Isolation and identification of spirorenone metabolites from the monkey (Macaca fascicularis)

The plasma level of spirorenone was determined 3 h after 1, 8, 22 and 46 daily oral administrations of 20 mg/kg to two female and two male monkeys ($\text{Macaca fascicularis}$). A fifth animal, female, was treated with eight daily doses of tritium-labelled drug and was completely bled from the carotid vein 4 h after the last administration in order to isolate and identify plasma metabolites.After repeated daily doses of spirorenone the mean plasma level of unchanged drug was 711 ± 213 ng/ml. In the plasma of the fifth animal four radioactively labelled compounds could be detected after extraction and subsequent HPLC separation. Mass spectrometric identification of three of the substances indicated 1,2-dihydrospirorenone, hydroxy-1,2-dihydrospirorenone and the unchanged drug itself.     

Synthesis and biological activity of brassinolide and its 22β,23β-isomer: Novel plant growth-promoting steroids

Brassinolide (2α,3α,22α, 23α-tetrahydroxy-24α-methyl-B-homo-7-oxa-5α-cholestan-6-one), a novel plant growth-promoting steroid isolated from rape pollen, and its hitherto unknown 22β, 23β-isomer were synthesized from a C-24 epimeric 60:40 mixture of 22-dehydrocampesterol (24α-methyl) and brassicasterol (24β-methyl) from oysters. The method of synthesis favored the formation of the 22β, 23β-isomer by better than 4:1. Comparative plant growth-promoting capabilities of brassinolide, both natural and synthetic, and its three side chain $\text{cis}$-glycolic isomers in the bean second internode bioassay showed that the natural and synthetic brassinolides were equally active and caused splitting of the internode at the 0.1 μg level. The least active was the 22β,23β-isomer of brassinolide. The isomers with the 22α, 23α and 24β, and the 22β, 23β and 24β configurations were highly active and were required at about 10 times the concentration of brassinolide to cause the same physiological response. In the bean first internode bioassay, an auxin-induced growth test system which employs isolated bean plant segments, the isomer with 22β, 23β and 24β configuration caused a greater response than brassinolide. Two of the four tetrahydroxy ketones obtained in the synthesis of the isomers were also active in both assays.     

Disproportionate rDNA replication does occur in diploid tissue in Drosophila hydei

Summary The rDNA content in Drosophila hydei has been compared in wild-type and in two translocation genotypes possessing only one nucleolus organizer. In highly polyploid salivary glands where rDNA is underreplicated, an independent polytenization of the rDNA occurs resulting in about the same rDNA level in each genotype independently of the number of nucleolous organizers present in the genome. Thus, the situation in the salivary glands of D. hydei is similar to that in D. melanogaster (Spear and Gall 1973).In tetraploid thoracic muscle where rDNA is not underreplicated, the rDNA percentage in the two translocation genotypes is also considerably increased, although the wild-type level is not completely attained. This result shows that rDNA replication is independently controlled even in a non-underreplicating tissue.In larval diploid brain the situation in the two translocation stocks is dissimilar: in one genotype the rDNA content remains unaltered whereas in the other it is increased. This demonstrates for the first time that a gene compensation does occur in a diploid tissue.Supported by a grant from the Deutsche Forschungsgemeinschaft (Ku 282/7)     

On the role of IS1 in the formation of hybrids between the bacteriophage P1 and the R plasmid NR1

Summary The genomes of bacteriophage P1 derivatives carrying drug resistance genes derived from an R plasmid NR1 were analysed by restriction cleavage and be DNA-DNA hybridization. Two representatives of a class of oversized P1CmSmSu phages were identified as P1 carrying the entire r-determinant of NR1 together with its two flanking, directly repeated IS1. In one case the r-determinant insertion is carried at the site of the residential IS1 of P1, in the other case it is transposed into another region of the P1 genome. Models postulate that the first type resulted from reciprocal recombination within IS1 elements and that the formation of the second type of P1-R hybrid depended both on IS1 mediated transposition and reciprocal recombination. Plaque forming P1Cm or P1CmSm phages are explained as IS1 mediated deletion derivatives of P1CmSmSu, although an alternative model postulates that sometimes P1Cm phages might result from two consecutive transposition events of only one IS1 without involving reciprocal recombination. Secondary P1 derivatives carrying only one IS1 at the site of the original r-determinant or of Cm insertions into P1 must have been produced by reciprocal recombination between the two IS1 flanking the insertions. An implication from this study, that any genetic material carried adjacent to an IS1 element may undergo passive transposition, is discussed.     

