Glucocorticoid hormones increase the activity of γ-glutamyltransferase in a highly differentiated hepatoma cell line

γ-Glutamyltransferase activity was detected in the plasma membrane of the highly differentiated hepatoma cell line Fao, (0.93 mU/mg cell protein). Dexamethasone (1 μM) provoked a 2–3-fold increase in the activity of the enzyme in the presence of fetal calf serum. Maximal induction occurred 48–72 h after addition of the glucocorticoid to the cell culture medium. The hormonal specificity was demonstrated by the relative potencies of several glucocorticoids and sex steroids: hydrocortisone and corticosterone increased γ-glutamyltransferase activity while tetrahydrocorticosterone and all sex steroids tested were ineffective. The effect of dexamethasone on γ-glutamyltransferase activity was specific since the activities of several other plasma membrane enzymes were not modified. The mechanism of the dexamethasone-induced increase in γ-glutamyltransferase activity was neither by modification of the affinity of the enzyme for its substrates nor by alteration of the subcellular distribution of the enzyme. This increase was prevented by cycloheximide and actinomycin D. The data presented are consistent with a specific glucocorticoid receptor-mediated induction of γ-glutamyltransferase activity in Fao cells. The kinetic parameters of the induction process by glucocorticoids are very similar to those found in adult rat liver. These results suggest that the Fao cell line is a very convenient system for the study of the molecular mechanisms of glucocorticoid effects on differentiated cells.     

Evidence for the mannosylation of a non-histone protein in monkey liver chromatin

Summary Mannose is incorporated in monkey liver chromatin by the means of a nuclear membrane mannosyl-transferase.¹⁴C-labelled chromatin is dissociated either by sulfuric acid or 6 M urea and 0.4 M GuCl. The fractions then enriched in non-histone¹⁴C-labelled proteins are excluded from Ultro-gel AcA 202, their analysis in SDS-polyacrylamide gel electrophoresis shows that radioactivity fits with one major protein band, confirming the presence of at least a non-histone protein labelled with mannose in monkey liver chromatin, with an apparent molecular weight of 13 000.     

Characteristics of development and variations in ecdysteroid levels in Clitumnus embryos deprived of their cephalic endocrine glands

Microsurgical suppression of presumptive endocrine areas in very young embryos of Clitumnus resulted in an arrest in development, which occurred at different stages according to the operation. In most cases however. the duration of embryonic life was not reduced significanly. The repercussions of each operation type on the ecdysteroid content of the embryos were determined using a radioimmunoassay technique.Consideration of the experimental results provides some indication on the part played by each endocrine formation during the last part of embryonic development that is between dorsal closure and hatching time. Our experiments offer original evidence that the corpora allata of the embyro are of utmost importance at the stage VII_3t, in initiating the normal expression of larval characteristics.     

Genetic and biochemical characterization of mutants of Saccharomyces cerevisiae blocked in six different steps of heme biosynthesis

Summary Heme-deficient mutants of Saccharomyces cerevisiae have been isolated from two isogenic strains with the use of an enrichment method based on photodynamic properties of Zn-protoporphyrin. They defined seven non-overlapping complementation groups. A mutant representative of each group was further analysed. Genetic analysis showed that each mutant carried a single nuclear recessive mutation. Biochemical studies showed that the observed accumulation and/or excretion of the different heme synthesis precursors by the mutant cells correlated well with the enzymatic deficiencies measured in acellular extracts. Six of the seven mutants were blocked in a different enzyme activity: 5-aminolevulinate synthase, porphobilinogen synthase, uroporphyrinogen I synthase, uroporphyrinogen decarboxylase, coproporphyrinogen III oxidase and ferrochelatase. The other mutant had the same phenotype as the mutant deficient in ferrochelatase activity. However, it possessed a normal ferrochelatase activity when measured in vitro, so this mutant was assumed to be deficient in protoporphyrinogen oxidase activity or in the transport and/or reduction of iron.The absence of PBG synthesis led to a total lack of uroporphyrinogen I synthase activity. The absence of heme, the end product, led to an important increase of coproporphyrinogen III oxidase activity, while the activity of 5-aminolevulinate synthase, the first enzyme of the pathway, was not changed. These results are discussed in terms of possible modes of regulation of heme synthesis pathway in yeast.     

