Measurement by radioimmunoassay of casein content in rabbit mammary gland during pregnancy and after prolactin stimulation in organ culture

A specific homologous radioimmunoassay was developed to measure rabbit beta-casein in rabbit mammary gland with a sensitivity of 0.5 ng/ml protein. It was used to measure casein concentration during pregnancy and in organ culture of mammary gland explants. Casein was detectable in virgin mammary glands, showed a small increase during the first half of pregnancy, increased more than 20-fold between Days 21 and 27, and diminished somewhat on the first days of lactation. After 24 hr of culture, mammary gland explants had no detectable casein, but the addition of increasing concentrations of prolactin to a culture medium which contained insulin (5 micrograms/ml) and cortisol (0.5 microgram/ml) induced a regular increase in the casein content of the tissue. Casein started to increase when 10 ng/ml of prolactin was present and maximal values were achieved for 100 ng/ml of the hormone.     

Peroxidative membrane damage in human erythrocytes induced by a concerted action of iodoacetate, vanadate and ferricyanide

Human erythrocytes incubated without substrate in the presence of iodoacetate (0.2 mM), vanadate (0.5 mM) and ferricyanide (5 mM) form aqueous membrane leaks of equivalent radii of 0.5-0.8 nm leading to complete colloid-osmotic lysis within 180 min. All three components are indispensable for the effect. Inosine but not glucose markedly enhances the rate of hemolysis. These effects are due to oxidative damage, as indicated by concomitant destruction of polyunsaturated fatty acids and suppression of both effects by radical scavengers. Hemoglobin is not oxidized under these conditions. GSH and membrane SH levels remain almost normal, and no crosslinking or irreversible aggregation of membrane proteins is observed. In the absence of O2 no membrane damage can be observed. It is proposed that radical formation originates from reduction of O2 by NADPH, analogous to processes described in microsomal membranes. NADH seems not to be involved, since leak formation occurs in spite of the blockage of NADH formation by iodoacetate. Vanadate and ferricyanide are probably required to amplify the peroxidative reaction sufficiently to overcome the cellular antioxidative capacity.     

Random sample planning in morphometry

This report tries to explain principles and presuppositions of planning sample sizes. The aim is estimation of the optimal, that means minimally necessary sample size, justifying an investigation ethically and economically. The problems are complicated because of the necessity of nested sampling or samples within samples. We describe the most important presuppositions using an example of mean value estimation. These are: establishing of demands for exactness, of error probability, and knowledge of variance of the parameters. It is explained in detail by means of binomially distributed variables as we find them in the point counting method.     

Uptake of GABA by rat brain synaptic vesicles isolated by a new procedure.

Uptake of GABA was demonstrated in rat brain synaptic vesicles which were prepared by a new and efficient procedure. The uptake activity co-purified with the synaptic vesicles during the isolation procedure. The purity of the vesicle fraction was rigorously examined by analysis of marker enzymes and marker proteins and also by immunogold electron microscopy using antibodies against p38 (synaptophysin). Contamination by other cellular components was negligible, indicating that GABA uptake by the synaptic vesicle fraction is specific for synaptic vesicles and not due to the presence of other structure possessing GABA uptake or binding activities. GABA uptake was ATP dependent and similar to the uptake of glutamate, which was assayed for a comparison. Both uptake activities were independent of sodium. They were inhibited by the uncoupler carbonyl cyanide 4-(trifluoromethoxy)phenylhydrazone, indicating that the energy for the uptake is provided by an electrochemical proton gradient. This gradient is generated by a proton ATPase of the vacuolar type as suggested by the effects of various ATPase inhibitors on neurotransmitter uptake and proton pumping. Competition experiments revealed that the transporters for GABA and glutamate are selective for the respective neurotransmitters.     

Yield and nitrogen uptake of rapessed (Brassica campestris L.) with ammonium and nitrate

Water culture, growth chamber, greenhouse and field experiments were conducted to compare the effect of NH₄−N and NO₃−N on yield and N uptake of rapeseed (Brassica campestris L.). In water culture, the yields of 28-day old rapeseed plants grown at 14 μg N ml⁻¹ were double with NO₃ compared to NH₄, but N uptake was little affected. There was no such effect when concentration was reduced to 3.5 or 7 μg N ml⁻¹. The yield and N uptake of 26-day old rapeseed grown on six soils (pH 4.6 to 6.5) in pots in a growth chamber were much greater with NO₃ than with NH₄, although N concentration was more in the NH₄- than the NO₃-grown plants. In a greenhouse experiment with rapeseed grown on 12 potted soils, the N uptake of applied N was greater with NO₃ than with NH₄ on all soils. Averages were 63% with NH₄ and 78% with NO₃. However, NH₄-fixation capacities of the soils were only weakly correlated with yield from the two sources of N (r=0.48) and the relation was similar with N uptake. In contrast to the behavior of water culture, growth chamber and greenhouse experiments, the 33 field experiments did not show consistent difference in seed yield with NH₄ and NO₃ applied at time of seeding. In nine field experiments where band application was used for Ca(NO₃)₂, (NH₄)₂ SO₄, NH₄ NO₃, yield tended to be greatest for (NH₄)₂SO₄. However, in 19 experiments on acid soils with and without lime, yields in most cases were similar with (NH₄)₂SO₄ and NH₄ NO₃. Nitrification inhibitors were added to spring banded NH₄-based fertilizers in five experiments, but the yields were not influenced. Scientific Paper No. 558, Lacombe Research Station, Agriculture Canada.     

Organization of the micronuclear genome of oxytricha nova

In the hypotrichous ciliated protozoan Oxytricha nova, approximately 95% of the micronuclear genome, including all of the repetitive DNA and most of the unique sequence DNA, is eliminated during the formation of the macronuclear genome. We have examined the interspersion patterns of repetitive and unique and eliminated and retained sequences in the micronuclear genome by characterizing randomly selected clones of micronuclear DNA. Three major classes of clones have been defined: (1) those containing primarily unique, retained sequences; (2) those containing only unique, eliminated sequences; and (3) those containing only repetitive, eliminated sequences. Clones of type one and three document two aspects of organization observed previously: clustering of macronuclear destined sequences and the presence of a prevalent repetitive element. Clones of the second type demonstrate for the first time that eliminated unique sequence DNA occurs in long stretches uninterrupted by repetitive sequences. To further examine repetitive sequence interspersion, we characterized the repetitive sequence family that is present in 50% of the clones (class three above). A consensus map of this element was obtained by mapping approximately 80 phage clones and by hybridization to digests of micronuclear DNA. The repeat element is extremely large (approximately 24 kb) and is interspersed with both macronuclear destined sequences and eliminated unique sequences.