasmalogens. M. elsdenii contained cyclopropane fatty acyl and alk-1-enyl chains in addition to the mono-unsaturated and saturated chains previously reported. As cells entered the stationary phase of growth at 30 and 42.5°C, there was a reciprocal increase in the proportion of cyclopropane acyl chains and decrease in the unsaturated moieties. The total proportion of cyclopropane and unsaturated acyl and alk-1-enyl chains was more than 65% at all growth temperatures studied, and there was no discernible increase in the sum of these moieties at the lower growth temperatures.     

Application of highly purified anti-galactocerebroside antibody to the analysis of lipid-lipid or lipid-protein interactions. Significance of lipid matrix

The significance of the lipid matrix in the reaction of liposomal antigen, antibody and complement (Ag-Ab-C) was analyzed using purified anti-glactocerebroside antibody and synthetic lipids and the following results were obtained. 1. For the optimal Ag-Ab-C reaction it was necesary that galactocerebroside (galxCMH) and lecithin molecules were well dispersed by virtue of cholesterol (Chol) and that the molar ratio of cholesterol to the sum of galactocerebroside and lecithin was more than one. 2. The Ag-Ab-C reactivity changed depending upon the chain length of lecithin, and the maximal reaction was observed in the case of dilauroylphosphatidylcholine. When the fatty acyl chain of lecithin was either shorter or longer than that of dilaurosylphosphatidylcholine, the reactivity was reduced. 3. The Ag-Ab-C reactivity was increased by elongation of the fatty acyl chain of galactocerebroside and an abrupt change was found to be around the carbon number 8 to 10 of the fatty acyl chain. 4. The Ag-Ab-C reactivity was elevated by the increase in unsaturation of fatty acyl moiety. 5. There is a tendency that an increase in the charge of the lipid matrix leads to the reduction of the Ag-Ab-C reactivity. 6. The results suggest that the physicochemical properties of lipids and especially the lipid-lipid interaction in the hydrophobic region of the lipid matrix play an important role in the Ag-Ab-C reaction.     

High rates of [1-14C]acetate incorporation into the lipid of isolated spinach chloroplasts.

Spinach chloroplasts, isolated by techniques yielding preparations with high O2- evolving activity, showed rates of light-dependent acetate incorporation into lipids 3-4 fold higher than any previously reported. Incorporation rates as high as 500 nmol of acetate/h per mg of chlorophyll were measured in buffered sorbitol solutions containing only NaHCO3 and [1-14C]acetate, and as high as 800 nmol/h per mg of chlorophyll when 0.13 mM-Triton X-100 was also included in the reaction media. The fatty acids synthesized were predominantly oleic (70-80% of the total fatty acid radioactivity) and palmitic (20-25%) with only minor amounts (1-5%) of linoleic acid. Linolenic acid synthesis was not detected in the system in vitro. Free fatty acids accounted for 70-90% of the radioactivity incorporated and the remainder was shared fairly evenly between 1,2-diacylglycerols and polar lipids. Oleic acid constituted 80-90% of the free fatty acids synthesized, but the diacylglycerols and polar lipids contained slightly more palmitic acid than oleic acid. Triton X-100 stimulated the synthesis of diacylglycerols 3-6 fold, but stimulated free fatty acid synthesis only 1-1.5-fold. Added glycerol 1-phosphate stimulated both the synthesis of diacylglycerols and palmitic acid relative to oleic acid, but did not increase acetate incorporation into total chloroplast lipids. CoA and ATP, when added separately, stimulated acetate incorporation into chloroplast lipids to variable extents and had no effect on the types of lipid synthesized, but when added together resulted in 34% of the incorporated acetate appearing in long-chain acyl-CoA. Pyruvate was a much less effective precursor of chloroplast fatty acids than was acetate.     

