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Introduction La fievre mediterraneenne familiale (FMF) peut se compliquer d’amylose AA, mais la raison pour laquelle uniquement certains patients vont developper cette complication grave n’est a ce jour pas totalement elucidee. Il existe cependant des arguments epidemiologiques et experimentaux en faveur de l’implication de facteurs environnementaux, et notamment du micro biote intestinal. Notre objectif etait d’evaluer la composition du microbiote intestinal et ses correlations avec des marqueurs inflammatoires dans la FMF compliquee ou non d’amylose AA. Materiels et methodes Le microbiote intestinal a ete etudie a partir d’un prelevement de selles par le sequencage du gene de l’acide ribonucleique 16S sur une plate-forme Illumina MiSeq. Les patients inclus etaient : atteints de FMF sans amylose AA (n = 34), atteints d’amylose AA secondaire a une FMF (n = 7), atteints d’amylose AA secondaire a une autre cause que la FMF (n = 19) et compares a des temoins sains (n = 26). Les associations entre les taxa bacteriens et les donnees cliniques et paracliniques etaient evaluees a l’aide de la methode statistique d’association multivariee avec modeles lineaires (MaAsLin). Des dosages seriques de bio marqueurs d’inflammation etaient effectues : les interleukines (IL), 1β-6, le Tumor Necrosis Factor-α et des adipokines (leptine, adiponectine) etaient doses par methode immunoenzymatique ; l’activite indoleamine 2,3-dioxygenase (IDO) etait mesuree via le rapport kynurenine sur tryptophane, doses tous deux par chromatographie liquide a haute performance. Resultats Les patients atteints de FMF sans amylose AA etaient majoritairement porteurs d’une mutation non ambigue de MEFV a l’etat homozygote ou heterozygote composite (91 %), tous recevaient de la colchicine quotidiennement et 2 etaient egalement traites par biotherapie anti-IL-1. Les patients atteints de FMF compliquee d’amylose AA secondaire etaient tous homozygotes pour une mutation non ambigue de MEFV (M694 V/M694 V, n = 6, ou M680I/M680I, n = 1). Les causes d’amylose AA chez les 19 patients sans FMF etaient respectivement : une autre maladie genetique rare (n = 4), un rhumatisme inflammatoire (n = 4), une obesite (n = 3), une cause inconnue a ce jour (n = 4) et a des causes diverses (maladie de Crohn, seropositivite VIH, maladie de Waldenstrom, syndrome de Schnitzler, n = 1 chacun). Une dysbiose etait observee d’une part chez les patients atteints de FMF sans amylose et d’autre part chez les patients avec amylose AA quelle que soit la cause, en comparaison a des temoins sains. Cette dysbiose etait caracterisee par une diminution de la diversite α et une alteration significative de la composition du microbiote intestinal. Dans chaque groupe, les bacteries appartenaient majoritairement aux phyla Firmicutes, Bacteroidetes, Proteobacteria et Actinobacteria. Parmi les patients atteints de FMF, chez ceux presentant une amylose AA, il a ete observe q’une sur representation de 2 unites operationnelles taxonomiques appartenant a l’ordre des Clostridiales, et a une concentration serique d’adiponectine et d’activite IDO significativement plus elevees (p Conclusion La presence d’une dysbiose chez les patients atteints de FMF ou d’amylose AA suggere un lien fonctionnel entre le microbiote intestinal et ces pathologies, et ouvre la possibilite de recourir a des techniques de manipulation du microbiote intestinal. Par ailleurs, les taux seriques d’adiponectine et d’activite IDO pourraient etre utilises en tant que bio marqueur diagnostique dans l’amylose AA.

