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Effect of androgen deficiency on the human meibomian gland and ocular surface.

The purpose of this study was to determine whether the chronic use of antiandrogen medications leads to meibomian gland dysfunction, altered lipid profiles in meibomian gland secretions, decreased tear film stability, and evaporative dry eye. Subjects taking antiandrogen therapy for prostatic indications, as well as age-related controls, were asked to complete a questionnaire that assessed dry eye symptoms and then were given a complete anterior segment examination. Moreover, meibomian gland secretions were obtained from each eye and analyzed by high-performance liquid chromatography/mass spectrometry for the relative content of cholesterol, cholesterol esters, wax esters, diglycerides, triglycerides, and specific molecular species in the diglyceride fraction. Our results demonstrate that patients taking antiandrogen treatment, compared with age-related controls, had a: 1) significant increase in the frequency of appearance of tear film debris, an abnormal tear film meniscus, irregular posterior lid margins, conjunctival tarsal injection, and orifice metaplasia of the meibomian glands; 2) significant increase in the degree of ocular surface vital dye staining; 3) significant decrease in the tear film breakup time and quality of meibomian gland secretions; and 4) significant increase in the frequency of light sensitivity, painful eyes, and blurred vision. In addition, the use of antiandrogen pharmaceuticals was associated with significant changes in the relative amounts of lipids in meibomian gland secretions. Our findings indicate that chronic androgen deficiency is associated with meibomian gland dysfunction and dry eye.

Aged↗

Effects of menopause and estrogen on cervical epithelial permeability.

The objective of the study was to characterize the effect and explore the mechanisms by which menopause affects paracellular permeability of cultured human cervical epithelium. The experimental system was cultures of human ectocervical epithelial (hECE) cells on filters. Assays included flux measurements of cell-impermeable molecules and determinations of transepithelial electrical conductance. hECE cells of postmenopausal women formed epithelia with lower paracellular permeability than hECE cells of premenopausal women. Treatment with estrogen increased paracellular permeability, but levels remained lower compared to cultures from premenopausal women. Lowering extracellular calcium or treatment with cytochalasin-D, conditions that decrease the tight junctional resistance (R(TJ)), increased the permeability, and the relative effects were greater in cells of premenopausal women than in postmenopausal women. Treatment of cells with sn-1,2-dioctanoyl diglyceride, an agent that increases the R(TJ), decreased the permeability, and the relative effect was greater in cells of postmenopausal women than in cells of premenopausal women. Estrogen had no significant effect on the responses to low calcium, cytochalasin-D, or sn-1,2-dioctanoyl diglyceride. Hydrostatic and hypertonic gradients in the subluminal to luminal direction, conditions that decrease the resistance of the lateral intercellular space increased the permeability to a similar degree in cultures of cells from premenopausal and from postmenopausal women. Pretreatment with estrogen augmented the increases in permeability in response to hydrostatic and hypertonic gradients. In cells exposed to low extracellular calcium, hydrostatic gradients had an additive increase in permeability. By extrapolation it was determined that in cultures of postmenopausal women R(TJ) contributes 97% to the total paracellular resistance, whereas in hECE cultures of premenopausal women the R(TJ) contributes only 84%. These results indicate that after menopause the transcervical paracellular permeability decreases significantly; this can explain the decrease in lubrication of the cervix and vagina in postmenopausal women. Part of the effect is due to lack of estrogen, and it can be reversed by treatment with the hormone. However, most of the effect is unrelated to estrogen and is caused by an increase in R(TJ).

Adult↗

Evaluation of the in-vitro digestion profiles of long and medium chain glycerides and the phase behaviour of their lipolytic products.

