Chicken riboflavin-binding protein. cDNA sequence and homology with milk folate-binding protein

The Rd gene is expressed in the livers and oviducts of laying hens and codes for the riboflavin-binding protein (RfBP) of egg yolk and egg white. A lambda gt11 cDNA library derived from chicken oviduct poly(A)+ RNA was screened with polyclonal rabbit antiserum to chicken RfBP. Positive clones were isolated and rescreened with a mixed oligonucleotide probe corresponding to residues 20-25 of the mature protein. The largest cDNA clone (969 base pairs) was subcloned into plasmid pIBI21, and the nucleotide sequence was determined by the dideoxynucleotide method. This clone contained the entire coding region for RfBP. The published amino acid sequence of the mature protein was confirmed. In addition, the following 17-residue signal peptide was deduced: Met-Leu-Arg-Phe-Ala-Ile-Thr-Leu-Phe-Ala-Val-Ile-Thr-Ser-Ser-Thr-Cys. Unexpectedly, the nucleotide sequence codes for 2 adjacent arginine residues at the carboxyl terminus that are not observed in the mature protein. The amino acid sequence of RfBP is homologous with bovine milk folate-binding protein. Eight of the nine pairs of cysteines involved in disulfide bonds in RfBP are conserved in folate-binding protein, as are all of the tryptophan residues. Sequence identity between homologous regions of these two vitamin-binding proteins is more than 30%.     

Physiological and Biochemical Mechanisms Involved in the Response to Abscisic Acid in Maize Coleoptiles

The role of abscisic acid (ABA) in controlling growth and developmenthas been studied in maize (Zea mays L.) coleoptile segments.Application of ABA reduces the elongation rate by about 50%and affects ion fluxes. In particular, proton extrusion is decreasedwhile potassium efflux is greatly enhanced. Apparently, ABAdoes not: seem to influence calcium influx from the apoplastinto the cytosol, but more likely it influences its efflux.Alteration of cytosolic calcium concentration may also be obtainedby increasing its release from internal stores. This possibilitymight be sustained by the increased hydrolysis of phosphatidylinositolupon ABA application. Change in the balance of ion fluxes shouldresult from regulation of transport mechanisms at the membranelevel and should produce changes in the transmembrane electricalpotential. The H⁺- ATPase and the ATP-dependent calcium transportactivities are both influenced by the treatment with ABA, –55%and –40%, respectively. Under these conditions [Ca²⁺]_cytand pH_cyt can be modified and, as a consequence of their regulation,they may play an important role in mediating the physiologicaland biochemical effects of ABA, acting as second intracellularmessengers. ¹Research supported by National Research Council of Italy, SpecialProject RAISA, Sub-Project N. 2, Paper n. 2782.     

Qualité et interprétation des examens du bilan biologique de thrombophilie constitutionnelle Cas particulier des principaux inhibiteurs: antithrombine, protéine C et protéine S

Biological screening for hereditary thrombophilia must be performed with constant concern for quality of the results and the interpretation. Different guidelines are available common to most laboratory tests, common to hemostasis tests, thrombophilia screening or specific for each test. These different steps are discussed in this paper with a special focus on the diagnosis of antithrombin, protein C and protein S deficiencies.     

Synthetic approaches to 6-substituted 9-(3-azido-3,4-dideoxy-β-d,l-erythro-pentopyranosyl)purines

Methyl 3-azido-2-O-benzoyl-3,4-dideoxy-β-dl-erythro-pentopyranoside (6) was synthesized through two routes in five steps from methyl 2,3-anhydro-4-deoxy-β-dl-erythro-pentopyranoside (1). The first route proceeded via selective azide displacement of the 3-tosyloxy group of methyl 4-deoxy-2,3-di-O-tosyl-α-dl-threo-pentopyranoside, followed by detosylation and benzoylation. The second route consisted, with a better overall yield, in the azide displacement of the mesyloxy group of methyl O-benzoyl-4-deoxy-3-O-methylsulfonyl-α-dl-threo-pentopyranoside (10), obtained by benzylate opening of 1, followed by benzoylation, debenzylation, and mesylation. Compound 6 was transformed into its glycosyl chloride, further treated by 6-chloropurine to give the nucleoside 9-(3-azido-2-O-benzoyl-3,4-dideoxy-β-dl-erythro-pentopyranosyl)-6-chloropurine (13). When treated with propanolic ammonia, 13 yielded 9-(3-azido-3,4-dideoxy-β-dl-erythro-pentopyranosyl)adenine.     

Potentiation of the antagonistic effect of ACTH11-24 on steroidogenesis by synthesis of covalent dimeric conjugates

Covalent dimerization of the adrenocorticotropin fragment ACTH11-24 increases its antagonistic potency on the ACTH-induced steroidogenesis in isolated bovine fasciculata/reticularis cells by 3 orders of magnitude when the C-termini are linked via a 10 A long spacer. This strong potentiation, probably mediated by cross-linking of the receptors, was shown to be dependent on the point of attachment of the monomeric fragment to the spacer, thus providing information about the position of the binding site in the hormonal segment and about the distance of the receptors on the cell surface.     

T wave changes in humans and dogs during experimental dives.

Electrocardiogram (ECG) analysis was performed in three human divers studied at 21 and 23.5 ATA while they breathed various gas mixtures containing H2 and/or He (COMEX HYDRA IX experiment) and in five dogs exposed to 91 ATA of He-O2 or He-N2-O2. In all cases, the O2 partial pressure was slightly higher than its physiological value. These human and animal studies reveal that elevated pressure of different inert gases did not change the resting heart rate or its respiratory fluctuation. However, the T wave amplitude increased in proportion to the gas density in the three divers; this was also found in four of the five dogs studied. Changes in peak T wave configurations were also observed in the dog experiments. Positional changes in QRS or T vectors cannot explain these T wave changes.     

Functional Effects of Adiponectin on Endothelial Progenitor Cells

Adiponectin is an adipokine whose plasma levels are inversely correlated to metabolic syndrome components. Adiponectin protects against atherosclerosis and decreases risks in myocardial infarction. Endothelial progenitor cells (EPCs) are a heterogeneous population of circulating cells involved in vascular repair and neovascularization. EPCs number is reduced in patients with cardiovascular disease. We hypothesize that the positive effects of adiponectin against atherosclerosis are explained in part by its interactions with EPCs. Cells were obtained from healthy volunteers' blood by mononuclear cell isolation and plating on collagen‐coated dishes. Three sub‐populations of EPCs were identified and characterized using flow cytometry. EPCs' expression of adiponectin receptors, AdipoR1, and AdipoR2 was evaluated by quantitative PCR. The effects of recombinant adiponectin on EPCs' susceptibility to apoptosis were assessed. Finally, expression of neutrophil elastase by EPCs and activity of this enzyme on adiponectin processing were assessed. Quantitative PCR analysis of EPCs mRNAs showed that AdipoR1 mRNA is expressed at higher levels than AdipoR2. Expression of AdipoR1 protein was confirmed by western blot. Adiponectin significantly increased survival of two sub‐populations of EPCs in conditions of serum deprivation. Such effect could not be demonstrated in the third EPCs sub‐population. We also demonstrated that EPCs, particularly one sub‐population, express neutrophil elastase. Neutrophil elastase activity was confirmed in EPCs' conditioned media. Adiponectin protects some EPCs sub‐populations against apoptosis and therefore could modulate EPCs ability to induce repair of vascular damage. Neutrophil elastase activity of EPCs could locally modulate adiponectin activity by its involvement in the generation of the globular form of adiponectin.