Enzyme localization in proteins separated by paper electrophoresis in human seminal plasma

Résumé Les transaminases -glutamo-oxaloacétique (GOT) et glutamo-pyruvique (GPT) ont été déterminées dans les protéines du liquide spermatique de l'homme après séparation par électrophorèse sur papier. L'amylase se trouve dans toutes les fractions, mais la GOT et la GPT sont localisées respectivement dans la globuline -2 et l'albumine.     

Storage of 5-hydroxytryptamine in human blood platelets

Zusammenfassung Menschliche Blutplättchen zeigten die höchste 5-Hydroxytryptamin (5HT)- und Adenosin-5-Triphosphat (ATP)-Konzentration in einer subzellulären Fraktion, die elektronenoptisch aus Vesikeln von unterschiedlicher Dichte bestand. Diese waren deutlich verschieden von anderen subzellulären Organellen, z.B. den -Granula. Inkubation der Plättchen mit 5 HT erhöhte die Elektronendichte und den 5 HT-Gehalt der Vesikel, veränderte aber die -Granula nicht, Es wird geschlossen, dass in menschlichen Blutplättchen ein Teil des 5 HT zusammen mit ATP in spezifischen subzellulären Organellen gespeichert ist.     

Exogenous fibronectin modifies the aggregation of collagen-stimulated human platelets

Fibronectins (FN) are adhesive glycoproteins whose role in platelet aggregation is unclear. Addition of 3, 6 and 12 micrograms/ml of human plasma FN in vitro to isolated human platelets, which had been freed from plasma FN by gel filtration and subsequently stimulated with collagen, inhibited the last stage of platelet aggregation. With 3 and 6 micrograms/ml of FN a shortening of the lag-time was also observed. These data showed that FN may play a role in platelet-collagen interaction as well as in platelet-platelet interaction.     

Exogenous fibronectin modifies the aggregation of collagen-stimulated human platelets

Summary Fibronectins (FN) are adhesive glycoproteins whose role in platelet aggregation is unclear. Addition of 3, 6 and 12 g/ml of human plasma FN in vitro to isolated human platelets, which had been freed from plasma FN by gel filtration and subsequently stimulated with collagen, inhibited the last stage of platelet aggregation. With 3 and 6g/ml of FN a shortening of the lag-time was also observed. These data showed that FN may play a role in platelet-collagen interaction as well as in platelet-platelet interaction.     

Influence of coronary dilators on the uptake of serotonin by human platelets and adrenaline by chromaffine granules

Zusammenfassung In einer unspezifischen, physikalisch-chemischen Wirkung von vier oberflächenaktiven Coronardilatatoren auf die Membranen von menschlichen Thrombozyten und isolierten chromaffinen Granula wird die Ursache für die Hemmung der Aufnahme von Serotonin bzw. Adrenalin erblickt.     

Application of the ion-thinning methods to the high-resolution study of human dental enamel crystals]

Ion thinning technique applied to high resolution T.E.M. study of human dental enamel crystallites makes it possible to visualize a great number of dislocations. Dislocations proved not to be due to artifacts induced by the ultramicrotome diamond knife. They are related to the crystal apartite structure.     

Effect of lactic acid on the aggregation of human platelets induced by ADP adrenaline and collagen

Zusammenfassung Nachweis, dass die Milchsäure die durch ADP und Adrenalin induzierte Aggregation der menschlichen Thrombozyten hemmt und keinen Einfluss auf die Kollagen-induzierte Plättchenaggregation in vitro hat. Die Hemmwirkung ist unspezifisch, kommt sie infolge der pH-Verschiebung in saurer Richtung zustande.

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The author wishes to thank Dr.H. Holmsen and Dr.N. O. Solum for valuable advice and criticism.     