An evaluation of the in-vitro digestion profile and phase behaviour of the common formulation lipids Miglyol 812 (medium chain triglyceride, MCT), Capmul MCM (C8/C10 monoglyceride/ diglyceride mixture), soybean oil (long chain triglyceride, LCT) and Maisine 35-1 (C18 monoglyceride/diglyceride mixture), is described. Experiments were conducted using titrimetric, high-performance thin-layer chromatographic (HPTLC) and ultracentrifugational techniques under model fasted and post-prandial intestinal conditions. The rate and extent of digestion of the medium chain lipids was greater than the corresponding long chain lipids, and independent of bile salt concentration, with complete conversion to monoglyceride and fatty acid occurring after 30 min digestion. The long chain lipid digests separated into an oily phase (containing undigested triglyceride and diglyceride), an aqueous phase (containing bile salt, fatty acid and monoglyceride) and a pellet phase (containing approximately 5 mm of fatty acid, presumably as an insoluble soap) after ultracentrifugation. Higher proportions of long chain fatty acid and monoglyceride were dispersed into the aqueous phase with increasing bile salt concentrations. In contrast, medium chain lipolytic products separated only into an aqueous phase and a pellet fraction in a bile-salt-independent manner. The digestion of both the C8/C10 and C18 monoglyceride/diglyceride lipid mixtures was more rapid than the corresponding triglyceride, especially at early time points. This investigation provides insight into the relative digestion kinetics of medium chain and long chain lipids and provides information regarding the phase behaviour of their lipolytic products under conditions modelled on those expected after oral administration. The data also provide a background for improved understanding of the potential utility of long chain and medium chain lipid-based formulations.

Caprylates↗

Effect of streptococcal lipids on Ehrlich ascite tumor cells.

The lipids extracted from group A hemolytic streptococci (strain Su, Blackmore and C203U) were examined for their antitumor effect against Ehrlich ascites carcinoma in mice. Total lipids were extracted from streptococcal cells according to the method of Folch et al, and separated into 9 lipid fractions by thin-layer chromatography, using various solvent systems. Three fractions were compound lipids (diphosphatidyl glycerols, monoglucosyl diglycerides and diglucosyl diglycerides), and the remaining 6 fractions were neutral lipids such as free fatty acids, glycerides, sterols and sterol esters. For biological testing, the lipid fractions suspended in physiological saline containing Tween 20 (0.02%) were incubated with Ehrlich tumor cells at 37 degrees C for 90 min, and the cell mixture was given intraperitoneally into mice thereafter. Among 9 lipid fractions, free fatty acids and monoglycerides from the streptococci examined were highly active in suppressing the depressing the development of ascites carcinoma in mice. Diphosphatidyl glycerols from two strains of streptococci (BLackmore and C203U) were also effective in suppressing the tumor growth in mice. However, the other lipid fractions had little effect on the tumor growth.

Animals↗

On a quantitative determination of antibodies to lipids and proteins.

The amount of precipitating antibodies in monospecific lipid- and protein antisera was determined by quantitative precipitation reactions according to the methods of Heidelberger and Kendall. Antisera were obtained by immunization of rabbits. Fragments of the thylakoid membrane from Antirrhinum chloroplasts were used as antigens for the binding of antibodies. These fragments had a diameter of 100 A. They were composed of 51% proteins and 39% lipids. It was found that antisera to the lipids mono-, tri- and digalactosyl diglyceride, sulfoquinovosyl diglyceride and phosphatidyl glycerol contained 20 to 92 micrograms precipitating antibodies per ml serum. In antisera to the coupling factor of photophosphorylation, cytochrome f, a polypeptide with the apparent MW 24000 and a serum to the total protein of the lamellar system 23 to 99 micrograms were found. In chloroplast antisera the amount of all precipitating lipid- and protein antibodies amounted to 765 micrograms per ml. The described antisera originated from blood that had been withdrawn from the test animals 4 to 10 week after antigen injection.

Animals↗

The lipids of buffalo spermatozoa and seminal plasma.