1. The influence of lipopolysaccharide (LPS)-induced sepsis on the various mast cell phenotypes of rat dura mater were examined both by immunohistochemical and biochemical methods. 2. Three different populations of mast cells were identified in control rats: connective tissue type mast cells (CTMC) which contain rat mast cell protease1 (RMCP1), histamine, serotonin and heparin, mucosal type mast cells (MMC) which contain RMCP2, histamine and serotonin, and intermediate type which contains both RMCP1 and RMCP2 and probably various proportions of amines and heparin. 3. LPS (25 mg kg(-1) i.p.) caused changes in the proportions of the various types of mast cells. The number of MMC and intermediate type mast cells significantly increased and the number of mast cells immunopositive for both heparin and serotonin significantly decreased. Biochemical analysis showed that the histamine concentration of dura increased while its serotonin concentration decreased. 4. While vasoactive intestinal peptide (VIP) (25 ng kg(-1) i.p.) appears to potentiate LPS effects on dura mater mast cells, non-selective inhibition of nitric oxide (NO) synthase by N(g)-nitro-L-arginine methyl ester (L-NAME) (30 mg kg(-1) i.p.) did not influence sepsis-induced mast cell changes. 5. These findings suggest that mast cells of dura mater may play a role in brain protection during sepsis.

Resume Introduction La production d'oxyde nitrique (NO) augmente lors d'exercices aerobies. Plusieurs facteurs (stress de cisaillement, catecholamines et hypoxie) connus pour activer une production accrue de NO sont reunis a l'exercice de sprint. Notre objectif a ete de mesurer dans le sang les produits stables derives du NO en reponse a l'epreuve de Wingate. Synthese des faits Un exercice de sprint augmente la concentration plasmatique de produits derives du NO d'environ 25 % chez des sujets actifs des deux sexes. Conclusion L'augmentation de produits derives du NO est inferieure a celle observee lors d'exercices aerobies et ne differe pas entre hommes et femmes.

Tryptophan (TRP) is the precursor of melatonin, the primary secretory product of the pineal gland. Hepatic heme deficiency decreases the activity of liver tryptophan pyrrolase, leading to increased plasma TRP and serotonin. As a paradox, patients with attacks of acute intermittent porphyria (AIP), exhibit low nocturnal plasma melatonin levels. This study using a rat experimental model was designed to produce a pattern of TRP and melatonin production similar to that in AIP patients. Pineal melatonin production was measured in response to: (a) a heme synthesis inhibitor, succinylacetone, (b) a heme precursor, delta-aminolevulinic acid (Ala), (c) a structural analogue of Ala, gamma-aminobutyric acid. Studies were performed in intact rats, perifused pineal glands, and pinealocyte cultures. Ala, succinylacetone, and gamma-aminobutyric acid significantly decreased plasma melatonin levels independently of blood TRP concentration. In the pineal gland, the key enzyme activities of melatonin synthesis were unchanged for hydroxyindole-O-methyltransferase and decreased for N-acetyltransferase. Our results strongly suggest that Ala overproduced by the liver acts by mimicking the effect of gamma-aminobutyric acid on pineal melatonin in AIP. They also support the view that Ala acts as a toxic element in the pathophysiology of AIP.

A serotonin N -acetyl transferase (NAT) activity was detected in the brain of Pieris brassicae . The maximal head NAT activity appeared to occur during the wandering-pharate phase. The haemolymph level of serotonin, the selected NAT substrate, presented a maximum on the 3rd day of the 5th instar for both short- and long-photoperiod animals with a longer increase in short-photoperiod animals. The circadian rhythm of head NAT, determined on the same day, appeared bimodal and was affected by pteridines only when they were injected at “specific” times. At the last larval stage, the effect of either serotonin, N -acetylserotonin or melatonin was characterized by inactivity immediately after the injection, leading to death by starvation. In “underchilled” chrysalids, serotonin induced a diapause breakdown while melatonin led to mortality. These data suggest, besides the role of juvenile hormone, ecdysone, and polypeptidic hormones, a role for indolamines during the photoperiodic-dependent development of Pieris .

