SUMMARY The possibility that the small intestine may represent a reservoir for Clostridium difficile was studied, using segments of human jejunum collected at necropsy. Our results (three of 100 specimens positive for C difficile culture) support the hypothesis that C difficile can be found in human jejunum and that it adheres to the normal mucosa as a resident bacterium. These findings suggest that gastrointestinal disease caused by C difficile has an endogenous origin.The possibility that the small intestine might represent a reservoir for disease caused by Clostridium dificile was suggested by Taylor et al, when they isolated C difficile from a jejunal aspirate of a patient with chronic colitis.' This hypothesis was confirmed by our experience with a case of pseudomembranous enteritis with spared colon, in which we isolated C difficile from the patient's ileum obtained atTo elucidate these findings we carried out a study to verify the rate of isolation of C difficile from human small intestine using segments of jejunum that had been obtained at necropsy. Material and methods COLLECTION OF SPECIMENSOver six months one hundred segments of proximal jejunum were collected within 48 hours from 100 patients who had died. The specimens were about 10 cm long and macroscopically free from lesions. Each segment was placed in a sterile Petri dish and immediately sent to the bacteriology laboratory.The subjects had died from different diseases, none of them had had diarrhoea or other gastrointestinal symptoms in life. The mean age was 70 years (range 52-86); 90% of the patients had received treatment with antibiotics-that is, ,B-lactam antibiotics alone, or in conjunction with aminoglycosides. PROCESSING OF THE SPECIMENSTo remove the bacteria that were not firmly attached to mucosa each segment was carefully washed with 10cc of a sterile saline solution using a vortex mixer for 10 minutes.3 This procedure was repeated three Accepted for publication 18 March 1986 861 times for each sample, changing the container and the washing liquid each time. After this the segment was stretched and the mucosa removed with a sterile Iancet; the material obtained was used to inoculate a cycloserine-cefoxitin-fructose selective agar (CCFA) plate.4 The plates were screened for colonies characteristic of C difficile; all the cultures were incubated for at least five days before being discarded. The three washings from each segment were centrifuged for 10 minutes at 5000 rpm and the sediments were used to inoculate a CCFA plate. ResultsWithin 48 hours the cultures from the mucosa were positive for C difficile in three cases. Prolonged incubation of the other samples did not yield any additional positive results. None of the centrifugated washings yielded C difficile. The ages of the culture positive patients were 63, 60, and 74 years; all of them had received treatment with antibiotics.

Catecholamine (CA) secretion from the adrenal medullary tissue is a key step of the adaptive response triggered by an organism to cope with stress. Whereas molecular and cellular secretory processes have been extensively studied at the single chromaffin cell level, data available for the whole gland level are much scarcer. We tackled this issue in rat by developing an easy to implement experimental strategy combining the adrenal acute slice supernatant collection with a high-performance liquid chromatography-based epinephrine and norepinephrine (NE) assay. This technique affords a convenient method for measuring basal and stimulated CA release from single acute slices, allowing thus to individually address the secretory function of the left and right glands. Our data point that the two glands are equally competent to secrete epinephrine and NE, exhibiting an equivalent epinephrine:NE ratio, both at rest and in response to a cholinergic stimulation. Nicotine is, however, more efficient than acetylcholine to evoke NE release. A pharmacological challenge with hexamethonium, an α3-containing nicotinic acetylcholine receptor antagonist, disclosed that epinephrine- and NE-secreting chromaffin cells distinctly expressed α3 nicotinic receptors, with a dominant contribution in NE cells. As such, beyond the novelty of CA assays from acute slice supernatants, our study contributes at refining the secretory behavior of the rat adrenal medullary tissue, and opens new perspectives for monitoring the release of other hormones and transmitters, especially those involved in the stress response.

We report for the first time a novel electrochemical treatment applied to a glassy carbon electrode (GCE) during pnitrophenol (PNP) oxidation and dedicated to the limitation of electrode passivation by nitrophenol compounds oxidation. We propose an electrochemical process of direct phenol oxidation by starting the electrolysis at a very low potential, À1.2 V/SCE, in order to generate a soluble monomer, p-aminophenol, on the electrode surface. Then, paminophenol elaborated on the electrode surface in the place of oligomers, gives benzoquinone as a by-product and no film formation was observed. Furthermore, the presence of a p-NiTSPc (film of nickel tetrasulfonated phtalocyanine) coating permitted to increase two times the electrode sensitivity without passivation, too.