Lipids were extracted from the spermatozoa and seminal plasma of buffalo and were analysed by column and thin-layer chromatography. Buffalo spermatozoa (10(9) cells) contained 1-15 mg total lipids, 0-285 mg neutral lipids, 0-396 mg glycolipids, 0-547 mg phospholipids and 0-015 mg gangliosides. The corresponding values for 1 ml seminal plasma were: 1-50, 0-439, 0-581, 0-594 and 0-010 mg. The sperm neutral lipids were comprised of 3-2% hydrocarbons, 4-5% cholesterol esters, 46-7% triglycerides, 17-5% 1,3-diglycerides, 3-1% 1,2-diglycerides, 17-6% cholesterol, 0-6% free fatty acids and 2-4% monoglycerides. The corresponding values for seminal plasma were: 4-2, 6-2, 33-0, 19-8, 4-2, 18-5, 3-8 and 4-7%. In the sperm phospholipids, there was 30-4% phosphatidyl choline; 19-4% phosphatidal choline; 10-8% phosphatidyl ethanolamine; 3-4% phosphatidal ethanolamine; 11-3% sphingomyelin; 1-5% phosphatidyl serine; 0-6% phosphatidyl inositol; 3-9% lysophosphatidyl choline; 4-4% lysophosphatidyl ethanolamine; 0-7% lysophosphatidyl serine; 5-5% diphosphatidyl glycerol and 0-3% phosphatidic acid. The corresponding values for seminal plasma were: 21-7, 17-3, 11-7, 4-1, 13-1, 2-8, 3-9, 5-6, 1-0, 7-4 and 0-5%. Comparative data for bull spermatozoa and seminal plasma were also obtained.

Animals↗

The expression of hormone-sensitive lipase in clonal beta-cells and rat islets is induced by long-term exposure to high glucose.

Hormone-sensitive lipase (HSL) is expressed and enzymatically active in beta-cells and has been proposed to be involved in the generation of the lipid-derived signal that seems to be necessary for glucose-stimulated insulin secretion. In this study, we investigated whether the expression of HSL in INS-1 cells and in rat islets is affected by exposure to high glucose concentrations. Incubation of INS-1 cells in 25 mmol/l glucose for 16 and 32 h induced HSL protein expression twofold, whereas no effect was observed after 4 and 8 h of incubation. The HSL activity, defined as the diglyceride lipase activity inhibited by anti-rat HSL antibodies, constituted approximately 25% of total diglyceride lipase activity and was induced to a similar extent as HSL protein levels. The glucose effect at 16 h on HSL protein expression level was confirmed in freshly isolated rat islets. Exposure of INS-1 cells to different glucose concentrations for 16 h showed that the inductive effect on HSL protein levels was maximum at 20 mmol/l glucose (2- to 2.5-fold). Northern blot analysis demonstrated a more than threefold elevation of HSL mRNA levels. The induction was blocked by actinomycin D, and the half-life of the transcript seemed to be unchanged by high glucose, suggesting a transcriptional nature of the glucose effect on HSL gene expression. The nonmetabolizable glucose analog 2-deoxyglucose, which has no mitogenic effect, induced HSL approximately 1.3-fold, whereas mannose was similar to glucose, stimulating HSL expression 1.7- to 2-fold. The results suggest that HSL is involved in the beta-cell responses to hyperglycemia and also in generating the lipid signal that is needed in stimulus-secretion coupling.

Animals↗

Comparison of the biosynthesis and composition of polyglycerophosphatides and phosphatidylinositols in mitochondria and microsomes isolated from neonatal and adult rat heart and liver.

