Comptes rendus des seances de la Societe de biologie et de ses filiales最新文献

Angiotensin (Ang) II and AngIII are two peptide effectors of the brain renin-angiotensin system that participate in the control of blood pressure and increase water consumption and vasopressin release. In an attempt to delineate the respective roles of these peptides in the regulation of vasopressin secretion, their metabolic pathways and their effects on vasopressin release were identified in vivo. For this purpose, we used recently developed selective inhibitors of aminopeptidase A (APA) and aminopeptidase N (APN), two enzymes that are believed to be responsible for the N-terminal cleavage of AngII and AngIII, respectively. Mice received [3H]AngII intracerebroventricularly (i.c.v.) in the presence or absence of the APA inhibitor, EC33 ((S)-3-amino-4-mercapto-butylsulfonate de sodium) or the APN inhibitor, EC27 ((S)-2-amino-pentan-1,5-dithiol). [3H]AngII and [3H]AngIII levels were evaluated from hypothalamus homogenates by HPLC. EC33 increased the half-life of [3H]AngII 2.6-fold and completely blocked the formation of [3H]AngIII, whereas EC27 increased the half-life of [3H]AngIII 2.3-fold. In addition, the effects of EC33 and EC27 on Ang- induced vasopressin release were studied in mice. AngII was injected i.c.v. in the presence or absence of EC33, and plasma vasopressin levels were estimated by RIA. While vasopressin levels were increased 2-fold by AngII, EC33 inhibited AngII-induced vasopressin release in a dose-dependent manner. In contrast, EC27 injected alone increased in a dose-dependent manner vasopressin levels. The EC27-induced vasopressin release was completely blocked by the coadministration of the Ang receptor antagonist (Sar1-Ala8) AngII. These results demonstrate for the first time that i) APA and APN are involved in vivo in the metabolism of brain AngII and AngIII, respectively, and that ii) the action of AngII on vasopressin release depends upon the prior conversion of AngII to AngIII. This shows that AngIII behaves as one of the main effector peptides of the brain renin-angiotensin system in the control of vasopressin release.

Fructo-oligosaccharides (FOS) are new food ingredients that are able to beneficially affect the host by selectively stimulating the growth and/or activity of colonic bifidobacteria (concept of prebiotics). A commercial enzyme preparation was found to possess a high fructosyltransferase activity and could be used as a biocatalyst for the industrial production of FOS from sucrose. Under optimum conditions (pH: 5.5, temperature: 55 degrees C and 7 units of fructosyltransferase activity per gram sucrose), in presence of glucose (competitive inhibitor) the actual yield reached the theoretical value (up to 50%). Actually, FOS that are commercially available for their prebiotic properties belong to inulin type with low degree of polymerisation (DP:3 to 10). Our FOS were identified by both HPLC and 13C-NMR spectrometry as neo-FOS type (neo-kestose, neo-nystose and neo-fructofuranosylnystose), a new structure which is very close to inulin type (same linkage between fructosyl units). The neo-FOS may act as a prebiotic factor due to their structural similarity with inulin type.

Antidepressants are used since 40 years. All presently used antidepressants have a slow onset of action and do not improve all patients; thus, there is an absolute need for new antidepressants. A variety of animal models, often based upon the monoaminergic theory of depressive disorders, has been used to screen the current antidepressants. In fact, the main focus of most of these animal models has been to predict the antidepressant potential i.e. to establish predictive validity. However, the evaluation of such animal models should also consider face validity, i.e. how closely the model resembles the human condition, and this should help to identify innovating medicines. Antidepressants, when taken by a healthy person, induce nothing more than side effects, unrelated to an action on mood, whereas they alleviate depressive symptomatology in depressed patients. We have speculated that genetically selected animal models would be closer to the human clinical situation than models based on standard laboratory strains. We have depicted here that marked differences exist between strains of mice in the amount of immobility i.e. "spontaneous helplessness" observed in the tail suspension test, a method used to screen potential antidepressants. We have studied the behavioural characteristics of mice selectively bred for spontaneous high or low immobility scores in the tail suspension test. Hopefully, these selectively bred lines will provide a novel approach to investigate behavioural, neurochemical and neuroendocrine correlates of antidepressant action.