Fasting gastrinemia, fundic argyrophil cell density, mast cell number, basal fundic histamine content and histidine decarboxylase activity were determined in 20 antrectomized patients and 20 control subjects. Fasting gastrinemia and fundic argyrophil cell density were significantly lower in antrectomized patients than in controls, whereas fundic mast cell number, basal histamine content, and histidine decarboxylase activity did not differ significantly between the two groups. In antrectomized patients the basal fundic histamine content appears related to the fundic mast cell number, as a consequence of the reduced effect of gastrin on argyrophil cells.

Previously, we reported that rabbit cerebral arteries contain mast cells that frequently establish close contacts with parasympathetic-like nerve fibers. Here we have examined the possible function of this link by comparing the effects of carbachol and compound 48/80 on mast cell morphology and on the serotonin (5-HT) and histamine content of these arteries. In vivo, 2 micrograms/min of compound 48/80 or 1 micrograms/min of carbachol was infused for 30 min into one internal carotid artery of pentobarbital anesthetized rabbits, the contralateral artery being infused with vehicle. In vitro, the action of 10(-6) M carbachol was tested on isolated middle cerebral artery trees (MCAs) in the presence or absence of 10(-7) M atropine. The effects of carbachol were also tested in vitro on sympathectomized arteries. The 5-HT and histamine contents of all MCAs were measured by radioenzymatic assay, and fragments were prepared for electron microscopy. No histamine was detectable in any artery studied. The 5-HT content of arteries from control animals and those perfused with vehicle (in vivo) or incubated in the physiological solution (in vitro) was 250-300 pmol/mg protein. Both compound 48/80 and carbachol reduced this amount by approximately 50% and induced a marked degranulation of mast cells. Both secretion and degranulation were dramatically blocked in vitro by atropine. No difference in the 5-HT content was observed between intact and sympathectomized arteries under any condition. We conclude that a large proportion of rabbit cerebrovascular 5-HT is stored in mast cells and that cholinergic nerve activation could theoretically release this pool by acting on mast cell muscarinic receptors.

We examined the effect of pioglitazone, a peroxisome proliferator-activated receptor-gamma (PPARgamma) agonist of the thiazolidinedione class, on dopaminergic nerve cell death and glial activation in the 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) mouse model of Parkinson's disease. The acute intoxication of C57BL/6 mice with MPTP led to nigrostriatal injury, as determined by tyrosine hydroxylase (TH) immunocytochemistry, and HPLC detection of striatal dopamine and metabolites. Damage to the nigrostriatal dopamine system was accompanied by a transient activation of microglia, as determined by macrophage antigen-1 (Mac-1) and inducible nitric oxide synthase (iNOS) immunoreactivity, and a prolonged astrocytic response. Orally administered pioglitazone (approximately 20 mg/kg/day) attenuated the MPTP-induced glial activation and prevented the dopaminergic cell loss in the substantia nigra pars compacta (SNpc). In contrast, there was little reduction of MPTP-induced dopamine depletion, with no detectable effect on loss of TH immunoreactivity and glial response in the striatum of pioglitazone-treated animals. Low levels of PPARgamma expression were detected in the ventral mesencephalon and striatum, and were unaffected by MPTP or pioglitazone treatment. Since pioglitazone affects primarily the SNpc in our model, different PPARgamma-independent mechanisms may regulate glial activation in the dopaminergic terminals compared with the dopaminergic cell bodies after acute MPTP intoxication.

