Artifactual phosphate binding due to impurities in [32P]orthophosphate

Many commercial preparations of [32P]orthophosphate contain radioactive impurities that interfere with binding and transport studies in biological systems. One type of impurity is micro-particulate whereas another may be pyrophosphate. Methods of removing these impurities from radiolabeled orthophosphate solutions are described.     

Congenital dyserytropoietic anaemia, type II (HEMPAS) in three siblings

These siblings of a Czech family aged 21, 19 and 6 years, respectively, with congenital dyserythropoietic anemia, type II, (HEMPAS) are reported. In two elder siblings ferrokinetic studies revealed a rapid plasma 59Fe clearance, markedly decreased erythrocyte incorporation and shortened 51Cr red-cell survival. Direct anti-globulin test was found positive in one of them. Further investigations revealed low values of blood plasma cholesterol, total lipids, beta-lipoproteins, beta-carotine and vitamin E and A as well as low values of the prothrombin complex. Liver biopsy demonstrated siderosis and disseminated intravascular coagulation in the liver in both patients. The possible reasons for these humoral aberrations are discussed.     

The interpretation of a moving retinal image

It is shown that from a monocular view of a rigid, textured, curved surface it is possible, in principle, to determine the gradient of the surface at any point, and the motion of the eye relative to it, from the velocity field of the changing retinal image, and its first and second spatial derivatives. The relevant equations are redundant, thus providing a test of the rigidity assumption. They involve, among other observable quantities, the components of shear of the retinal velocity field, suggesting that the visual system may possess specialized channels for computing these components.     

N-anthraniloylation converts peptide p-nitroanilides into fluorogenic substrates of proteases without loss of their chromogenic properties

By simple substitution of an N-acyl group for the anthraniloyl(o-aminobenzoyl) group, chromogenic p-nitroanilide substrates are converted into highly sensitive fluorogenic substrates of proteases. The fluorescence of the anthraniloyl group is completely quenched by the p-nitroanilide moiety in the intact substrates and is released during their enzymatic hydrolysis. The approach is exemplified by the synthesis of anthraniloyl-Phe p-nitroanilide, anthraniloyl-Lys p-nitroanilide, and anthraniloyl-Gly-Gly-Phe p-nitroanilide as substrates for chymotrypsin, trypsin, and alkaline mesentericopeptidase, respectively. The kinetic parameters of these substrates are slightly better than those of similar derivatives bearing other acyl groups, suggesting that the enhanced sensitivity is completely due to the method of measurement. Since the conversion does not affect the chromogenic properties of the substrates, the same compounds can be used as usual p-nitroanilide substrates as well.     

Localization of ribosomal genes and nucleolar activity in lymphocytes of swine (Sus scrofa domestica) stimulated by phytohemagglutinins

The Pig chromosomes that contain rDNA sites displayed a polymorphism in the distribution of the genes among the nucleolar organizers located on pairs Nos. 8 and 10. Two, or more often three, active sites were observed in the chromosomes of lymphocytes stimulated with phytohemagglutinin. Only 5% of the metaphases showed a 4th small active site. At the onset of stimulation most cells contained one-two nucleoli; four nucleoli were never observed. After prolonged stimulation, the number of nuclei containing three nucleoli increased. A 4th small nucleolus appeared in a few cells, presumably formed by activation of the smallest rDNA site.     

Substratum topography and cell traction on sulphuric acid treated bacteriological-grade plastic

Treatment of bacteriological-grade plastic with concentrated sulphuric acid is a well known technique which increases the wettability of the surface and renders it suitable for eukaryotic cell adhesion. We have noticed that these substrata present a distinctive surface topography in the presence of a serum supplement under normal culture conditions. The adsorbed serum layer is comprised of fine furrows and ridges and the influence of adherent cells on this layer leads to minute tears and distortions in the direction of the corrugations. This provides a novel system for the investigation of cell spreading and locomotion by scanning electron microscopy.