Studies on thymocyte subpopulations in guinea pigs. Differences in cell volume and proliferative ability in vitro of two populations separated by density gradient centrifugation with Percoll

Thymus cells from guinea pigs were separated according to buoyant density by centrifugation with PVP-coated colloidal silica particles (Percoll). By creating an S-shaped density gradient, two populations (referred to as peak-I and peak-II cells) were obtained which differed in size as well as ability to spontaneous proliferation in vitro. Peak I contained low density cells of large size and was highly enriched with DNA-synthesizing cells. These continued to proliferate in culture for at least 30 h as demonstrated by mitotic studies in the intervals 0-10 and 20-30 h. In order to grow in vitro, however, the cycling cells of peak I depended on the medium (RPMI 1640) being supplemented with L-alanine. The high density cells of peak II constituted 70% of the thymocytes and had a small and uniform volume. This population was depleted of proliferating cells. The simple and rapid separation of these two major populations is considered a useful first step for the further characterization of thymocyte subpopulations. We suggest that peak I primarily includes proliferating precursor cells from the cortex as well as mature, immunocompetent cells. Peak II consists largely of small cortical cells.     

5S ribosomal gene clusters in wheat: pulsed field gel electrophoresis reveals a high degree of polymorphism

Summary The long-range structure of 5S rRNA gene clusters has been investigated in wheat (Triticum aestivum L.) by means of pulsed field gel electrophoresis. Using aneuploid stocks, 5S rRNA gene clusters were assigned to sites on chromosomes 1B, 1D, 513 and 5D. Cluster sizes were evaluated and the copy number of 5S DNA repeats was estimated at 4700-5200 copies for the short repeating unit (410 bp) and about 3100 copies for the long repeat (500 bp) per haploid genome. A comparison of wheat cultivars revealed extremely high levels of polymorphism in the 5S rRNA gene clusters. With one restriction enzyme digest all varieties tested gave unique banding patterns and, on a per fragment basis, 21-fold more polymorphism was detected among cultivars for 5S DNA compared to standard restriction fragment length polymorphisms (RFLPs) detected with single copy clones. Experiments with aneuploid stocks suggest that the 5S rRNA gene clusters at several chromosomal sites contribute to this polymorphism. A number of previous reports have shown that wheat cultivars are not easily distinguished by isozymes or RFLPs. The high level of variation detected in 5S rRNA gene clusters therefore offers the possibility of a sensitive fingerprinting method for wheat. 5S DNA and other macro-satellite sequences may also serve as hypervariable Mendelian markers for genetic and breeding experiments in wheat.     

Hyperprolactinaemia and testosterone production. Observations in 2 men on long-term dialysis

2 men had received maintenance dialysis for more than 5 years because of uraemia. Gradual impairment of libido had led to total impotence of more than 6 months duration. Concurrently hyperprolactinaemia occurred. Bromocriptine was given for 2 and 4 months, respectively. Potency was not restored, but in 1 patient the blood transfusion requirements were halved and general well-being and strength increased. In both men, the serum prolactin levels normalized. Inversely, the low plasma testosterone levels increased during the treatment period.     

The presence of 5 alpha-sitostanol in the serum of a patient with phytosterolemia, and its biosynthesis from plant steroids in rats with bile fistula

The presence of 5 alpha-sitostanol (24-ethyl-5 alpha-cholestan-3 beta-ol) in serum of a patient with the rare genetic disease phytosterolemia was confirmed. This study aimed at clarifying the pathway(s) for the formation of 5 alpha-sitostanol, by use of rats with bile fistula. 5 alpha-Sitostanol was formed only slowly from sitosterol, but readily from 24-ethyl-4-cholesten-3-one. Some conversion was also obtained with 7 alpha-hydroxysitosterol as precursor. In view of the low rate of 7 alpha-hydroxylation of sitosterol, however, a pathway from sitosterol to 5 alpha-sitostanol involving 7 alpha-hydroxysitosterol as intermediate is probably of small physiological importance. Intestinal microorganisms are not essential for the above conversions, since the 5 alpha-sitostanol was found in bile from bile fistula rats. 5 alpha-Sitostanol was converted to water soluble metabolites (bile acids) much more slowly than was cholestanol (5 alpha-cholestan-3 beta-ol), and was accumulated serum to a much larger extent.     

Laser light-scattering characterization of mitochondrial complex III-Triton X-100-phospholipid mixed micelles

Bovine-heart mitochondrial complex III was purified in the presence of Triton X-100, and the size and shape of the resulting protein-surfactant-phospholipid mixed micelles were investigated by laser light-scattering. The protein appears to be present in the form of a dimer, irrespective of temperature (between 25 and 40 degrees C) and protein concentration (between 0.5 and 5 mg/ml). The molecular weight of the micelle increases with temperature from 600 000 (25 degrees C) to 692 000 (40 degrees C). The variation of the solvent second virial coefficient in this temperature range suggests that, with increasing temperature, some of the free surfactant molecules become integrated in the mixed micelles. The average quadratic radius of gyration of these is of 42 +/- 5 nm, corresponding in our case to an ellipsoidal shape.