The level of biosynthesis and the composition of polyglycerophosphatides (phosphatidylglycerol, phosphatidyglycerolphosphate, and diphosphatidylglycerol or cardiolipin) and phosphatidylinositols were examined in mitochondria and microsomes, respectively, isolated from neonatal and adult rat heart and liver. Biosynthesis of [3H]polyglycerophosphatides [( 3H]phosphatidylglycerol and [3H]phosphatidylglycerolphosphate) was 4.5 times higher in neonatal than in adult heart mitochondria, whereas in the respective liver mitochondria this synthesis was only 15% higher in neonatal mitochondria. The biosynthesis of [3H]phosphatidylinositol was twice as high in neonatal as in adult heart microsomes, but very similar in the respective liver microsomes. The major biosynthesized polyglycerophosphatide was [3H]phosphatidylglycerol. The accumulation of [3H]phosphatidylglycerolphosphate depended on the origin of the mitochondria. Under our experimental conditions [3H]phosphatidylinositol was the only synthesized phosphoinositide in all microsomes. The biosynthesis of cardiolipin depended on the origin of the mitochondria and was highest in adult rat liver mitochondria and lowest in adult heart mitochondria. In all cases the biosynthesized [14C,3H] cardiolipin from [14C]phosphatidylglycerol and [3H]CDP-diglycerides had a ratio of 14C/3H around unity. The biosynthesis of [3H]CDP-diglycerides, the key precursor for the biosynthesis of phosphatidylglycerol, phosphatidylinositol, and cardiolipin, was 30% higher in neonatal than in adult heart microsomes and very similar in the respective liver microsomes. The subcellular localization of the enzymes required for the biosynthesis of the lipids and liponucleotides examined was found to be the same in membranes isolated from neonatal and adult rat heart and liver.

Aging↗

Lipid composition of winged bean (Psophocarpus tetragonolobus).

The lipids were extracted from the winged bean (Psophocarpus tetragonolobus) seed with water-saturated n-butanol. Lipids were separated into groups by preparative TLC on silica gel G. The amount of each lipid type was determined by analysis of the fatty acid constituents in each lipid type. Glyceride was the major lipid accounting for 89.6% of the total, followed by an unknown lipid 4%, free fatty acid of 2.3%, 1,3-diglyceride, 1,2-diglyceride and steryl ester as 1% each and finally a polar lipid as 0.2%. The results show that winged bean oil should be suitable for edible purposes. Triglycerides showed a similar profile of fatty acids to those of whole lipid: the major fatty acids were palmitic (10.9%), stearic (4.5%), oleic (37.1%), linoleic (19.0%), eicosenoic (3.6%), behenic (18.5%) and lignoceric (4.2%) acids. Compared to soybean oil, winged bean oil contained long chain fatty acids and a fairly small amount of linolenic acid which is favorable regarding oil stability against autoxidation.

Chemical Phenomena↗

Investigations concerning the mode of action of 3,4-dihydroxybutyl-1-phosphonate on Escherichia coli.

Experiments were performed to evaluate the ability of the enzymes of Escherichia coli involved in glycerol 3-phosphate metabolism to recognize phosphonic acid analogues of the natural substrate. Neither the catabolic membrane-bound glycerol-3-phosphate dehydrogenase nor the acyl coenzyme A: glycerol-3-phosphate acyltransferase can use 3,4-dihydroxybutyl-1-phosphnate or 2,3-dihydroxypropyl-1-phosphonate are inhibitors of the reduction of dihydroxyactone phosphate as substrates. The 4-carbon phosphonic acid analogue does not exhibit inhibitory activity for either of these enzymes. While the 3-carbon phosphonic acid analogue has no inhibitory effect upon the catabolic dehydrogenase, it does appear to have a slight but reproducible inhibitory effect on the acyltransferase. Glycerol 3-phosphate and 3,4-dihydroxybutyl-1-phosphonate by glycerol 3-phosphate:NAD (P) oxidoreductase. rac-2,3-Dihydroxypropyl-1-phosphonate does not appear to be recognized by this enzyme. The apparent K-i for snglycerol 3-phosphate is 19 muM and for D-3,4-dihydroxybutyl-1-phosphonate it is 42 muM. In addition the glycerol 3-phosphate:NAD(P) oxidoreductase catalyzes the reduction of 4-hydroxy-3-oxobutyl-1-phosphonate (apparent K-m of 182 muM), a phosphonic acid analogue of dihydroxyacetone phosphate. 3,4-Dihydroxybutyl-1-phosphonate is both a competitive inhibitor (apparent Ki of 740 muM) and a substrate (apparent K-m of 450 muM) for the CDP-diglyceride: glycerol 3 phosphate phosphatidyltransferase but it has no effect upon CDP-diglyceride:L-serine phosphatidyltransferase. The relationship ofthese in vitro studies to in vivo investigations is discussed.

