Silvana Rapelli

Comparison between Extra Virgin Olive Oil and Oleic Acid Rich Sunflower Oil: Effects on Postprandial Lipemia and LDL Susceptibility to Oxidation

The aim of our study was to determine whether the minor polar components of virgin olive oil could have favorable effects (1) on fasting and postprandial lipid profile and (2) on low-density lipoprotein (LDL) composition and susceptibility to oxidation in vitro. Ten normolipidic subjects were included in a crossover study (two diet periods of 3 weeks) and received either virgin olive oil (OO diet) or oleic acid rich sunflower oil. An oral fat load was performed at the end of each period. The plasma lipid levels were not significantly different after both diets in the fasting and postprandial states. A few minor variations of the LDL composition were observed only in the postprandial lipemia, and they were different after both diets. The LDL oxidation susceptibility was evaluated by the formation of conjugated dienes. With LDL isolated in the fasting state, the diene production decreased (p = 0.0573) only after the OO diet. The dienes determined at time 0 and the maximal dienes obtained during the oxidation reaction decreased (p = 0.0145 and p = 0.0184, respectively) only after the OO fat load. Nevertheless, the diene production decrease was not significant (p = 0.0848). Our results suggest a mild effect of minor components of virgin olive oil related to a decrease of LDL susceptibility to oxidation; further analyses are necessary to give clear conclusions about their role.

Vitamin D Metabolites Activate the Sphingomyelin Pathway and Induce Death of Glioblastoma Cells

Summary 1α,25-dihydroxyvitamin D3 was previously shown to induce cell death in brain tumour cell lines when added to the medium at micromolar concentration. In this paper we show that Cholecalciferol, a poor ligand of the vitamin D receptor, also induces cell death of HU197 human glioblastoma cell line and early passages cultures derived from a recurrent human glioblastoma. This finding suggests that the effects of vitamin D metabolites on brain tumour cells are at least partially independent from the activation of the classic nuclear receptor pathway. Vitamin D metabolites have been shown to activate the sphingomyelin pathway inducing an increase in cellular ceramide concentration. We determined the levels of sphingomyelin ceramide and ganglioside GD3 in Hu197 cells after treatment with cholecalciferol. A significant increase in ceramide concentration and a proportional decrease in sphingomyelin was already present after 6 hours of cholecalciferol treatment when no morphological changes were visible in the cultures. Treatment with ceramides (N-acetyl-sphingosine or natural ceramide from bovine brain) of the same cells also induces cell death. Similarly, treatment of the same cells with bacterial Sphingomyelinase also results in cell death. The demonstration of an increase in intracellular ceramide after cholecalciferol treatment and the ability of ceramide to induce cell death suggest that the sphingomyelin pathway may be implicated in the effect of vitamin D metabolites on human glioblastoma cells. Inhibition of ceramide biosynthesis by fumonisin B1 treatment did not alter the dose response curve of HU197 cells to cholecalciferol. Insensitivity to fumonisin B1 together with a decrease in sphingomyelin content after cholecalciferol treatment indicate that activation of sphingomyelinase should be responsible for the increase in intracellular ceramide concentration.

Lipids of Xenopus laevis Spermatozoa

Xenopus laevis sperm lipid composition has been studied. The cholesterol content of Xenopus spermatozoa is 194 μ/mg DNA. Their content of glycolipids and phospholipids (measured as inorganic phosphorus) is respectively 40 and 27 μ/mg DNA. The phospholipid pattern is quite homogeneous and all the principal molecular species are present. In all the examined samples, a glycolipid with low mobility, not yet structurally identified, is present. Finally, using as a probe filipin, we have observed cholesterol distribution on the Xenopus sperm plasma membrane by freeze‐fracture. In agreement with the chemical data here presented, Xenopus spermatozoa are heavily labelled by filipin. The filipin‐cholesterol complexes seem to be distributed on the entire sperm plasma membrane and appear as protuberances on the P face, suggesting that most of the cholesterol reside in the inner leaflet of the membrane.

