Inversal relationship between basal adenylcyclase activity and maximal degree of stimulation in membranes of red blood cells from rats.

The degree of stimulation of adenylcyclase activity, in membranes from immature red blood cells from rats, brought about by isoprenaline, guanylyl-imidodiphosphate and sodium fluoride is strongly dependent on the basal activity of the enzyme. The inversal relationship between basal activity and the maximal degree of stimulation by (--) isoprenaline, shows an apparent seasonal dependence.     

Inversal relationship between basal adenylcyclase activity and maximal degree of stimulation in membranes of red blood cells from rats

Summary The degree of stimulation of adenylcyclase activity, in membranes from immature red blood cells from rats, brought about by isoprenaline, guanylyl-imidodiphosphate and sodium fluoride is strongly dependent on the basal activity of the enzyme. The inversal relationship between basal activity and the maximal degree of stimulation by (–) isoprenaline, shows an apparent seasonal dependence.This work was supported by a grant from the Deutsche Forschungsgemeinschaft.     

Recombination of integral and peripheral protein fractions from human red cell membrane with homologous lipids

Summary Integral and peripheral protein fractions from human red cell membranes were recombined with a total red cell lipid extract and with homologous lipids in varying mixtures, by dialysis from 2-chlorethanol solutions. The 2 protein fractions were compared for lipid binding capacity and for selectivity towards individual lipids.     

Conversion of glycine max seed agglutinins from nonspecific to anti-(A + B).

The seeds of glycine max contain agglutinins which are typically nonspecific in their reactivity. Our investigations show that the phytagglutinins in GM can be converted from nonspecific to anti-(A + B) after the lectin is absorbed with horse red cells. The anti-A and anti-B fractions can be further separated by suitably absorbing the lectin with human red cells. The lectin absorbed with horse red cells or with group-0 human red cells shows an A-stressed activity.     

Conversion of glycine max seed agglutinins from nonspecific to anti-(A+B)

Summary The seeds of glycine max contain agglutinins which are typically nonspecific in their reactivity. Our investigations show that the phytagglutinins in GM can be converted from nonspecific to anti-(A+B) after the lectin is absorbed with horse red cells. The anti-A and anti-B fractions can be further separated by suitably absorbing the lectin with human red cells. The lectin absorbed with horse red cells or with group-0 human red cells shows an A-stressed activity.     

Differential binding of vitamin E to sarcoplasmic reticulum from fast and slow muscles of the rabbit

Summary The distribution of [³H]-tocopherol between the intracellular membranes of fast white and slow red muscles was studied in vivo. In vitro experiments confirmed a higher incorporation of the vitamin into the reconstructed sarcoplasmic reticulum membranes from slow muscle.This work was supported by funds from the Consiglio Nazionale delle Ricerche to the C.S. Biologia e Fisiopatologia Muscolare and to the Istituto di Patologia Generale, Università di Bologna, and in part by a grant from the Muscular Dystrophy Association to Prof. Aloisi.     

Altered microviscosity of in vivo lipid-manipulated membranes inTetrahymena pyriformis: A fluorescence study

Summary By determination using fluorescence polarization measurements with 1,6-diphenyl 1,3,5-hexatriene, ergosterolreplaced pellicle and microsome membranes ofTetrahymena cells become less fluid, whereas those membranes from chimyl alcohol-fed cells are more fluid, when compared with the control native membranes.Acknowledgments. We wish to express thanks to Central Scientific Commerce, Inc., Tokyo for permission to use an Elscint Microviscosimeter MV-Ia. This work was in part supported by grant from the Ministry of Education.     

Role of tropomyosin in the sensitivity of (Na+ + K+) ATPase to ouabain]

EDTA treatment of isolated plasma membranes from MF2S cells increased 1,000 fold the sensitivity of (Na+ + K+) ATPase activity to ouabain. The original sensitivity of the enzyme to the drug is recovered after addition of tropomyosin together with Ca++ ions to the treated membranes.     

31P-NMR study on nucleotides and intracellular pH of hereditary spherocytes.

As determined by 31p-NMR spectroscopy, intracellular pH of hereditary spherocytes was lower (pH 6.7-6.9) than that of normal red cells. The level of adenosine diphosphate in hereditary spherocytes was found to be persistently high. The metabolism of nucleotides and other phosphoryl compounds in human red blood cells have been studied in detail by 31p-MNR spectroscopy. However, to our knowledge, there seems to be no report describing the result of 31p-NMR spectroscopy on red blood cells from hereditary spherocytosis.     

Species difference in the effects of proteolytic enzymes on red cell membrane

Summary Pronase and -chymotrypsin digested the major glycoprotein in the human and mouse red cell membranes and in SDS gel electrophoresis the glycoprotein disappeared accompanied by the appearance of a new band of lower mol.wt. However in the membranes of sheep, rat and rabbit, no digestion was demonstrated. The effects of pronase on anion permeability were almost identical for human and animal erythrocytes.     