Abscisic Acid Translocation and Metabolism in Soybeans following Depodding and Petiole Girdling Treatments

It was found earlier that depodding and girdling treatments which obstruct translocation, result in increased leaf AbA levels and partial stomatal closure. In the present work (±) [2-¹⁴C]abscisic acid (AbA) was introduced into leaves and the mass, and radioactivity of AbA and AbA-metabolites were analyzed following translocation obstruction to determine whether the increased AbA was due to higher rates of synthesis, or lower rates of catabolism or export. The (±) [2-¹⁴C]AbA was introduced into soybean (Merr.) leaves by injection into the petiole region. AbA and AbA-metabolites (phaseic acid [PA], dihydrophaseic acid [DPA], AbA-conjugate, and an unknown metabolite) were separated with preparative high performance liquid chromatography. Methyl esters of AbA (free and that released after hydrolysis of AbA-conjugate), PA and DPA were determined with gas chromatography using electron capture detection.     

Activities and multiplicity of phenolase from spinach chloroplasts during leaf ageing

Phenolase activity in spinach leaves homogenates depends on the stage of development of leaves and on the kind of homogenization procedure. Under constant experimental conditions it is low in non-senescent leaves. With the onset of senescence there is a 15–20-fold increase in soluble activity in the supernatants of broken chloroplasts as well as an increase in activation of latent phenolase in fractions containing thylakoids. This rise in activity is due to an increase in particular multiple forms, differing for supernatants and membrane sediments. Phenolase from spinach lacks monophenolase and laccase activities.     

In vitro stimulation of human granulopoiesis by purified protein derivative (PPD)

Summary The effect of purified protein derivative (PPD) on human granulopoiesis was studied in an in vitro semisolid culture system of human bone marrow in which PPD was incorporated into the leukocyte feeder layers. We observed that preincubation of the feeder layers with PPD was necessary to induce a significant rise of agar culture colony-forming units (CFU-c) with a maximum of 3 days' preincubation and a dose of 200 g for 10 ⁶ leukocytes. A similar effect was obtained when a conditioned medium from PPD-stimulated leukocytes was used instead of feeder layers. We have found a significant correlation between the skin test response of the leukocyte donors to PPD and the colony-stimulating activity of their leukocytes exposed to PPD: these results suggest that PPD could stimulate human granulopoiesis by an indirect effect on CSF-producing mononuclear cells.     

Calcitonin-responsive adenylate cyclase in cultured F9 embryonal carcinoma cells

Hormonal responsiveness of the adenylate cyclase system of cultured F9 teratocarcinoma cells was investigated. Of numerous hormones tested only calcitonin, (−)isoproterenol, and prostaglandin E₁, stimulate F9 adenylate cyclase activity. Of the active hormones, calcitonin is the most potent stimulator of cAMP formation. Treatment of intact F9 cells with calcitonin results in a time- and hormone concentration-dependent increase in the intracellular concentration of cAMP. cAMP accumulation is enhanced within 5 min after addition of 60 nM synthetic salmon calcitonin to intact F9 cells. These results raise the possibility that calcitonin may play a regulatory role in early embryonic development.     

Decouverte de Septorella brachycera grambast et de Septorella ultima Grambast (Charophytes,Clavatoracees) dans le Maestrichtien superieur des Petites Pyrenees. Consequences stratigraphiques

The discovery of an assemblage of Clavatoraceae including Septorella brachycera and S. ultima in the “Marnes d'Auzas” (upper Maestrichtian) of the Petites Pyrénées constitutes the first element of comparison between the upper Cretaceous of the Petites Pyrénées and Provence, but creates a problem with regard to the subdivision of the provencial Rognacian, based, in part, on the superposition of these two species.