Altered membrane lipid composition in a human meningosarcoma

SummaryIn a sample of meningosarcoma, obtained at the time of surgery, the amount of total gangliosides and phospholipids was examined, together with the cholesterol content and the distribution of different ganglioside and phospholipid species. The phosphatidylinositol, phosphatidylinositol-4-phosphate, phosphatidylinositol-4, 5-bisphosphate and phosphatidylcholine fatty acid composition was also analyzed. The ganglioside pattern in the meningosarcoma was different from the previously reported pattern in meningiomas of different histological origin, showing a higher concentration of GD3, indicating that the so-called b pathway of ganglioside biosynthesis was the preferred one in this type of tumor; moreover the percentage content of polysialylated gangliosides was very low. Cholesterol and phospholipid content was lower than in meningiomas; the phosphatidylcholine increase and the sphingomyelin decrease would indicate a lower membrane microviscosity, a characteristic of tumor cells. Phosphoinositide and phosphatidylcholine fatty acid analysis revealed a considerable amount of docosahexaenoic acid. This abnormal presence of this fatty acid could lead to the production, after receptor stimulation, of a diacylglycerol containing docosahexaenoic acid, which, in turn, could be responsible for an altered activation pattern of protein kinase C, in this way promoting carcinogenesis.

A new procedure for gangliosidic N-acetylneuraminic acid analysis in serum

In this paper a method is presented which is suitable for the extraction, purification and analysis of serum gangliosides. The advantage in comparison with other previously published procedures is the complete extraction of sialoglycolipids without contamination of sialoglycoproteins and/or sialoglycopeptides. The method could be used as a second-level test for the diagnosis and follow-up of cancer patients, and also could be potentially used for pharmaco-kinetic studies after ganglioside treatment.

Studies on glycoprotein metabolism in different types of sphingolipidosis

Abnormalities in glycoprotein composition have been demonstrated in brain tissue from patients with GM1 gangliosidosis and Niemann-Pick disease and in spleen from patients with Gauchers disease. The mechanism producing these glycoprotein abnormalities is probably different in each disease.

Fatty acid pattern of the different phosphoinositide fractions in human meningiomas

Previous studies showed no differences in the phospholipid content of human meningiomas compared to normal leptomeninges, but only a higher unsaturation degree in the individual phospholipid fractions of tumors. Inasmuch as phosphoinositides play a role in the membrane responsiveness to numerous effectors, we studied the fatty acid pattern of the different phosphoinositide fractions of 14 human meningiomas of different histological origin. The fatty acid analysis revealed remarkable differences among the histological types, and, above all, among the different phosphoinositide fractions of a single tumor class. The phosphoinositides derived from transitional meningiomas appeared to be the most saturated ones, because of their low arachidonic acid content. Furthermore, in all the meningiomas, long chain polyunsaturated fatty acids were present only in the phosphatidylinositol fractions and the polyphosphorylated compounds appeared to be significantly different from the corresponding monophosphorylated ones. The possible significance of the different fatty acid distribution in the three phosphoinositide classes is discussed.

Presence of glycoproteins containing the polylactosamine structure in brain and liver of GM1 gangliosidosis patients

The material derived from defective degradation of glycoproteins, which accumulates in brain and liver of a patient with GM1 gangliosidosis type I, was investigated, and the structure of the main storage compounds determined. For comparison, brain and liver of a patient with GM1 gangliosidosis type II were also analyzed. Analysis of the glycopeptides obtained after pronase digestion of the defatted residue indicates the storage of glycoprotein-like material in type I, but not in type II. Treatment with endo-beta-galactosidase showed that the stored material contained N-acetyllactosamine repeating units. Two major oligosaccharides, OS I and OS II, were isolated after the enzyme treatment, whose structures are: GlcNAc beta 1----3 Gal (OS I) and Gal beta l----4GlcNAc beta 1----3 Gal (OS II). Treatment with exo-beta-galactosidase transformed the trisaccharide OS II into the disaccharide OS I, indicating that the deficiency of beta-galactosidase in GM1 gangliosidosis type I, but not in type II, also affects glycoprotein catabolism, leading to the accumulation of glycopeptides containing terminal beta-galactosyl residues and N-acetyllactosamine repeating units. These results indicate the severe impairment in the catabolism of glycoconjugates with beta-linked galactose in type I, although this impairment is not as pronounced in type II.

Carbohydrate, Protein and Lipid Metabolism in the Skin; Biochemical and Molecular Aspects

Human skin is one of the more heavy extended organs of the body, which is completely wrapped up by the skin itself; therefore, this one constitutes a physical barrier at the interface between the body and the environment.