Infrared and time-resolved fluorescence spectroscopic studies of the polymorphic phase behavior of phosphatidylethanolamine/diacylglycerol lipid mixtures

Fourier transform infrared (FTIR) and time-resolved fluorescence spectroscopy have been employed to examine the structural dynamics of lipid fatty acyl chains and lipid/water interfacial region of a binary lipid mixture containing unsaturated phosphatidylethanolamine (PE) and diacylglycerol (DG). Infrared vibrational frequencies of the CH2 symmetric stretching and the C = O stretching bands of the lipids were measured at different lipid compositions and temperatures. For 0% DG, the lamellar gel to lamellar liquid crystalline (L beta-L alpha) and the L alpha to inverted hexagonal (L alpha-HII) phase transitions were observed at approximately 15 degrees and 55 degrees C, respectively. As the DG content increased gradually from 0% to 15%, the L alpha-HII phase transition temperature decreased drastically while the L beta-L alpha phase transition temperature decreased only slightly. At 10% DG, a merge of these two phase transitions was noticed at approximately 10 degrees C. For the composition study at 23 degrees C, the L alpha-HII transition occurred at approximately 6-10% DG as indicated by abrupt increases in both the CH2 and C = O stretching frequencies at those DG contents. Using time-resolved fluorescence spectroscopy, abrupt decreases in both the normalized long time residual and the initial slope of the anisotropy decay function of lipid probes, 1-palmitoyl-2-[[2-[4-(6-phenyl-trans-1,3,5- hexatrienyl)phenyl]ethyl]carbonyl]-3-sn-phosphatidylcholine, in these PE/DG mixtures were observed at the L alpha-HII phase transition. These changes in the anisotropy decay parameters suggested that the rotational dynamics and orientational packing of the lipids were altered at the composition-induced L alpha-HII transition, and agreed with a previous temperature-induced L alpha-HII transition study on pure unsaturated PE (Cheng (1989) Biophys. J. 55, 1025-1031). The fluorescence lifetime of water soluble probes, 8,1-anilinonapthalenes sulfonate acid, in PE/DG mixtures increased abruptly at the L alpha-HII phase transition, suggesting that the conformation and hydration of the lipid/water interfacial region also undergo significant changes at the L alpha-HII transition.     

Free Fatty Acids, Neutral Glycerides, and Phosphoglycerides in Transient Focal Cerebral Ischemia

Abstract: Cerebral ischemia is known to cause an increase in levels of free fatty acids (FFAs) and diacylglycerols (DGs), although the mechanism(s) leading to these changes is not well understood. In this study, we examined FFA and DG levels along with those of other lipids in rats during and after transient focal cerebral ischemia induced by temporary occlusion of the right middle cerebral artery (MCA) and both common carotid arteries. During the duration of ischemia (15–60 min), there was a time-dependent increase (two- to 10-fold) in FFA levels in the right MCA cortex, whereas levels of DG and other lipids were not altered appreciably. FFA levels in right MCA cortex returned to near control values after reperfusion. However, following a 60-min ischemic insult, there was a second phase of FFA level increase that was evident after 16 h. The FFAs accumulated during the ischemia period were different from those after reperfusion, suggesting differences in mechanisms for their release. During the second phase of FFA release, there were increases in levels of DGs and triacylglycerols (TGs) with unusually high proportions of 20:4(n-6) and 22:6(n-3). The increases in FFA, DG, and TG levels were marked by a decrease in content of phosphoglycerides (PGs). It is interesting that the increases in levels of FFAs and neutral glycerides accounted only for 10% of the total PGs depleted. The lipid changes during this reperfusion period correlated well with the development of cortical infarct. Because FFAs are potent inhibitors of mitochondrial respiratory function, the time-dependent FFA accumulation during the ischemia period may be an important determinant for the extent of ischemia-induced injury after reperfusion.     

Biochemical changes induced by accelerated aging in sunflower seeds. I. Lipid peroxidation and membrane damage

When stored at 42°C and 100% relative humidity for 1 to 8 days, sunflower seeds (Helianthus annuus L. cv. Rodeo) aged prematurely and lost 25% of their initial viability. A ten-fold increase in conjugated dienes as well as a decrease of unsaturated fatty acids in diacylglycerol and polar lipids fractions were observed after 8 days of accelerated aging, demonstrating the occurrence of lipid peroxidation in prematurely aged sunflower seeds. However, the viability remained relatively high. The absence of membrane damage in seeds and of lipid peroxidation in isolated microsqmes suggested that lipid peroxidation concerned mainly lipid reserves. These results suggest that, at least within the first 8 days of treatment, the lipid reserve in sunflower seeds might act as a detoxifying trap, protecting membranes from excessive damage.     

