Quantitative Nasal Culture: a Tool in Antibiotic Research

The use of the quantitative nasal culture was investigated as a means of evaluation of new antimicrobial drugs in man. Cyclacillin was somewhat more active in vitro than penicillin G against penicillin G-resistant organisms. Cyclacillin was highly effective in suppressing staphylococci susceptible to penicillin G in nasal carriers but did not suppress staphylococci resistant to penicillin G. Although in previous studies by others cyclacillin was effective in treating mice infected with penicillin G-resistant staphylococci, in the present studies cyclacillin was not effective in suppressing nasal penicillin G-resistant staphylococci in man at doses which markedly suppressed penicillin G-sensitive organisms.     

Deoxyribonucleic Acid Replication in Simian Virus 40-Infected Cells IV. Two Different Requirements for Protein Synthesis During Simian Virus 40 Deoxyribonucleic Acid Replication

The replication of simian virus 40 (SV40) deoxyribonucleic acid (DNA) was inhibited by 99% 2 hr after the addition of cycloheximide to SV40-infected primary African green monkey kidney cells. The levels of 25S (replicating) and 21S (mature) SV40 DNA synthesized after cycloheximide treatment were always lower than those observed in an infected untreated control culture. This is consistent with a requirement for a protein(s) or for protein synthesis at the initiation step in SV40 DNA replication. The relative proportion of 25S DNA as compared with 21S viral DNA increased with increasing time after cycloheximide treatment. Removal of cycloheximide from inhibited cultures allowed the recovery of viral DNA synthesis to normal levels within 3 hr. During the recovery period, the ratio of 25S DNA to 21S DNA was 10 times higher than that observed after a 30-min pulse with (3)H-thymidine with an infected untreated control culture. The accumulation of 25S replicating SV40 DNA during cycloheximide inhibition or shortly after its removal is interpreted to mean that a protein(s) or protein synthesis is required to convert the 25S replicating DNA to 21S mature viral DNA. Further evidence of a requirement for protein synthesis in the 25S to 21S conversion was obtained by comparing the rate of this conversion in growing and resting cells. The conversion of 25S DNA to 21S DNA took place at a faster rate in infected growing cells than in infected confluent monolayer cultures. A temperature-sensitive SV40 coat protein mutation (large-plaque SV40) had no effect on the replication of SV40 DNA at the nonpermissive temperature.     

Linear, Single-Stranded Deoxyribonucleic Acid Isolated from Kilham Rat Virus

Kilham rat virus (KRV) was grown in a rat nephroma cell line and was purified by two isopycnic centrifugations in cesium chloride. The virus contains single-stranded deoxyribonucleic acid (DNA) with a molecular weight of approximately 1.6 x 10(6). The DNA was extracted from the virion by both phenol extraction and by 2% sodium dodecyl sulfate at 50 C. KRV DNA, extracted by both procedures, was observed in an electron microscope by using a cytochrome c or diethylaminoethyldextran monolayer. The DNA was also exposed to exonuclease I, an enzyme which hydrolyzes specifically linear, single-stranded DNA. Hydrolysis of 70 to 80% of the DNA was observed. Both the enzymatic and the electron microscope studies support the conclusion that extracted KRV DNA is a single-stranded, linear molecule. The length of the DNA was measured in the electron microscope and determined to be 1.505 +/- 0.206 mum.     

THE PERMEABILITY OF PINCHED-OFF NERVE ENDINGS TO SODIUM, POTASSIUM AND CHLORIDE AND THE EFFECTS OF GRAMICIDIN

Abstract—

• 1 The passive permeability of synaptosomes to ions was measured by a light-scattering technique. Synaptosomes were freely permeable to acetate, HCO₃^_, SCN^?and NH₄⁺ and were impermeable to choline and SO₄^2-.
• 2 Gramicidin D selectively increased the permeability of synaptosomes to Na⁺ and K⁺ ions.
• 3 The relative permeabilities of Na⁺, K⁺ and Cl^?, measured in the presence of a number of more permeant counter-ions, was in the ratio 1:19:12. These values are discussed in terms of the source of the resting potential.

     

BRAIN RECOVERY FROM ESSENTIAL FATTY ACID DEFICIENCY IN DEVELOPING RATS 1

Abstract— Rats were supplied from before birth with an essential fatty acid (EFA) deficient, a control, or an EFA deficient-control combination diet for various periods up to 6 months. It was found that EFA deficiency resulted in brain weights decreased in comparison with control values throughout development. The brain weight/body weight relationship, however, expressed by Donaldson's equation was generally maintained in animals fed either totally deficient or control diets. Animals deficient even during the brain's most actively growing period were able to recover completely on restoration of the control diet for a sufficiently long period. Fatty acid alterations in brain ethanolamine phosphoglyceride (EPG) during EFA deficiency were extensive. Acids of the ω6 family (18:2, 20:2, 20:3, 20:4, 22:3, 22:4 but not 22:5) were reduced from control figures. In the w9 family 20:3 and 22:3 were especially elevated whereas 22:6 ω3 levels were similar to those of the controls, finally decreasing only after a lengthy period of EFA deprivation. Mean unsaturation contents, as expressed by the proposed unsaturation index notation (Ul_mol) agreed closely in EPG fatty acids of deficient and control rats at a particular age. On substitution of the control for the deficient diet the ω6 family rebounded in a manner such that values for 20:4, 22:4, and 22:5 exceeded comparable figures in control animals. Concomitantly the ω9 family receded below control levels, and ω3 acids remained or returned to normal. This overadjustment in ω6 and ω9 families continued even after a prolonged period on the control diet.     

Factors affecting the respiration of rabbit sperm measured with an oxygen electrode

The respiration of rabbit sperm was measured by a Clarke type electrode which has two advantages over the conventional Warburg technique, greater sensitivity, and no necessity for a carbon dioxide-free atmosphere. It was not necessary to resaturate the sample chamber of the oxygen monitor with air, down to about 30% desaturation.Rabbit seminal plasma had a measurable oxygen uptake (0.6 μl/hr/ml) but this was much less than for human seminal plasma (4.3 μl/hr/ml). Hoderate dilution of the sperm and storing the semen at 0°C after slow cooling had no effect on oxygen uptake. Unlike those of most other species, rabbit sperm were also little affected by deliberate exposure to cold shock and the respiration before and after rapid cooling to 0°C was about the same. On the other hand very brief exposure of rabbit sperm to 65°C abolished motility and greatly reduced the respiration rate. Bicarbonate (6 mM) stimulated the oxygen uptake of freshly collected samples of rabbit sperm after washing. Increasing the phosphate concentration of the medium to 80 mM did not greatly depress the oxygen uptake.