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Biomedical subjects

L Laurent

Publications and source records attributed to L Laurent.

8 recordsLinked to original sources

[Genetic studies of relationship between Mediterranean and Atlantic populations of loggerhead turtle Caretta caretta with mitochondrial marker].

The loggerhead turtle Caretta caretta is an endangered species in the Mediterranean. Therefore, the definition of the Mediterranean population, and their relationships to the Atlantic population is of fundamental importance. For this purpose, we have sequenced a portion of the mitochondrial cytochrome b gene to generate genetic markers. Results indicate that the Mediterranean nesting female population is genetically isolated from the Atlantic nesting female population, but loggerhead turtles of Atlantic origin were found in the West Mediterranean basin. This entry of Atlantic loggerheads in the Mediterranean confirms earlier speculations and presents special conservation problems. The Spanish swordfish longline fishery which incidentally captures large numbers of loggerheads in the West Mediterranean basin has therefore an impact on the Atlantic population. These data demonstrate the international nature of marine turtle conservation.

Animals

Structure and biochemical composition of desmosomes and tonofilaments isolated from calf muzzle epidermis.

Complexes of plasma membrane segments with desmosomes and attached tonofilaments were separated from the stratum spinosum cells of calf muzzle by means of moderately alkaline buffers of low ionic strength and mechanical homogenization. These structures were further fractionated by the use of various treatments including sonication, sucrose gradient centrifugation, and extraction with buffers containing high concentrations of salt, urea, citric acid, or detergents. Subfractions enriched in desmosome-tonofilament-complexes and tonofilament fragments were studied in detail. The desmosome structures such as the midline, the trilaminar membrane profile, and the desmosomal plaque appeared well preserved and were notably resistant to the various treatments employed. Fractions containing desmosome-tonofilament complexes were invariably dominated by the nonmembranous proteins of the tonofilaments which appeared as five major polypeptide bands (apparent molecular weights: 48,000; 51,000; 58,000; 60,000; 68,000) present in molar ratios of approx. 2:1:1:2:2. Four of these polypeptide bands showed electrophoretic mobilities similar to those of prekeratin polypeptides from bovine hoof. However, the largest polypeptide (68,000 mol wt) migrated significantly less in polyacrylamide gels than the largest component of the hoof prekeratin (approximately 63,000 mol wt). In addition, a series of minor bands, including carbohydrate-containing proteins, were identified and concluded to represent constituents of the desmosomal membrane. The analysis of protein-bound carbohydrates (total 270 microgram/mg phospholipid in desmosome-enriched subfractions) showed the presence of relatively high amounts of glucosamine, mannose, galactose, and sialic acids. These data as well as the lipid composition (e.g., high ratio of cholesterol to phospholipids, relatively high contents of sphingomyelin and gangliosides, and fatty acid pattern) indicate that the desmosomal membrane is complex in protein and lipid composition and has a typical plasma membrane character. The similarity of the desmosome-associated tonofilaments to prekeratin filaments and other forms of intermediate-sized filaments is discussed.

Animals

A competitive receptor binding assay for platelet-activating factor (PAF): quantification of PAF in rat brain.

A radioreceptor assay (RRA) was developed using rabbit platelet membrane preparations to quantify platelet-activating factor (PAF) and lyso-PAF, the deacylated derivative of PAF, in a variety of tissues and biological fluids. We examined PAF and lyso-PAF levels in different rat brain areas with regard to the many proven and postulated actions of PAF in brain functions. Human saliva was selected to check the validity of this RRA. The samples were extracted with methanol/chloroform/water and purified by high-performance liquid chromatography on a 5-microns Nucleosil Si column (overall recovery: 78%). Sample extracts were acetylated before chromatography to assay lyso-PAF. PAF itself was assayed in non-aceylated samples. A competitive binding assay was performed using aliquots of platelet membrane preparation and tritiated PAF. The minimum detectable amount of PAF was 144 pg per tube and the receptor was highly specific for PAF. In human saliva, we confirm the presence of PAF and lyso-PAF within the range expected. Moreover there was a good correlation between the RRA and the aggregation assay (r = 0.976). A defined cocktail of protease inhibitors allowed storage of platelet membrane preparations for at least 3 months at -20 degrees C with no change in binding properties. In the brain we observed the prevalent presence of lyso-PAF and large variations in PAF and lyso-PAF concentrations between the different brain areas analyzed. PAF was undetectable in the hypothalamus but the lyso-PAF concentration was 2.5 micrograms/g wet tissue. The PAF concentration in the cortex varied from 0 to 16 ng/g wet tissue while that of lyso-PAF was 0.7 micrograms/g wet tissue. Moreover the amount of lyso-PAF varied between the different brain areas analyzed. The hippocampus contained the highest amount (7 micrograms/g wet tissue), and relatively high levels were found in the hypothalamus, medulla oblongata and corpus striatum. The cerebellum and cortex contained the lowest levels of lyso-PAF. These findings show that PAF is present in the central nervous system mainly in its inactive form, lyso-PAF, and suggest that its effects as a modulator of brain function may be dependent on deacetylation, rather than synthesis.

Animals