Brain Lipid Extract Research Articles

D-Amphetamine binding to brain lipid.

The interaction of d-amphetamine with several brain tissue components has been investigated. A brain lipid extract and a number of individual phospholipids were found to bind d-amphetamine when measured by means of a hexane-buffer partition coefficient technique. Phosphatidylserine showed the greatest binding and phosphatidylethanolamine and dipalmitoyllecithin were also effective. Phospholipid binding of d-amphetamine was shown to be related to fatty acid composition. Cholesterol and ganglioside did not bind. It is suggested that phospholipid binding of d-amphetamine may play a role in the pharmacological action of this drug.

Determination of less than a nanomol of cerebrosides by high performance liquid chromatography with gradient elution analysis

A gradient elution high performance liquid chromatography method with detection at 230 nm for the analysis of perbenzoylated cerebrosides containing hydroxy and nonhydroxy fatty acids is described. The quantitative range of the method is 0.5-10 nmol of cerebrosides. The limit of detection for injected sample is about 10 picomol. The analysis time by liquid chromatography is less than 5 minutes and prior purification of the cerebrosides from other lipids in brain lipid extracts is not necessary. The cerebrosides are first perbenzoylated with 50 micronl of 10% benzoyl chloride in pyridine and then separated on a chromatographic column of Zipax. The gradient elution is with 2.8-5.5% of dioxane in hexane. This gradient elution micromethod is at least 10 times more sensitive than isocratic elution methods with detection at 280 nm. The method is applicable to other biological materials containing minute amounts of cerebrosides if the glycolipid fraction is first isolated from the lipid extracts. A further fourfold increase in the sensitivity is achieved by replacing air in the reference cell of the detector by gradient elution solvent.

In Vivo Studies on the Introduction of the 4-t-Double Bond of the Sphingenine Moiety of Rat Brain Ceramides

Abstract Erythro-dl-[3-3H]sphinganine was injected directly into the brains of 14- to 15-day-old rats. After 2 hours, 2.0% of the injected radioactivity could be recovered in the ceramide fraction of the brain lipid extract. The ceramide material was fractionated by thin layer chromatography into the classes of N-acylsphinganine and N-acylsphingenine, containing approximately equal amounts of radioactivity. Free 3H-labeled long chain bases were recovered after alkaline hydrolysis of the two ceramide classes and confirmed to be sphinganine and sphingenine. The presence of tritium in the unsaturated base sphingenine indicates a direct conversion of sphinganine to sphingenine or an acylated sphinganine to acylated sphingenine. No free [3H]sphingenine could be demonstrated in the lipid extract indicating either that it is present in very small amounts, due perhaps to rapid acylation, or that it is not produced as the free base but rather through acylated sphinganine. A time study over 145 min of the incorporation of [3-3H]sphinganine into ceramides showed an initial rapid production of N-acyl-[3H]sphinganine, which stayed at a steady level for the duration, while N-acyl-[3H]sphingenine appeared more slowly but was the predimonate ceramide at the final set time. This suggests N-acylsphinganine may be the precursor of N-acylsphingenine. When N-[1-14C]stearoyl-[3-3H]sphinganine, with an isotope ratio of 2.22 (3H:14C), was the injected substrate, ceramide containing [3H]sphingenine and 14C fatty acid could be isolated. The observed isotone ratio for the N-acylsphingenine was 2.52, indicating an 87% retention of 14C compared to 3H. The similarity of ratios suggests a direct conversion of the injected ceramide to the isolated compound; the small loss of 14C due perhaps to some hydrolysis followed by resynthesis.

Ceramide, triglyceride, and sterol ester in normal human whole brain at different ages.

AbstractSeparation of ceramides from other lipids by two dimensional thin layer chromatography, characterization of ceramide in human brain lipid extracts, and the levels of ceramide, triglyceride, and sterol ester in human brain at different ages (25 week fetus and postnatal ages of 1 day, 3 weeks, 6, 8 and 22 months, and 6, 8.5, 10, 18, 33, 55 and 98 years) are described.

Intravenous Infusion of a Standardised Coagulation Active Phospholipid Complex in Uremic Coagulation Deficiency

SummaryInfusion for 90 min of 10 ml brain lipid extract brought thrombin generation and prothrombin consumption levels in uremic patients to normal for 3 h. After quick injection of the same doses (within 3 min) the effect lasted less than l1/2h. Control subjects produced hypercoagulability with the same treatment. The results of these clinical experiments, obtained with 12 subjects, were shown by analysis of variance to be statistically significant.

Long-chain alcohols in mammalian tissues

Long-chain alcohols have been detected in lipid extracts of bovine and porcine brain and heart muscle. They were found to occur at levels of approximately 0.002% (w/w) of the total lipids. Hexadecanol, octadecanol, octadecenol and, in the bovine tissues, docosanol were identified as major constituents.