We described a double-site enzyme-linked immunosorbent assay (ELISA) to measure polysialic acid neural cell adhesion molecule (PSA-NCAM) level in CSF. Immunocapture of PSA-bearing molecules is first effected by means of a monoclonal antibody (anti-MenB), directed against sialic acid polymers and adsorbed into plastic wells. Linked PSA-NCAM is then revealed by means of a second antibody, directed against an aminoacid sequence of NCAM and labelled with peroxydase. The lowest amount of PSA-NCAM detectable was estimated to be 0.11 microgram/l. This value was considered as the threshold for positivity. PSA-NCAM level was measured using this method in CSF from 29 patients with medulloblastoma. CSF had been collected at different times following tumor excision and stored at--80 degrees C. At the same times, cytological examination in CSF (medulloblastoma metastatic cells) and craniospinal imaging (tomographic scan or MRI) had been performed. PSA-NCAM was never detected in control CSF. For patients in remission, beyond the post-operative period of 1 or 2 months, 18 on 21 exhibited a PSA-NCAM level below the threshold value. For refractory patients, so classified according to the positivity of cytology and/or imaging, whatever the time after the tumor excision, PSA-NCAM was always positive (23/23), while either cytology or imaging were positive less frequently (16/23 for both). For relapses, PSA-NCAM was more frequently positive (6/7) than cytology and imaging (1/7 and 5/7, respectively). We concluded that PSA-NCAM positivity in CSF may be a reliable marker to detect the invasive or metastatic feature of medulloblastoma.

Among the 315 protein toxins elicited by gram positive and gram negative bacteria so far characterized, about 50 toxins are currently considered as totally or partially, responsible of the pathological manifestations and/or lethality resulting from host infection or intoxication (contaminated food) by relevant toxinogenic bacteria. A certain number of criteria are required for the assessment of indisputable involvement of a toxin or an array of toxins (from the same bacteria) in infectious diseases: 1) The bacterial microorganism clearly identified as the pathogenic agent of the disease produces component(s) considered as toxin(s); 2) The administration to appropriate animal(s) of the toxin(s) separated from the relevant bacteria or produced by genetic engineering from a heterologous tox+ recombinant bacterial strain produces symptoms and pathophysiological disorders that mimic those observed in the natural disease or at least those elicited in experimental animals by the cognate toxin-producing bacteria; 3) The in vitro incubation of the isolated toxin(s) with appropriate animal organs, tissues or cells elicits certain pathophysiological, biochemical or metabolic manifestions observed in the host infected with the relevant toxinogenic bacteria; 4) Toxin concentration in the organism of the host infected by the toxinogenic bacteria should be compatible with the characteristics of the relevant disease. The toxins of pathogenic interest exhibit a variety of effects in bacterial diseases. Bacteria that colonize a wound or mucosal surface but do not invade target cells can produce toxins that act locally or enter the bloodstream and attack internal organs (e.g. Corynebacterium diphtheriae, Vibrio cholerae, ...). Bacteria growing in a wound can produce toxins that destroy host tissue and kill phagocytes in the immediate vicinity of the bacteria, thus facilitating bacterial growth and spread. On the basis of the above mentioned criteria, the following bacterial diseases among many others are toxin-associated (toxinoses): diphtheria, tetanus, botulism, whooping cough, diarrhea, bloody diarrhea, hemolytic uremic syndrome, cholera, scarlet fever, toxic shock syndrome, gas gangrene, B. fragilis diarrhea, anthrax, pseudomembranous colitis.

