Mathematical model of autoimmunity]

A mathematical model of autoimmunity is developed. This model is a system of two nonlinear differential equations, which describe the concentration dynamics of tissue cells and agressive lymphocytes. An analysis of the solutions shows that this model reproduces general behaviour of autoimmune diseases.     

A fluorescence study of the binding of cytochrome C to mixed-phospholipid microvesicles : evidence for a preferred orientation of the bound protein.

Kinetic and equilibrium experiments are reported on the binding of the fluorescent probe 1,8-anilino-naphtalene sulfonate (ANS) to microvesicles of natural lecithin containing 10 per cent of an anionic phospholipip (90 : 10 mixtures). Kinetics discriminated between fast binding to the outer leaflet of the bilayer and apparently slow binding to the inner leaflet controlled by the diffusion of the probe across the bilayer. The equilibrium distribution of ANS between the two leaflets was not dependent on the nature of the anionic species and the spectral properties of bound ANS were identical in all cases investigated. A hyperbolic saturation was observed allowing to propose an affinity scale for the binding of ANS to mixtures of lecithin with phosphatidic acid, phosphatidylinositol, and cardiolipin. The effects on binding of ionic strength and sodium dodecylsulfate were also considered. The binding of horse heart ferricytochrome c to ANS-labelled microvesicles was studied quantitatively making use of the quenching of the probes fluorescence by the heme. Perrin-F?rster energy transfer could be analysed on the basis of a simple model of the physical arrangement of the system which was elaborated from published data referring to ANS and cytochrome c binding to phospholipids. Experimental and theoretical computed values of the quenching efficiency were compared and led to conclude in favor of a preferred orientation of the heme crevice fully accessible from the external space at the lipid interface.     

Role of membrane-bound calcium in changes in the ATPase activity, permeability and structural state of human erythrocyte membranes]

A study was conducted on the reconstituted erythrocytes obtained by the method of fast reversible hemolysis. The concentration of free Ca2+ ions in the reconstituted erythrocytes was supported by Ca-EGTA and Ca-nitrate buffers. Oubain-uninhibited ATPase component with a high affinity for Ca2+ (K0.5=4 micron) and alteration of passive and active K+-permeability in the region of free Ca2+ concentration up to 10 micron could be determined only when the content of membrane-bound Ca+ varied. Depletion of the inner side of the membrane of reconstituted erythrocyte is accompanied by alteration of hydrophobic character of the hydrocarbon region of the membrane. It is suggested that Ca+-induced alterations in the structure of the erythrocyte membrane may be a direct cause of the alterations in ATPase activity with a high Ca2+ affinity and permeability for univalent cations.     

New indications for the participation of group P plasmids R in the transfer of chromosomal genes in intergenera crosses]

Use of E. coli strains with phenotypes Rec+ and Rec- asrecipients in intergenera crosses confirmed the supposition put forward by the authors formerly that new chromosomal markers in transconjugantes originated due to Psuedomonas aeruginosa. These chromosomal markers were transferred together with plasmid R conditioning the conjugation, and maintained without being built-into E. coli chromosome. Between the arg+ marker and the plasmid R18 there existed labile physical connection demonstrable only under definite conditions of recombinant selection.     

Isolation and some properties of phospholipase D from Bacillus subtilis G-22]

A method of isolating highly purified phospholipase D from Bac. subtilis G-22 is described. It includes ammonium sulphate fractionation, thermal denaturation, chromatography on lipoprotein bound with sepharose 6B and AH-sepharose 4B. The enzyme is 130-fold purified, its yield exceeds 90.0%, its specific activity is 164 units per mg of protein. The homogeneity of the enzyme is demonstrated by polyacrylamide gel electrophoresis, ultracentrifugation, isoelectric focusing and N-terminal amino acid determination by means of dinitrophenylation and dancylation. Proline is found to be N-terminal amino acid. The molecular weight of the enzyme, as determined from gel filtration through Sephadex G-100, is 21500 +/- 300, its sedimentation constant is 1.4S, isoelectric point is at pH 4.2. The molecular weight calculated from amino acid composition, is 21000--22000. Polypeptide chain contains of 196--205 amino acid residues. Phospholipase D develops its maximal activity at pH 8.5 and does not contain free SH-groups. Benzylsulphofluoride does not inhibit the enzyme activity. Phospholipase D is activated by Cd2+, Co2+, Zn2+, Ca2+ and is inhibited by EDTA, pIi50 being about 2.6.