Lipid class and fatty acid composition of rat brain and sciatic nerve in alloxan diabetes.

Quantitative analysis of major lipid classes and the fatty acids carried out on lipid extracts of brain from young and adult alloxan diabetic rats revealed decreases in total lipid, cholesterol, the more polar cerebroside, and phospha-tidylethanolamine in brain from young diabetic rats. No significant changes in lipid class composition were found in brains from adult rats with alloxan diabetes. No significant changes in fatty acids were detected in the total brain lipids of young or adult rats as a result of alloxan diabetes. In contrast, similar studies on the lipids of sciatic nerve revealed an increase in cholesterol balanced by a decrease of triglycerides in young rats and a decrease of total lipid and the more polar cerebroside only in adult rats with alloxan diabetes. Compared to normal rats, total lipids from sciatic nerves of young alloxan diabetic rats showed relative decreases in palmitate (16:0), oleate (18:1), and linoleate (18:2) with increases in stearate (18:0), eicosanoate (20:0), eicosenoate (20:1), arachidonate (20:4), docosanoate (22:0), docosapentaenoate (22:5), docosahexaenoate (22:6), lignocerate (24:0). In lipid extracts from sciatic nerves of adult diabetic rats, only oleate (18:1) was increased and linoleate (18:2) decreased compared to normal.

The fatty acid and aldehyde composition of the major phospholipids of mouse brain

Phospholipid classes were separated from mouse brain lipid extracts by preparative thin-layer chromatography (TLC). Methyl esters were prepared from the intact phospholipids by direct transesterification at room temperature in the presence of silica gel by using 0.5M: NaOH-methanol in order to prevent interference by aldehydes or derivatives. Dimethyl acetal derivatives of phosphoglyceride alkenyl ethers (alkenyl moiety with a double bond in 1,2-position relative to oxygen linkage) were prepared, using 5% concentrated HCl in methanol, followed by preparative TLC for isolation.The major phospholipids present were ethanolamine phosphoglycerides (EPG) 39.8%, choline phosphoglycerides (CPG) 39.7%, serine phosphoglycerides (SPG) 15.0%, and sphingomyelin (Sph) 5.4%. One-fifth of the total phospholipids (PL) were in the form of plasmalogens, mainly EPG. Choline and serine plasmalogens were present in trace quantities. The major aldehyde components of the plasmalogens were 16ratio0, 18ratio0, and 18ratio1.The EPG were rich in long-chain poly-unsaturated fatty acids, including 28.8% of 22ratio6 and 17.0% of 20ratio4, but contained only 7.2% of 16ratio0. In contrast, the CPG contained 39.6% of 16ratio0, and 31.0% of 18ratio1 with a small content of polyunsaturated fatty acids. The SPG exhibited a still different pattern containing 38.2% of 18ratio0, 23.2% of 18ratio1, 24.3% of 22ratio6, 2.9% of 16ratio0, and 3.8% of 20ratio4.

The quantitative determination of cholesterol in brain lipid extracts using gas—liquid chromatography

The quantitative determination of cholesterol in brain lipid extracts using gas—liquid chromatography

Effects of brain lipid extracts on reproduction in the laboratory rat

Pregnant rats were given daily subcutaneous injections of lipid extracts from different regions of bovine brain. 1. 1. Extracts from whole hypothalamus administered for 4 days beginning on the day of mating did not interfere with the rate of embryonic development trough Day 5 of pregnancy. 2. 2. When treatment continued through Day 17 of pregnancy, then was discontinued and the animals allowed to litter, the number of normal offspring born was significantly reduced compared with their controls. 3. 3. The active principle(s) were present in extracts from the anterior hypothalamus but not from posterior hypothalamus or cerebral cortex.

Distribution and Properties of Anaphylactic and Venom-Induced Slow-Reacting-Substance and Histamine in Guinea Pigs

