[Signals of danger inhibit exploration-induced activation of the nitrergic system of the nucleus accumbens].

In Sprague-Dawley rats it was shown by means of in vivo microdialysis combined with HPLC analysis that exploratory behavior in a new chamber produced an increase in the extracellular level of citrulline (an NO co-product) in the medial n. accumbens, which was prevented by intra-accumbal infusions of 7-nitroindazole (0.5 mM), a neuronal NO synthase inhibitor. When presented during exploratory activity, a tone previously pared with footshock, firstly, inhibited the exploration and, secondly, prevented the exploration-induced increase in accumbal extracellular citrulline level. These changes did not occur in control animals (the same procedure but without footshock or unpaired tone and footshock presentation). The data obtained indicate for the first time that the n. accumbens could be implicated in fear transfer to exploratory behavior and show the involvement of the nitrergic system inhibition in this process.

[Circulating citrulline levels: a biomarker for intestinal functionality assessment]. / La citrullinémie: un biomarqueur de la fonctionnalité intestinale.

Circulating citrulline is emerging as an innovating biomarker candidate for assessment of intestinal function. Amino acid synthesized by enterocytes of intestinal mucosal, citrulline is not included in proteins or nutrition products and is a precursor for production of arginine by the kidney. Plasma citrulline is, in clinical situation, an established biomarker of enterocyte functional metabolic mass (trophicity) in children and in adult patients due to its high relation to active functional small bowel remnant length in intestinal diseases (short bowel, extensive enteropathies, intestinal toxicity of chemotherapy and radiotherapy. Plasma citrulline concentration (30-50  µmol/L), independent of nutritional status, if less than 10  µmol/L can give an objective threshold for nutrition parenteral use in case of intestinal failure due to enterocyte abnormalities or lack. Its regular dosage allows the monitoring of intestinal function except in case of significant renal failure.

Improving adaptation to weaning: effect of intermittent suckling regimens on piglet feed intake, growth, and gut characteristics.

Daily separation of sows and piglets during lactation, intermittent suckling (IS), improves feed intake and postweaning adaptation in piglets. The aim of the current study was to determine how, in piglets that have been subjected to IS, age at weaning and the duration of the preceding IS period contribute to postweaning adaptation through effects on feed intake, growth, and gut characteristics. All piglets had ad libitum access to creep feed from d 7. Litters were subjected to conventional weaning (CW) or to 1 of 3 IS regimens. In CW, litters (n = 29) had continuous access to the sow until weaning (d 26, d 0 = farrowing). During IS, litters had access to the sow between 1600 and 0600 h. Litters in the IS treatments were subjected to IS 1) from d 19 onward and weaned at d 26 (IS19-7D, n = 33), 2) from d 19 onward and weaned at d 33 (IS19-14D, n = 28), or 3) from d 26 onward and weaned at d 33 (IS26-7D, n = 33). The IS19-7D regimen resulted in a relative growth check within the first 2 d after weaning similar to CW litters (72 +/- 13 and 90 +/- 7%, respectively), but in a greater piglet growth (P = 0.014) and feed intake (P = 0.001) between d 2 and 7 postweaning. Moreover, IS19-7D was not associated with a (further) reduction in villus height as observed at d 2 postweaning in CW litters. In IS piglets weaned after an extended lactation (d 33), a markedly smaller weaning-associated relative growth check was observed shortly postweaning (11 +/- 18 and 32 +/- 19% for IS19-14D and IS26-7D litters, respectively). In these litters, feed intake and growth within the first 2 d after weaning were slightly greater when piglets were subjected to IS for 2 wk (IS19-14D) rather than for 1 wk (IS26-7D; P = 0.032 and P = 0.037 for feed intake and growth, respectively). Irrespective of duration of IS, weaning at d 33 with IS was not associated with a reduction in villus height. Irrespective of treatment, plasma citrulline concentrations were reduced at d 2 and 8 postweaning compared with the values at weaning (P < or = 0.01). No correlation was observed between postweaning plasma citrulline concentrations and postweaning small intestinal villus height. This study indicates that 1 wk of IS before weaning at d 26 of lactation improves feed intake and growth between d 2 and 7 postweaning and does not result in a reduction of villus height as observed in CW piglets, although it did not prevent a profound growth check shortly after weaning. However, combining 1 wk of IS with an extended lactation improved postweaning adaptation markedly in terms of growth, feed intake, and gut characteristics. Increasing the duration of IS from 1 to 2 wk slightly improved growth and feed intake shortly after weaning, but the contribution to postweaning adaptation seemed to be relatively small compared with extending lactation.