Characterization of the seed and leaf lipids of high and low linolenic acid flax genotypes

The total seed lipids of four flax (Linum usitatissimum) genotypes, differing markedly in their acyl composition, were extracted and fractionated using column, preparative, and thin-layer chromatography. In the total lipid extract of seeds, the lower linolenate content of the cultivar Glenelg (39.1% compared to that of cv. Croxton (50.5%) was associated with a higher oleate content. Further reductions in linolenate content in the induced mutants of cv. Glenelg, M1722 (17.2%) and "Zero" (1.9%) were accompanied by equivalent increases in linoleate but only minor increases in oleate. Similar changes were observed in the major triacylglycerol fraction of the simple lipids (fatty acid esters of glycerol and sterols), but there was considerable heterogeneity for acyl composition in the minor simple lipid components, including both diacylglycerols and sterol esters, and the complex lipids (glycolipids and phospholipids). The induced mutations substantially reduced linolenate content of all lipid fractions but in no case was it eliminated. Maturation of "Zero" seed at 15/10 degrees C (compared to 24/19 degrees C) increased linoleate and decreased stearate and oleate contents in all lipid fractions. In contrast to seed lipids, the acyl composition of the leaf lipids of the mutant genotypes was the same as those of their parent.     

Etioplast Development in Dark-grown Leaves of Zea mays L

The ultrastructure of etioplasts and the acyl lipid and the fatty acid composition of sequential 2-centimeter sections cut from the base (youngest) to the top (oldest) of nonilluminated 5-day-old etiolated leaves of Zea mays L., and the acyl lipid and fatty acid composition of the etioplasts isolated from them have been investigated. There is a 2.5-fold increase in the size of the plastids from the base to the tip of the leaf, and an increase both in the size of the prolamellar body and in the length of lamellae attached to it. The etioplasts in the bundle sheath and mesophyll cells of the older, but not the younger leaf tissue, are morphologically distinct. The monogalactosyl and digalactosyldiglycerides, phosphatidylcholine, phosphatidylglycerol, and phosphatidylinositol were the only detectable acyl lipids in the isolated etioplast fractions. Together with phosphatidylethanolamine these were also the major acyl lipids in the whole leaf sections. With increasing age of the leaf tissue, increases occurred in two of the major plastid lipids, monogalactosyldiglyceride and phosphatidylglycerol, while the levels of essentially nonplastid lipids remained constant or declined slightly. The monogalactosyldiglyceride to digalactosyldiglyceride ratio increased from 0.4 to 1.1 in the tissue sections of increasing age and from 0.7 to 1.2 in the etioplasts isolated from them. Similarly, the galactolipid to phospholipid ratio increased from 0.8 to 1.4 in the tissue and from 0.5 to 4.5 in the isolated plastids. In the latter, the proportions of phosphatidylglycerol (as a per cent of total phospholipid) increased from 20 to 41% with increasing age of plastids.

Linolenic acid was the major fatty acid in the total lipid of each of the etioplast fractions, but it was only the major fatty acid in the total lipid of the oldest leaf tissue. Its proportion in both total lipid extracts and individual lipids increased with age. The trans Δ³ hexadecenoic acid was absent from all lipids. The protochlorophyllide content of the tissue increased with age. The results are discussed in relation to the use of illuminated etiolated leaves for studying chloroplast development.

     

Absorption of dietary triacylglycerol by lipolysis and lipid resynthesis in the mesenteron of larvalAeshna cyanea (insecta,odonata)

Summary Voluntary uptake of triolein, margarine, and lipid-rich natural food (Tubifex) by fasting dragonfly larvae (Aeshna cyanea) led to heavy accumulations of lipid absorption droplets in the enterocytes within 2 days, while subsequent lipid clearance of the midgut epithelium took several weeks depending on the ingested lipid load. No endocytotic lipid uptake was observed after application of a molecular-dispersed fat dye. The smallest lipid droplets first appeared in the subapical groundplasm of the enterocytes and showed a reversible increase in size on their way towards the base. Lipid droplets were also observed at appropriate intervals after oral administration of oleic acid, after feeding margarine in the cold, and after injection of triolein into the isolated midgut.Comparative biochemical analysis after triolein feeding evidenced release of lipase and hydrolytic liberation of FA from TG in the midgut lumen, as well as time-dependent accumulations of TG in the midgut epithelium and of DG in the hemolymph.Oral injection of [¹⁴C] oleic acid was followed by its rapid absorption into the midgut epithelium, where it was utilized for the synthesis of MG and esterification to DG and TG. Discharge of radioactive lipid into the hemolymph occurred in the form of FA and DG, while the rectal fat body showed approximately equal labeling of the FA, DG, and TG fractions.Abbreviations AG acylglycerol - DG diacylglycerol - ER endoplasmic reticulum - FA fatty acid - MG monoacylglycerol - TG triacylglycerol Dedicated to Prof. Dr. Dr. R.Lehmensick, Bonn, in honor of his 85^th birthday.     