Summary A striking parallel between the capacity of specific antigen and cobra venom to give rise to slow-reacting-substance activity in tissues of sensitized guinea pigs was demonstrated. By either method of treatment lung was the major source of slow-reacting-substance. Aorta, ileum, spleen and uterus showed considerably less capacity to form slow-reacting-substance. Antigen and venom showed significant differences in their ability to cause histamine release from various tissues. Both materials caused histamine release from lung, but venom produced considerably more histamine release than antigen from heart, kidney and diaphragm. Of several tissue lipid extracts examined, only lung lipid extract produced SRS on treatment with venom. Typical SRS activity was found to be present in untreated lipid extracts of brain and small intestine. Histamine was found in lipid extracts of lung and was dialyzable. Anaphylactic SRS was not dialyzable until it had been extracted into 80% ethanol in water. SRS formed from chopped lung treated with venom, however, was found to be dialyzable without extraction into ethanol. On the other hand, SRS prepared from lung lipid extract treated with venom was not dialyzable before or after ethanol extraction. The distribution of ethanol extracted anaphylactic SRS between ether and water at different pH values showed that it was not completely extractable into ether even at pH 2 to 3. Gasliquid chromatography of anaphylactic SRS preparations following methanolysis did not show a pattern of fatty acids different from control samples. On the basis of these observations anaphylactic SRS does not appear to be a fatty acid. Four metabolites of histamine, N-acetylhistamine, imidazole-acetic acid, methylhistamine and methylimidazole-acetic acid, as well as histamine incubated with venom, failed to show SRS activity.

STUDIES ON BRAIN GANGLIOSIDES. II. ANALYSIS OF HUMAN BRAIN GANGLIOSIDE FRACTIONS OBTAINED BY PREPARATIVE THIN-LAYER CHROMATOGRAPHY.

Purified gangliosides prepared by the tetrahydrofuran procedure were chromatographed on thin layers of silica gel, using a solvent system of chloroform–methanol −7% ammonia, 55/40/10, v/v/v. A resorcinol detection spray revealed a complex reproducible pattern of five major and several minor components which was similar to that observed in crude lipid extracts of brain. The major components were isolated by a preparative thin-layer technique as a slow fraction containing three components, an intermediate, and a fast fraction. Analysis gave N-acetylneuraminic acid contents of 33.4%, 26.6%, and 15.8% for the slow, intermediate, and fast fractions and the following molar ratios of ceramide:glucose:galactose:hexosamine:N-acetylneuraminic acid: slow, 1.0:0.9:3.2:1.0:2.6; intermediate, 1.0:0.8:2.7:0.8:1.7; and fast, 1.0:0.8:2.2:0.7:0.8.

ACETYLCHOLINE-RELEASING MATERIAL IN NEURAL TISSUES.

WHILE examining cholinergic substances in sciatic nerve1, we noted that acetone extracts of nerve caused a slow contraction of the guinea pig ileum (Fig. 1A). Identical lipid extracts of brain had the same effect (Fig. 1B) ; similar activity could not be detected in kidney, intestine, lung or liver. A salient characteristic of the lipoidal material is that its action is completely blocked by atropine at a dose, 6 ng/ml., that blocked the action of acetylcholine. The lipoidal material causes a slower contraction than does acetylcholine (Fig. 1) ; the material is resistant to acetylcholinesterase, which destroys acetylcholine ; the material retains activity after heating at pH. 10 for 5 min in a boiling water bath, conditions which destroy acetylcholine; the material does not act on a strip of ileum that had been stored either at room temperature for 1 day or at 4° C for 3 days, whereas acetylcholine elicits the usual contractions ; finally, the material is not active on the anoxic ileum, which responds to acetylcholine (Table 1).

The separation of phosphatidyl ethanolamine and phosphatidyl serine by column chromatography

AbstractA method for the complete separation and apparent quantitative recovery of both phosphatidyl ethanolamine and phosphatidyl serine from lipid extracts of beef brain is presented. Evidence for the purity, quantitative recovery, and the suitability of the technique for the subsequent analysis of the fatty acid and fatty aldehyde composition of the phospholipids is described.

The identification of lysolecithin in lipid extracts of brain

The identification of lysolecithin in lipid extracts of brain

Lipid Extracts of Platelets and Brain as Substitutes for Platelets in Coagulation Tests

Lipid Extracts of Platelets and Brain as Substitutes for Platelets in Coagulation Tests

Relation Between Cerebroside and Neutral Fat Contents of Blood Plasma, Erythrocytes and Stroma

That cerebrosides may exist in small amounts in blood has been shown by the development and application of a titrimetric microprocedure for this group of lipids.1, 2, 3 This method, slightly modified, has been employed on lipid extracts of blood, stroma and brain. The data indicate that in erythrocytes, stroma, and brain, much of the lipid which formerly was designated as neutral fat∗ is cerebroside, whereas in plasma only negligible amounts of cerebroside are present. Furthermore, there are indications that the cerebrosides may comprise a significant fraction of the blood lipids in certain pathological conditions.Method. Procedure for hydrolysis: The procedure of Kirk2 was modified to facilitate hydrolysis of the cerebrosides and filtration of the hydrolysate. Aliquot samples of the lipid extract and blanks for initial and total reduction values2 are measured into 10 cc glass-stoppered graduated cylinders. The solvents are evaporated carefully in a boiling water bath. To each of the samples for initial r...