Direct Measurement of Potential Difference across the Human Red Blood Cell Membrane

The electrical potential difference across the human red cell membrane has been measured directly. A biological amplifier with neutralized input capacity was used. Human red cells in modified Ringer solution were impaled individually with 3 M KCl-filled glass microelectrodes. Movements of the microelectrodes were effected by Leitz micromanipulators. Results showed a potential difference of -8.0 ± 0.21 (SEM) mv, the inside being negative with respect to the outside. This value is approximately that calculated by using the Nernst equation considering the intracellular and extracellular chloride concentrations.

As a control, similar measurements were made on nylon microcapsules containing hemoglobin. The measured potential of -0.52 ± 0.02 (SEM) mv, which agreed very well with the value calculated on the basis of Donnan equilibrium, was much smaller in magnitude as compared to the results for the red cell, and there was evidence of fixed charges on the microcapsule membrane. There was no evidence of this in the case of the red cell.

     

COMPARISON OF THE FATTY ACIDS OF LIPIDS OF SUBCELLULAR BRAIN FRACTIONS

Abstract— Rat brain grey and white matter were fractionated to yield myelin, nerve terminal, synaptic vesicle, nerve terminal 'ghost', and microsomal fractions of white and grey matter. Ester-type glycolipids were found in all fractions except myelin, while cerebrosides occurred in significant concentrations only in myelin and white microsomes. Comparison of the fatty acid profile of the ethanolamine- and serine-containing phospholipids showed marked differences between myelin and the particles from grey matter, while the microsomes of white matter were of intermediate composition. Docosahexaenoic acid, a minor acid in myelin, was a major fatty acid in microsomes of grey and white matter. The fatty acid composition of sphingomyelin was distinctly different in the fractions derived from grey and white matter, clustering about stearate and nervonate in the latter, but only about stearate in the grey. Marked differences in the positional distribution of fatty acids were seen within phosphatidyl choline from myelin and nerve terminals. Ribonucleic acid was found in nerve terminal and synaptic vesicle fractions. The sphingosine found in the ganglioside from microsomes of both grey and white matter was similar with respect to distribution of the C₁₈ and C₂₀ homologues.
The possibility is discussed that microsomes furnish characteristic lipids for the synthesis or renewal of specific membranes, and that these lipids are accumulated somewhat before being released.     

Effect of Pregnancy on Sebum Excretion

Sebum excretion rates from forehead skin were measured serially during and after pregnancy in 10 normal women. Only minor fluctuations occurred during the middle and last trimesters of pregnancy, but there was a pronounced decrease in the postpartum period. Probably a powerful sebotrophic factor is present in pregnancy, but its nature is conjectural.     

Fractionation of deoxyribonucleic acid from phage-infected bacteria

1. DNA has been isolated in 90% yield from T5-infected cultures of Escherichia coli ;pulse'-labelled with [(3)H]thymidine. It had a buoyant density in caesium chloride solution identical with the DNA of mature T5 phage, and no components of unusual buoyant density were detected. 2. The DNA preparation was resolved into two major components of differing specific activity on a column of kieselguhr coated with methylated serum albumin. The DNA of high specific activity could be eluted from the column only with 2n-ammonia, and the firm binding did not appear to be due to an artifact of preparation. 3. A similar fractionation into two DNA components of differing specific activity was observed when the ;pulse'-labelled culture was lysed with sodium dodecyl sulphate and the lysate rocked with phenol. The DNA of high specific activity was found in the interface precipitate between the phenol and aqueous layers. 4. The amounts of DNA in the two fractions were measured at different times after infection and the radioactivity content of each was determined at various times after a short ;pulse' of [(3)H]thymidine. The interface fraction contained the replicating phage DNA, and the DNA from mature phage particles appeared in the aqueous fraction. 5. Analogous results were obtained with T2-infected E. coli. In the presence of chloramphenicol the DNA in the interface fraction was not converted into DNA extractable into the aqueous layer. Since chloramphenicol prevents the condensation of DNA into phage heads, it is suggested that any DNA in extended configuration is trapped inside the rigid-layer framework of the cell wall. 6. Treatment with lysozyme released much of the DNA from the interface precipitate. This DNA was firmly bound by the chromatographic column and had the same buoyant density in caesium chloride solution as normal T5-phage DNA. Sucrose-gradient sedimentation studies showed that it was heterogeneous and that as much as 60% sedimented faster than T5-phage DNA.     

Oxidation of pyrimidine nucleosides and nucleotides by osmium tetroxide

1. Pyrimidine nucleosides such as thymidine, uridine or cytidine are oxidized readily at 0° by osmium tetroxide in ammonium chloride buffer. There is virtually no oxidation in bicarbonate buffer of similar pH. Oxidation of 1-methyluracil yields 5,6-dihydro-4,5,6-trihydroxy-1-methyl-2-pyrimidone. 2. Osmium tetroxide and ammonia react reversibly in aqueous solution to form a yellow 1:1 complex, probably OsO₃NH. A second molecule of ammonia must be involved in the oxidation of UMP since the rate of this reaction is approximately proportional to the square of the concentration of unprotonated ammonia. 3. 4-Thiouridine reacts with osmium tetroxide much more rapidly than does uridine. The changes of absorption spectra are different in sodium bicarbonate buffer and in ammonium chloride buffer. They occur faster in the latter buffer and, under suitable conditions, cytidine is a major product. 4. Polyuridylic acid is oxidized readily by ammoniacal osmium tetroxide, but its oxidation is inhibited by polyadenylic acid. Pyrimidines of yeast amino acid-transfer RNA are oxidized more slowly than the corresponding mononucleosides, especially the thymine residues. Appreciable oxidation can occur without change of sedimentation coefficient.