Acyltransferases↗

Glycogen, protein and lipid content of adult Fasciola gigantica (Trematoda).

The concentration of carbohydrates, proteins, and lipids in the whole worm, gut, and body musculature of Fasciola gigantica was investigated from worms collected from cattle slaughtered in the Jos abattoir. Protein accounted for the highest concentration of the constituents constituting 62.59 +/- 0.86% of the dry weight of the fluke. This was followed by lipid with 28.15 +/- 0.86% of the dry weight while carbohydrate (glycogen) was 6.29 +/- 0.11% of the dry weight. Thin-layer chromatography revealed the presence of six distinct bands of neutral lipids, namely: monoglycerides, 1,2-diglycerides, 1,3-diglycerides, free fatty acids, triglycerides and sterol esters from both whole worm homogenate and fractions of the gut and muscle tissues. In the case of phospholipids, lysophosphatidyl choline, sphingomyeline, phosphatidyl choline and phosphatidyl-ethanol amine were recovered. When the free fatty acids were subjected to further fractionation, palmitic, stearic, as well as oleic acids were recovered in appreciable quantities indicating that lipids are being catabolized in these trematodes.

Animals↗

Human plasma platelet-activating factor acetylhydrolase. Oxidatively fragmented phospholipids as substrates.

Human plasma platelet-activating factor (PAF) acetylhydrolase hydrolyzes the sn-2 acetyl residue of PAF, but not phospholipids with long chain sn-2 residues. It is associated with low density lipoprotein (LDL) particles, and is the LDL-associated phospholipase A2 activity that specifically degrades oxidatively damaged phospholipids (Stremler, K. E., Stafforini, D. M., Prescott, S. M., Zimmerman, G. A., and McIntyre, T. M. (1989) J. Biol. Chem. 264, 5331-5334). To identify potential substrates, we synthesized phosphatidylcholines with sn-2 residues from two to nine carbon atoms long, and found the V/k ratio decreased as the sn-2 residue was lengthened: the C5 homolog was 50%, the C6 20%, while the C9 homolog was only 2% as efficient as PAF. However, the presence of an omega-oxo function radically affected hydrolysis: the half-life of the sn-2 9-aldehydic homolog was identical to that of PAF. We oxidized [2-arachidonoyl]phosphatidylcholine and isolated a number of more polar phosphatidylcholines. We treated these with phospholipase C, derivatized the resulting diglycerides for gas chromatographic/mass spectroscopic analysis, and found a number of diglycerides where the m/z ratio was consistent with a series of short to medium length sn-2 residues. We treated the polar phosphatidylcholines with acetylhydrolase and derivatized the products for analysis by gas chromatography/mass spectroscopy. The liberated residues were more polar than straight chain standards and had m/z ratios from 129 to 296, consistent with short to medium chain residues. Therefore, oxidation fragments the sn-2 residue of phospholipids, and the acetylhydrolase specifically degrades such oxidatively fragmented phospholipids.

1-Alkyl-2-acetylglycerophosphocholine Esterase↗

[DNA-bound lipids from eukaryotic (loach spermatozoa, pigeon erythrocytes) and prokaryotic (E. coli B, phage T2) cells].