Electrophoretic and biochemical studies of human aldehyde dehydrogenase isozymes in various tissues

Four major ALDH isozymes have been identified in human tissues using starch gel electrophoresis and isoelectric focusing. The isozyme bands have been termed as ALDH I, II, III and IV according to their decreasing electrophoretic migration and increasing isoelectric point. The isozymes have been partially purified via preparative isoelectric focusing. Kinetic characteristics of ALDH I and II were found to be quite similar to ALDH enzyme 2 and enzyme 1 described earlier by Greenfield and Pietruszko (Biochem Biophys Acta, $\text{483}$ 35–45 1977). ALDH III and IV showed a very high Km for propionaldehyde (1.0–1.5 mM at pH 9.5) and were not inhibited by disulfiram at pH 9.5. A variant phenotype of ALDH which lacked in isozyme I was detected in various tissues from Japanese individuals. Comparative kinetic properties of normal and variant enzyme are given.     

The metabolic interactions between Paramecium bursaria Ehrbg. and Chlorella spec. in the Paramecium bursaria-symbiosis

In an organism (strain C 1/2 from Dr. P. R. Hayes, Leeds) regarded as a typical representative of the genus Flavobacterium, flexirubin-type pigments have been identified. The Flavobacterium pigments contain structural elements of both, the pigments of the genus Flexibacter and the pigments of the genus Cytophaga. As flexirubin-type pigments seem to have a rather restricted distribution among bacteria, and have formerly proved to be useful chemosystematic markers for the flexibacteria, this new observation may indicate that there is a relatively close phylogenetic relationship between this type of flavobacteria and the Cytophaga-Flexibacter group.     

Hereditary motor endplate disease (med) of the mouse: Observations on dissociated myogenic cells and their development in culture

Summary Myogenic cells from mice homozygous for the lethal mutation motor endplate disease (med/med) were grown in culture. Like muscle cells taken from wild type (+/?) litter mates they fused to form myotubes which contracted, developed cross striations, and exposed acetylcholine receptors (AChR) on their surface. However, a decrease of 30% in the number of mononucleated cells per unit fresh weight of muscle was observed as early as 2–3 days postnatal, i.e., at least one week prior to the onset of physiological symptoms. Hence, in addition to influencing the functional maintenance of motor endplates, the med gene seems to control early events in muscle development.     

Biliprotein assembly in the disc-shaped phycobilisomes of Rhodella violacea electron microscopical and biochemical analyses of C-phycocyanin and allophycocyanin aggregates

C-phycocyanin and allophycocyanin from the red alga Rhodella violacea were investigated by electron microscopy and biochemical methods using samples taken from the same fractions.The molecular weights of the native biliprotein aggregates C-phycocyanin and allophycocyanin are about 139,000 (140,000) and 130,000 (145,000) as revealed by calibrated gel chromatography, gradient gel electrophoresis and morphological measurements on the basis of an average protein packing density. These molecular weights are direct evidence for a trimeric aggregation form ()₃ of these biliproteins. Independently, their monomers were determined to be about 34,400 (C-phycocyanin) and 33,900 (allophycocyanin).C-phycocyanin and allophycocyanin are ringshaped, six-membered, biliprotein aggregates with dimensions of about 10.2×3.0 nm and 10.0×3.0 nm, respectively. The aggregates are made up of six subunits, 3 and 3, which are assumed to be associated in alternating positions. They are arranged in regular hexagons in C₆ symmetry. Hexameric aggregates ()₆, so far only isolated for C-phycocyanin, originate by face to face association of two trimeric aggregates.     

A fast and simple radiometric assay for adenosine deaminase using reversed-phase thin-layer chromatography

A new quantitative radiometric assay for adenosine deaminase is described. The reaction conditions are similar to those used in other radioassays and are shown to result in an activity which increases linearly with time and with enzyme concentration. An original feature of the technique resides in the use of reversed-phase thin-layer chromatography to separate adenosine from inosine. The separation is complete, fast, and reproducible. Both compounds can be recovered almost quantitatively from the plates. The assay is very simple and allows the determination of up to 36 samples in 3 h.     

Decreased cAMP responsiveness to glucagon in livers from ethynyl estradiol treated rats.

The effects of glucagon on the concentration and output of cAMP were studied in liver slices and in perfused livers from female rats and from animals treated with ethynyl estradiol (15 μg/kg daily for 14 days). The basal content of cAMP in liver slices, or of cAMP released into the perfusion medium in the absence of glucagon, was unaffected by prior treatment of the animal with estrogen. When glucagon was added to the medium, the concentration of cAMP in liver slices was 2.29 ± 0.32 and 1.10 ± 0.11 pmol cAMP/mg wet weight from control and ethynyl estradiol treated rats, respectively. When glucagon was added, the output of cAMP by perfused livers was maximal at 20 minutes with livers from either control or ethynyl estradiol treated rats. Output of cAMP by the perfused liver, when glucagon was added to the medium, was 8.76 ± 0.69 and 1.84 ± 0.08 nmol/g by livers from control and ethynyl estradiol treated rats, respectively. This effect was the same whether animals had been fasted for 12 hours previously, or were allowed free access to food until sacrifice. Clearly, as measured by cAMP accumulation, prior treatment of the rat with ethynyl estradiol reduced the sensitivity of the hepatic cAMP response to glucagon.