To quantify the inflammatory cell response in rat air pouch pseudosynovial membrane during monosodium urate monohydrate (MSU) crystal-induced inflammation.In the rat air-pouch model, we used a computer-assisted histomorphometric method to quantify cell distributions, based on cell linear densities, in histologic sections of membranes from pouches injected with MSU or saline. The volume, white blood cell (WBC) count, and histamine content of the pouch exudates were determined at several time points.Injection of 10 mg of MSU crystals into the pouch produced an acute exudate. After peaking at 24 hours, the exudate volume and WBC count decreased spontaneously over the next 3 days, simulating the self-limited course of acute gout. Membrane thickness followed a parallel course. Membrane polymorphonuclear cell (PMN) linear densities were closely correlated with exudate WBC counts, suggesting PMN recruitment from the subintimal synovial membrane. Both monocyte/macrophage and mast cell linear densities increased in the subintimal layer 2 hours after crystal injection (P = 0.038 and P = 0.03, respectively, versus controls), whereas PMN linear densities showed 2 peaks, one at 4 hours and the other 24 hours. The exudate histamine content peaked 6 hours after crystal injection, when mast cell linear densities were minimal in the membranes, suggesting mast cell degranulation.An increase in monocyte/macrophage and mast cell densities in the membrane preceded the PMN influx in the pouch membrane and exudate, suggesting that mast cells may be involved in the early phase of MSU crystal-induced inflammation, at least in this rat model.

1. The influence of lipopolysaccharide (LPS)-induced sepsis on the various mast cell phenotypes of rat dura mater were examined both by immunohistochemical and biochemical methods. 2. Three different populations of mast cells were identified in control rats: connective tissue type mast cells (CTMC) which contain rat mast cell protease1 (RMCP1), histamine, serotonin and heparin, mucosal type mast cells (MMC) which contain RMCP2, histamine and serotonin, and intermediate type which contains both RMCP1 and RMCP2 and probably various proportions of amines and heparin. 3. LPS (25 mg kg(-1) i.p.) caused changes in the proportions of the various types of mast cells. The number of MMC and intermediate type mast cells significantly increased and the number of mast cells immunopositive for both heparin and serotonin significantly decreased. Biochemical analysis showed that the histamine concentration of dura increased while its serotonin concentration decreased. 4. While vasoactive intestinal peptide (VIP) (25 ng kg(-1) i.p.) appears to potentiate LPS effects on dura mater mast cells, non-selective inhibition of nitric oxide (NO) synthase by N(g)-nitro-L-arginine methyl ester (L-NAME) (30 mg kg(-1) i.p.) did not influence sepsis-induced mast cell changes. 5. These findings suggest that mast cells of dura mater may play a role in brain protection during sepsis.

Objective: Growth regulatory properties of nitric oxide (NO) in cultured endothelial cells is controversial. The aim of our study was to investigate the effect of l-arginine, the endogenous NO precursor, and l-NAME, an inhibitor of NO synthase on the reendothelialization process after angioplasty. Methods: Fifty-five New Zealand White rabbits underwent denudation of the left iliac artery. After injury the rabbits were randomized in three groups: l-arginine 2.25% (l-arginine, n =19); N G-nitro-l-arginine methyl ester 15 mg/kg/day (l-NAME, n =19); and placebo (controls, n =17). Treatment was solubilized in drinking water. Reendothelialization was evaluated at 4 weeks by macroscopic evaluation of Evans blue staining and endothelial-specific immunostaining (CD-31) on cross sections. Intimal hyperplasia was evaluated by morphometric analysis. Results: Despite a significant increase in plasma arginine ( P =0.001) and a reduction in intimal hyperplasia ( P =0.003) with l-arginine, neither agent had a significant effect on reendothelialization at 4 weeks (controls=36±4%, l-arginine=43±3%, l-NAME=33±4%; NS). Conclusion: These results suggest that, in spite of previously demonstrated effects on neointimal hyperplasia, the NO pathway does not influence the regrowth of macrovascular endothelial cells in vivo.