Sympathetic hyperactivation, a common feature of obesity and metabolic syndrome, is a key trigger of hypertension. However, some obese subjects with autonomic imbalance present a dissociation between sympathetic activity-mediated vasoconstriction and increased blood pressure. Here, we aimed to determine in a rat model of metabolic syndrome whether the endothelium endothelial nitric oxide (NO) synthase (eNOS)-NO pathway contributes to counteract the vasopressor effect of the sympathetic system. Rats were fed a high-fat and high-sucrose (HFS) diet for 15 wk. Sympathovagal balance was evaluated by spectral analysis of heart rate variability and plasmatic catecholamine measurements. Blood pressure was measured in the presence or absence of N-nitro-l-arginine methyl ester (l-NAME) to inhibit the contribution of eNOS. Vascular reactivity was assessed on isolated aortic rings in response to α-adrenergic agonist. The HFS diet increased sympathetic tone, which is characterized by a higher low on the high-frequency spectral power ratio and a higher plasmatic concentration of epinephrine. Despite this, no change in blood pressure was observed. Interestingly, HFS rats exhibited vascular hyporeactivity (-23.6%) to α-adrenergic receptor stimulation that was abolished by endothelial removal or eNOS inhibition (l-NAME). In addition, eNOS phosphorylation (Ser) was increased in response to phenylephrine in HFS rats only. Accordingly, eNOS inhibition in vivo revealed higher blood pressure in HFS rats compared with control rats (147 vs. 126 mmHg for mean blood pressure, respectively). Restrain of adrenergic vasopressor action by endothelium eNOS is increased in HFS rats and contributes to maintained blood pressure in the physiological range. NEW & NOTEWORTHY Despite the fact that prohypertensive sympathetic nervous system activity is markedly increased in rats with early metabolic syndrome, they present with normal blood pressure. These observations appear to be explained by increased endothelial nitric oxide synthase response to adrenergic stimulation, which results in vascular hyporeactivity to α-adrenergic stimulation, and therefore blood pressure is preserved in the physiological range. Listen to this article's corresponding podcast at http://www.physiology.org/doi/10.1152/ajpheart.00217.2017 .

L’excès de phosphore (P) dans les eaux de retenues est responsable d’une prolifération de cyanobactéries. Cela s’observe fréquemment dans la plupart des pays européens. En France, le lac de Ribou, localisé à Cholet, est un cas typique (Maine-et-Loire) illustrant cette problématique. La présence de P est principalement liée aux rejets des stations d’épuration (STEP), situées sur le bassin versant, non équipées de processus de déphosphatation et également liées aux pratiques agricoles et aux élevages de bovins. Ainsi, la concentration moyenne de P total entrant dans le lac de Ribou par le Trézon sur l’année 2006 a été de 0,34 mg•L-1, ce qui en fait une eau de qualité « passable » et de nature « mésotrophe » alors que la retenue de Ribou a un caractère eutrophe. De plus, en matière de flux, il se déverse dans le lac de Ribou environ 18,7 tonnes de P•an-1. Sortir de l’eutrophisation reviendrait à abaisser cette valeur à 1 tonne de P•an-1, soit une concentration moyenne de P dans les eaux d’alimentation de la retenue de 0,03 mg•L-1. Pour sortir de cette situation, un plan de gestion dans le cadre du Schéma d’Aménagement de Gestion de l’Eau (SAGE) a démarré en 2006. Il comprend un ensemble de 29 mesures avec, en particulier, la mise en place de contrats agro-environnementaux permettant de réduire les surfaces en cultures au profit de surfaces en prairies, la réalisation de diagnostics agro-environnementaux sur 170 exploitations, l’organisation de journées de formation pour la plantation et l’entretien des haies, la prise en compte du phosphore dans les projets de modernisation des six STEP du bassin versant (non-rejet, déphosphatation, etc.), la mise en place de bandes enherbées, le suivi mensuel de la qualité de l’eau brute sur 16 points de prélèvement du bassin versant, l’organisation de réunions techniques auprès des acteurs concernés permettant la promotion de méthodologies visant à réduire le phosphore et la promotion de la « désintensification » des systèmes de production agricole. Enfin, parmi ces mesures, l’aménagement de zones humides tampons est envisagé dans le but de diminuer d’un facteur 10 les apports actuels d’ici 2010. Ainsi, une démarche de restauration a été mise en oeuvre. En effet, dans un premier temps, un diagnostic du lac de Ribou par analyse en composante principale sur un ensemble de données physico-chimiques a montré que la station la plus anthropisée correspond à la station TR qui doit faire l’objet de priorité dans le cadre d’un aménagement futur. Dans un second temps, au cours de l’année 2006, des inventaires floristiques ont été conduits sur cette zone de confluence entre le Trézon et le lac de Ribou, en appliquant la méthode des quadrats selon l’échelle de Braun-Blanquet. Nous avons mis en évidence 21 espèces végétales en mai et 12 en octobre. Parmi celles-ci, les espèces Juncus effusus, Phalaris arundinacea, Salix caprea, Rorippa amphibia, et Lemna minor ont été répertoriées.Ces inventaires ont permis, d’une part, de diagnostiquer l’état écologique du milieu aquatique à travers l’observa...

