Biomedical subjects
Publications and source records attributed to F Rosati.
A procedure is described for the detection of anabolic steroids in urine by using capillary gas chromatography-mass spectrometry. The method is suitable for doping control and has a sensitivity as low as 1 ppb. The excretion mode and the metabolism of ten commercial anabolic steroids is reported.
In this paper we describe a mild procedure which results in the extraction of a glycoprotein fraction from the vitelline coat (VC) of Ciona intestinalis while leaving behind the bulk of the VC components. When acting upon the spermatozoa this fraction inhibits sperm binding to the VC and fertilization and elicits sperm activation including the acrosome reaction. SDS-PAGE shows that it contains the same (fucosyl) glycoprotein components previously recognized in the total extracts of VC. It is suggested that this material contains the sperm receptors or those components of the receptors that are essential for their Chinese function.
We have studied the binding of spermatozoa to the receptor sites on the vitelline coat (VC) of glycerol-treated eggs (ghost eggs) of the Ascidian, Ciona intestinalis (Protochordate). Glycerol treatment cytolyses the egg without affecting the ability of the VC to bind spermatozoa in a species-specific manner; however, in this system binding is not followed by the acrosome reaction. The ghost eggs are metabolically inert. As a base line for our analysis, we have studied the concentration-dependent heat evolved and oxygen consumption of spermatozoa when diluted in sea water. The process has been analyzed on the basis of equations derived by Liquori and Tripiciano to describe cell growth. Upon binding to the ghost eggs, the spermatozoa produce an explosive heat evolution (excess heat) which is not accompanied by oxygen consumption. The excess heat produced plotted against sperm concentration (at constant egg concentrations) gives an asymmetric bell-shaped curve. This is interpreted as being due to the competitive effect of sperm agglutination at a high sperm concentration. It is concluded that only spermatozoa that attach singly (monomeric spermatozoa) to the egg undergo metabolic activation.
We have studied the differentiation of the oocyte vitelline coat (VC) and jelly coat (JC) of the starfish, Astropecten aurantiacus. The precursor material of both envelopes is secreted by the oocyte while the follicle cells do not appear to participate in the secretory process. The first indication of differentiation of the VC is the deposition of a fine fibrillar material between the microvilli which emerge from the oocyte surface. External to this, a more loosely organized material becomes the precursor of the JC. At this time both layers are periodic acid-Schiff (PAS)-positive. In a later stage, the material between the microvilli acquires a more compact organization, looses its PAS-positivity while acquiring fucose binding protein (FBP) affinity. On the contrary, the JC remains PAS-positive and FBP-negative. In the full grown oocytes the VC is made up of densely packed fibrils oriented tangentially to the oocyte surface and is tightly bound to the microvilli. The observations are discussed in connection with the problem of the role of the egg envelopes in sperm-egg recognition and in the induction of the acrosome reaction.
In this paper we argue that the mechanisms underlying the segregation of the somatic and germ cell lines are basically similar. The interaction of the genome with specific cytoplasmic factors is responsible for the restriction of their developmental potencies in the somatic cell lines, and for the prevention of such a restriction in the germ cell line. In particular, it is suggested that meiosis is part of the differentiation program of the germ cells.
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Separated T and B lymphocytes from human peripheral blood were studied using the freeze-fracture technique. Quantitative analysis performed on density and size of intramembranous particles (IMPs) present on both fracture faces of the plasma membrane has revealed remarkable differences between cells belonging to the two main lymphocyte populations. In particular: (a) both fracture faces of the cytoplasmic membrane of B lymphocytes exhibit larger particles than T lymphocytes; (b) the mean densities, on both protoplasmic (PF) and external (EF) fracture faces, in B lymphocytes are lower than in T lymphocytes; (c) in B cells the partition ratio of particles between PF and EF is reversed with respect to T cells; (d) on both fracture faces of B lymphocytes, the IMP densities present a normal distribution while on T cells, density values show bimodal distributions indicating the existence of two cell subsets differing in particle density.
Receptors for IgM were detected on peripheral blood human eosinophils by a rosette technique with ox red blood cells coated with the IgM fraction of the specific immunserum. Between 14% and 43% (mean 27%) FcmuR positive cells were found after an overnight incubation period at 37 degrees C by using this technique. The specificity of the receptors for IgM was assessed by studying the inhibitory capacity of purified human IgM in the rosette assay. From an ultrastructural point of view, the EAM rosette-forming cells are mature eosinophilic granulocytes characterized by a nucleus with a variable number of lobes and a certain number of "first type" granules partially or totally devoid of their content.
The sperm receptors of the ascidian oocyte are located at the outer surface of the vitelline coat (formerly called the chorion). The fucose residues are the receptor's most important components for sperm recognition and binding. We asked whether the fucosyl-containing glycoproteins of the vitelline coat are a product of the oocyte, the follicle cells, or the test cells. Ovaries of Ciona intestinalis were injected with L-[3H]fucose and the progress of its incorporation was followed by using autoradiography and sodium dodecyl sulfate/polyacrylamide gel electrophoresis of the injected gonads and of the isolated vitelline coats. We found that incorporation of fucose begins within the vitellogenic oocytes, and fucose slowly accumulates in the differentiating vitelline coat. At no time could fucose incorporation be detected in the follicle cells or in the test cells. Sodium dodecyl sulfate/polyacrylamide gel electrophoresis of vitelline coats prepared from the injected ovaries showed fucose incorporation into the same three glycoproteins present in vitelline coats from mature oocytes and identified by their affinity for 125I-labeled fucose-binding protein [Pinto, M. R., De Santis, R., D'Alessio, G. & Rosati, F. (1981) Exp. Cell Res. 132, 289-295]. A radioactive band not found in the mature oocyte was also present.
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A new procedure, based on centrifugation through multiple discontinuous gradients of polyvinylpyrrolidone-coated silica particles has been developed for the separation of almost pure eosinophils from blood samples of healthy donors. When this technique is employed for the purification of eosinophils from the peripheral blood of patients with parasitic diseases, two cell fractions almost pure in eosinophil granulocytes are harvested at different densities. Preliminary results, concerning the characterization of these cell fractions, indicate that in the terms of detectable membrane receptors and K cell activity, two distinct eosinophil subpopulations can be recognized.
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A freeze-fracture study has been carried out on human peripheral blood lymphocytes (hPBL) from healthy donors. Lymphocytes were frozen either from 37 degrees C or 4 degrees C. Quantitative analysis performed on density and size of intramembranous particles (IMPs) present on both fracture faces of the plasma membrane has revealed: a) a difference in size between IMPs on the external face (EF) and those on the protoplasmic face (PF); b) a remarkable influence of temperature either on size or density of the IMPs; c) the existence at 4 degrees C of two lymphocyte populations differing in IMP density.
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The aim of this study was to investigate the possible formation of spontaneous rosettes between human eosinophils and sheep erythrocytes. In patients with parasitic diseases a percentage of 15.7 +/- 6 (range 6-23%), eosinophils were able to form rosettes. Each rosette showed as few as four to eight erythrocytes attached per eosinophil. Two types of interaction were observed at the electron microscope: point attachments and large surface contacts. In patients with non-parasite induced eosinophilia the formation of rosettes is decidedly below the above-mentioned percentage (less than 2%) or totally absent. It follows that during parasitic infections with blood eosinophilia, whatever the mechanism responsible, the formation of spontaneous rosettes could be diagnostic significance and might reflect qualitative or quantitative modifications in the eosinophil population.