Using thin-layer chromatography, some specific DNA-bound neutral lipids and phospholipids of loach spermatozoa, pigeon erythrocytes, E. coli B and phage T2 cells were studied. These lipids are represented by loosely and firmly bound components. The content of neutral lipids in the above DNAs (per mg of DNA) is 10.6, 4.8, 7.81 and 1.43 micrograms, respectively; that of phospholipids is 4.31, 1.28, 1.14 and 0.54 micrograms, respectively. The eucaryotic DNA-bound lipids are highly deficient of free cholesterol, phosphatidylcholine, phosphatidylinositol and phosphatidylserine but are rich in cardiolipin, phosphatidylethanolamine, cholesterol esters, diglycerides and free fatty acids. The quantitative and qualitative composition of DNA-bound lipids of loach spermatozoa changes during the transition from the superhelical to the relaxed conformation of DNA. Procaryotic DNA-bound neutral lipids are also represented by the free cholesterol, diglyceride and free fatty acid fractions, whereas the DNA-bound phospholipids of procaryotes consist of only two fractions, i.e., cardiolipin and phosphatidylethanolamine. The role of DNA-bound lipids in the structural and functional organization of eucaryotic and procaryotic genomes is discussed.

Animals↗

Control of membrane lipid synthesis in Escherichia coli during growth and during the stringent response.

The regulation of phospholipid synthesis in cells of Escherichia coli was studied in vivo during growth and during the stringent response to amino acid starvation. Strains harboring the hybrid plasmid pLC44-14 (Clark, L., and Carbon, J. (1976) Cell 9, 91-99), which had increased levels of glycerophosphate acyltransferase, were used to study the involvement of this enzyme in the control of phospholipid synthesis. In addition, regulation was studied by measuring the levels of three early intermediates of phospholipid synthesis:phosphatidic acid, CDP-diglyceride, and dCDP-diglyceride. The liponucleotides were measured by a new enzymatic method which allows determinations to be made on crude lipid extracts. Results from experiments on growing cells are consistent with regulation of membrane lipid synthesis occurring in fatty acid synthesis or at the level of glycerophosphate acylation, but not at any later step. Experiments on the inhibition of lipid synthesis during the stringent response make it possible to rule out explanations which involve the inhibition of a single enzyme; enzymes both before and after the liponucleotides in phospholipid synthesis must be affected.

Amino Acids↗

Guanine nucleotide effects on catecholamine secretion from digitonin-permeabilized adrenal chromaffin cells.

The nonhydrolyzable GTP analogue guanosine 5'-(beta, gamma-imido)triphosphate (GMP-PNP) produced an ATP-dependent but Ca2+-independent stimulation of [3H]norepinephrine release from permeabilized chromaffin cells. This stimulation of secretion was 25-35% of the secretion induced by 10 microM Ca2+. A similar Ca2+-independent stimulation was produced by other non-hydrolyzable GTP analogues. No effect was seen with a variety of other nucleotides, including GTP. The GMP-PNP effect was specifically inhibited by low concentrations of guanine nucleotides. Addition of cAMP did not mimic the Ca2+-independent GMP-PNP effect, but did slightly enhance Ca2+-dependent secretion. Pretreatment with pertussis toxin had no effect on Ca2+-dependent secretion or on the GMP-PNP effect. There was no detectable diglyceride or inositol phosphate produced during GMP-PNP treatment, and addition of diglyceride and inositol trisphosphate did not induce secretion. Guanosine 5'-(beta-thio)diphosphate (GDP-beta-S), in addition to its ability to inhibit the GMP-PNP effect, partially inhibited Ca2+-dependent secretion. At 10 microM free Ca2+, the effects of GMP-PNP and Ca2+ were nonadditive. In fact, secretion in the presence of both GMP-PNP and 10 microM Ca2+ was slightly less than secretion due to Ca2+ alone. These data suggest that a guanine nucleotide-dependent process interacts in some way with one or more components of the normal Ca2+-dependent secretory pathway. However, it may not be an intrinsic part of the mechanism underlying Ca2+-dependent secretion.

