Further studies on gel filtration of urinary substances.

Urine specimens from healthy children were concentrated and filtered through Sephadex G-100 and G-25. The eluate was recovered in fractions of 5 ml each. Total hexoses, galactose, total proteins and total uronic acids were determined on each fraction. Total hexoses and galactose were eluted in 6 distinct peaks. The uronic acid containing GAG were eluted in 4 small peaks on G-25 and 1 peak on G-100. This peak is eluted always with high amounts of galactose containing glycoproteins.     

Further studies on gel filtration of urinary substances

Summary Urine specimens from healthy children were concentrated and filtered through Sephadex G-100 and G-25. The eluate was recovered in fractions of 5 ml each. Total hexoses, galactose, total proteins and total uronic acids were determined on each fraction. Total hexoses and galactose were eluted in 6 distinct peaks. The uronic acid containing GAG were eluted in 4 small peaks on G-25 and 1 peak on G-100. This peak is eluted always with high amounts of galactose containing glycoproteins.This work was partially supported by research grants from the CONICET, República Argentina.     

Analyses immunoélectrophorétiques des fractions protéiques du sérum humain séparées par électrophorèse en gel d'amidon

Summary Human plasma components, separated by starch gel electrophoresis, are eluted and concentrated by lyophilisation. Immuno-electrophoretic analysis shows that certain of these fractions are immunologically heterogenous. The authors point out that this immunological criterium should be used before employing an eluted fraction, even if it appears electrophoretically homogenous.      

5-Hydroxytryptamine binding to butanol extracts from myelin fragments

Summary The myelin fraction of rat brain stem was treated with butanol-water mixtures, and the extracted proteolipids were separated by Sephadex LH₂₀ column chromatography. 2 peaks of proteolipids eluted in chloroform-methanol 4/1 showed the binding capacity for C¹⁴·5-HT. This finding suggests the necessity of the more careful investigations for the probability of proteolipids as receptor proteins in the central nervous system.     

Separation of modulator-dependent protein kinase (type I) from cyclic GMP-dependent protein kinase in mouse testes

A new type of enzyme, modulator-dependent protein kinase (type I) (M-PKI), was successfully isolated from the cytosol fraction of mouse testes. It was eluted slightly after the peak of cyclic GMP-dependent protein kinase (G-PK) by Sephadex G-200 gel filtration. Unlike either cyclic AMP-dependent protein Kinase (A-PK) or G-PK, its maximal activity depended exclusively on the presence of crude protein kinase modulators (PKM) or partially purified stimulatory modulator (PKMs).     

Neue mehrfache Untersuchungen am gleichen Kammerwasser menschlicher Augen

Summary The normal human aequous has a volume of 0.12–0.20 cm³ and its protein content is from 10 to 20 mg%. We ascertained the specific weight by the falling drop method in a mixture of xylol and brombenzene. In order to estimate the protein content, we placed a series of drops from the human aequous on filter paper with the Agla pipette; they were then tinted with amido-black and the optical density of the eluted spots was read in the Beckman spectrophotometer. The remnants of 2–3 specimens of human aequous were then brought together and concentrated there until the protein concentration is just sufficient to allow a separation into the various fractions by paper-electrophoresis.      

Isolation, from anti-dinitrophenyl sera taken at the beginning of immunization, of a protein combining with the hapten and which is not related to immunoglobulins]

A protein, not antigenically related to the immunoglobulins, has been isolated by affinity chromatography from the serum of rabbits immunized with dinitrophenylated human gamma-globulin. Its concentration in the serum reaches maximum during the first three days which follow the injection. On "Sephadex G-200", it is eluted after the ribonuclease. Its electrophoretic mobility is slower than that of the IgG.     

Collagen breakdown by gingival collagenase and elastase

Summary The granule fraction of human polymorphonuclear leucocytes (PMNs), the concentrated product of gingival washing from 2 human volunteers and the culture fluid of samples of human gingiva were incubated with neutral salt soluble collagen from rat skin and the patterns of collagen degradation were studied by SDS polyacrylamide gel electrophoresis. Collagenase from human gingiva cleaved the collagen molecules in a fashion similar to that of the PMN granule fraction. Collagen was also attacked by elastase from human PMNs and, to a lesser extent, by elastase from the gingival washings.Supported by a grant of Swiss National Fund for Scientific Research No. 3.604.0.75.Acknowledgments. We thank Prof. M. Baggiolini, Wander S.A., Bern for his helpful advice. We are very grateful to Mrs I. Condacci and Miss E. Andersen for their skillful technical assistance.     

Hemagglutination by simian adenovirus 7]

Simian adenovirus 7, either complete virus or its capsid subunits, agglutinates Rat (Sprague-Dowley) red blood cells in the presence of heterotypical antiserum. Haemagglutination takes place at 4 degrees C and room temperature. The antigen could not be eluted and its haemagglutinin properties are heat-stable. The reaction is specific. It is inhibited by homologous antiserum only. This property and its characteristics permit a camparison of this strongly oncogenic adenovirus to the human adenovirus of subgroup III of Rosen.     