Neonicotinoid insecticides are nicotine-derived molecules which exert acute neurotoxic effects over the insect central nervous system by activating nicotinic acetylcholine receptors (nAChRs). However, these receptors are also present in the mammalian central and peripheral nervous system, where the effects of neonicotinoids are faintly known. In mammals, cholinergic synapses are crucial for the control of vascular tone, blood pressure and skeletal muscle contraction. We therefore hypothesized that neonicotinoids could affect cholinergic networks in mammals and sought to highlight functional consequences of acute intoxication in rats with sub-lethal concentrations of the highly used acetamiprid (ACE) and clothianidin (CLO). In this view, we characterized their electrophysiological effects on rat α3β4 nAChRs, knowing that it is predominantly expressed in ganglia of the vegetative nervous system and the adrenal medulla, which initiates catecholamine secretion. Both molecules exhibited a weak agonist effect on α3β4 receptors. Accordingly, their influence on epinephrine secretion from rat adrenal glands was also weak at 100 M, but it was stronger at 500 M. Challenging ACE or CLO together with nicotine (NIC) ended up with paradoxical effects on secretion. In addition, we measured the rat arterial blood pressure (ABP) in vivo by arterial catheterization. As expected, NIC induced a significant increase in ABP. ACE and CLO did not affect the ABP in the same conditions. However, simultaneous exposure of rats to both NIC and ACE/CLO promoted an increase of ABP and induced a biphasic response. Modeling the interaction of ACE or CLO on α3β4 nAChR is consistent with a binding site located in the agonist pocket of the receptor. We present a transversal experimental approach of mammal intoxication with neonicotinoids at different scales, including in vitro, ex vivo, in vivo and in silico. It paves the way of the acute and chronic toxicity for this class of insecticides on mammalian organisms

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Advanced material with antibacterial properties would be a promising way to improve the disinfection process in food plants. Our objective was to combine the bactericidal effect of TiO 2 with the mechanical strength of TiN coatings. A TiO 2 rutile film was obtained after annealing of a supplied 316 stainless steel with a TiN coating. This TiO 2 upperlayer displays a photocatalytic activity under UV light exposure. The substrates with the TiN coating and the TiO 2 upperlayer are more hydrophobic than the 316 control. The adhesion of either Listeria or Pseudomonas, on 316-TiN is characterized by the presence of clusters of cells, while the oxidation of the TiN surface leads to a more hydrophilic layer where cells are individualized. After UV illumination of the adherent cells and subsequent growth, the residual bacterial population present on 316-TiO 2 is lower than that present on the 316-TiN. The bactericidal effect is more important on Listeria than on Pseudomonas.