Xl-fli, the Xenopus homologue of the fli-1 gene, is expressed during embryogenesis in a restricted pattern evocative of neural crest cell distribution.

The Xenopus laevis fli cDNA, belonging to the ets family of transcription factors, was isolated from a library prepared from unfertilized eggs. It encodes a polypeptide with extensive homology to murine and human Fli proteins. The long 3'-untranslated region contains five nuclear polyadenylation signals and three cytoplasmic polyadenylation elements, as well as many A/T rich elements. Two polyadenylated transcripts appear at the early neurula and accumulate up to the tadpole stage. In situ hybridization reveals an expression in territories invaded by neural crest cells. In the head region, fli is expressed in the peri-ocular zone, in the branchial buds and at the level of the brain floor. In the trunk, a metamerized expression is detected in the dorsum. At a lower level, the tailbud and the peri-cardiac region also appear positive.

Lipids and lipoproteins of malnourished children during early renutrition: apolipoprotein A-IV as a potential index of recovery.

Twenty-six children with marasmus and 27 with kwashiorkor were compared with 23 control children of matching ages. Kwashiorkor was characterized by increased phospholipids (NS), low (P < 0.01) apolipoprotein (apo) B-rich LDL, and near normal apo A-I and HDL-C. In children with marasmus apo B (P < 0.02) LDL-C (NS), apo A-I (P < 0.01), and HDL-C (P < 0.001) decreased. Fifteen children in each group were followed for 2 wk. Control values were progressively reached after 2 wk. In the younger children final apo B was higher than in control subjects (P < 0.03) but apo A-I was identical. Apo A-IV, assayed because it correlates with the functional state of intestine, was near normal in children with kwashiorkor and decreased with treatment. In children with marasmus apo A-IV decreased by 50%, increased with treatment in older children, but further diminished in younger children. After 2 wk apo A-IV was significantly lower in all patients than in control subjects. Apo A-IV, by remaining depressed after other variables normalized, seems a good index of nutritional status.

Cell cycle-specific variation of intracellular plasminogen activator activity in cultured human alveolar epithelial carcinoma and rat hepatoma cells.

Plasminogen activator activity was demonstrated in two carcinoma cell lines: A549 cells derived from a human alveolar epithelial carcinoma; and ZHC cells derived from a rat hepatoma. Both cells had intracellular plasminogen activator activity throughout their cell cycles and in each case this activity reached a maximum. For A549 cells the maximal activity took place either during the G2 phase or in the course of the S to G2 transition, suggesting that plasminogen activator might play a role in cell division. For ZHC cells, the maximal activity occurred at the start of the S phase, suggesting that in these cells plasminogen activator might be involved in DNA replication.

In vitro evidence that hypothyroidism modifies glucocorticoid receptors.

We have previously reported that hypothyroidism is accompanied by modifications of the cytosolic rat liver glucocorticoid receptors. In the present study we have used molybdate, which stabilizes the glucocorticoid receptor from hypothyroid rats, allowing its characterization. We confirm a decrease in the number of glucocorticoid binding sites in the liver of hypothyroid rats as compared to that in normal rats (8000 per cell versus 23,000 per cell for normal rats). At 22 degrees C and in the absence of molybdate, thermal activation shows the same kinetics in both complexes from normal and hypothyroid rats, though activated hormone-receptor complexes from hypothyroid rats are progressively degraded (t 1/2 = 230 min). At the same temperature the dissociation rate of native complexes from hypothyroid rats is slower (k-1 = 8 X 10(-3) min-1) than normal (k-1 = 16 X 10(-3) min-1). Competitive dissociation studies using [3H]dexamethasone indicate that native complexes from hypothyroid rats show altered affinities for agonist and antagonist. Hypothyroidism decreases the number of rat liver glucocorticoid receptors and alters their properties, as evidenced by their greater instability and differences in steroid binding. These effects may be due to regulating factors and/or slight, probably post-transcriptional, modifications of the binding protein.

Cell cycle variations of dinucleoside polyphosphates in synchronized cultures of mammalian cells.

Zajdela hepatoma culture cells (ZHC) and mouse embryo fibroblasts (Swiss 3T3) were synchronized in G1 or S phase by serum deprivation and aphidicolin treatment, respectively, to study the variations in adenylyl nucleotide (Ap4X) pool size during the progress of the cell cycle. Only minor variations, which never exceeded a factor of 2, were observed when the Ap4X concentrations were expressed on a cellular basis. The variations were found to be strictly parallel to the ATP variations. Upon release from an aphidicolin block, the minor variations of Ap4X followed DNA synthesis and preceded cytokinesis. When the nucleotide content was compared with the amount of proteins, the faint specific cell cycle changes were almost completely damped when the cells were synchronized by serum deprivation, but remained practically unchanged in the case of aphidicolin synchronization. These results suggest that the observed variations could reflect the accumulation of some nucleotides before cell division. It is not clear yet whether the variation in Ap4X concentration is significant by itself or is simply a phenomenon resulting from changes in the ATP pool.

Effects of thyroidectomy on heart and liver rat tRNAs: study of chromatographic and electrophoretic behaviour.

Modifications of tRNAs in various physiological or experimental conditions are well documented. We have compared isoacceptor tRNAs extracted from target organs (heart and liver) from thyroidectomized rats to those of control animals. Nine liver aminoacyl-tRNAs and eight heart aminoacyl-tRNAs from thyroidectomized and control rats were analysed by RPC-5 chromatography. Quantitative differences were demonstrated in the relative proportions of the various liver tRNA isoacceptors for glycine, lysine, methionine, phenylalanine and serine and of the heart isoacceptor tRNAs for glycine, lysine, methionine, phenylalanine and valine. A qualitative variation was noted only for tRNATyr from the heart and liver of thyroidectomized rats. Isoacceptor tRNAs were obtained at a high resolution using a two-dimensional polyacrylamide gel electrophoresis. Isoacceptor tRNAs corresponding to 14 amino acids for the liver and 12 amino acids for the heart were identified. Although for most of the tRNAs examined the number of isoacceptors remained unchanged, the number of spots corresponding to tRNAGlu and tRNAHis from the liver and tRNAAla from the heart was different after thyroidectomy. Furthermore the change in electrophoretic behaviour of tRNATyr from the liver of thyroidectomized rats suggests a structural modification of one of the isoacceptors in relation to the change in thyroid status. Thus, thyroid hormones appear to induce some modification of the isoacceptor tRNAs in their target organs.

Impairment of enzyme induction by glucocorticoids in Zajdela hepatoma cells.

Whereas glucocorticoids induce TAT, TRP, GPT in liver and only TAT in HTC cells, no hormonal effect on the synthesis of these enzymes was found in Zajdela hepatoma cells grown in vivo as an ascitic tumor, or in vitro as layer cultures. Although these cells remain uninducible, the hormone penetrates normally, but a strong decrease of the specific binding of cytosol and nuclear proteins with the hormone was observed. The impairment at the level of the hormone receptors could account for the non-inducibility of enzyme synthesis in ZHC cells.