A tale of two citrullines--structural and functional aspects of myelin basic protein deimination in health and disease.

Myelin basic protein (MBP) binds to negatively charged lipids on the cytosolic surface of oligodendrocyte membranes and is responsible for adhesion of these surfaces in the multilayered myelin sheath. The pattern of extensive post-translational modifications of MBP is dynamic during normal central nervous system (CNS) development and during myelin degeneration in multiple sclerosis (MS), affecting its interactions with the myelin membranes and with other molecules. In particular, the degree of deimination (or citrullination) of MBP is correlated with the severity of MS, and may represent a primary defect that precedes neurodegeneration due to autoimmune attack. That the degree of MBP deimination is also high in early CNS development indicates that this modification plays major physiological roles in myelin assembly. In this review, we describe the structural and functional consequences of MBP deimination in healthy and diseased myelin.

The role of citrullinated proteins suggests a novel mechanism in the pathogenesis of multiple sclerosis.

The pathogenesis of MS is unknown. In our studies, we have demonstrated an important role for citrullinated myelin basic protein (MBP). The accompanying loss of positive charge compromises the ability of MBP to interact with the lipid bilayer. The conversion of arginine to citrulline in brain is carried out by an enzyme peptidyl arginine deiminase (PAD) 2. The amount of PAD 2 in brain was increased in MS normal-appearing white matter. The mechanism responsible for this increase involved hypomethylation of the promoter region in the PAD 2 gene in MS, but no change (compared to normal) was found in thymus tissue DNA from the same MS patients. In addition, no change was observed in other neurological diseases, including Alzheimer's, Parkinson's, and Huntington's. We propose that citrullinated MBP, resulting from elevated levels of PAD 2 represents an important biochemical pathway in the pathogenesis of MS.

Accessibility of endothelial and inducible nitric oxide synthase to the intracellular citrulline-arginine regeneration pathway.

This study investigates our hypothesis that argininosuccinate synthase (AS), the rate-limiting enzyme for arginine (L-arg) regeneration from citrulline (L-cit), plays a pivotal role in supplying L-arg to endothelial (eNOS), but not inducible (iNOS) nitric oxide synthase, for nitric oxide (NO) production. Transgenic rat blood-brain barrier (TR-BBB) endothelial cells were used as a model to elucidate the accessibility of the L-arg compartments for NOS isozymes. NO production via eNOS or iNOS, with or without alpha-methyl-DL-aspartic acid (MDLA), an AS inhibitor, was measured by a fluorometric method. NO production via eNOS was activated by the calcium ionophore A23187, while via iNOS was induced by cytokines. AS activity was assayed by the amount of argininosuccinate regenerated from radioactive aspartic acid from cell extracts. Upon increased AS activity (5.9-fold) in cells grown in L-arg-free/L-cit-supplemented medium, A23187-activated NO production also significantly increased, however cytokine-induced NO production was not detected. A23187-activated NO production was observed not only in L-arg containing medium, but also L-arg-free and L-arg-free/L-cit-supplemented medium, and was abolished by MDLA regardless of medium type. Cytokine-induced NO production was only observed in L-arg containing medium, not in L-arg-free or L-arg-free/L-cit-supplemented medium, and it was not inhibited by MDLA in the L-arg containing medium. Our results indicate that extracellular L-arg was the only L-arg pool for cytokine-induced NO production and intracellular L-arg regenerated from L-cit via AS pathway was the major L-arg pool for A23187-activated NO production in TR-BBB endothelial cells. Therefore, modulation of AS activity could be a promising strategy to selectively alter NO production via eNOS, but not iNOS.

Transcriptional profiling of extracellular amino acid sensing in Saccharomyces cerevisiae and the role of Stp1p and Stp2p.