Red cell metabolism in red and grey kangaroos

Glucose utilization, lactate production and glutathione regeneration were measured in the red blood cells of 2 species of Australian Marsupials, Eastern grey Kangaroo (Macropus gigantus) and red kangaroo (Macropus rufus), and were found to be significantly lower in the red blood cells from grey than that of red kangaroos.     

Functional and pharmacological induced structural changes of the cystic fibrosis transmembrane conductance regulator in the membrane solved using SAXS

The cystic fibrosis transmembrane conductance regulator (CFTR) chloride channel is a membrane-integral protein that belongs to the ATP-binding cassette superfamily. Mutations in the CFTR gene cause cystic fibrosis in which salt, water, and protein transports are defective in various tissues. To investigate the conformation of the CFTR in the membrane, we applied the small-angle x-ray scattering (SAXS) technique on microsomal membranes extracted from NIH/3T3 cells permanentely transfected with wild-type (WT) CFTR and with CFTR carrying the ΔF508 mutation. The electronic density profile of the membranes was calculated from the SAXS data, assuming the lipid bilayer electronic density to be composed by a series of Gaussian shells. The data indicate that membranes in the microsome vesicles, that contain mostly endoplasmic reticulum membranes, are oriented in the outside-out conformation. Phosphorylation does not change significantly the electronic density profile, while dephosphorylation produces a significant modification in the inner side of the profile. Thus, we conclude that the CFTR and its associated protein complex in microsomes are mostly phosphorylated. The electronic density profile of the ΔF508-CFTR microsomes is completely different from WT, suggesting a different assemblage of the proteins in the membranes. Low-temperature treatment of cells rescues the ΔF508-CFTR protein, resulting in a conformation that resembles the WT. Differently, treatment with the corrector VX-809 modifies the electronic profile of ΔF508-CFTR membrane, but does not recover completely the WT conformation. To our knowledge, this is the first report of a direct physical measurement of the structure of membranes containing CFTR in its native environment and in different functional and pharmacological conditions.     

Physiological dissection of various effects of ruthenium red dye on Paramecium cells.

Polycationic dye ruthenium red, but not alcian blue, if externally applied to Paramecium cells quickly inhibits their phagocytosis. Ruthenium red combined with the cell surface diminishes frequency and duration of ciliary reversals and gradually inactivates the Ca++ gating mechanism. This effect persists for 1-3 after ruthenium red removal from the culture medium.     

Änderung des Glukosetransports frischer menschlicher Erythrozyten bei längerer Inkubation

Summary Incubation of fresh defibrinated blood at 37°C increases the volume of red cells and decreases their permeation rate for glucose. The kinetics of sugar transport show direct proportionality between rate and the osmotic content of red cells. The measured volume changes and the changes of osmotic content calculated from the glucose permeation curves are in good agreement. Adenosine reconstitutes the increased volume and the decreased permeation rate in red cells. Therefore the decrease in permeation rate is explained by the increase in volume.     

Red cell metabolism in red and grey kangaroos

Summary Glucose utilization, lactate production and glutathione regeneration were measured in the red blood cells of 2 species of Australian Marsupials, Eastern grey kangaroo (Macropus gigantus) and red kangaroo (Macropus rufus), and were found to be significantly lower in the red blood cells from grey than that of red kangaroos.The author wishes to thank Mrs T. Stephens, M. A. Gruca and A. Mulley for help in blood collection and laboratory work. This work was supported by a grant from the National Health and Medical Research Council of Australia.     

31P-NMR study on nucleotides and intracellular pH of hereditary spherocytes

Summary As determined by³¹p-NMR spectroscopy, intracellarar pH of hereditary spherocytes was lower (pH 6.7–6.9) than that of normal red cells. The level of adenosine diphosphate in hereditary spherocytes was found to be persistently high. The metabolism of nucleotides and other phosphoryl compounds in human red blood cells have been studied in detail by³¹p-NMR spectroscopy^1–3. However, to our knowledge, there seems to be no report describing the result of³¹p-NMR spectroscopy on red blood cells from hereditary spherocytosis.     

Étude de la compétition antigénique chez des souris thymectomisées à la naissance

Summary The injection of horse red blood cells into the mouse 4 days before immunization with sheep red blood cells produces an inhibition of the primary response against sheep red blood cells. If the animal is thymectomized at birth, this antigenic competition does not occur. These findings do not permit a choice between the two hypotheses about antigenic competition.     

Galactose-l-phosphate uridyl transferase activity in red cells of various animal species

Summary Monkey red cells were chosen as controls in tests of human red cell galactose-l-phosphate uridyl transferase, after comparing activities and isozyme patterns of the enzyme from several domestic or laboratory-bred species.     

Inclusion of Gross virus cell surface associated antigen in liposomes]

The Gross virus associated cell surface antigen GCSAa was extracted from (C58NT)D cells by solubilization of membranes with detergent and partially purified. This antigen was entraped in liposomes. Absorption experiments of the cytotoxic activity towards EmaleG2 cells of a W/Fu anti (C58NT)D serum showed the presence of the antigen at the surface of sensibilized liposomes.