Heterologous immunoadsorption in the purification of enzymes: N-acetyl-β-glucosaminidase

Summary An immunoadsorbent column was prepared using a specific antibody toN-acetyl--glucosaminidase of human origin. Although no precipitating activity of the antisera was found with mouse or rat liver extracts, enzyme was easily eluted from the column which provided about 50fold, single-step purifications of these heterologous enzymes.     

Alkaline phosphatase in human lymphocyte subpopulations.

The levels of membrane alkaline phosphatase have been measured on different lymphocyte fractions from human peripheral blood separated on bovine serum albumin discontinuous gradients. The peak in enzyme activity was observed in a non-T-, non-B-cell fraction, rich in "null" lymphocytes; the lowest values were found in the fraction with the highest proportion of T-cells.     

On the complementarity of long repeated sequences in DNA to hnRNA

Long repeated sequences containing up to 18,000 base pairs were found in a human DNA fraction isolated with the nuclease S1-dioxane method. Hybridization studies showed that the long repeats contained a greater proportion of sequences complementary to hnRNA than short repeats. They also exhibited homology to the latter, as shown by cross-hybridization experiments.     

Alkaline phosphatase in human lymphocyte subpopulations

Summary The levels of membrane alkaline phosphatase have been measured on different lymphocyte fractions from human peripheral blood separated on bovine serum albumin discontinuous gradients. The peak in enzyme activity was observed in a non-T-, non-B-cell fraction, rich in null lymphocytes; the lowest values were found in the fraction with the highest proportion of T-cells.     

Anticomplementary fraction from the poisonous secretion of the paratoid gland of the toad (Bufo marinus paracnemis Lutz)

Fractionation of the poisonous secretion of the toad Bufo marinus paracnemis Lutz, by dialysis and chromatography on QAE-Sephadex, led to the isolation of a fraction which was adsorbed to the ion exchanger. This fraction, when incubated with human serum, yielded an anticomplementary effect that was evaluated by measuring the kinetics of lytic activity on sensitized sheep red cells (classical pathway) and unsensitized rabbit cells (alternative pathway).     

Hemagglutination of neuraminidase-treated human erythrocytes by Leishmania enriettii infected guinea-pig sera.

Sera from guinea-pigs infected with the protozoan parasite Leishmania enriettii showed higher hemagglutination (HA) titres for neuraminidase treated human erythrocytes than those of normal guinea-pig sera. This HA activity was associated mostly with the 19S fraction of the immune serum and could be absorbed out with an antigenic fraction of the parasite membrane. Antigenic determinants involved in this HA reaction consisted of, at least, beta-D-galactosyl or lactosyl residues.     

Immunologische Eigenschaften der menschlichen mit Heparin präzipitierten Plasmafraktion

Summary The plasmatic fraction, which is precipitated by cooling human heparin plasma, has been found to contain the constituent identical with-globulin and also another component with the fibrinogen of Cohns' fraction I immunologically identical. With the help of the agar-gelose diffusion method, no qualitative immunological differences have been found with regard to the antigenous composition of the plasmatic fraction isolated in cases of various system diseases.     

Novel and sensitive noncompetitive enzyme immunoassay for kassinin in rat plasma

A novel and sensitive noncompetitive enzyme immunoassay for kassinin is described. Kassinin was biotinylated using sulfosuccinimidyl-6-(biotinamido)hexanoate. The biotinylated kassinin was trapped on anti-kassinin IgG-coated polystyrene balls and, after washing to eliminate other biotinylated substances, was eluted with HCl. The biotinylated kassinin eluted was reacted with anti-kassinin Fab'-peroxidase conjugate and trapped onto streptavidin-coated polystyrene balls. Peroxidase activity bound to the polystyrene balls was assayed by fluorimetry. The detection limit of kassinin was 0.13 pg (0.1 fmol)/tube or 0.065 microgram/l of rat plasma, which was 750-fold or 15-fold lower than that for competitive radioimmunoassay.     

Regional and subcellular distribution of superoxide dismutase in brain.

Superoxide dismutase has been found to be widely distributed and of approximately the same specific activity in all regions of human brain examined. It is not reduced during degeneration of the basal ganglia in Huntington's Chorea. After subcellular fractionation of human and guinea-pig cerebral cortex, the highest specific activity of the enzyme was found in the soluble fraction.     

Dynamics of excretion of urinary chemosignals in the house mouse (Mus musculus) during the natural estrous cycle

The volatile fraction of urinary metabolites was investigated chromatographically at five different stages of the natural estrous cycle. A very substantial endocrine dependency has been noted for 11 compounds: 4 ketones, 2 acetate esters, 3 dihydrofuran isomers, dehydro-exo-brevicomin, and 2,5-dimethylpyrazine. The compounds were structurally verified through combined gas chromatography/mass spectrometry.