S. cerevisiae responds to the presence of amino acids in the environment through the membrane-bound complex SPS, by altering transcription of several genes. Global transcription analysis shows that 46 genes are induced by L-citrulline. Under the given conditions there appears to be only one pathway for induction with L-citrulline, and this pathway is completely dependent on the SPS component, Ssy1p, and either of the transcription factors, Stp1p and Stp2p. Besides the effects on amino acid permease genes, an ssy1 and an stp1 stp2 mutant exhibit a number of other transcriptional phenotypes, such as increased expression of genes subject to nitrogen catabolite repression and genes involved in stress response. A group of genes involved in the upper part of the glycolysis, including those encoding hexose transporters Hxt4p, Hxt5p, Hxt6p, Hxt7p, hexokinase Hxk1p, glyceraldehyde 3-phosphate dehydrogenase Tdh1p and glucokinase (Glk1p), shows increased transcription levels in either or both of the mutants. Also, most of the structural genes involved in trehalose and glycogen synthesis and a few genes in the glyoxylate cycle and the pentose phosphate pathway are derepressed in the ssy1 and stp1 stp2 strains.

La arginina en su contexto metabólico y fisiológico / Arginine on its metabolic and physiological context

La L-arginina es uno de los aminoácidos más versátiles desde el punto de vista metabólico y fisiológico. Entre sus funciones conocidas se incluyen: sustrato de la biosíntesis de proteínas y de péptidos bioactivos, participación en la detoxificación de amonio, liberación de hormonas, y biosíntesis de poliaminas y creatina. Las funciones de la arginina se han visto recientemente incrementadas con el descubrimiento de su papel como sustrato precursor del óxido nítrico, un efector multifuncional implicado en la vasodilatación, neurotransmisión, y con actividad antimicrobiana y antitumoral. Desde el punto de vista nutricional, la arginina es un aminoácido "condicionalmente esencial" para los mamíferos. Sus requerimientos se cubren gracias al aporte dietético y la síntesis endógena, siendo ésta última en el hombre, al parecer, suficiente para cubrir las necesidades fisiológicas habituales pero insuficiente en períodos o condiciones de elevada demanda del aminoácido. Tal situación sugiere un potencial uso terapéutico para la arginina que está comenzando a ser analizado. Esta revisión pretende resumir los conocimientos actuales sobre la síntesis de arginina y sus principales destinos metabólicos (AU)

L-Citrulline, the by-product of nitric oxide synthesis, decreases vascular smooth muscle cell proliferation.

Endothelium injury plays an important role in atherosclerosis. Damage to the endothelium results in vascular smooth muscle cell proliferation. Natriuretic peptides present a potent antimitogenic action, mediating their biological effects via the binding of guanylate cyclase-linked atrial natriuretic peptide (ANP) receptor and the production of cyclic GMP. In a previous study, we demonstrated that L-citrulline, the by-product of nitric oxide synthesis, could relax rabbit aortic rings by stimulating the guanylate cyclase-linked ANP receptor. In this work, we investigated the effect of L-citrulline on vascular smooth muscle cell proliferation. L-Citrulline (10(-8) M) significantly decreased rat aortic (A10 cell line) vascular smooth muscle proliferation. The percentage of inhibition exerted by L-citrulline on days 3, 5, and 7 of the proliferation curve was 20.0 +/- 0.5%, 37.5 +/- 8.3%, and 28. 5 +/- 7.2%, respectively. In addition, L-citrulline also inhibited serum-induced DNA synthesis, measured as 5-bromo-2'-deoxyuridine incorporation. 5-Bromo-2'-deoxyuridine incorporation into nuclei of vehicle-treated cells was 40.5 +/- 2.4%, whereas in L-citrulline-treated cells the percentage decreased to 36.0 +/- 4.1%, 29.1 +/- 2.0% (P <.01, n = 4), 30.5 +/- 2.4% (P <.05, n = 4), and 23.1 +/- 0.5% (P <.001, n = 4) for 10(-10), 10(-9), 10(-8), and 10(-7) M, respectively. Zaprinast, a phosphodiesterase type V inhibitor, enhanced 5-bromo-2'-deoxyuridine incorporation in serum-stimulated cells. Moreover, L-citrulline inhibition of serum-stimulated DNA synthesis was abolished by HS-142-1 (10(-5) M), an ANP receptor antagonist. In another group of experiments, L-citrulline was shown to increase intracellular cyclic GMP levels from 2.1 +/- 0.2 pmol of cGMP/mg protein to 4.1 +/- 0.1 for L-citrulline (10(-8) M) (P <.001, n = 3). These findings suggest that L-citrulline decreases vascular smooth muscle cell proliferation in the A10 cell line by acting on DNA synthesis by mechanisms that involve the ANP receptor.