[The isolation of thrombocytes from the whole blood of dogs with a density gradient (Ficoll-metrizoate)].
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Biomedical subjects
Publications and source records attributed to C Niemann.
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The administration of 20 mg/kg of acetylsalicylic acid in 18 clinically healthy dogs resulted in a thrombocytopathy with lengthened capillary bleeding time and irreversible aggregation inhibition. Through the set up of individual dilution series, one could conclude the proportional percentage of aggregation functional transfused thrombocytes. The capillary bleeding time was shortened after the intravenous injection of prednisolone (5 mg/kg) without measurable influence on the thrombocytes. Compared to the singular use of cortisone or blood transfusion alone, the effect on capillary bleeding time became magnified when one combined transfusion and corticosteroids. The fresh blood conserves (12 hours) were, with respect to the haemostyptical properties, superior to blood stored for 5 days. The component of aggregation-functional thrombocytes was, due to the prophylactic cortisone application prior to transfusion, not increased.
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A method for the cultivation of organ explants from abdominal mammary glands of virgin mice has been established. In a serum-free medium containing aldosterone, prolactin, insulin, and cortisol (APIH medium) mammary gland development was documented by lobuloalveolar morphogenesis. The hormonal requirements for in vitro expression of beta-casein and of the mammary-derived growth inhibitor (MDGI) were tested. To this end, a full length cDNA coding for mouse MDGI was prepared displaying strong homologies to a mouse heart fatty acid binding protein, which is also expressed in the mammary gland. MDGI and beta-casein transcripts were found to be absent in the mammary tissue from primed virgin mice, and were induced upon culture of mammary explants in the APIH medium. An immunohistochemical analysis with specific antibodies against MDGI and casein revealed a different pattern of expression for the two proteins. In the APIH medium, MDGI was expressed mainly in differentiating alveolar cells of the lobuloalveolar structures, whereas beta-casein was present in both ductules and alveoli. The relationship between functional differentiation and MDGI expression was further studied in explants from glands of late-pregnant mice. At this stage of development, MDGI is found both in ducts and in alveoli. If explants were cultured with epidermal growth factor (EGF) and insulin, the lobuloalveolar structure was still present, whereas MDGI disappeared. Reinduction of MDGI expression was achieved by subsequent PIH treatment. Independent on developmental stage, EGF strongly inhibits MDGI mRNA expression. It is concluded that MDGI-expression is associated with functional differentiation in the normal gland.