Genetic inactivation of monoamine oxidase A (MAOA) in C3H/HeJ mice causes a complete absence of barrels in the somatosensory cortex, and similar alterations are caused by pharmacological inhibition of MAOA in wild type mice. To determine when and how MAOA inhibition affects the development of the barrel field, the MAOA inhibitor clorgyline was administered to mice of the outbred strain OF1 for various time periods between embryonic day 15 (E15) and postnatal day 7 (P7), and the barrel fields were analyzed with cytochrome oxidase and Nissl stains in P10 and adult mice. High-pressure liquid chromatography measures of brain serotonin (5-HT) showed three- to eightfold increases during the periods of clorgyline administration. Perinatal mortality was increased and weight gain was slowed between P3 and P6. Clorgyline treatments from E15 to P7 or from P0 to P7 disrupted the formation of barrels in the anterior snout representation and in parts of the posteromedial barrel subfield (PMBSF). Treatments from P0 to P4 caused similar although less severe barrel field alterations. Clorgyline treatments only during embryonic life or starting on P4 caused no detectable abnormalities. In cases with barrel field alterations, a rostral-to-caudal gradient of changes was noted: Rostral barrels of the PMBSF were most frequently fused and displayed an increased size tangentially. Thus, MAOA inhibition resulting in increased brain levels of 5-HT affects barrel development during the entire first postnatal week, with a sensitive period between P0 and P4. The rostral-to-caudal gradient of changes in the barrel field parallels known developmental gradients in the sensory periphery and in the maturation thalamocortical afferents. The observed barrel fusions could correspond to a default in the initial segregation of thalamic fibers or to a continued, exuberant growth of these fibers that overrides the tangential domain that is normally devoted to individual whiskers.

Brain microdialysis was used to study changes in the glutamate and aspartate extracellular concentrations in the striatum of conscious rats submitted to 30 minutes cerebral ischaemia, using the four-vessel occlusion model. Perfusion of the N-methyl-D-aspartate (NMDA) receptor channel blockers, dizocilpine (MK-801; 75 microM) and Mg2+ (2.5 mM), inhibited the ischaemia-induced accumulation of glutamate and aspartate. The AMPA/kainate receptor antagonist, 2,3-dihydroxy-6-nitro-7-sulfamylbenzo (F) quinoxaline (NBQX; 15 microM and 450 microM) had no effect on glutamate and aspartate levels during ischaemia. On the other hand, omission of Ca2+ from the perfusing solution did not alter the increases in glutamate and aspartate induced by ischaemia. These results suggest that the glutamate and aspartate accumulation in four-vessel occlusion ischaemia is mediated by activation of NMDA receptors in a Ca2+ independent manner.

Introduction Notre objectif est de comprendre les mecanismes qui regulent le developpement et la fonction des cellules β pour apprehender les places respectives des anomalies de masse et de fonction β dans l’etiologie du diabete de type 2. Nos travaux precedents ont montre que les glucocorticoides (GC) inhibent le developpement des cellules β et que l’invalidation conditionnelle chez la souris du recepteur des GC (GR) dans les precurseurs pancreatiques (souris GR-Prec) aboutissait a un doublement de masse β. Le but de cette etude etait de comprendre les mecanismes impliques dans l’accroissement de cellules β. Materiels et methodes Nous avons etudie chez les souris GR-Prec et temoins adultes la tolerance au glucose sous regime normal ou hyperlipidique, mesure la masse et la proliferation des cellules beta et defini le transcriptome des ilots par micropuces a ARN. Resultats Les souris GR-Prec presentent une augmentation de la masse beta associee paradoxalement a une intolerance au glucose due a une alteration de l’insulinosecretion sous regime normal ou hyperlipidique. L’analyse du transcriptome des ilots de souris GR-Prec a identifie les genes Tph1 et 2 (Tryptophane hydroxylase), enzyme de synthese de serotonine, neurotransmetteur recemment implique dans la regulation de masse et fonction beta. Chez les souris GR-Prec, l’augmentation de Tph1 et 2 entraine une augmentation du contenu en serotonine dans les ilots. L’absence de signalisation GC stimule donc le systeme serotonine des cellules beta. A l’inverse, l’activation de la voie des GC dans des cellules beta in vitro diminue l’expression des genes Tph1 et Tph2 et la synthese de serotonine et contre-regule la stimulation de serotonine induite par la prolactine et l’exendine-4. Conclusion Les GC sont des puissants inhibiteurs de la synthese de serotonine dans les cellules beta, suggerant que la serotonine est un intermediaire des effets des GC sur la masse et fonction beta.