Adenosine Triphosphate↗

Estradiol- and testosterone-induced alterations in phosphatidylcholine and triglyceride synthesis in hepatic endoplasmic reticulum.

Pathways of phosphatidylcholine and triglyceride biosynthesis were studied in hepatic endoplasmic reticulum from castrated and noncastrated male rats pretreated with estradiol or testosterone. In vitro measurements of hepatic microsomal enzymes which catalyze phosphatidylcholine biosynthesis revealed a significant increase in the specific activity of the enzyme governing phosphatidylcholine biosynthesis by the sequential methylation of phosphatidylethanolamine in the estradiol-treated castrate animals. The specific activity of phosphorylcholine-glyceride transferase was decreased by estradiol treatment in both castrate and noncastrate animals. The specific activity of diglyceride acyltransferase, which catalyzes triglyceride biosynthesis, was decreased by estradiol pretreatment in both castrate and noncastrate animals and was increased by testosterone in the castrate animals. The changes in specific activity of the enzymes governing phosphatidylcholine biosynthesis may account for the previously noted increased in vivo incorporation of methyl groups of l-methionine into hepatic phosphatidylcholine in female and estradiol-treated animals; the data suggest that in female and estradiol-treated rats a greater proportion of hepatic phosphatidylcholine is synthesized by the stepwise methylation of phosphatidylethanolamine. The decrease in diglyceride acyltransferase specific activity seen after estradiol administration may account for the lipotropic-like effect of estradiol.

Acyltransferases↗

Molecular species of mono-, di-, and triphosphoinositides of bovine brain.

The mono-, di-, and triphosphoinositides of bovine brain were isolated by chromatography on columns of DEAE-cellulose, alumina, and silicic acid. The major molecular species in each phosphoinositide class were identified and quantitatively estimated by combined thin-layer and gas-liquid chromatography of the component diglycerides, which were released by hydrolysis with a specific brain phosphodiesterase. The diglycerides were treated with pancreatic lipase, and the positional distribution of the fatty acids was determined. Over 27 molecular species were identified, and these accounted for about 95% of each phosphoinositide class, but the 1-stearate 2-arachidonate derivative contributed more than 40% of the total in each class. The other molecular species also were qualitatively and quantitatively similar in the three phosphoinositide classes. All the long-chain and polyunsaturated acids were confined to the 2-position and were preferentially paired with stearic acid in the 1-position. Oleic acid in the 2-position was about equally divided between species with palmitic and stearic acids in the 1-position. These results suggest that the mono-, di-, and triphosphoinositides of the bovine brain have similar compositions and that the various molecular species may be metabolically related.

Animals↗

Four-directional-development thin-layer chromatography of lipids using trimethyl borate.

Solvent mixtures containing trimethyl borate virtually eliminated the pronounced interconversion of 1,2- and 1,3-dipalmitins during their resolution by thin-layer chromatography on Silica Gel G. With trimethyl borate, an average of 1-2% of 1,2-dipalmitin was converted to 1,3-dipalmitin. A four-directional-development TLC procedure incorporating trimethyl borate resolves cholesteryl glucoside, ceramides, monogalactosyl diglyceride, 1- and 2-monopalmitin, palmitic acid, cholesterol, 1,2- and 1,3-dipalmitin, tripalmitin, methyl palmitate, cholesteryl palmitate, beta-carotene and some of its degradation products, squalene, and tetracosane. Digalactosyl diglyceride, phosphatidic acid, phosphatidylglucose, cerebrosides, and other phospholipids remain near the origin. A mixture containing triolein, 1,2- and 1,3-diolein, 1- and 2- monoolein, oleic acid, and cholesterol was resolved in one dimension. A similar series of palmitic-containing neutral lipids was also resolvable in one dimension. These procedures were applied to the TLC of human sera lipids.

Acetates↗