Ectoprotein kinase-mediated phosphorylation of FAT/CD36 regulates palmitate uptake by human platelets

Glycoprotein IV (FAT/CD36) has been shown to be phosphorylated by a cAMP-dependent, platelet membrane-bound ectokinase. In this study, we demonstrate that ectophosphorylation of FAT/CD36 regulates initial palmitate uptake. This is the first time that short-term regulation of the activity of a long-chain fatty acid carrier could be shown. Phosphorylation of FAT/CD36 was paralleled by a significant decrease in initial palmitate uptake by morphologically and functionally intact platelets. Maximum inhibition of palmitate uptake was achieved at 0.5 nM extracellular ATP, being significantly decreased to 72% compared to the control. Inhibition of palmitate uptake was abolished by co-incubation with the specific protein kinase A inhibitor peptide PKI or with β,γ-methylene-ATP, and was reversible upon addition of alkaline phosphatase. An extracellular ATP concentration above 5 μM completely prevented the ectophosphorylation-mediated inhibition of palmitate uptake. We conclude that FAT/CD36-mediated palmitate uptake by human platelets is short-term regulated via cAMP-dependent ectophosphorylation of FAT/CD36. Received 18 July 2002; received after revision 29 August 2002; accepted 19 September 2002 RID="*" ID="*"Corresponding author.     

Visualisation of lectin binding sites on the surface of human platelets using lectins adsorbed to gold granules

Summary Washed human platelets have been incubated with the lectins WGA, ConA and RCA₁, adsorbed to differentsized gold particles. Plasma membrane receptors for each lectin were then located by scanning and transmission electron microscopy.Acknowledgments. We would like to thank Mlle Dominique Dupuis for technical assistance. This work was supported in part by grant No. CRL 78.5.128.1 from INSERM.     

Identification of proteins in human prostate tumor material by two-dimensional gel electrophoresis and mass spectrometry

Protein patterns in cells collected from benign prostatic tissues and prostate carcinomas were analyzed using two-dimensional polyacrylamide gel electrophoresis and mass spectrometry. Polypeptide expression was evaluated by computer-assisted image analysis (PDQUEST). Proteins expressed by prostate tumors were identified via in-gel digestion and subsequent matrix-assisted laser desorption/ionization mass spectrometry. In addition to cytoskeletal and mitochondrial proteins, a 40-kDa protein was identified as prostatic acid phosphatase (PAP). PAP expression decreased approximately twofold between benign and malignant tissue. Increased expression of heat shock protein 70 and decreased expression of tropomyosin 1 were also observed in the malignant tissue. The analysis of prostate material by two-dimensional gel electrophoresis and mass spectrometry shows that particular proteins are of interest as markers of disease. Received 18 December 2000; accepted 4 January 2001     

Podocalyxin enhances the adherence of cells to platelets

Podocalyxin (PODXL) is a mucin protein of the CD34 family expressed in kidney glomerular podocytes, vascular endothelium, progenitor bone marrow and tumor cells. It is assumed that PODXL plays an anti-adherent role in kidney podocytes. CHO cells stably expressing human PODXL (CHO-PODXL) or human tumor cells (Tera-1) inherently expressing PODXL showed increased adherence to platelets. The adherence of cells was inhibited (70%) by blockers of platelet P-selectin, prevented by the soluble ectodomain of human PODXL (PODXL-Δ) or by the arginine-glycine-aspartate (RGDS) peptide and partially impeded by inhibition of integrin αVβ3/αVβ5, suggesting a coordinated action of P-selectin and integrins. Colocalization of platelet P-selectin and PODXL expressed on CHO cells was demonstrated by confocal immunofluorescence. No adherence to platelets was observed when PODXL was expressed in glycomutant CHO cells deficient in sialic acid. Received 14 August 2007; received after revision 12 September 2007; accepted 13 September 2007     

The identification of chlorpromazine metabolites in human blood by gas liquid chromatography

Zusammenfassung Chlorpromazin und 5 sciner Metaboliten wurden gaschromatographisch in menschlichem Plasma identifiziert. Zwei dieser Metaboliten wurden auch in Erythrozyten aufgefunden. Die Blutproben wurden von Paticnten genommen, welche täglich 100–250 mg Chlorpromazin erhielten. Chlorpromazin und die nicht konjugierten Metabolite scheinen vom Plasma innerhalb weniger Stunden nach Verabreichung einer Dosis Chlorpromazin zu verschwinden. Die gebundenen Metabolite konnten jedoch zu jeder Zeit nachgewiesen werden. Das metabolische Bild kann von einem Patienten zum anderen wechseln.