Rabbit leptomeningeal arteries contain granular cells resembling mast cells that frequently contact autonomic and sensory nerve profiles. In the present in vitro study, we determined whether these cells could be stimulated by substance P (SP) and calcitonin gene-related peptide (CGRP), which are stored and released by sensory C fibers. Immunohistochemistry of the middle cerebral artery showed that 5-HT was stored only in mast cell-like granules. This pool of 5-HT decreased in a dose-dependent manner when exogenous SP and CGRP were added to the incubation solution or when endogenous neuropeptides were released from nerve terminals by capsaicin. The simultaneous administration of CGRP and SP induced a dramatic exocytosis and a 5-HT release significantly greater than the sum of the individual effects of the two neuropeptides. We conclude that, as in classical connective tissue mast cells, the amine content of these granular cells can be released by a degranulation process induced by neuropeptides. The effects of capsaicin suggest that this phenomenon can be triggered by axon reflex of C fibers. The data also provide the first evidence of a synergistic action of SP and CGRP on mast cell degranulation.

A comparison was made in the blood levels of various cell types and biochemical substances and in lymphocyte antigens between 107 healthy sheep from a flock contaminated with scrapie (HC sheep) and 93 sheep from a noncontaminated flock (NC sheep), which served as a control population. Significant differences between the two groups of sheep were found in some of the levels, as had previously been found with lymphocyte antigens. The HC sheep, which included genetically resistant animals, could be distinguished from the NC sheep by their lower levels of various white cells, a noticeable decrease in urea, a moderate decrease in Mg2+ and Mn2+ ions, beta- and gamma-globulins, serotonin and vitamin B12, a strong increase in uric acid and a moderate increase in K+, Cl-, HCO3-, Zn2+, and Al3+ ions, as well as in total lipids and in the albumin to globulin ratio. In this HC population, the only enzyme with an increased level was aldolase; the levels of the other 7 enzymes measured were lowered. The observed modifications were considered to be signs of latent disturbances in the leukocyte system and in hepatic and renal functions, in spite of apparent resistance. Eleven lymphocyte antigens were studied. These antigens are not independent of the blood levels of the various substances measured, but are often correlated in a statistically significant manner with some of them. In the HC sheep, the lymphocyte antigens correlated with the modified levels in the blood were different from those in the control population.(ABSTRACT TRUNCATED AT 250 WORDS)

A new procedure for serum inorganic sulphate determination is described. The method is simple, accurate and highly reproducible. It is based on radioisotopic dilution of 35S-sulphate by protein-free, phosphate-free serum. The decrease in initial 35S-sulphate specific activity (corresponding to added serum sulphate) is determined in the supernatant after partial precipitation of sulphate by barium ion. Serum sulphate is computed thereafter from a standard curve. Recovery of added sulphate from dialysed or undialysed serum was 100%. The intra- and interassay coefficients of variation were 3.5% and 4.1%, respectively. The serum sulphate concentration measured by this method in 93 normal subjects was 400 +/- 90 mumol/l (mean +/- SD), which agreed with the values reported in the literature. Serum sulphate did not correlate with sex (p greater than 0.40, n = 93), but a significant correlation with age was observed (r = 0.25, p less than 0.02, n = 93).

Pruritus is frequently observed in hemodialyzed patients; its etiology seems to be multifactorial. Increased plasma histamine levels have been reported in chronic renal failure. As histamine is a potent inducer of pruritus. The authors investigated its concentrations and kinetics in plasma of stable hemodialyzed patients.

Significantly (p less than 0.001) decreased whole blood unconjugated serotonin levels were detected in AIDS patients as compared to patients with advanced cancers and to healthy individuals.