Under certain environmental conditions, marine and freshwater phytoplankton may produce phycotoxins inhibitors of serine/thréonine protein phosphatases 1, 2A and 3. In the marine environment, dinoflagellates produce fatty polyethers: okadaic acid and its derivatives, the dinophysistoxins, which accumulate in shellfish and can cause diarrhetic shellfish poisoning (DSP) when ingested. In freshwater, the toxins are microcystins and nodularin, 7 or 5 amino acid cyclic peptides and are hepatotoxic. These toxins have caused massive poisoning of wild animals or domestic livestock and now are a health threat for humans through use of drinking and recreation water. Moreover, all these toxins are potent tumor promoters but belong to a new class, different from the TPA class, because they do not act on Protein Kinase C. Although the mutagenicity Ames test responds negatively, several results show their genotoxic potential, and therefore they are a health hazard through chronic exposition to low doses. Finally, okadaic acid, through its easy penetration in all cellular types can be used as a tool to study mechanisms involved in protein phosphorylation/dephosphorylation processes.

Melatonin receptors belong to the super-family of G protein-coupled receptors. They modulate a large spectrum of physiological functions including regulation of circadian rhythms and seasonal reproduction. Pharmacological evidence suggests the expression of two types of receptors, called Mel1 and Mel2. So far, only Mel1 receptors have been cloned and classified into three subtypes (Mel1A, Mel1B, Mel1C). Mel1 receptors are expressed in the brain, the retina and several other peripheral tissues. All Mel1 subtypes show comparable pharmacological profiles including inhibition of adenylyl cyclase. Cloning and expression of two allelic isoforms of the Mel1 receptor from Xenopus laevis has revealed another signalling pathway, inhibition of cGMP levels via the soluble guanylyl cyclase pathway. The two isoforms are differentially coupled to the cAMP and cGMP pathways indicating the existence of functional differences between melatonin receptors. Future research topics will include cloning of the Mel2 receptor, receptor regulation and the elucidation of melatonin receptor's function in peripheral tissues.

Melatonin, synthetized by the pineal gland, is the chemical messenger which allows seasonal animals to perceive day length changes. In the ewe, the nervous message, transformed into a hormonal one, triggers pulsatile activity of the LHRH neurons. About 40 days are necessary for melatonin to centrally stimulate the pulsatile LHRH activity. Its sites and mode of action are not completely elucidated, but a precise hypothalamic zone has been defined in which radioactive melatonin binds specifically and where cold melatonin delivered locally stimulates LHRH activity. In veterinary clinic, the most frequent mode of distribution is the sub-cutaneous implant, which induces an advance of the cyclical ovulatory activity of ewes and goats. The date of fertilization is advanced and fecundity of females is improved. It can be used alone, or in association with other hormonal treatments, or after an artificial photoperiodic treatment. Under these conditions, it allows a quantitative and qualitative increase in out-of-season sperm production in rams and he-goats. Such an implant is registered and marketed in France, UK, Greece, Australia and New-Zealand.

Regulation of homeostasic balance between cell proliferation and cell death, called apoptosis, is essential for development and maintenance of multicellular organisms. Recent research into the molecular mechanisms of apoptosis has revealed that apoptosis is a genetically and evolutionarily conserved process that can become deranged when the components of the cellular apoptotic machinery are mutated, perturbated by viral gene products or present in inappropriated quantities. Analysis of the regulatory apoptotic pathways has led to a better understanding of the etiology and pathogenesis of many human diseases, notably cancers, infectious diseases or autoimmune diseases. Our understanding of the regulation of apoptosis in health and disease is far from complete and the use of understanding into new therapeutic modalities has only begun to be approached.

We have shown that given cytokines are capable of inducing the expression of transcription factors of the Ets family in two very distinct cell types: 1) endothelial cells of blood vessels, but only during neovascularization, and 2) fibrocytic cells from stroma surrounding tumors, but only if these tumors bear characteristics of invasiveness. In such cases, the fibrocytic cells also express some metalloproteinases (collagenase 1, urkinase plasminogen activator, sometimes stromelysin1). In ex vivo reconstruction experiments, we demonstrate that the corresponding genes are directly up-regulated by the Ets family transcription factors, often associated with the transcription complex Jun/Fos. The proteinases are thought to dismantle the stroma and allow invasive tumors to proceed toward further expansion. We speculate that inactivation of the Ets factors could seriously hamper both neovascularization and tumor expansion.