Diacylglycerol transport in the insect fat body: evidence of involvement of lipid droplets and the cytosolic fraction

In this report we show the existence of a distinct pool of fat body diacylglycerol (DG) that can be distinguished from the bulk DG. This is a dynamic pool of DG that uses FA entering the fat body from the hemolymph, whereas the bulk DG uses the fatty acids stored in the fat body fat droplets. Using a dual labeling technique, it was possible to compare the effect of hormone-stimulated DG synthesis and secretion on the distribution of radiolabeled FA among the lipids of the dynamic pool (short-term radiolabeling), with the hormonal effect on the total complement of fat body lipids (long-term radiolabeling). We observed that, whereas DG represents 2% to 3% of the fat body lipid mass, about 20% of the short-term radiolabeled lipids are represented by DG. Stimulation of lipolysis produces a fast decrease in the fraction of short-term radiolabeled DG, whereas there is an increase in the mass of fat body DG. The subcellular distribution of bulk DG showed that its majority (62%) was in the fat cake whereas only 2.9% was in the cytosol. On lipolysis stimulation, the largest changes in specific activities of newly synthesized DG were detected in the cytosol and the fat cake, suggesting that newly synthesized DG localized in the lipid droplets and the cytosol is preferentially mobilized.     

Analysis of tumor-associated alkyldiacylglycerols and other lipids during radiation-induced thymic leukemogenesis.

Lipid composition of thymuses investigated during the development of thymic leukemogenesis induced by exposing C57BL/6J mice to gamma radiation led to the following conclusions. 1. Alkyldiacylglycerols, a class of lipids that are generally elevated in most neoplastic tissues, occurred only in small quantities (less than 1% of the total lipids) in the thymuses of both control and irradiated mice. However, we found a 3- to 8-fold increase in this fraction in thymic tumors of mice at 5 mo after irradiation when compared to controls of similar age. However, the small quantity of this lipid class in thymic leukemia and the fact that similar levels were found in some samples of involuted thymuses of mice 3 days after irradiation, suggests to us that the level of alkyldiacylglycerols is not sufficiently specific or sensitive for detecting early stages of thymic leukemogenesis. 2. Thymuses 3 days after irradiation and leukemic thymuses contain 2- to 3-fold greater quantities of cholesterol esters than control thymuses. No major differences were found in the distribution of acyl moieties in the cholesterol esters of the various thymus samples from the same aged mice except that in thymic tumors the quantity of 18:1 esters was increased by about 25% over that of the controls. The apparent lack of specificity of increased cholesterol esters for neoplasia indicates that its measurement would not provide a suitable indicator of early neoplastic transformation. 3. Acyl composition of the triacylglycerols of thymuses revealed an increase in the 18:1 and a decrease in the 18:2 acids at 3 days after irradiation when compared to the same aged controls. However, thymic tumors occurring at 5 mo after irradiation contained less 14:0, 16:0, and 16:1 acids and increased amounts of 18:1 and 18:2 acids esterified as triacylglycerols compared to controls. The fatty acid distribution in the phospholipid fraction of thymuses was not altered by the appearance of leukemia.     

Polyunsaturated Fatty Acids and Betaine Lipids from Pavlova lutheri

The polar lipids and fatty acids of the microalgae Pavlova lutheriwere analyzed. The principal polar lipid components were monogalactosyldiacylglycerol(MGDG), digalactosyldiacylglycerol (DGDG), sulfoquinovosyldiacylglycerol(SQDG), 1,2-diacylglyceryl-O-2'-hydroxymethyl-(N,N,N-trimethyl)-rß-alanine(DGTA) and 1,2-diacylglyceryl-3-O-carboxyhydroxymethylcholine(DGCC). Each polar lipid had a different set of combinationsof fatty acids, the most characteristic feature being the localizationof polyunsaturated fatty acids in the betaine lipids. The percentagesof polyunsaturated fatty acids in DGTA and DGCC were 70% and50%, respectively, and these fatty acids were localized at theC-2 position in the betaine lipids. An analysis of the incorporationof ¹⁴C-labelled compounds into the algal cells indicated theinvolvement of DGCC in acyl exchange. (Received October 17, 1994; Accepted October 2, 1995)